greenmedinfo--2020-08-16--Paeonol suppresses the effect of ox-LDL on mice vascular endothelial cells by regulating miR-338-3p/

Paeonol suppresses the effect of ox-LDL on mice vascular endothelial cells by regulating miR-338-3p/TET2 axis in atherosclerosis

Atherosclerosis is the common vascular disease .
Vascular smooth muscle cell proliferation and vascular endothelial cell ( VEC ) dysfunction are involved in the causes of atherosclerosis .
And oxidized low-density lipoprotein ( ox-LDL ) -induced vascular endothelial cells ( VECs ) are suitable models for studying atherosclerosis development .
Paeonol was reported to repress ox-LDL-induced VEC progression .
However , its detailed mechanism was not fully reported .
MicroRNAs ( miRNAs ) acted as regulators in multiple diseases .
Previous findings found that microRNA-338-3p ( miR-338-3p ) was overexpressed in Atherosclerosis process .
However , the function and underlying @ @ @ @ @ @ @ to be elucidated .
The purpose of this research was to reveal the role of miR-338-3p in paeonol-regulated ox-LDL-induced VEC progression .
Cell counting kit-8 ( CCK-8 ) and flow cytometry were employed to determine cell viability and apoptosis , respectively .
Moreover , the levels of IL-6 and IL-1β were analyzed using enzyme-linked immunosorbent assay , as well as the contents of reactive oxygen species , lactate dehydrogenase , and malonic dialdehyde were investigated using related kits .
Furthermore , quantitative real-time polymerase chain reaction was carried out to determine the expressionof @ @ @ @ @ @ @ to detect the level of tet methylcytosine dioxygenase 2 ( TET2 ) .
Besides , the interaction between miR-338-3p and TET2 was predicted by DIANA , and then confirmed by the dual-luciferase reporter assay and RNA immunoprecipitation assay .
Ox-LDL repressed mice VEC viability , and promoted apoptosis , inflammatory response , and oxidative injury .
Paeonol inhibited the effect of ox-LDL on the growth of the VECs .
Furthermore , paeonol regulated VEC development via downregulating miR-338-3p expression .
Interestingly , miR-338-3p targeted TET2 and inhibited TET2expression .
MiR-338-3p modulated ox-LDL-treated VEC @ @ @ @ @ @ @ demonstrated that paeonol attenuated the effect of ox-LDL on the development of mice VECs via modulating miR-338-3p/TET2 axis , providing a theoretical basis for the treatment of AS .

Atherosclerosis is the common vascular disease. Vascular smooth muscle cell proliferation and vascular endothelial cell (VEC) dysfunction are involved in the causes of atherosclerosis. And oxidized low-density lipoprotein (ox-LDL)-induced vascular endothelial cells (VECs) are suitable models for studying atherosclerosis development. Paeonol was reported to repress ox-LDL-induced VEC progression. However, its detailed mechanism was not fully reported. MicroRNAs (miRNAs) acted as regulators in multiple diseases. Previous findings found that microRNA-338-3p (miR-338-3p) was overexpressed in Atherosclerosis process. However, the function and underlying mechanism of miR-338-3p in ox-LDL-treated VECs needed to be elucidated. The purpose of this research was to reveal the role of miR-338-3p in paeonol-regulated ox-LDL-induced VEC progression. Cell counting kit-8 (CCK-8) and flow cytometry were employed to determine cell viability and apoptosis, respectively. Moreover, the levels of IL-6 and IL-1β were analyzed using enzyme-linked immunosorbent assay, as well as the contents of reactive oxygen species, lactate dehydrogenase, and malonic dialdehyde were investigated using related kits. Furthermore, quantitative real-time polymerase chain reaction was carried out to determine the expressionof miR-338-3p. Western blot assay was conducted to detect the level of tet methylcytosine dioxygenase 2 (TET2). Besides, the interaction between miR-338-3p and TET2 was predicted by DIANA, and then confirmed by the dual-luciferase reporter assay and RNA immunoprecipitation assay. Ox-LDL repressed mice VEC viability, and promoted apoptosis, inflammatory response, and oxidative injury. Paeonol inhibited the effect of ox-LDL on the growth of the VECs. Furthermore, paeonol regulated VEC development via downregulating miR-338-3p expression. Interestingly, miR-338-3p targeted TET2 and inhibited TET2expression. MiR-338-3p modulated ox-LDL-treated VEC growth through suppressing TET2 expression. We demonstrated that paeonol attenuated the effect of ox-LDL on the development of mice VECs via modulating miR-338-3p/TET2 axis, providing a theoretical basis for the treatment of AS.


greenmedinfo--2020-08-16--P. edulis extracts showed antifungal and antioxidant activity.

The leaves ofwere subjected to physicochemical analysis , such as ion content , extractives , and structural molecules .
The @ @ @ @ @ acetate extracts were submitted to phytochemical analyzes by GC-MS , HPLC-MS , and spectrophotometry .
@ @ @ @ @ DPPH , ABTS and TAA methods ) potential , antimicrobial ( MIC method ) action @ @ @ @ @ ( flow cytometry analysis ) were performed .
The extracts showed a moderate antioxidant capacity @ @ @ @ @ several metabolites , mainly phenols , such as caffeic acid , p-coumaric acid and luteolin @ @ @ @ @ ethanolic extracts showed antifungal activity .
In addition , the extracts did not affect splenocytes @ @ @ @ @ promoted the production of IL-6 , IL-10 , IL-17 and TNF-α cytokines.extracts showed antifungal and @ @ @ @ @ to induce immunostimulatory action in splenocyte cultures .

The leaves ofwere subjected to physicochemical analysis, such as ion content, extractives, and structural molecules. The hexanic, ethanolic and ethyl acetate extracts were submitted to phytochemical analyzes by GC-MS, HPLC-MS, and spectrophotometry. In addition, antioxidant (DPPH, ABTS and TAA methods) potential, antimicrobial (MIC method) action, cytotoxicity and immunostimulant activity (flow cytometry analysis) were performed. The extracts showed a moderate antioxidant capacity and revealed the presence of several metabolites, mainly phenols, such as caffeic acid, p-coumaric acid and luteolin. The ethyl acetate and ethanolic extracts showed antifungal activity. In addition, the extracts did not affect splenocytes viability at 12.5 μg/mL and promoted the production of IL-6, IL-10, IL-17 and TNF-α cytokines.extracts showed antifungal and antioxidant activity and were able to induce immunostimulatory action in splenocyte cultures.


greenmedinfo--2020-08-16--Paeoniflorin attenuates chronic constriction injury-induced neuropathic pain by suppressing spinal N

Paeoniflorin attenuates chronic constriction injury-induced neuropathic pain by suppressing spinal NLRP3 inflammasome activation.

Neuropathic pain remains one of the most common pain conditions worldwide .
Accumulating evidence shows that activation of the NOD-like receptor protein 3 ( NLRP3 ) inflammasome contributes to the pathogenesis of neuropathic pain , although the role of the NLRP3 inflammasome in neuropathic pain has not yet been fully elucidated .
In animal models of neuropathic pain , paeoniflorin ( PF ) was shown to have analgesic , anti-inflammatory , and antidepressant effects .
However , the role of the NLRP3 inflammasome in the analgesic properties of PF has not yet been studied .
In this study , we @ @ @ @ @ @ @ NLRP3 inflammasome in the spinal cord was involved in the development of neuropathic pain and whether PF could be an effective treatment for this type of pain .
We found that activation of the NLRP3 inflammasome mediated the development of neuropathic pain following chronic constriction injury of the sciatic nerve and that PF attenuated neuropathic pain by inhibiting NLRP3 inflammasome activation .
Moreover , PF enhanced the translocation of the transcription factor nuclear factor erythroid 2-related factor 2 into the nucleus and suppressed nuclear factor-kappa B activity in the spinal cord .
These @ @ @ @ @ @ @ potential therapeutic agent for neuropathic pain .

Neuropathic pain remains one of the most common pain conditions worldwide. Accumulating evidence shows that activation of the NOD-like receptor protein 3 (NLRP3) inflammasome contributes to the pathogenesis of neuropathic pain, although the role of the NLRP3 inflammasome in neuropathic pain has not yet been fully elucidated. In animal models of neuropathic pain, paeoniflorin (PF) was shown to have analgesic, anti-inflammatory, and antidepressant effects. However, the role of the NLRP3 inflammasome in the analgesic properties of PF has not yet been studied. In this study, we aimed to confirm whether activation of the NLRP3 inflammasome in the spinal cord was involved in the development of neuropathic pain and whether PF could be an effective treatment for this type of pain. We found that activation of the NLRP3 inflammasome mediated the development of neuropathic pain following chronic constriction injury of the sciatic nerve and that PF attenuated neuropathic pain by inhibiting NLRP3 inflammasome activation. Moreover, PF enhanced the translocation of the transcription factor nuclear factor erythroid 2-related factor 2 into the nucleus and suppressed nuclear factor-kappa B activity in the spinal cord. These results suggest that PF may be a potential therapeutic agent for neuropathic pain.


greenmedinfo--2020-08-16--Paeoniflorin ameliorates depressive-like behavior in prenatally stressed offspring by restoring the

Paeoniflorin ameliorates depressive-like behavior in prenatally stressed offspring by restoring the HPA axis- and glucocorticoid receptor- associated dysfunction.

BACKGROUND : Prenatal stress ( PS ) can increase the risk of nervous , endocrine and metabolic diseases and induce depression in offspring .
Paeoniflorin ( PA ) is an amorphous glucoside isolated from the aqueous extract of roots of the peony plant ( Paeonia lactiflora Pall . )
and exerts various pharmacological effects in the nervous system .
METHODS : Male prenatally stressed offspring were used to investigate the antidepression-like effects and possible mechanism of PA. We measured animal behavior , HPA axis , Nissil staining , and Ng expression .
Additionally , we assessed the modulation of hippocampal @ @ @ @ @ @ @ and SNARE complex expression by western blotting .
RESULTS : The results showed that administration of PA ( 15 , 30 , and 60 mg/kg/day , i.g . )
for 28 days markedly increased sucrose intake and decreased the immobility time and the total number of crossings , center crossings , rearing , and grooming in male PS offspring .
Moreover , PA significantly reduced the serum corticosterone ( CORT ) , adrenocorticotropin ( ACTH ) , corticotropin-releasing hormone ( CRH ) and hippocampal glutamate ( Glu ) levels in male PS offspring @ @ @ @ @ @ @ of GR nuclear translocation .
Furthermore , PA markedly increased neurogranin ( Ng ) protein expression in the hippocampus CA3 region in offspring .
PA also markedly decreased hippocampal Glu by inhibiting SNAP25 , VAMP2 , Syntaxin1a and related protein expression ; SNARE complex formation ; and EAAT2/3 , NR1 , NR2A , and FKBP5 protein expression .
CONCLUSIONS : Taken together , the results of this study show that PA has antidepression-like effects in male PS offspring , partially due to the HPA axis , GR dysfunction and Glu transport system .

BACKGROUND: Prenatal stress (PS) can increase the risk of nervous, endocrine and metabolic diseases and induce depression in offspring. Paeoniflorin (PA) is an amorphous glucoside isolated from the aqueous extract of roots of the peony plant (Paeonia lactiflora Pall.) and exerts various pharmacological effects in the nervous system.
METHODS: Male prenatally stressed offspring were used to investigate the antidepression-like effects and possible mechanism of PA. We measured animal behavior, HPA axis, Nissil staining, and Ng expression. Additionally, we assessed the modulation of hippocampal glucocorticoid receptors (GR) nuclear translocation and SNARE complex expression by western blotting.
RESULTS: The results showed that administration of PA (15, 30, and 60 mg/kg/day, i.g.) for 28 days markedly increased sucrose intake and decreased the immobility time and the total number of crossings, center crossings, rearing, and grooming in male PS offspring. Moreover, PA significantly reduced the serum corticosterone (CORT), adrenocorticotropin (ACTH), corticotropin-releasing hormone (CRH) and hippocampal glutamate (Glu) levels in male PS offspring, which were stimulated by an increase of GR nuclear translocation. Furthermore, PA markedly increased neurogranin (Ng) protein expression in the hippocampus CA3 region in offspring. PA also markedly decreased hippocampal Glu by inhibiting SNAP25, VAMP2, Syntaxin1a and related protein expression; SNARE complex formation; and EAAT2/3, NR1, NR2A, and FKBP5 protein expression.
CONCLUSIONS: Taken together, the results of this study show that PA has antidepression-like effects in male PS offspring, partially due to the HPA axis, GR dysfunction and Glu transport system.


greenmedinfo--2020-08-16--A review on the pharmacokinetics of paeoniflorin and its anti-inflammatory and immunomodulatory effe

A review on the pharmacokinetics of paeoniflorin and its anti-inflammatory and immunomodulatory effects.

Increasing pharmacological evidence supports that paeoniflorin , a water-soluble monoterpene glycoside isolated from Paeonia lactiflora Pall .
( Shaoyao in @ @ @ @ @ wide range of medicinal properties including anti-inflammatory , antioxidant , antithrombotic , anticonvulsive , analgesic @ @ @ @ @ hepatoprotective , antidepressant-like , antitumoral , and immune-regulatory activities ; as well as enhancing cognition @ @ @ @ @ In addition to pharmacodynamic studies , information on pharmacokinetics is also significant for the further @ @ @ @ @ .
The present review focuses on the absorption , distribution , metabolism , and excretion @ @ @ @ @ pharmacological activities of paeoniflorin on inflammation and immune function .
According to the findings obtained @ @ @ @ @ vivo , a broad application prospect has been opened for paeoniflorin .
However , further @ @ @ @ @ the direct molecular mechanisms and key targets underlying the beneficial effects of paeoniflorin on inflammation @ @ @

Increasing pharmacological evidence supports that paeoniflorin, a water-soluble monoterpene glycoside isolated from Paeonia lactiflora Pall. (Shaoyao in Chinese), has a wide range of medicinal properties including anti-inflammatory, antioxidant, antithrombotic, anticonvulsive, analgesic, cardioprotective, neuroprotective, hepatoprotective, antidepressant-like, antitumoral, and immune-regulatory activities; as well as enhancing cognition and attenuating learning impairment. In addition to pharmacodynamic studies, information on pharmacokinetics is also significant for the further development and utilization of paeoniflorin. The present review focuses on the absorption, distribution, metabolism, and excretion of paeoniflorin, especially main pharmacological activities of paeoniflorin on inflammation and immune function. According to the findings obtained both in vitro and in vivo, a broad application prospect has been opened for paeoniflorin. However, further studies are needed to clarity the direct molecular mechanisms and key targets underlying the beneficial effects of paeoniflorin on inflammation and immunity.


greenmedinfo--2020-08-16--The results suggest that paeoniflorin can simultaneously exert antidepression and prokinetic effects

The results suggest that paeoniflorin can simultaneously exert antidepression and prokinetic effects.

Paeoniflorin , an organic compound extracted from the roots of the white peony ( ) plant , has previously been shown to exert antidepression and prokinetic effects .
The traditional Chinese prescription Si-Ni-San , of which paeoniflorin is a constituent , is often used in treating depression and functional gastrointestinal disorders .
The effectiveness of Si-Ni-San has been shown to be less effective in a paeoniflorin-deleted form .
The present study further investigates whether paeoniflorin alone is as effective as herbal prescriptions in which the compound is a constituent , specifically any antidepressive and prokinetic effect on rats subjected to @ @ @ @ @ @ @ .
The FST was used to establish the depression model .
Sprague-Dawley rats were administrated with 10 mg/kg paeoniflorin by gastrogavage three times before the behavioral test and gastrointestinal motility tests , respectively .
In antidepression studies , fluoxetine was used as the positive control .
In order to determine the effect of paeoniflorin on the gastrointestinal movement , mosapride was used as the positive control .
Plasma and hippocampus monoamine , hypothalamic-pituitary-adrenal axis , plasma brain-derived neurotrophic factor ( BDNF ) , superoxide dismutase ( SOD ) , methane dicarboxylic aldehyde ( @ @ @ @ @ @ @ and hippocampus nitric oxide ( NO ) were assessed using an enzyme-linkedimmunosorbent assay ( ELISA ) .
Gastrointestinal ( GI ) motility was measuredand .
Rats subjected to FST showed decreased gastric emptying and intestinal transit , decreased plasma and hippocampus 5-hydroxytryptamine , norepinephrine , dopamine , ghrelin , motilin , and reduced plasma BDNF and SOD as well as increased plasma and hippocampus corticotropin-releasing hormone , adrenocorticotropic hormone , corticosterone , plasma MDA , and hippocampus NO .
Paeoniflorin reversed these symptoms in a similar manner to fluoxetine and mosapride , respectively.
@ @ @ @ @ @ @ of healthy rats dose-dependently .
The results suggest that paeoniflorin can simultaneously exert antidepression and prokinetic effects via polypharmacology .

Paeoniflorin, an organic compound extracted from the roots of the white peony () plant, has previously been shown to exert antidepression and prokinetic effects. The traditional Chinese prescription Si-Ni-San, of which paeoniflorin is a constituent, is often used in treating depression and functional gastrointestinal disorders. The effectiveness of Si-Ni-San has been shown to be less effective in a paeoniflorin-deleted form. The present study further investigates whether paeoniflorin alone is as effective as herbal prescriptions in which the compound is a constituent, specifically any antidepressive and prokinetic effect on rats subjected to a forced swimming test (FST). The FST was used to establish the depression model. Sprague-Dawley rats were administrated with 10 mg/kg paeoniflorin by gastrogavage three times before the behavioral test and gastrointestinal motility tests, respectively. In antidepression studies, fluoxetine was used as the positive control. In order to determine the effect of paeoniflorin on the gastrointestinal movement, mosapride was used as the positive control. Plasma and hippocampus monoamine, hypothalamic-pituitary-adrenal axis, plasma brain-derived neurotrophic factor (BDNF), superoxide dismutase (SOD), methane dicarboxylic aldehyde (MDA), ghrelin, motilin, and hippocampus nitric oxide (NO) were assessed using an enzyme-linkedimmunosorbent assay (ELISA). Gastrointestinal (GI) motility was measuredand. Rats subjected to FST showed decreased gastric emptying and intestinal transit, decreased plasma and hippocampus 5-hydroxytryptamine, norepinephrine, dopamine, ghrelin, motilin, and reduced plasma BDNF and SOD as well as increased plasma and hippocampus corticotropin-releasing hormone, adrenocorticotropic hormone, corticosterone, plasma MDA, and hippocampus NO. Paeoniflorin reversed these symptoms in a similar manner to fluoxetine and mosapride, respectively., paeoniflorin can stimulate the jejunal contract of healthy rats dose-dependently. The results suggest that paeoniflorin can simultaneously exert antidepression and prokinetic effects via polypharmacology.


greenmedinfo--2020-08-16--Vitexin can act as a potential blocker of the STAT3 signaling cascade and mitigate the survival as w

Vitexin can act as a potential blocker of the STAT3 signaling cascade and mitigate the survival as well as invasion of HCC cells.

Hepatocellular carcinoma ( HCC ) is a major malignancy that stands second in terms of global cancer-related mortality .
STAT3 has been described as a latent transcription factor that promotes tumorigenesis .
This study was designed to examine the effect of vitexin on STAT3 signaling and important hallmarks of cancer .
HCC cells were employed to decipher the impact of vitexin on activation of STAT3 signaling using Western blotting , EMSA , immunocytochemistry , and reporter assay .
The combinational apoptotic effects of vitexin with approved anti-cancer drugs was examined by live-dead assay , and its anti-invasive potential was studied @ @ @ @ @ @ @ in cell-based assays were verified using in silico analysis .
Vitexin effectively inhibited sustained activation of JAK1 , JAK2 , Src , and STAT3 in HCC cells .
Vitexin downregulated DNA binding ability , reduced the nuclear pool of STAT3 , and diminished epidermal growth factor ( EGF ) -driven STAT3 gene expression .
Interestingly , treatment with tyrosine phosphatase inhibitor altered the vitexin-induced STAT3 phosphorylation , and the attenuation of STAT3 by vitexin was found to be driven through the upregulation of PTPεC .
The combinational studies indicated that vitexin can exhibit @ @ @ @ @ @ @ .
It also suppressed the CXCL12-induced cell invasion .
The results of cell-based assays are supported by in silico analysis as the vitexin displayed favorable interaction with kinase domain of JAK2 protein .
Overall , this study demonstrated that vitexin can act as a potential blocker of the STAT3 signaling cascade and mitigate the survival as well as invasion of HCC cells .

Hepatocellular carcinoma (HCC) is a major malignancy that stands second in terms of global cancer-related mortality. STAT3 has been described as a latent transcription factor that promotes tumorigenesis. This study was designed to examine the effect of vitexin on STAT3 signaling and important hallmarks of cancer. HCC cells were employed to decipher the impact of vitexin on activation of STAT3 signaling using Western blotting, EMSA, immunocytochemistry, and reporter assay. The combinational apoptotic effects of vitexin with approved anti-cancer drugs was examined by live-dead assay, and its anti-invasive potential was studied using matrigel assay. The results obtained in cell-based assays were verified using in silico analysis. Vitexin effectively inhibited sustained activation of JAK1, JAK2, Src, and STAT3 in HCC cells. Vitexin downregulated DNA binding ability, reduced the nuclear pool of STAT3, and diminished epidermal growth factor (EGF)-driven STAT3 gene expression. Interestingly, treatment with tyrosine phosphatase inhibitor altered the vitexin-induced STAT3 phosphorylation, and the attenuation of STAT3 by vitexin was found to be driven through the upregulation of PTPεC. The combinational studies indicated that vitexin can exhibit substantial apoptotic effects with doxorubicin and sorafenib. It also suppressed the CXCL12-induced cell invasion. The results of cell-based assays are supported by in silico analysis as the vitexin displayed favorable interaction with kinase domain of JAK2 protein. Overall, this study demonstrated that vitexin can act as a potential blocker of the STAT3 signaling cascade and mitigate the survival as well as invasion of HCC cells.


greenmedinfo--2020-08-16--Vitexin ameliorates chronic stress plub high fat diet-induced nonalcoholic fatty liver disease by in

Vitexin ameliorates chronic stress plub high fat diet-induced nonalcoholic fatty liver disease by inhibiting inflammation.

Evidences showed that chronic stress ( CS ) can aggravate the situation of nonalcoholic fatty liver disease ( NAFLD ) @ @ @ @ @ the major components in hawthorn , which is widely used to reduce blood lipid .
@ @ @ @ @ explore the therapeutic effects and potential mechanisms of vitexin on chronic stress mice with high-fat @ @ @ @ @ The results showed that 5-week vitexin administration ( 40 mg/kg , i.g . )
could @ @ @ @ @ , alleviate lipid metabolism , and inhibit liver inflammation in CSHFD mice .
In addition @ @ @ @ @ macrophage infiltration , obviously down-regulated the mRNA and protein expressions of hepatic SREBP-1c , FAS @ @ @ @ @ found that vitexin treatment could significantly inhibit the expressions of TLR4/NF-κB signaling in CSHFD mice @ @ @ @ @ vitexin could ameliorate chronic stress combined with high-fat diet induced NAFLD , and its mechanisms @ @ @ @ @ TLR4/NF-κB signaling and reduce fatty acid synthesis proteins .

Evidences showed that chronic stress (CS) can aggravate the situation of nonalcoholic fatty liver disease (NAFLD). Vitexin is one of the major components in hawthorn, which is widely used to reduce blood lipid. This study was aimed to explore the therapeutic effects and potential mechanisms of vitexin on chronic stress mice with high-fat diet (CSHFD). The results showed that 5-week vitexin administration (40 mg/kg, i.g.) could obviously reduce hepatic fat deposition, alleviate lipid metabolism, and inhibit liver inflammation in CSHFD mice. In addition, vitexin significantly reduced hepatic macrophage infiltration, obviously down-regulated the mRNA and protein expressions of hepatic SREBP-1c, FAS, ACC.Moreover, we also found that vitexin treatment could significantly inhibit the expressions of TLR4/NF-κB signaling in CSHFD mice. This results suggested that vitexin could ameliorate chronic stress combined with high-fat diet induced NAFLD, and its mechanisms is closely related to inhibit TLR4/NF-κB signaling and reduce fatty acid synthesis proteins.


greenmedinfo--2020-08-16--Withaferin A ameliorates ovarian cancer-induced cachexia and proinflammatory signaling.

BACKGROUND : Ovarian cancer is the fifth leading cause of cancer-related deaths amongst women in the United States .
Cachexia is the primary cause of death in approximately 30 % of cancer patients , and is often evidenced in ovarian cancer patients .
We tested the steroidal lactone Withaferin A to examine if it could ameliorate ovarian cancer-induced cachexia .
METHODS : Six-week-old severely immunodeficient female mice were xenografted with the ovarian cancer cell line A2780 followed by treatment with Withaferin A or vehicle .
Changes in functional grip strength were assessed on a weekly basis .
Postmortem , H @ @ @ @ @ @ @ muscle sections and immunofluorescent immunohistochemistry was performed on skeletal muscle and tumor sections .
The levels of NF-κB-related proinflammatory cytokines were assessed in the xenografted tumors and in resident host skeletal muscle .
RESULTS : Xenografting of the A2780 cell line resulted in a significant rate of mortality , which was attenuated by a therapeutic dosage of Withaferin A .
Mice that received vehicle treatment following xenografting exhibited functional muscle decline over the course of the study .
The therapeutic dosage Withaferin A treatment attenuated this reduction in grip strength , whereas the @ @ @ @ @ @ @ to be toxic/lethal and demonstrated a further decline in functional muscle strength and an increased rate of mortality on par with vehicle treatment .
At a histological level , the vehicle treated tumor-bearing mice exhibited a profound reduction in myofibrillar cross-sectional area compared to the vehicle treated tumor-free control group .
The atrophic changes induced by the xenografted tumor were significantly ameliorated by treatment with Withaferin A .
The combination of functional muscle weakening and induction of myofibrillar atrophy corroborate a cachectic phenotype , which was functionally rescued by Withaferin A .
Further @ @ @ @ @ @ @ type conversion observed in the settings of cancer-induced cachexia .
In both host resident skeletal muscle and the xenografted tumors , we report an increase in NF-κB-related proinflammatory cytokines that was reversed by Withaferin A treatment .
Finally , we demonstrated that Withaferin A significantly downregulates cytosolic and nuclear levels of phospho-p65 , the active canonical NF-κB transcription factor , in xenografted tumors .
CONCLUSIONS : Cumulatively , our results demonstrate a previously overlooked role of Withaferin A in a xenograft model of ovarian cancer .
We propose mechanisms by which Withaferin A @ @ @ @ @ @ @ an attenuation of the cachectic phenotype in an i.p .
xenograft model of ovarian cancer .

BACKGROUND: Ovarian cancer is the fifth leading cause of cancer-related deaths amongst women in the United States. Cachexia is the primary cause of death in approximately 30% of cancer patients, and is often evidenced in ovarian cancer patients. We tested the steroidal lactone Withaferin A to examine if it could ameliorate ovarian cancer-induced cachexia.
METHODS: Six-week-old severely immunodeficient female mice were xenografted with the ovarian cancer cell line A2780 followed by treatment with Withaferin A or vehicle. Changes in functional grip strength were assessed on a weekly basis. Postmortem, H&E staining was performed on skeletal muscle sections and immunofluorescent immunohistochemistry was performed on skeletal muscle and tumor sections. The levels of NF-κB-related proinflammatory cytokines were assessed in the xenografted tumors and in resident host skeletal muscle.
RESULTS: Xenografting of the A2780 cell line resulted in a significant rate of mortality, which was attenuated by a therapeutic dosage of Withaferin A. Mice that received vehicle treatment following xenografting exhibited functional muscle decline over the course of the study. The therapeutic dosage Withaferin A treatment attenuated this reduction in grip strength, whereas the supratherapeutic dosage of Withaferin A was found to be toxic/lethal and demonstrated a further decline in functional muscle strength and an increased rate of mortality on par with vehicle treatment. At a histological level, the vehicle treated tumor-bearing mice exhibited a profound reduction in myofibrillar cross-sectional area compared to the vehicle treated tumor-free control group. The atrophic changes induced by the xenografted tumor were significantly ameliorated by treatment with Withaferin A. The combination of functional muscle weakening and induction of myofibrillar atrophy corroborate a cachectic phenotype, which was functionally rescued by Withaferin A. Further, treatment completely abolished the slow-to-fast myofiber type conversion observed in the settings of cancer-induced cachexia. In both host resident skeletal muscle and the xenografted tumors, we report an increase in NF-κB-related proinflammatory cytokines that was reversed by Withaferin A treatment. Finally, we demonstrated that Withaferin A significantly downregulates cytosolic and nuclear levels of phospho-p65, the active canonical NF-κB transcription factor, in xenografted tumors.
CONCLUSIONS: Cumulatively, our results demonstrate a previously overlooked role of Withaferin A in a xenograft model of ovarian cancer. We propose mechanisms by which Withaferin A reduces NF-κB-dependent pro-inflammatory cytokine production leading to an attenuation of the cachectic phenotype in an i.p. xenograft model of ovarian cancer.


greenmedinfo--2020-08-16--Withaferin-A attenuates the ageing-mediated motor degenerative changes in the substantia nigra and s

Withaferin-A attenuates the ageing-mediated motor degenerative changes in the substantia nigra and striatum of aged rats.

Withaferin-A ( WA ) was evaluated for its neuroprotective efficacy on the dopamine ( DA ) neurons of the substantia nigra ( SN ) and striatum ( ST ) in aged rats .
Wistar albino rats were divided into group I , young ( 3 months old ) ; group II , aged ( 24 months old ) ; group III , aged rats supplemented with WA ( 50 mg/kg bodyweight once per day for 30 days ) , and group IV , young rats supplemented with WA ( 50 mg/kg bodyweight ) .
At the end of the experiment @ @ @ @ @ @ @ various motor behavior analyses , and were sacrificed by transcardial perfusion .
The brains were dissected out and subjected to various analyses , including histological , histomorphometrical , and immunolocalization of the tyrosine hydroxylase ( TH ) enzyme .
The data of rotarod analysis ( p < 0.001 ) showed a significant motor impairment in aged rats ( number of falls 10.2± 0.86 ) and reduction in retention time ( 31.23 ± 2.56 s ) compared to young controls ( 2.41 ± 0.35 and 84.05 ± 5.15 s ) .
The stride length @ @ @ @ @ @ @ ) in aged rats ( 4.21± 0.57 and 4.38 ± 0.61 cm ) when compared to young control rats ( 6.98 ± 0.25 and 7.13 ± 0.70 cm ) .
The histomorphometric data of the aged animals showed a significant reduction in the neuronal diameter ( p < 0.001 ) , density ( p < 0.001 ) , and volume ( p < 0.001 ) in the SN of aged rats when compared to young rats .
Immunohistology demonstrated a marked reduction in the levels of TH enzyme in both the SN and @ @ @ @ @ @ @ young rats .
Both structural and functional impairments were reversed in the aged animals after the supplementation of WA ( p < 0.001 ) .
The present study clearly indicates that WA attenuates the ageing-mediated motor degenerative changes in the SN and ST of aged rats and ascertains its neuroprotective potential .

Withaferin-A (WA) was evaluated for its neuroprotective efficacy on the dopamine (DA) neurons of the substantia nigra (SN) and striatum (ST) in aged rats. Wistar albino rats were divided into group I, young (3 months old); group II, aged (24 months old); group III, aged rats supplemented with WA (50 mg/kg bodyweight once per day for 30 days), and group IV, young rats supplemented with WA (50 mg/kg bodyweight). At the end of the experiment period, the animals were subjected to various motor behavior analyses, and were sacrificed by transcardial perfusion. The brains were dissected out and subjected to various analyses, including histological, histomorphometrical, and immunolocalization of the tyrosine hydroxylase (TH) enzyme. The data of rotarod analysis (p<0.001) showed a significant motor impairment in aged rats (number of falls 10.2± 0.86) and reduction in retention time (31.23 ± 2.56 s) compared to young controls (2.41 ± 0.35 and 84.05 ± 5.15 s). The stride length was significantly reduced (p<0.001) in aged rats (4.21± 0.57 and 4.38 ± 0.61 cm) when compared to young control rats (6.98 ± 0.25 and 7.13 ± 0.70 cm). The histomorphometric data of the aged animals showed a significant reduction in the neuronal diameter (p<0.001), density (p<0.001), and volume (p<0.001) in the SN of aged rats when compared to young rats. Immunohistology demonstrated a marked reduction in the levels of TH enzyme in both the SN and ST of aged animals when compared to young rats. Both structural and functional impairments were reversed in the aged animals after the supplementation of WA (p<0.001). The present study clearly indicates that WA attenuates the ageing-mediated motor degenerative changes in the SN and ST of aged rats and ascertains its neuroprotective potential.


greenmedinfo--2020-08-16--Withaferin A suppresses breast cancer cell proliferation.

Withaferin A ( WFA ) , a C , C-epoxy steroidal lactone isolated from the medicinal plant Withania somnifera ( L. ) Dunal , inhibits growth of tumor cells in different cancer types .
However , the mechanisms underlying the effect of WFA on tumor cells are not fully understood .
In the present study , we evaluated the blockade of TASK-3 channels by WFA in TASK-3-expressing HEK-293 cells .
Explore if the WFA-mediated TASK-3 blockade can be used as a pharmacological tool to decrease the cell viability in cancer cells .
A combination of functional experiments ( patch-clamp , @ @ @ @ @ @ @ ) and molecular docking analysis were used to get insights into the mechanism by which the inhibition of TASK-3 by WFA affects the growth and viability of cancer cells .
Withaferin A was found to inhibit the activity of TASK-3 channels .
The inhibitory effect of Withaferin A on TASK-3 potassium currents was dose-dependent and independent of voltage .
Molecular modeling studies identified putative WFA-binding sites in TASK-3 channel involved the channel blockade .
In agreements with the molecular modeling predictions , mutation of residues F125 to A ( F125A ) , L197 @ @ @ @ @ @ @ the double mutant F125A-L197 V markedly decreased the WFA-induced inhibition of TASK-3 .
Finally , the cytotoxic effect of WFA was tested in MDA-MB-231 human breast cancer cells transfected with TASK-3 or shRNA that decreases TASK-3 expression .
Together , our results show that the cytotoxiceffect of WFA on fully transformed MDA-MB-231 cells depends on the expression of TASK-3 .
Herein , we also provide insights into the mechanism of TASK-3 inhibition by WFA .

Withaferin A (WFA), a C,C-epoxy steroidal lactone isolated from the medicinal plant Withania somnifera (L.) Dunal, inhibits growth of tumor cells in different cancer types. However, the mechanisms underlying the effect of WFA on tumor cells are not fully understood. In the present study, we evaluated the blockade of TASK-3 channels by WFA in TASK-3-expressing HEK-293 cells. Explore if the WFA-mediated TASK-3 blockade can be used as a pharmacological tool to decrease the cell viability in cancer cells. A combination of functional experiments (patch-clamp, gene downregulation, overexpression and pharmacological inhibition) and molecular docking analysis were used to get insights into the mechanism by which the inhibition of TASK-3 by WFA affects the growth and viability of cancer cells. Withaferin A was found to inhibit the activity of TASK-3 channels. The inhibitory effect of Withaferin A on TASK-3 potassium currents was dose-dependent and independent of voltage. Molecular modeling studies identified putative WFA-binding sites in TASK-3 channel involved the channel blockade. In agreements with the molecular modeling predictions, mutation of residues F125 to A (F125A), L197 to V (L197 V) and the double mutant F125A-L197 V markedly decreased the WFA-induced inhibition of TASK-3. Finally, the cytotoxic effect of WFA was tested in MDA-MB-231 human breast cancer cells transfected with TASK-3 or shRNA that decreases TASK-3 expression. Together, our results show that the cytotoxiceffect of WFA on fully transformed MDA-MB-231 cells depends on the expression of TASK-3. Herein, we also provide insights into the mechanism of TASK-3 inhibition by WFA.


greenmedinfo--2020-08-16--Identification of bioactive molecule from Withania somnifera (Ashwagandha) as SARS-CoV-2 main protea

Identification of bioactive molecule from Withania somnifera (Ashwagandha) as SARS-CoV-2 main protease inhibitor.

SARS-CoV-2 is the causative agent of COVID-19 and has been declared as pandemic disease by World Health Organization .
Lack of targeted therapeutics and vaccines for COVID-2019 have triggered the scientific community to develop new vaccines or drugs against this novel virus .
Many synthetic compounds and antimalarial drugs are undergoing clinical trials .
The traditional medical practitioners widely use Indian medicinal plant ( Ashwagandha ) natural constituents , called withanolides for curing various diseases .
The main protease ( M ) of SARS-CoV-2 plays a vital role in disease propagation by processing the polyproteins which are required for its @ @ @ @ @ @ @ significant target for drug discovery .
In the present study , we evaluate the potential of 40 natural chemical constituents of Ashwagandha to explore a possible inhibitor against main protease of SARS-CoV-2 by adopting the computational approach .
The docking study revealed that four constituents of Ashwagandha ; Withanoside II ( -11.30 Kcal/mol ) , Withanoside IV ( -11.02 Kcal/mol ) , Withanoside V ( -8.96 Kcal/mol ) and Sitoindoside IX ( -8.37 Kcal/mol ) exhibited the highest docking energy among the selected natural constituents .
Further , MD simulation study of 100 @ @ @ @ @ @ @ affinity andhydrogen-bonding interactions with the protein active site and indicates its stability in the active site .
The binding free energy score also correlates with the highest score of -87.01 ± 5.01 Kcal/mol as compared to other selected compounds .
In conclusion , our study suggests that WithanosideV in Ashwagandha may be serve as a potential inhibitor against Mof SARS-CoV-2 to combat COVID-19 and may have an antiviral effect on nCoV .
Communicated by Ramaswamy H. Sarma .

SARS-CoV-2 is the causative agent of COVID-19 and has been declared as pandemic disease by World Health Organization. Lack of targeted therapeutics and vaccines for COVID-2019 have triggered the scientific community to develop new vaccines or drugs against this novel virus. Many synthetic compounds and antimalarial drugs are undergoing clinical trials. The traditional medical practitioners widely use Indian medicinal plant(Ashwagandha) natural constituents, called withanolides for curing various diseases. The main protease (M) of SARS-CoV-2 plays a vital role in disease propagation by processing the polyproteins which are required for its replication. Hence, it denotes a significant target for drug discovery. In the present study, we evaluate the potential of 40 natural chemical constituents of Ashwagandha to explore a possible inhibitor against main protease of SARS-CoV-2 by adopting the computational approach. The docking study revealed that four constituents of Ashwagandha; Withanoside II (-11.30 Kcal/mol), Withanoside IV (-11.02 Kcal/mol), Withanoside V (-8.96 Kcal/mol) and Sitoindoside IX (-8.37 Kcal/mol) exhibited the highest docking energy among the selected natural constituents. Further, MD simulation study of 100 ns predicts Withanoside V possess strong binding affinity andhydrogen-bonding interactions with the protein active site and indicates its stability in the active site. The binding free energy score also correlates with the highest score of -87.01 ± 5.01 Kcal/mol as compared to other selected compounds. In conclusion, our study suggests that WithanosideV in Ashwagandha may be serve as a potential inhibitor against Mof SARS-CoV-2 to combat COVID-19 and may have an antiviral effect on nCoV. Communicated by Ramaswamy H. Sarma.


greenmedinfo--2020-08-16--RNA-seq reveals novel mechanistic targets of withaferin A in prostate cancer cells.

Withaferin A ( WA ) is a promising phytochemical exhibiting in vitro and in vivo anticancer activities against prostate and other cancers , but the mechanism of its action is not fully understood .
In this study , we performed RNA-seq analysis using 22Rv1 human prostate cancer cell line to identify mechanistic targets of WA .
Kyoto Encyclopedia of Genes and Genomes pathway analysis of the differentially expressed genes showed most significant enrichment of genes associated with metabolism .
These results were validated using LNCaP and 22Rv1 human prostate cancer cells and Hi-Myc transgenic mice as models .
The intracellular @ @ @ @ @ @ @ acids and neutral lipids were decreased significantly following WA treatment in both cells , which was accompanied by downregulation of mRNA ( confirmed by quantitative reverse transcription-polymerase chain reaction ) and protein levels of key fatty acid synthesis enzymes , including ATP citrate lyase , acetyl-CoA carboxylase 1 , fatty acid synthase and carnitine palmitoyltransferase 1A .
Ectopic expression of c-Myc , but not constitutively active Akt , conferred a marked protection against WA-mediated suppression of acetyl-CoA carboxylase 1 and fatty acid synthase protein expression , and clonogenic cell survival .
WA was @ @ @ @ @ @ @ fatty acid synthesis in comparison with known modulators of fatty acid metabolism including cerulenin and etomoxir .
Intraperitoneal WA administration to Hi-Myc transgenic mice ( 0.1 mg/mouse , three times/week for 5 weeks ) also resulted in a significant decrease in circulating levels of total free fatty acids and phospholipids , and expression of ATP citrate lyase , acetyl-CoA carboxylase 1 , fatty acid synthase and carnitine palmitoyltransferase 1A proteins in the prostate in vivo .

Withaferin A (WA) is a promising phytochemical exhibiting in vitro and in vivo anticancer activities against prostate and other cancers, but the mechanism of its action is not fully understood. In this study, we performed RNA-seq analysis using 22Rv1 human prostate cancer cell line to identify mechanistic targets of WA. Kyoto Encyclopedia of Genes and Genomes pathway analysis of the differentially expressed genes showed most significant enrichment of genes associated with metabolism. These results were validated using LNCaP and 22Rv1 human prostate cancer cells and Hi-Myc transgenic mice as models. The intracellular levels of acetyl-CoA, total free fatty acids and neutral lipids were decreased significantly following WA treatment in both cells, which was accompanied by downregulation of mRNA (confirmed by quantitative reverse transcription-polymerase chain reaction) and protein levels of key fatty acid synthesis enzymes, including ATP citrate lyase, acetyl-CoA carboxylase 1, fatty acid synthase and carnitine palmitoyltransferase 1A. Ectopic expression of c-Myc, but not constitutively active Akt, conferred a marked protection against WA-mediated suppression of acetyl-CoA carboxylase 1 and fatty acid synthase protein expression, and clonogenic cell survival. WA was a superior inhibitor of cell proliferation and fatty acid synthesis in comparison with known modulators of fatty acid metabolism including cerulenin and etomoxir. Intraperitoneal WA administration to Hi-Myc transgenic mice (0.1 mg/mouse, three times/week for 5 weeks) also resulted in a significant decrease in circulating levels of total free fatty acids and phospholipids, and expression of ATP citrate lyase, acetyl-CoA carboxylase 1, fatty acid synthase and carnitine palmitoyltransferase 1A proteins in the prostate in vivo.


greenmedinfo--2020-08-16--Withaferin A alleviates traumatic brain injury induced secondary brain injury via suppressing apopto

Withaferin A alleviates traumatic brain injury induced secondary brain injury via suppressing apoptosis in endothelia cells and modulating activation in the microglia.

Traumatic brain injury ( TBI ) is a major public health concern with high rates of morbidity and mortality worldwide .
Currently used medications , though effective , are also associated with several adverse effects .
Development of effective neuroprotective agents with fewer side-effects would be of clinical value .
Previous studies have shown that withaferin compounds have a potential neuroprotective effect in nervous system disorders .
However , the effect of withaferin compounds , especially withaferin A ( WFA ) , on traumatic brain injury is unclear .
In the present study , both in vivo and in vitro @ @ @ @ @ @ @ could exert a neuroprotective effect after TBI and were used to explore the associated mechanisms .
The results showed that WFA significantly improved neurobehavioral function in a dose-dependent fashion and alleviated histological alteration of injury to tissues in TBI mice .
In vitro models of TBI revealed that dose-dependent WFA treatment increased the viability of SH-SY5Y cells .
In addition , WFA treatment could attenuate blood-brain barrier disruption and brain edema via suppressing apoptosis in endothelial cells .
Furthermore , both our in vivo and in vitro results reveal that WFA treatment could @ @ @ @ @ @ @ ( IL-1β , IL-6 , and TNF-α ) , which correlate with an overall reduction in microglial activation .
These data suggest that the neuroprotection by WFA is , at least in part , related to regulation of microglial activation and inhibition of vascular endothelial cell apoptosis .
Taken together , these findings support further investigation of WFA as a promising therapeutic agent for promoting functional recovery after traumatic brain injury .

Traumatic brain injury (TBI) is a major public health concern with high rates of morbidity and mortality worldwide. Currently used medications, though effective, are also associated with several adverse effects. Development of effective neuroprotective agents with fewer side-effects would be of clinical value. Previous studies have shown that withaferin compounds have a potential neuroprotective effect in nervous system disorders. However, the effect of withaferin compounds, especially withaferin A (WFA), on traumatic brain injury is unclear. In the present study, both in vivo and in vitro models were used to assess whether WFA could exert a neuroprotective effect after TBI and were used to explore the associated mechanisms. The results showed that WFA significantly improved neurobehavioral function in a dose-dependent fashion and alleviated histological alteration of injury to tissues in TBI mice. In vitro models of TBI revealed that dose-dependent WFA treatment increased the viability of SH-SY5Y cells. In addition, WFA treatment could attenuate blood-brain barrier disruption and brain edema via suppressing apoptosis in endothelial cells. Furthermore, both our in vivo and in vitro results reveal that WFA treatment could significantly reduce levels of several neuroinflammation cytokines (IL-1β, IL-6, and TNF-α), which correlate with an overall reduction in microglial activation. These data suggest that the neuroprotection by WFA is, at least in part, related to regulation of microglial activation and inhibition of vascular endothelial cell apoptosis. Taken together, these findings support further investigation of WFA as a promising therapeutic agent for promoting functional recovery after traumatic brain injury.


greenmedinfo--2020-08-16--Synergistic effect of 5-fluorouracil and withaferin-A induces endoplasmic reticulum stress-mediated

Synergistic effect of 5-fluorouracil and withaferin-A induces endoplasmic reticulum stress-mediated autophagy and apoptosis in colorectal cancer cells.

The development of chemo-resistance against 5-fluorouracil ( 5-FU ) in tumor cells is one of the main debacles in colorectal cancer ( CRC ) patients .
A recent combination of 5-FU with oxaliplatin or cetuximab drastically improves the survival rate in CRC patients ; however , the toxicity issue can not be evaded completely .
Thus , searching for novel drug combinations with high specificity and low toxicity is seemingly important .
Owing to the less undesirable effects of natural products on normal cells , here we investigated the synergistic antitumor effect of withaferin-A ( WA ) in combination with @ @ @ @ @ @ @ combination of WA and 5-FU induces a significant antiproliferative effect and modulates endoplasmic reticulum ( ER ) stress in favor of cell death in colorectal cancer ( CRC ) cells .
Mechanistically , the combination upregulates the expression of ER stress sensors ( BiP , PERK , CHOP , ATF-4 , and eIF2α ) and executes PERK axis mediated apoptosis in CRC cells .
Additionally , the combined treatment of WA and 5-FU mediated ER stress induces autophagy and apoptosis , which were confirmed by immunoblotting , acridine orange ( AO ) staining @ @ @ @ @ @ @ In contrast , inhibition of ER stress with salubrinal significantly decreases both autophagic and apoptotic cell populations .
Moreover , pharmacological inhibition of either autophagy or apoptosis by their respective inhibitors 3-methyladenine ( 3-MA ) or carbobenzoxy-valyl-alanyl-aspartyl- [ O-methyl ] -fluoro-methyl ketone ( Z-VAD-FMK ) decreases their respective population of cells but could not affect either of the population significantly .
Finally , the combination attenuates the expression of β-catenin pathway associated proteins and arrests cell cycle at the GM phase in CRC cells .
In summary , the combination of WA and @ @ @ @ @ @ @ stress-mediated induction of autophagy and apoptosis , inhibiting theβ-catenin pathway and arresting the cell cycle at a GM phase in CRC cells .

The development of chemo-resistance against 5-fluorouracil (5-FU) in tumor cells is one of the main debacles in colorectal cancer (CRC) patients. A recent combination of 5-FU with oxaliplatin or cetuximab drastically improves the survival rate in CRC patients; however, the toxicity issue cannot be evaded completely. Thus, searching for novel drug combinations with high specificity and low toxicity is seemingly important. Owing to the less undesirable effects of natural products on normal cells, here we investigated the synergistic antitumor effect of withaferin-A (WA) in combination with 5-FU. Our results demonstrate that the combination of WA and 5-FU induces a significant antiproliferative effect and modulates endoplasmic reticulum (ER) stress in favor of cell death in colorectal cancer (CRC) cells. Mechanistically, the combination upregulates the expression of ER stress sensors (BiP, PERK, CHOP, ATF-4, and eIF2α) and executes PERK axis mediated apoptosis in CRC cells. Additionally, the combined treatment of WA and 5-FU mediated ER stress induces autophagy and apoptosis, which were confirmed by immunoblotting, acridine orange (AO) staining and annexin-V FITC by flow cytometry. In contrast, inhibition of ER stress with salubrinal significantly decreases both autophagic and apoptotic cell populations. Moreover, pharmacological inhibition of either autophagy or apoptosis by their respective inhibitors 3-methyladenine (3-MA) or carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoro-methyl ketone (Z-VAD-FMK) decreases their respective population of cells but could not affect either of the population significantly. Finally, the combination attenuates the expression of β-catenin pathway associated proteins and arrests cell cycle at the GM phase in CRC cells. In summary, the combination of WA and 5-FU decreases cell viability by inducing ER stress-mediated induction of autophagy and apoptosis, inhibiting theβ-catenin pathway and arresting the cell cycle at a GM phase in CRC cells.


greenmedinfo--2020-08-16--These results suggest that Withaferin-A may prove a potent lead molecule in oral cancer treatment.

PURPOSE : Oral cancer is one of the common oral cavity and pharynx cancers and its treatment is limited by early metastasis , late diagnosis and emergence of drug resistance .
Herein we investigated the anticancer effects of Withaferin-A against the cisplatin-resistant SCC-4 human oral cancer cells .
METHODS : The proliferation rate of the human oral cancer cells was evaluated by CCK-8 assay .
Apoptotic cell death was studied by acridine orange ( AO ) / ethidium bromide ( EB ) staining as well as by flow cytometry using annexin V/propidium iodide ( PI ) staining .
Cell cycle @ @ @ @ @ @ @ Reactive oxygen species ( ROS ) examination was carried out by flow cytometry .
Protein expressions were determined by immunoblotting .
RESULTS : The results of the MTT assay showed that Withaferin-A caused decrease in the proliferation of SCC-4 cells and exhibited an IC50 of 14µM .
The anticancer effects of Withaferin-A were mainly due to the induction of apoptosis which was linked with upsurge of Bax and depletion of BCl-2 .
Annexin V/PI assay showed that the apoptotic cells were 0.75 , 5.8 , 12.4 and 22.66 % at 0,7,14 and 28 @ @ @ @ @ @ @ increase in the ROS and decrease in the MMP levels of the SCC-4 cells .
Western blot analysis showed that the expression of LC3B II increased while of p62 decreased remarkably upon treatment with Withaferin-A , suggestive of autophagic cell death .
Cell cycle analysis by flow cytometry showed that Withaferin-A caused increase in the G2/M phase cells triggering arrest of cancer cells at the G2/M checkpoint of the cell cycle .
Finally , western blot analysis showed that Withaferin-A blocked the MAPK/RAS/RAF signalling pathway in the SCC-4 cells .
CONCLUSIONS : These @ @ @ @ @ @ @ potent lead molecule in oral cancer treatment and warrants further investigations .

PURPOSE: Oral cancer is one of the common oral cavity and pharynx cancers and its treatment is limited by early metastasis, late diagnosis and emergence of drug resistance. Herein we investigated the anticancer effects of Withaferin-A against the cisplatin-resistant SCC-4 human oral cancer cells.
METHODS: The proliferation rate of the human oral cancer cells was evaluated by CCK-8 assay. Apoptotic cell death was studied by acridine orange (AO) / ethidium bromide (EB) staining as well as by flow cytometry using annexin V/propidium iodide (PI) staining. Cell cycle analysis was performed by flow cytometry. Reactive oxygen species (ROS) examination was carried out by flow cytometry. Protein expressions were determined by immunoblotting.
RESULTS: The results of the MTT assay showed that Withaferin-A caused decrease in the proliferation of SCC-4 cells and exhibited an IC50 of 14µM. The anticancer effects of Withaferin-A were mainly due to the induction of apoptosis which was linked with upsurge of Bax and depletion of BCl-2. Annexin V/PI assay showed that the apoptotic cells were 0.75, 5.8, 12.4 and 22.66% at 0,7,14 and 28 µM concentrations of Withaferin-A. Withaferin-Aalso caused increase in the ROS and decrease in the MMP levels of the SCC-4 cells. Western blot analysis showed that the expression of LC3B II increased while of p62 decreased remarkably upon treatment with Withaferin-A, suggestive of autophagic cell death. Cell cycle analysis by flow cytometry showed that Withaferin-A caused increase in the G2/M phase cells triggering arrest of cancer cells at the G2/M checkpoint of the cell cycle. Finally, western blot analysis showed that Withaferin-A blocked the MAPK/RAS/RAF signalling pathway in the SCC-4 cells.
CONCLUSIONS: These results suggest that Withaferin-A may prove a potent lead molecule in oral cancer treatment and warrants further investigations.


greenmedinfo--2020-08-16--Synergistic combinations of paclitaxel and withaferin A against human non-small cell lung cancer cel

Synergistic combinations of paclitaxel and withaferin A against human non-small cell lung cancer cells.

Platinum-taxane combination chemotherapy still represents the standard of care for advanced non-small cell lung cancer ( NSCLC ) with no targetable driver mutations .
However , the efficacy of these drugs has plateaued at 10-14 months primarily due to dose-limiting toxicity , chemoresistance , and metastasis .
Here , we explored the effects of withaferin A ( WFA ) alone and in combination with paclitaxel ( PAC ) on the growth , proliferation , migration , and invasion of human NSCLC cells .
We show that the sensitivity of H1299 and A549 cells to concomitant treatment with PAC and WFA @ @ @ @ @ @ @ or WFA alone .
Using the combination index and dose-reduction index , we demonstrated that various combinations ( 1:40 , 1:20 , 1:10 ) of PAC to WFA , respectively , were highly synergistic .
In addition , PAC+WFA co-treatment synergistically inhibited colony formation , migration , invasion and increased the induction of apoptosis in H1299 and A549 cells .
Interestingly , the synergism of PAC and WFA was not schedule-dependent but was enhanced when cells were pretreated with WFA indicating a chemo-sensitizing effect .
Importantly , WFA was active against both PAC-sensitive @ @ @ @ @ @ @ ) cells bothand .
Mechanistically , WFA inhibits the proliferation of NSCLC cells via thiol oxidation .
The effects of WFA were inhibited in the presence of N-acetyl cysteine and other thiol donors .
Taken together , our results demonstrate the efficacy of WFA alone or alongside PAC on NSCLC cells and provide a strong rationale for further detailed testing in clinically relevant models for the development of PAC+WFA combination as an alternative therapeutic strategy for advanced NSCLC .

Platinum-taxane combination chemotherapy still represents the standard of care for advanced non-small cell lung cancer (NSCLC) with no targetable driver mutations. However, the efficacy of these drugs has plateaued at 10-14 months primarily due to dose-limiting toxicity, chemoresistance, and metastasis. Here, we explored the effects of withaferin A (WFA) alone and in combination with paclitaxel (PAC) on the growth, proliferation, migration, and invasion of human NSCLC cells. We show that the sensitivity of H1299 and A549 cells to concomitant treatment with PAC and WFA was greater than that of either PAC or WFA alone. Using the combination index and dose-reduction index, we demonstrated that various combinations (1:40, 1:20, 1:10) of PAC to WFA, respectively, were highly synergistic. In addition, PAC+WFA co-treatment synergistically inhibited colony formation, migration, invasion and increased the induction of apoptosis in H1299 and A549 cells. Interestingly, the synergism of PAC and WFA was not schedule-dependent but was enhanced when cells were pretreated with WFA indicating a chemo-sensitizing effect. Importantly, WFA was active against both PAC-sensitive (TS-A549) and PAC-resistant (TR-A549) cells bothand. Mechanistically, WFA inhibits the proliferation of NSCLC cells via thiol oxidation. The effects of WFA were inhibited in the presence of N-acetyl cysteine and other thiol donors. Taken together, our results demonstrate the efficacy of WFA alone or alongside PAC on NSCLC cells and provide a strong rationale for further detailed testing in clinically relevant models for the development of PAC+WFA combination as an alternative therapeutic strategy for advanced NSCLC.


greenmedinfo--2020-08-16--Overview of the anticancer activity of withaferin A, an active constituent of the Indian ginseng Wit

Overview of the anticancer activity of withaferin A, an active constituent of the Indian ginseng Withania somnifera.

Cancer is still considered a '' hopeless case '' , besides all of the advancements in oncology research .
On the other hand , the natural products , as effective lead molecules , have gained significant interest for research due to the absence of toxic and harmful side effects usually associated with conventional treatment methods .
Medicinal properties of herbal plants are strongly evidenced in traditional medicine from ancient times .
In the context above , withaferin A ( WA ) was identified as the active principle of the plant Withania somnifera , its molecule being reported to have excellent @ @ @ @ @ @ @ cell lines .
Furthermore , the in silico approaches in the medicinal chemistry of WA revealed the biological targets and gave momentum for the research that leads to many amazing pharmacological activities of WA which are not yet explored .
This includes a broad spectrum of anticancer actions manifested in different organs ( breast , pancreas , colon ) , melanoma and B cell lymphoma , etc .
This review is an extensive survey of the most recent anticancer studies reported for WA , along with its mechanism of action and details about @ @ @ @ @ @ @ .

Cancer is still considered a"hopeless case", besides all of the advancements in oncology research. On the other hand, the natural products, as effective lead molecules, have gained significant interest for research due to the absence of toxic and harmful side effects usually associated with conventional treatment methods. Medicinal properties of herbal plants are strongly evidenced in traditional medicine from ancient times. In the context above, withaferin A (WA) was identified as the active principle of the plant Withania somnifera, its molecule being reported to have excellent anticancer and tumour inhibition activities in various cell lines. Furthermore, the in silico approaches in the medicinal chemistry of WA revealed the biological targets and gave momentum for the research that leads to many amazing pharmacological activities of WA which are not yet explored. This includes a broad spectrum of anticancer actions manifested in different organs (breast, pancreas, colon), melanoma and B cell lymphoma, etc. This review is an extensive survey of the most recent anticancer studies reported for WA, along with its mechanism of action and details about its in vitro and/or in vivo behaviour.


greenmedinfo--2020-08-16--Low concentration of withaferin A inhibits oxidative stress-mediated migration and invasion in oral

Low concentration of withaferin A inhibits oxidative stress-mediated migration and invasion in oral cancer cells.

Withaferin A ( WFA ) has been reported to inhibit cancer cell proliferation based on high cytotoxic concentrations .
However , the low cytotoxic effect of WFA in regulating cancer cell migration is rarely investigated .
The purpose of this study is to investigate the changes in migration and mechanisms of oral cancer Ca9-22 cells after low concentrations of WFA treatment .
WFA under 0.5μM at 24 h treatment shows no cytotoxicity to oral cancer Ca9-22 cells ( ~95 % viability ) .
Under this condition , WFA triggers reactive oxygen species ( ROS ) production and inhibits 2D ( @ @ @ @ @ @ @ ( transwell ) and Matrigel invasion .
Mechanically , WFA inhibits matrix metalloproteinase ( MMP ) -2 and MMP-9 activities but induces mRNA expression for a group of antioxidant genes , such as nuclear factor , erythroid 2-like 2 ( ) , heme oxygenase 1 ( ) , glutathione-disulfide reductase ( ) , and NAD ( P ) H quinone dehydrogenase 1 ( ) ) in Ca9-22 cells .
Moreover , WFA induces mild phosphorylation of the mitogen-activated protein kinase ( MAPK ) family , including extracellular signal-regulated kinases 1/2 ( ERK1/2 ) @ @ @ @ @ @ @ , and p38 expression .
All WFA-induced changes were suppressed by the presence of ROS scavenger-acetylcysteine ( NAC ) .
Therefore , these results suggest that low concentration of WFA retains potent ROS-mediated anti-migration and -invasion abilities for oral cancer cells .

Withaferin A (WFA) has been reported to inhibit cancer cell proliferation based on high cytotoxic concentrations. However, the low cytotoxic effect of WFA in regulating cancer cell migration is rarely investigated. The purpose of this study is to investigate the changes in migration and mechanisms of oral cancer Ca9-22 cells after low concentrations of WFA treatment. WFA under 0.5μM at 24 h treatment shows no cytotoxicity to oral cancer Ca9-22 cells (~95% viability). Under this condition, WFA triggers reactive oxygen species (ROS) production and inhibits 2D (wound healing) and 3D cell migration (transwell) and Matrigel invasion. Mechanically, WFA inhibits matrix metalloproteinase (MMP)-2 and MMP-9 activities but induces mRNA expression for a group of antioxidant genes, such as nuclear factor, erythroid 2-like 2 (), heme oxygenase 1 (), glutathione-disulfide reductase (), and NAD(P)H quinone dehydrogenase 1 ()) in Ca9-22 cells. Moreover, WFA induces mild phosphorylation of the mitogen-activated protein kinase (MAPK) family, including extracellular signal-regulated kinases 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), and p38 expression. All WFA-induced changes were suppressed by the presence of ROS scavenger-acetylcysteine (NAC). Therefore, these results suggest that low concentration of WFA retains potent ROS-mediated anti-migration and -invasion abilities for oral cancer cells.


greenmedinfo--2020-08-16--Withaferin A demonstrates real potential as a therapeutic agent to treat or prevent the spread of CO

Withaferin A demonstrates real potential as a therapeutic agent to treat or prevent the spread of COVID-1.

The outbreak and continued spread of the novel coronavirus disease 2019 ( COVID-19 ) is a preeminent global health threat that has resulted in the infection of over 11.5 million people worldwide .
In addition , the pandemic has claimed the lives of over 530,000 people worldwide .
Age and the presence of underlying comorbid conditions have been found to be key determinants of patient mortality .
One such comorbidity is the presence of an oncological malignancy , with cancer patients exhibiting an approximate two-fold increase in mortality rate .
Due to a lack of data , no consensus has @ @ @ @ @ @ @ the diagnosis and treatment of cancer patients .
Interestingly , two independent research groups have discovered that Withaferin A ( WFA ) , a steroidal lactone with anti-inflammatory and anti-tumorigenic properties , may bind to the viral spike ( S- ) protein of SARS-CoV-2 .
Further , preliminary data from our research group has demonstrated that WFA does not alter expression of ACE2 in the lungs of tumor-bearing female mice .
Downregulation of ACE2 has recently been demonstrated to increase the severity of COVID-19 .
Therefore , WFA demonstrates real potential as a @ @ @ @ @ @ @ spread of COVID-19 due to the reported interference in viral S-protein to host receptor binding and its lack of effect on ACE2 expression in the lungs .

The outbreak and continued spread of the novel coronavirus disease 2019 (COVID-19) is a preeminent global health threat that has resulted in the infection of over 11.5 million people worldwide. In addition, the pandemic has claimed the lives of over 530,000 people worldwide. Age and the presence of underlying comorbid conditions have been found to be key determinants of patient mortality. One such comorbidity is the presence of an oncological malignancy, with cancer patients exhibiting an approximate two-fold increase in mortality rate. Due to a lack of data, no consensus has been reached about the best practices for the diagnosis and treatment of cancer patients. Interestingly, two independent research groups have discovered that Withaferin A (WFA), a steroidal lactone with anti-inflammatory and anti-tumorigenic properties, may bind to the viral spike (S-) protein of SARS-CoV-2. Further, preliminary data from our research group has demonstrated that WFA does not alter expression of ACE2 in the lungs of tumor-bearing female mice. Downregulation of ACE2 has recently been demonstrated to increase the severity of COVID-19. Therefore, WFA demonstrates real potential as a therapeutic agent to treat or prevent the spread of COVID-19 due to the reported interference in viral S-protein to host receptor binding and its lack of effect on ACE2 expression in the lungs.


greenmedinfo--2020-08-16--Withania somnifera root extract inhibits fatty acid synthesis in prostate cancer cells.

Prior research argues for a role of increasedfatty acid synthesis in pathogenesis of prostate adenocarcinoma , which remains a leading cause of cancer-associated mortality in American men .
A safe and effective inhibitor of fatty acid synthesis is still a clinically unmet need .
Herein , we investigated the effect of ethanol extract ofroot ( WRE ) standardized for one of its components ( withaferin A ) on fatty acid synthesis using LNCaP and 22Rv1 human prostate cancer cells.is a medicinal plant used in the Ayurvedic medicine practiced in India .
Western blotting and confocal microscopy revealed a statistically significant @ @ @ @ @ @ @ acid metabolism enzymes including ATP citrate lyase ( ACLY ) , acetyl-CoA carboxylase 1 ( ACC1 ) , fatty acid synthase ( FASN ) , and carnitine palmitoyltransferase 1A ( CPT1A ) in WRE-treated cells compared with solvent control .
The mRNA levels of , , , andwere also lower in WRE-treated cells in comparison with control .
Consequently , WRE treatment resulted in a significant decrease in intracellular levels of acetyl-CoA , total free fatty acids , and neutral lipid droplets in both LNCaP and 22Rv1 cells .
WRE exhibited greater potency @ @ @ @ @ @ @ concentration than cerulenin and etomoxir .
Exposure to WRE results in downregulation of c-Myc and p-Akt ( S473 ) proteins in 22Rv1 cell line .
However , overexpression of only c-Myc conferred protection against clonogenic cell survival and lipogenesis inhibition by WRE .
In conclusion , these results indicate that WRE is a novel inhibitor of fatty acid synthesis in human prostate cancer cells .

Prior research argues for a role of increasedfatty acid synthesis in pathogenesis of prostate adenocarcinoma, which remains a leading cause of cancer-associated mortality in American men. A safe and effective inhibitor of fatty acid synthesis is still a clinically unmet need. Herein, we investigated the effect of ethanol extract ofroot (WRE) standardized for one of its components (withaferin A) on fatty acid synthesis using LNCaP and 22Rv1 human prostate cancer cells.is a medicinal plant used in the Ayurvedic medicine practiced in India. Western blotting and confocal microscopy revealed a statistically significant decrease in protein levels of key fatty acid metabolism enzymes including ATP citrate lyase (ACLY), acetyl-CoA carboxylase 1 (ACC1), fatty acid synthase (FASN), and carnitine palmitoyltransferase 1A (CPT1A) in WRE-treated cells compared with solvent control. The mRNA levels of,,, andwere also lower in WRE-treated cells in comparison with control. Consequently, WRE treatment resulted in a significant decrease in intracellular levels of acetyl-CoA, total free fatty acids, and neutral lipid droplets in both LNCaP and 22Rv1 cells. WRE exhibited greater potency for fatty acid synthesis inhibition at equimolar concentration than cerulenin and etomoxir. Exposure to WRE results in downregulation of c-Myc and p-Akt(S473) proteins in 22Rv1 cell line. However, overexpression of only c-Myc conferred protection against clonogenic cell survival and lipogenesis inhibition by WRE. In conclusion, these results indicate that WRE is a novel inhibitor of fatty acid synthesis in human prostate cancer cells.


greenmedinfo--2020-08-16--Effect of L-carnitine supplementation on lipid profiles of patients with liver disease.

Results of previous studies regarding the effect of L-carnitine on lipid profiles in the patients with liver diseases are contradictory .
This meta-analysis was performed to assess the effect of L-carnitine on serum levels of low-density lipoprotein ( LDL ) , high-density lipoprotein ( HDL ) , triglyceride ( TG ) , and total cholesterol ( TC ) in overweight patients with liver diseases .
A systematic search was carried out using the Web of Science , PubMed , Scopus , and Cochrane library databases to identify articles published before April 2019 investigating the effects of L-carnitine supplementation on patients @ @ @ @ @ @ @ language or time limitation for the studies .
A meta-analysis was carried out using both the random and fixed effects model where appropriate , and I2 index were used to evaluate heterogeneity .
These results indicated that L-carnitine supplementation significantly reduces blood levels of TC and TG in patients with liver disease , whereas carnitine had no effect on the levels of HDL and LDL .
The reducing effect of L-carnitine on both TC and TG was found following long-term carnitine supplementation ( ≥24 weeks ) , supplementation with doses less than or @ @ @ @ @ @ @ patients with chronic hepatitis C. This meta-analysis indicates the beneficial effect of L-carnitine on TC and TG in overweight patients with liver disease , particularly patients with chronic hepatitis C , in both long-term and low doses .

Results of previous studies regarding the effect of L-carnitine on lipid profiles in the patients with liver diseases are contradictory. This meta-analysis was performed to assess the effect of L-carnitine on serum levels of low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglyceride (TG), and total cholesterol (TC) in overweight patients with liver diseases. A systematic search was carried out using the Web of Science, PubMed, Scopus, and Cochrane library databases to identify articles published before April 2019 investigating the effects of L-carnitine supplementation on patients with liver disease. There was no language or time limitation for the studies. A meta-analysis was carried out using both the random and fixed effects model where appropriate, and I2 index were used to evaluate heterogeneity. These results indicated that L-carnitine supplementation significantly reduces blood levels of TC and TG in patients with liver disease, whereas carnitine had no effect on the levels of HDL and LDL. The reducing effect of L-carnitine on both TC and TG was found following long-term carnitine supplementation (≥24 weeks), supplementation with doses less than or equal to 2,000 mg/d, and in patients with chronic hepatitis C. This meta-analysis indicates the beneficial effect of L-carnitine on TC and TG in overweight patients with liver disease, particularly patients with chronic hepatitis C, in both long-term and low doses.


greenmedinfo--2020-08-16--Paeoniflorin inhibits IgE-mediated allergic reactions by suppressing the degranulation of mast cells

Paeoniflorin inhibits IgE-mediated allergic reactions by suppressing the degranulation of mast cells though binding with FcɛRI alpha subunits.

Paeoniflorin ( PF ) , a monoterpene glycoside isolated from the aqueous extract of the Chinese herb Radix Paeoniae Alba , has been used for treating various inflammatory diseases .
In this study , we aimed to investigate the anti-allergic activities of PF .
The anti-anaphylactic activity of PF was investigated using mast cell ( MC ) degranulation assay as well as Cainflux in vitro and skin swelling and extravasation assays in vivo .
The results showed that PF inhibited MC degranulation ( histamine release from 74.5 ± 4.95 ng/ml to 58.7 ± 6.06 ng/ml ) and Cainflux challenged by @ @ @ @ @ @ @ PF reduced the degree of swelling and Evans blue exudation in mice paws .
Furthermore , PF dose-dependently reduced serum inflammatory mediator release in mice sensitized with ovalbumin for 48 h by inhibiting MC degranulation .
Molecular docking study revealed that PF bound better with the α subunit of FcɛRI than with the β subunit .
SPR revealed that PF had a strong affinity interaction with FcɛRI α subunit and the Kvalue was ( 7.08 ± 0.97 ) e-6 M. Our findings revealed that PF inhibited anaphylactic responses in vivo and in vitro @ @ @ @ @ @ @ novel FcɛRI inhibitor for preventing MC-related allergic diseases .

Paeoniflorin (PF), a monoterpene glycoside isolated from the aqueous extract of the Chinese herb Radix Paeoniae Alba, has been used for treating various inflammatory diseases. In this study, we aimed to investigate the anti-allergic activities of PF. The anti-anaphylactic activity of PF was investigated using mast cell (MC) degranulation assay as well as Cainflux in vitro and skin swelling and extravasation assays in vivo. The results showed that PF inhibited MC degranulation (histamine release from 74.5 ± 4.95 ng/ml to 58.7 ± 6.06 ng/ml) and Cainflux challenged by DNP-BSA in vitro. In addition, PF reduced the degree of swelling and Evans blue exudation in mice paws. Furthermore, PF dose-dependently reduced serum inflammatory mediator release in mice sensitized with ovalbumin for 48 h by inhibiting MC degranulation. Molecular docking study revealed that PF bound better with the α subunit of FcɛRI than with the β subunit. SPR revealed that PF had a strong affinity interaction with FcɛRI α subunit and the Kvalue was (7.08 ± 0.97) e-6 M. Our findings revealed that PF inhibited anaphylactic responses in vivo and in vitro, and it can be considered a novel FcɛRI inhibitor for preventing MC-related allergic diseases.


greenmedinfo--2020-08-16--Prevention of acute graft-versus-host-disease by Withaferin a via suppression of AKT/mTOR pathway.

Acute Graft versus Host Disease ( aGVHD ) is a frequent and serious complication in patients receiving allogeneic bone marrow transplantation ( allo-BMT ) and often requires rigorous prophylaxis .
The current treatment regimens for aGVHD are associated with several side effects which necessitates the development of novel interventions that prevent aGVHD without precluding graft-versus-tumor effects .
In the present study , we show that treatment of donor graft with plant steroidal lactone Withaferin A ( WA ) prior to transplantation markedly reduced aGVHD mediated damage in target organs without compromising the graft-versus.-tumor activity of the transplanted lymphocytes .
WA @ @ @ @ @ @ @ of donor lymphocytes .
This was attributed to the ability of WA to inhibit early signaling events in T-cell activation including lymphoblast formation and activation of AKT/mTOR pathway .
Mortality and morbidity related to allo-transplantation was significantly reduced in recipients of WA treated donor splenocytes compared to recipient of vehicle treated donor splenocytes .
Further , WA treatment did not have any effect on reconstitution of lymphoid and myeloid lineages in recipients , resulting in stable and complete donor chimerism .
In agreement with previous reports showing the effectiveness of WA in a @ @ @ @ @ @ @ data further establishes that WA is able to attenuate aGVHD in an MHC-mismatched high dose chemo-conditioned murine model without compromising engraftment .
This study provides compelling scientific basis for possible application of WA for prevention and treatment of aGVHD in patients receiving allo-BMT .

Acute Graft versus Host Disease (aGVHD) is a frequent and serious complication in patients receiving allogeneic bone marrow transplantation (allo-BMT) and often requires rigorous prophylaxis. The current treatment regimens for aGVHD are associated with several side effects which necessitates the development of novel interventions that prevent aGVHD without precluding graft-versus-tumor effects. In the present study, we show that treatment of donor graft with plant steroidal lactone Withaferin A (WA) prior to transplantation markedly reduced aGVHD mediated damage in target organs without compromising the graft-versus.-tumor activity of the transplanted lymphocytes. WA abrogated post-transplant cytokine storm associated with allo-activation of donor lymphocytes. This was attributed to the ability of WA to inhibit early signaling events in T-cell activation including lymphoblast formation and activation of AKT/mTOR pathway. Mortality and morbidity related to allo-transplantation was significantly reduced in recipients of WA treated donor splenocytes compared to recipient of vehicle treated donor splenocytes. Further, WA treatment did not have any effect on reconstitution of lymphoid and myeloid lineages in recipients, resulting in stable and complete donor chimerism. In agreement with previous reports showing the effectiveness of WA in a mouse model of partial chimerism, our data further establishes that WA is able to attenuate aGVHD in an MHC-mismatched high dose chemo-conditioned murine model without compromising engraftment. This study provides compelling scientific basis for possible application of WA for prevention and treatment of aGVHD in patients receiving allo-BMT.


greenmedinfo--2020-08-16--Withanone and Withaferin-A are predicted to interact with transmembrane protease serine 2 (TMPRSS2)

Withanone and Withaferin-A are predicted to interact with transmembrane protease serine 2 (TMPRSS2) and block entry of SARS-CoV-2 into cells.

Coronavirus disease 2019 ( COVID-19 ) initiated in December 2019 in Wuhan , China and became pandemic causing high fatality and disrupted normal life calling world almost to a halt .
Causative agent is a novel coronavirus called Severe Acute Respiratory Syndrome Coronavirus 2 ( SARS-CoV-2/2019-nCoV ) .
While new line of drug/vaccine development has been initiated world-wide , in the current scenario of high infected numbers , severity of the disease and high morbidity , repurposing of the existing drugs is heavily explored .
Here , we used a homology-based structural model of transmembrane protease serine 2 ( TMPRSS2 @ @ @ @ @ @ @ required for entry of virus to the target host cell .
Using the strengths of molecular docking and molecular dynamics simulations , we examined the binding potential of Withaferin-A ( Wi-A ) , Withanone ( Wi-N ) and caffeic acid phenethyl ester to TPMRSS2 in comparison to its known inhibitor , Camostat mesylate .
We found that both Wi-A and Wi-N could bind and stably interact at the catalytic site of TMPRSS2 .
Wi-N showed stronger interactions with TMPRSS2 catalytic residues than Wi-A and was also able to induce changes in its allosteric @ @ @ @ @ @ @ effect of Wi-N on TMPRSS2 expression in MCF7 cells and found remarkable downregulation of TMPRSS2 mRNA in treated cells predicting dual action of Wi-N to block SARS-CoV-2 entry into the host cells .
Since the natural compounds are easily available/affordable , they may even offer a timely therapeutic/preventive value for the management of SARS-CoV-2 pandemic .
We also report that Wi-A/Wi-N content varies in different parts of Ashwagandha and warrants careful attention for their use.Communicated by Ramaswamy H. Sarma .

Coronavirus disease 2019 (COVID-19) initiated in December 2019 in Wuhan, China and became pandemic causing high fatality and disrupted normal life calling world almost to a halt. Causative agent is a novel coronavirus called Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2/2019-nCoV). While new line of drug/vaccine development has been initiated world-wide, in the current scenario of high infected numbers, severity of the disease and high morbidity, repurposing of the existing drugs is heavily explored. Here, we used a homology-based structural model of transmembrane protease serine 2 (TMPRSS2), a cell surface receptor, required for entry of virus to the target host cell. Using the strengths of molecular docking and molecular dynamics simulations, we examined the binding potential of Withaferin-A (Wi-A), Withanone (Wi-N) and caffeic acid phenethyl ester to TPMRSS2 in comparison to its known inhibitor, Camostat mesylate. We found that both Wi-A and Wi-N could bind and stably interact at the catalytic site of TMPRSS2. Wi-N showed stronger interactions with TMPRSS2 catalytic residues than Wi-A and was also able to induce changes in its allosteric site. Furthermore, we investigated the effect of Wi-N on TMPRSS2 expression in MCF7 cells and found remarkable downregulation of TMPRSS2 mRNA in treated cells predicting dual action of Wi-N to block SARS-CoV-2 entry into the host cells. Since the natural compounds are easily available/affordable, they may even offer a timely therapeutic/preventive value for the management of SARS-CoV-2 pandemic. We also report that Wi-A/Wi-N content varies in different parts of Ashwagandha and warrants careful attention for their use.Communicated by Ramaswamy H. Sarma.


greenmedinfo--2020-08-16--Withaferin A attenuates ovarian cancer-induced cardiac cachexia.

Cachexia is a common multifactorial syndrome in the advanced stages of cancer and accounts for approximately 20-30 % of all cancer-related fatalities .
In addition to the progressive loss of skeletal muscle mass , cancer results in impairments in cardiac function .
We recently demonstrated that WFA attenuates the cachectic skeletal muscle phenotype induced by ovarian cancer .
The purpose of this study was to investigate whether ovarian cancer induces cardiac cachexia , the possible pathway involved , and whether WFA attenuates cardiac cachexia .
Xenografting of ovarian cancer induced cardiac cachexia , leading to the loss of normal heart @ @ @ @ @ @ @ heart weight .
Further , ovarian cancer induced systolic dysfunction and diastolic dysfunction Treatment with WFA preserved systolic function in tumor-bearing mice , but diastolic dysfunction was partially improved .
In addition , WFA abrogated the ovarian cancer-induced reduction in cardiomyocyte cross-sectional area .
Finally , treatment with WFA ameliorated fibrotic deposition in the hearts of tumor-bearing animals .
We observed a tumor-induced MHC isoform switching from the adult MHCα to the embryonic MHCβ isoform , which was prevented by WFA treatment .
Circulating Ang II level was increased significantly in the tumor-bearing @ @ @ @ @ @ @ .
Our results clearly demonstrated the induction of cardiac cachexia in response to ovarian tumors in female NSG mice.Further , we observed induction of proinflammatory markers through the AT1R pathway , which was ameliorated by WFA , in addition to amelioration of the cachectic phenotype , suggesting WFA as a potential therapeutic agent for cardiac cachexia in oncological paradigms .

Cachexia is a common multifactorial syndrome in the advanced stages of cancer and accounts for approximately 20-30% of all cancer-related fatalities. In addition to the progressive loss of skeletal muscle mass, cancer results in impairments in cardiac function. We recently demonstrated that WFA attenuates the cachectic skeletal muscle phenotype induced by ovarian cancer. The purpose of this study was to investigate whether ovarian cancer induces cardiac cachexia, the possible pathway involved, and whether WFA attenuates cardiac cachexia. Xenografting of ovarian cancer induced cardiac cachexia, leading to the loss of normal heart functions. Treatment with WFA rescued the heart weight. Further, ovarian cancer induced systolic dysfunction and diastolic dysfunction Treatment with WFA preserved systolic function in tumor-bearing mice, but diastolic dysfunction was partially improved. In addition, WFA abrogated the ovarian cancer-induced reduction in cardiomyocyte cross-sectional area. Finally, treatment with WFA ameliorated fibrotic deposition in the hearts of tumor-bearing animals. We observed a tumor-induced MHC isoform switching from the adult MHCα to the embryonic MHCβ isoform, which was prevented by WFA treatment. Circulating Ang II level was increased significantly in the tumor-bearing, which was lowered by WFA treatment. Our results clearly demonstrated the induction of cardiac cachexia in response to ovarian tumors in female NSG mice.Further, we observed induction of proinflammatory markers through the AT1R pathway, which was ameliorated by WFA, in addition to amelioration of the cachectic phenotype, suggesting WFA as a potential therapeutic agent for cardiac cachexia in oncological paradigms.


greenmedinfo--2020-08-17--Oxidative stress induced by piperine leads to apoptosis in Candida albicans.

: Candida albicans is a member of pathogens with potential drug resistance threat that needs novel chemotherapeutic strategies .
Considering the multifarious biological activities including bioenhancer activity , anti-Candida potential of piperine was evaluated against planktonic/biofilm and hyphal growth of C. albicans alone or in combination as a synergistic agent with fluconazole .
Piperine inhibits planktonic growth at or less than 15 μg/ml , hyphae induction at 5 μg/ml concentration , and exhibits stage-dependent activity against biofilm growth of a fluconazole-resistant strain of C. albicans ( ATCC10231 ) .
Though piperine could n't kill inoculum completely at minimum inhibitory concentration @ @ @ @ @ @ @ at higher concentrations , as shown in apoptosis assay .
FIC index values indicate that piperine exhibits excellent synergistic activity with fluconazole against planktonic ( 0.123 ) and biofilm ( 0.215 ) growth of an FLC resistant strain .
Mode of anti-Candida activity was studied by identifying piperine responsive proteins wherein the abundance of 25 proteins involved in stress response , signal transduction and cell cycle were modulated ( 22 up and 3 down-regulated ) significantly in response to piperine ( MIC50 ) .
Modulation of the proteins involved suggests that piperine affects @ @ @ @ @ @ @ by cell cycle arrest and apoptosis in C. albicans .
Flow cytometry-based mitochondrial membrane potential ( MMP ) , cell cycle and apoptosis assay , as well as real-time quantitative polymerase chain reaction analysis of selected genes , confirms piperine induced oxidative stress ( TRR1 ) , cell cycle arrest and apoptosis ( CaMCA1 ) .
Based on our results , we conclude that piperine inhibits planktonic and difficult-to treat-biofilm growth of C. albicans by affecting membrane integrity thereby inducing oxidative stress and apoptosis .
LAY ABSTRACT : Piperine inhibit Candida albicans growth @ @ @ @ @ @ @ our study .
Piperine exhibits excellent synergistic potential with fluconazole The proteome analysis suggests that piperine induced membrane damage leads to oxidative stress followed by cell cycle arrest and apoptosis .

: Candida albicans is a member of pathogens with potential drug resistance threat that needs novel chemotherapeutic strategies. Considering the multifarious biological activities including bioenhancer activity, anti-Candida potential of piperine was evaluated against planktonic/biofilm and hyphal growth of C. albicans alone or in combination as a synergistic agent with fluconazole. Piperine inhibits planktonic growth at or less than 15 μg/ml, hyphae induction at 5 μg/ml concentration, and exhibits stage-dependent activity against biofilm growth of a fluconazole-resistant strain of C. albicans (ATCC10231). Though piperine couldn't kill inoculum completely at minimum inhibitory concentration (MIC), it is fungicidal at higher concentrations, as shown in apoptosis assay. FIC index values indicate that piperine exhibits excellent synergistic activity with fluconazole against planktonic (0.123) and biofilm (0.215) growth of an FLC resistant strain. Mode of anti-Candida activity was studied by identifying piperine responsive proteins wherein the abundance of 25 proteins involved in stress response, signal transduction and cell cycle were modulated (22 up and 3 down-regulated) significantly in response to piperine (MIC50). Modulation of the proteins involved suggests that piperine affects membrane integrity leading to oxidative stress followed by cell cycle arrest and apoptosis in C. albicans. Flow cytometry-based mitochondrial membrane potential (MMP), cell cycle and apoptosis assay, as well as real-time quantitative polymerase chain reaction analysis of selected genes, confirms piperine induced oxidative stress (TRR1), cell cycle arrest and apoptosis (CaMCA1). Based on our results, we conclude that piperine inhibits planktonic and difficult-to treat-biofilm growth of C. albicans by affecting membrane integrity thereby inducing oxidative stress and apoptosis.
LAY ABSTRACT: Piperine inhibit Candida albicans growth (planktonic and biofilm) significantly in our study. Piperine exhibits excellent synergistic potential with fluconazole The proteome analysis suggests that piperine induced membrane damage leads to oxidative stress followed by cell cycle arrest and apoptosis.


greenmedinfo--2020-08-17--Piperine suppresses the Wnt/β-catenin pathway and has anti-cancer effects on colorectal cancer cells

Piperine suppresses the Wnt/β-catenin pathway and has anti-cancer effects on colorectal cancer cells.

More than 94 % of colorectal cancer cases have mutations in one or more Wnt/β-catenin signaling pathway components .
Inactivating mutations in APC or activating mutations in β-catenin ( CTNNB1 ) lead to signaling overactivation and subsequent intestinal hyperplasia .
Numerous classes of medicines derived from synthetic or natural small molecules , including alkaloids , have benefited the treatment of different diseases , including cancer , Piperine is a true alkaloid , derived from lysine , responsible for the spicy taste of black pepper ( Piper nigrum ) and long pepper ( Piper longum ) .
Studies have shown that @ @ @ @ @ @ @ properties ; however , piperine molecular mechanisms of action are still not fully understood .
By using Wnt/β-catenin pathway epistasis experiment we show that piperine inhibits the canonical Wnt pathway induced by overexpression of β-catenin , β-catenin S33A or dnTCF4 VP16 , while also suppressing β-catenin nuclear localization in HCT116 cell line .
Additionally , piperine impairs cell proliferation and migration in HCT116 , SW480 and DLD-1 colorectal tumor cell lines , while not affecting the non-tumoral cell line IEC-6 .
In summary , piperine inhibits the canonical Wnt signaling pathway and displays @ @ @ @ @ @ @

More than 94% of colorectal cancer cases have mutations in one or more Wnt/β-catenin signaling pathway components. Inactivating mutations in APC or activating mutations in β-catenin (CTNNB1) lead to signaling overactivation and subsequent intestinal hyperplasia. Numerous classes of medicines derived from synthetic or natural small molecules, including alkaloids, have benefited the treatment of different diseases, including cancer, Piperine is a true alkaloid, derived from lysine, responsible for the spicy taste of black pepper (Piper nigrum) and long pepper (Piper longum). Studies have shown that piperine has a wide range of pharmacological properties; however, piperine molecular mechanisms of action are still not fully understood. By using Wnt/β-catenin pathway epistasis experiment we show that piperine inhibits the canonical Wnt pathway induced by overexpression of β-catenin, β-catenin S33A or dnTCF4 VP16, while also suppressing β-catenin nuclear localization in HCT116 cell line. Additionally, piperine impairs cell proliferation and migration in HCT116, SW480 and DLD-1 colorectal tumor cell lines, while not affecting the non-tumoral cell line IEC-6. In summary, piperine inhibits the canonical Wnt signaling pathway and displays anti-cancer effectson colorectal cancer cell lines.


greenmedinfo--2020-08-17--Piperine synergistically enhances the effect of temozolomide against temozolomide-resistant human gl

Piperine synergistically enhances the effect of temozolomide against temozolomide-resistant human glioma cell lines.

Temozolomide ( TMZ ) is an alkylating chemotherapy agent used in the clinical treatment of glioblastoma multiforme ( GBM ) @ @ @ @ @ ) is a naturally occurring pungent nitrogenous substance present in the fruits of peppers .
@ @ @ @ @ of PIP alone and in combination with TMZ in GBM cellsusingparameters such as cell proliferation @ @ @ @ @ activities , cell cycle kinetics , wound-healing ability , and loss of mitochondrial membrane potential @ @ @ @ @ with PIP and alow concentration of PIP-TMZ , inhibited cell growth , similar to TMZ.PIP-TMZ @ @ @ @ @ caspase-8/-9/-3 , MMP loss , and inhibition ofwound-healing motility .
Reverse transcription polymerase chain reaction @ @ @ @ @ ( andexpression upon treatment with a low concentration PIP-TMZ , suggesting an S to G1 @ @ @ @ @ insight into the apoptotic potential of the combination of a low concentration of PIP-TMZ , @ @ @ @ @ for its validation .

Temozolomide (TMZ) is an alkylating chemotherapy agent used in the clinical treatment of glioblastoma multiforme (GBM) patients. Piperine (PIP) is a naturally occurring pungent nitrogenous substance present in the fruits of peppers. We investigated the anti-cancer efficacies of PIP alone and in combination with TMZ in GBM cellsusingparameters such as cell proliferation, cellular apoptosis,caspase-8/-9/-3 activities, cell cycle kinetics, wound-healing ability, and loss of mitochondrial membrane potential (MMP). Treatment with PIP and alow concentration of PIP-TMZ, inhibited cell growth, similar to TMZ.PIP-TMZ promoted apoptosis by activation of caspase-8/-9/-3, MMP loss, and inhibition ofwound-healing motility. Reverse transcription polymerase chain reaction analysis showed significant inhibition of(andexpression upon treatment with a low concentration PIP-TMZ, suggesting an S to G1 arrest. Our findings provide insight into the apoptotic potential of the combination of a low concentration of PIP-TMZ, though furtherstudy will be needed for its validation.