greenmedinfo--2020-10-14--In-depth characterization of antibacterial activity of melittin against Staphylococcus aureus.

Skin infections caused by methicillin-resistant Staphylococcus aureus ( MRSA ) require the development of new and effective topical antibiotics .
In this context , melittin , the main component of apitoxin , has a potent antibacterial effect .
However , little is known regarding the anti-inflammatory potential this peptide in infection models , or its ability to induce clinically important resistance .
Here , we aimed to conduct an in-depth characterization of the antibacterial potential of melittin in vitro and evaluate the pharmaceutical potential of an ointment containing melittin for the treatment of non-surgical infections induced by MRSA .
The @ @ @ @ @ @ @ 0.12 to 4μM .
The antibacterial effect was mainly bactericidal and fast ( approximately 0.5 h after incubation ) and was maintained even in stationary cells and mature MRSA biofilms .
Melittin interacts synergistically with beta-lactams and aminoglycosides , and its ability to form pores in the membrane reversesthe resistance of vancomycin-intermediate Staphylococcus aureus ( VISA ) to amoxicillin , and vancomycin .
Its ability to induce resistance in vitro was absent , and melittin was stable in several conditions often associated with infected wounds .
In vivo , a ointment containing melittin @ @ @ @ @ @ @ pro-inflammatory cytokines , such as tumor necrosis factor-α , interleukin-6 ( IL-6 ) , and IL-1 beta .
Collectively , these data point to melittin as a potential candidate for topical formulations aimed at the treatment of non-surgical infections caused by MRSA .

Skin infections caused by methicillin-resistant Staphylococcus aureus (MRSA) require the development of new and effective topical antibiotics. In this context, melittin, the main component of apitoxin, has a potent antibacterial effect. However, little is known regarding the anti-inflammatory potential this peptide in infection models, or its ability to induce clinically important resistance. Here, we aimed to conduct an in-depth characterization of the antibacterial potential of melittin in vitro and evaluate the pharmaceutical potential of an ointment containing melittin for the treatment of non-surgical infections induced by MRSA. The minimum inhibitory concentration of melittin varied from 0.12 to 4μM. The antibacterial effect was mainly bactericidal and fast (approximately 0.5 h after incubation) and was maintained even in stationary cells and mature MRSA biofilms. Melittin interacts synergistically with beta-lactams and aminoglycosides, and its ability to form pores in the membrane reversesthe resistance of vancomycin-intermediate Staphylococcus aureus (VISA) to amoxicillin, and vancomycin. Its ability to induce resistance in vitro was absent, and melittin was stable in several conditions often associated with infected wounds. In vivo, a ointment containing melittin reduced bacterial load and the content of pro-inflammatory cytokines, such as tumor necrosis factor-α, interleukin-6 (IL-6), and IL-1 beta. Collectively, these data point to melittin as a potential candidate for topical formulations aimed at the treatment of non-surgical infections caused by MRSA.


greenmedinfo--2020-10-14--Melittin mitigated ulcerative colitis pathogenesis and could be considered as a potent and promising

Melittin mitigated ulcerative colitis pathogenesis and could be considered as a potent and promising therapeutic alternative treatment.

Ulcerative colitis ( UC ) is a chronic disease driven primarily by uncontrolled pervasive inflammatory responses affecting the colon and rectum .
Currently available medications carry multiple detrimental adverse effects , which have emphasized the mandatory need for safer and more efficient novel therapeutic alternatives .
Melittin is the main constituent of bee venom and exhibits potent anti-inflammatory properties .
The antiulcerogenic effect of oral melittin ( 40 μg/kg ) was explored in the current study using the acetic acid-induced colitis model .
Increase in body weight and decrease in colon mass index were observed in the melittin group .
@ @ @ @ @ @ @ damage .
Melittin administration has efficiently amended the elevated levels of the cytokines , tumor necrosis factor ( TNF-α ) and interleukin 6 ( IL-6 ) seen in the colitis group .
This was accompanied by inhibition of the upstream signaling molecules , Toll-like receptor 4 ( TLR4 ) , tumor necrosis factor receptor ( TNF-R ) -associated factor ( TRAF6 ) , mitogen-activated protein kinase 38 ( p38 MAPK ) , and nuclear factor kappaB ( NF-κB ) in the melittin group .
Moreover , treatment with melittin resulted in marked decrease @ @ @ @ @ @ @ PGE2 ) together with the enzymes involved in its synthesis , secretory phospholipase A2 ( sPLA2 ) and cyclooxygenase 2 ( COX-2 ) .
Additionally , melittin has attenuated acetic acid-induced oxidative stress as manifested by the significant diminishment in malondialdehyde ( MDA ) as well as the increase in superoxide dismutase ( SOD ) and reduced glutathione ( GSH ) levels .
Therefore , melittin mitigated UC pathogenesis and could be considered as a potent and promising therapeutic alternative for UC treatment .

Ulcerative colitis (UC) is a chronic disease driven primarily by uncontrolled pervasive inflammatory responses affecting the colon and rectum. Currently available medications carry multiple detrimental adverse effects, which have emphasized the mandatory need for safer and more efficient novel therapeutic alternatives. Melittin is the main constituent of bee venom and exhibits potent anti-inflammatory properties. The antiulcerogenic effect of oral melittin (40 μg/kg) was explored in the current study using the acetic acid-induced colitis model. Increase in body weight and decrease in colon mass index were observed in the melittin group. Microscopically, melittin ameliorated acetic acid-induced histological damage. Melittin administration has efficiently amended the elevated levels of the cytokines, tumor necrosis factor (TNF-α) and interleukin 6 (IL-6) seen in the colitis group. This was accompanied by inhibition of the upstream signaling molecules, Toll-like receptor 4 (TLR4), tumor necrosis factor receptor (TNF-R)-associated factor (TRAF6), mitogen-activated protein kinase 38 (p38 MAPK), and nuclear factor kappaB (NF-κB) in the melittin group. Moreover, treatment with melittin resulted in marked decrease in colonic level of prostaglandin E2 (PGE2) together with the enzymes involved in its synthesis, secretory phospholipase A2 (sPLA2) and cyclooxygenase 2 (COX-2). Additionally, melittin has attenuated acetic acid-induced oxidative stress as manifested by the significant diminishment in malondialdehyde (MDA) as well as the increase in superoxide dismutase (SOD) and reduced glutathione (GSH) levels. Therefore, melittin mitigated UC pathogenesis and could be considered as a potent and promising therapeutic alternative for UC treatment.


greenmedinfo--2020-10-14--Synergistic activity of melittin with mupirocin against methicillin-resistant S. Aureus.

Methicillin-Resistant ( MRSA ) biofilms are involved in various nosocomial infections , being in the limelight of academic research .
The current study aimed to determine the antimicrobial effects of melittin on planktonic and biofilm forms of .
Following the identification of MRSA and SCCmec types ( using PCR method ) , Minimum Inhibitory Concentration ( MIC ) , Minimum Bactericidal Concentration ( MBC ) , and fractional inhibitory concentration index ( FICi ) , for melittin and mupirocin were determined by broth microdilution assay .
Melittin anti-biofilm activity was determined , using a microtiter-plate test ( MtP ) and @ @ @ @ @ @ @ .
The quorum sensing inhibitory activity of½ MIC melittin was examined using a quantitative real-time RT-PCR method , and melittin cytotoxicity on Vero cells was examined by tetrazolium-based colorimetric ( MTT ) test .
The Results of our study showed that Geometric means of MIC values of the melittin and mupirocin were 4.4 and 14.22 μg/ml respectively .
The geometric mean of the FICi for both melittin-mupirocin was 0.75 .
Nobiofilm was formed andgene ( as a biofilm regulator ) expression down-regulated .
It seems that melittin can be useful in the treatment @ @ @ @ @ @ @ theexpression .
Furthermore , synergistic growth-inhibitory effects of mupirocin with melittin could be considered as a promising approach in the treatment of MRSA isolates .

Methicillin-Resistant(MRSA) biofilms are involved in various nosocomial infections, being in the limelight of academic research. The current study aimed to determine the antimicrobial effects of melittin on planktonic and biofilm forms of. Following the identification of MRSA and SCCmec types (using PCR method), Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC), and fractional inhibitory concentration index (FICi), for melittin and mupirocin were determined by broth microdilution assay. Melittin anti-biofilm activity was determined, using a microtiter-plate test (MtP) and scanning electron microscope (SEM) methods. The quorum sensing inhibitory activity of½ MIC melittin was examined using a quantitative real-time RT-PCR method, and melittin cytotoxicity on Vero cells was examined by tetrazolium-based colorimetric (MTT) test. The Results of our study showed that Geometric means of MIC values of the melittin and mupirocin were 4.4 and 14.22 μg/ml respectively. The geometric mean of the FICi for both melittin-mupirocin was 0.75. Nobiofilm was formed andgene (as a biofilm regulator) expression down-regulated. It seems that melittin can be useful in the treatment ofinfections (especially MRSA) by reducing theexpression. Furthermore, synergistic growth-inhibitory effects of mupirocin with melittin could be considered as a promising approach in the treatment of MRSA isolates.


greenmedinfo--2020-10-14--Melittin ameliorates inflammation in mouse acute liver failure via inhibition of PKM2-mediated Warbu

Melittin ameliorates inflammation in mouse acute liver failure via inhibition of PKM2-mediated Warburg effect.

Acute liver failure ( ALF ) is a fatal clinical syndrome with no special drug .
Recent evidence shows that modulation of macrophage to inhibit inflammation may be a promising strategy for ALF treatment .
In this study we investigated the potential therapeutic effects of melittin , a major peptide component of bee venom both in mice model of ALF and in LPS-stimulated macrophages in vitro , and elucidated the underlying mechanisms .
ALF was induced in mice by intraperitoneal injection of D-galactosamine/LPS .
Then the mice were treated with melittin ( 2 , 4 , and 8 mg/kg , @ @ @ @ @ @ @ treatment markedly improved mortality , attenuated severe symptoms and signs , and alleviated hepatic inflammation in D-galactosamine/LPS-induced ALF mice with the optimal dose being 4 mg/kg .
In addition , melittin within the effective doses did not cause significant in vivo toxicity .
In LPS-stimulated RAW264.7 macrophages , melittin ( 0.7 μM ) exerted anti-oxidation and anti-inflammation effects .
We showed that LPS stimulation promoted aerobic glycolysis of macrophages through increasing glycolytic rate , upregulated the levels of Warburg effect-related enzymes and metabolites including lactate , LDHA , LDH , and GLUT-1 , @ @ @ @ @ @ @ suppressed M2 isoform of pyruvate kinase ( PKM2 ) , thus disrupted the Warburg effect to alleviate inflammation .
Molecular docking analysis confirmed that melittin targetedPKM2 .
In LPS-stimulated RAW264.7 macrophages , knockdown of PKM2 caused similar anti-inflammation effects as melittin did .
In D-galactosamine/LPS-induced ALF mice , melittin treatment markedly decreased the expression levels of PKM2 and HIF-1α in liver .
This work demonstrates that melittin inhibits macrophage activation-mediated inflammation via inhibition of aerobic glycolysis by targeting PKM2 , which highlights a novel strategy of using melittin for ALF treatment .

Acute liver failure (ALF) is a fatal clinical syndrome with no special drug. Recent evidence shows that modulation of macrophage to inhibit inflammation may be a promising strategy for ALF treatment. In this study we investigated the potential therapeutic effects of melittin, a major peptide component of bee venom both in mice model of ALF and in LPS-stimulated macrophages in vitro, and elucidated the underlying mechanisms. ALF was induced in mice by intraperitoneal injection of D-galactosamine/LPS. Then the mice were treated with melittin (2, 4, and 8 mg/kg, ip). We showed that melittin treatment markedly improved mortality, attenuated severe symptoms and signs, and alleviated hepatic inflammation in D-galactosamine/LPS-induced ALF mice with the optimal dose being 4 mg/kg. In addition, melittin within the effective doses did not cause significant in vivo toxicity. In LPS-stimulated RAW264.7 macrophages, melittin (0.7 μM) exerted anti-oxidation and anti-inflammation effects. We showed that LPS stimulation promoted aerobic glycolysis of macrophages through increasing glycolytic rate, upregulated the levels of Warburg effect-related enzymes and metabolites including lactate, LDHA, LDH, and GLUT-1, and activated Akt/mTOR/PKM2/HIF-1α signaling. Melittin treatment suppressed M2 isoform of pyruvate kinase (PKM2), thus disrupted the Warburg effect to alleviate inflammation. Molecular docking analysis confirmed that melittin targetedPKM2. In LPS-stimulated RAW264.7 macrophages, knockdown of PKM2 caused similar anti-inflammation effects as melittin did. In D-galactosamine/LPS-induced ALF mice, melittin treatment markedly decreased the expression levels of PKM2 and HIF-1α in liver. This work demonstrates that melittin inhibits macrophage activation-mediated inflammation via inhibition of aerobic glycolysis by targeting PKM2, which highlights a novel strategy of using melittin for ALF treatment.


greenmedinfo--2020-10-14--Profiling anticancer and antioxidant activities of phenolic compounds present in black walnuts.

Our recent studies have demonstrated multiple health-promoting benefits from black walnut kernels .
These biological functions of black walnuts are likely associated with their bioactive constituents .
Characterization of phenolic compounds found in black walnut could point out underexplored bioactive activities of black walnut extracts and promote the development of novel applications of black walnut and its by-products .
In the present study , we assessed bioactivity profiles of phenolic compounds identified in the kernels of black walnuts using a high-throughput screening ( HTS ) approach .
Black walnut phenolic compounds were evaluated in terms of their total antioxidant capacity @ @ @ @ @ @ @ induction , and anticancer activities .
The anticancer activities were identified by evaluating the effects of the phenolic compounds on the growth of the tumorigenic alveolar epithelial cells ( A549 ) and non-tumorigenic lung fibroblast cells ( MRC-5 ) .
Out of 16 phenolic compounds tested , several compounds ( penta -- galloyl -- d-glucose , epicatechin gallate , quercetin , ( - ) -epicatechin , rutin , quercetin 3 -- d-glucoside , gallic acid , ( + ) -catechin , ferulic acid , syringic acid ) exerted antioxidant activities that were significantly @ @ @ @ @ @ @ used as a control .
Two phenolic compounds , penta -- galloyl -- d-glucose and quercetin 3 -- d-glucoside , exhibited antiproliferative activities against both the tumorigenic alveolar epithelial cells ( A549 ) and non-tumorigenic lung fibroblast cells ( MRC-5 ) .
The antioxidant activity of black walnut is likely driven not only by penta -- galloyl -- d-glucose but also by a combination of multiple phenolic compounds .
Our findings suggested that black walnut extracts possibly possess anticancer activities and supported that penta -- galloyl -- d-glucose could be a potential bioactive @ @ @ @ @ @ @ .

Our recent studies have demonstrated multiple health-promoting benefits from black walnut kernels. These biological functions of black walnuts are likely associated with their bioactive constituents. Characterization of phenolic compounds found in black walnut could point out underexplored bioactive activities of black walnut extracts and promote the development of novel applications of black walnut and its by-products. In the present study, we assessed bioactivity profiles of phenolic compounds identified in the kernels of black walnuts using a high-throughput screening (HTS) approach. Black walnut phenolic compounds were evaluated in terms of their total antioxidant capacity, antioxidant response element (ARE) induction, and anticancer activities. The anticancer activities were identified by evaluating the effects of the phenolic compounds on the growth of the tumorigenic alveolar epithelial cells (A549) and non-tumorigenic lung fibroblast cells (MRC-5). Out of 16 phenolic compounds tested, several compounds (penta--galloyl--d-glucose, epicatechin gallate, quercetin, (-)-epicatechin, rutin, quercetin 3--d-glucoside, gallic acid, (+)-catechin, ferulic acid, syringic acid) exerted antioxidant activities that were significantly higher compared to Trolox, which was used as a control. Two phenolic compounds, penta--galloyl--d-glucose and quercetin 3--d-glucoside, exhibited antiproliferative activities against both the tumorigenic alveolar epithelial cells (A549) and non-tumorigenic lung fibroblast cells (MRC-5). The antioxidant activity of black walnut is likely driven not only by penta--galloyl--d-glucose but also by a combination of multiple phenolic compounds. Our findings suggested that black walnut extracts possibly possess anticancer activities and supported that penta--galloyl--d-glucose could be a potential bioactive agent for the cosmetic and pharmaceutical industries.


greenmedinfo--2020-10-14--Ferulic acid influences Nrf2 activation to restore testicular tissue from cadmium-induced oxidative

Ferulic acid influences Nrf2 activation to restore testicular tissue from cadmium-induced oxidative challenge, inflammation, and apoptosis.

Here , we examined the protective effect of ferulic acid ( FA ) on cadmium chloride ( CdCl ) -mediated reproductive toxicity in male rats .
Animals were divided into four groups : control , FA ( 20 mg/kg ) , CdCl ( 6.5 mg/kg ) , and FA + CdCl .
CdCltreatment evoked a significant increase in testis cadmium concentration in addition to obvious increase in testosterone , luteinizing hormone , and follicle-stimulating hormone levels .
Moreover , CdCl-induced oxidative damage through exhausting the cellular defenses ( superoxide dismutase , catalase , glutathione peroxidase , glutathione reductase , and @ @ @ @ @ @ @ erythroid 2-related factor 2 ( Nrf2 ) expression accompanied by increases of malondialdehyde and nitric oxide contents .
Testicular inflammation was evident indicated by increased levels of interleukin-1β and tumor necrosis factor-α in CdCl-treated rats .
CdClexposure also decreased the expression of the proliferating cell nuclear antigen and augmented apoptotic events associated with prominent histopathological alterations .
However , FA coadministration mitigated the impaired hormonal level , apoptotic and inflammatory injuries elicited by CdCland maintained the oxidant/antioxidant balance in testicular tissue via Nrf2 activation .
PRACTICAL APPLICATIONS : Cadmium is an environmental toxicant @ @ @ @ @ @ @ reproductive toxicity .
However , antioxidant application has been found to protect against heavy metals-mediated toxic effects .
Here , we examined the potential protective efficacy of ferulic acid against cadmium-mediated testicular impairments through estimating the amount of cadmium in the testis , hormonal profile , oxidative status , inflammatory response , apoptotic and proliferating markers in addition to the histopathological alterations .
The obtained findings revealed that ferulic acid supplementation was able to abolish the testicular damages coupled with cadmium exposure .
The protective efficiency of ferulic acid may correlated with its @ @ @ @ @ @ @ activities ; suggesting that ferulic acid may be used to ameliorate cadmium-induced testicular deficits .

Here, we examined the protective effect of ferulic acid (FA) on cadmium chloride (CdCl)-mediated reproductive toxicity in male rats. Animals were divided into four groups: control, FA (20 mg/kg), CdCl(6.5 mg/kg), and FA + CdCl. CdCltreatment evoked a significant increase in testis cadmium concentration in addition to obvious increase in testosterone, luteinizing hormone, and follicle-stimulating hormone levels. Moreover, CdCl-induced oxidative damage through exhausting the cellular defenses (superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, and glutathione) and downregulating the nuclear factor erythroid 2-related factor 2 (Nrf2) expression accompanied by increases of malondialdehyde and nitric oxide contents. Testicular inflammation was evident indicated by increased levels of interleukin-1β and tumor necrosis factor-α in CdCl-treated rats. CdClexposure also decreased the expression of the proliferating cell nuclear antigen and augmented apoptotic events associated with prominent histopathological alterations. However, FA coadministration mitigated the impaired hormonal level, apoptotic and inflammatory injuries elicited by CdCland maintained the oxidant/antioxidant balance in testicular tissue via Nrf2 activation. PRACTICAL APPLICATIONS: Cadmium is an environmental toxicant and known to cause adverse effects including reproductive toxicity. However, antioxidant application has been found to protect against heavy metals-mediated toxic effects. Here, we examined the potential protective efficacy of ferulic acid against cadmium-mediated testicular impairments through estimating the amount of cadmium in the testis, hormonal profile, oxidative status, inflammatory response, apoptotic and proliferating markers in addition to the histopathological alterations. The obtained findings revealed that ferulic acid supplementation was able to abolish the testicular damages coupled with cadmium exposure. The protective efficiency of ferulic acid may correlated with its strong antioxidant, anti-inflammatory, and antiapoptotic activities; suggesting that ferulic acid may be used to ameliorate cadmium-induced testicular deficits.


greenmedinfo--2020-10-14--Ferulic acid delayed amyloid β-induced pathological symptoms by autophagy pathway.

The amyloidβ ( Aβ ) generation or aggregation plays a crucial role in Alzheimer 's disease ( AD ) .
Autophagy agonists , which function as the clearance of Aβ , could be the potential drug candidates against AD .
In staple food crops , ferulic acid ( FA ) is an enormously copious and almost ubiquitous phenolic antioxidant .
In the present study , FA significantly inhibited Aβ-induced pathological symptoms of paralysis and hypersensitivity to exogenous serotonin , meanwhile restrained Aβ monomers , oligomers , and deposits in AD C. elegans .
FA increased the expression of autophagy reporter @ @ @ @ @ @ @ , the autophagy inhibitors abolished the restrictive action of FA on the worm paralysis phenotype .
According to these results , FA triggered autophagy and ameliorated Aβ-induced pathological symptoms by the autophagy pathway .
Moreover , FA activated the HLH-30 transcription factor to nuclear localization , which acts upstream of autophagy in fasted animals , reduced the level of lipids , but affected nor the growth of E.coli OP50 , neither animal food intake behavior .
These suggest that FA induced a fasting-like effect to activate the autophagy pathway .
Additionally , FA @ @ @ @ @ @ @ disease worm .
Thus , FA could not only affect AD , broadly but also neurodegenerative diseases characterized by misfolded or aggregated proteins .

The amyloidβ (Aβ) generation or aggregation plays a crucial role in Alzheimer's disease (AD). Autophagy agonists, which function as the clearance of Aβ, could be the potential drug candidates against AD. In staple food crops, ferulic acid (FA) is an enormously copious and almost ubiquitous phenolic antioxidant. In the present study, FA significantly inhibited Aβ-induced pathological symptoms of paralysis and hypersensitivity to exogenous serotonin, meanwhile restrained Aβ monomers, oligomers, and deposits in AD C. elegans. FA increased the expression of autophagy reporter LGG-1 and enhanced autophagy flux. However, the autophagy inhibitors abolished the restrictive action of FA on the worm paralysis phenotype. According to these results, FA triggered autophagy and ameliorated Aβ-induced pathological symptoms by the autophagy pathway. Moreover, FA activated the HLH-30 transcription factor to nuclear localization, which acts upstream of autophagy in fasted animals, reduced the level of lipids, but affected nor the growth of E.coli OP50, neither animal food intake behavior. These suggest that FA induced a fasting-like effect to activate the autophagy pathway. Additionally, FA ameliorated poly Q aggregations in Huntington's disease worm. Thus, FA could not only affect AD, broadly but also neurodegenerative diseases characterized by misfolded or aggregated proteins.


greenmedinfo--2020-10-14--Quercetin, chrysin, caffeic acid and ferulic acid ameliorate cyclophosphamide-induced toxicities in

Quercetin, chrysin, caffeic acid and ferulic acid ameliorate cyclophosphamide-induced toxicities in SH-SY5Y cells.

Clinical use of cyclophosphamide ( CP ) causes apoptosis-induced cell death in the immune system , liver , heart and kidneys .
To prevent the cells against side effects of CP and its metabolites , increasing antioxidant defence mechanism of the body with supplemental antioxidants is important .
Therefore , there is a requirement for effective agents which could prevent the healthy cells from the harmful effects of drug-induced toxicities .
Several antioxidants have been used in protecting or alleviating CP-induced cell death .
However , no such study is reported in CP-induced SH-SY5Y cell toxicity .
The aim of @ @ @ @ @ @ @ effects of caffeic acid , chrysin , quercetin and ferulic acid against CP-induced toxicity in SH-SY5Y neuron cells .
In this study protective effects of quercetin , chrysin , caffeic acid and ferulic acid against CP-induced cell toxicity in SH-SY5Y cells was evaluated by cell proliferation assay , lipid peroxidation ( LPO ) analysis to decipher antioxidant capacity , tunel assay and qRT-PCR experiments to examine anti-apoptotic activities of the antioxidants .
The results showed that CP-induced cell toxicity in SH-SY5Y cells and treatments with the antioxidants suppressed the cell death .
Our @ @ @ @ @ @ @ cells via a decrease in LPO levels , downregulating the expression of Cas-3 , Cyt c and Bax and upregulating expression of anti-apoptotic gene Bcl-2 .
The use of antioxidant as nutritional supplements , and in particular of caffeic acid , chrysin , quercetin and ferulic acid , reduce apoptotic effects of CP in SH-SY5Y cells that could add insight into therapeutic approaches to CP-induced cell injuries .

Clinical use of cyclophosphamide (CP) causes apoptosis-induced cell death in the immune system, liver, heart and kidneys. To prevent the cells against side effects of CP and its metabolites, increasing antioxidant defence mechanism of the body with supplemental antioxidants is important. Therefore, there is a requirement for effective agents which could prevent the healthy cells from the harmful effects of drug-induced toxicities. Several antioxidants have been used in protecting or alleviating CP-induced cell death. However, no such study is reported in CP-induced SH-SY5Y cell toxicity. The aim of this study was to evaluate likelihood ameliorative effects of caffeic acid, chrysin, quercetin and ferulic acid against CP-induced toxicity in SH-SY5Y neuron cells. In this study protective effects of quercetin, chrysin, caffeic acid and ferulic acid against CP-induced cell toxicity in SH-SY5Y cells was evaluated by cell proliferation assay, lipid peroxidation (LPO) analysis to decipher antioxidant capacity, tunel assay and qRT-PCR experiments to examine anti-apoptotic activities of the antioxidants. The results showed that CP-induced cell toxicity in SH-SY5Y cells and treatments with the antioxidants suppressed the cell death. Our results suggests that these anti-oxidants protected SH-SY5Y cells via a decrease in LPO levels, downregulating the expression of Cas-3, Cyt c and Bax and upregulating expression of anti-apoptotic gene Bcl-2. The use of antioxidant as nutritional supplements, and in particular of caffeic acid, chrysin, quercetin and ferulic acid, reduce apoptotic effects of CP in SH-SY5Y cells that could add insight into therapeutic approaches to CP-induced cell injuries.


greenmedinfo--2020-10-14--Chrysin mitigates bleomycin-induced pulmonary fibrosis.

Pulmonary fibrosis is a chronic condition characterized by fibroblast proliferation , and the infiltration of inflammatory cells that can initiate local tissue hypoxia .
In this study the effect of chrysin ( 50 mg/kg/orally ) in a model of bleomycin ( BLM ) -induced pulmonary fibrosis was studied .
Chrysin managed to decrease mortality rate associated with BLM instillation and it managed to improve lung architecture and lung fibrosis by decreasing hydroxyproline content and transforming growth factor-β1 ( TGF-β1 ) protein expression .
Chrysin showed anti-inflammatory effect displayed by the decrease in inflammatory cells infiltrates , the decline in permeability @ @ @ @ @ @ @ in lactate dehydrogenase ( LDH ) activity .
Chrysin demonstrated potent antioxidant effect by decreasing lipid peroxidation , increasing antioxidant defense mechanisms by increasing superoxide dismutase ( SOD ) activity and reduced glutathione ( GSH ) content .
Additionally , the effect of chrysin on nitric oxide ( NOx ) content was assessed , where chrysin decreased NOx , increased the protein expression of endothelial nitric oxide synthase ( eNOS ) , and decreased inducible nitric oxide synthase ( iNOS ) protein expression .
Chrysin also succeeded in decreasing thioredoxin-interacting protein ( TXNIP @ @ @ @ @ @ @ system , showing potent antioxidant effect .
Finally , both tissueand bronchoalveolar lavage fluid contents of hypoxia inducible factor one alpha ( HIF1α ) were decreased by chrysin indicating that chrysin decreased local tissue hypoxia .
In conclusion , this study exposed a possible proof that chrysin could mitigate pulmonary fibrosis induced by BLM through its anti-inflammatory , antioxidant , antifibrotic effects and its effect in alleviating hypoxia .

Pulmonary fibrosis is a chronic condition characterized by fibroblast proliferation, and the infiltration of inflammatory cells that can initiate local tissue hypoxia. In this study the effect of chrysin (50 mg/kg/orally) in a model of bleomycin (BLM)-induced pulmonary fibrosis was studied. Chrysin managed to decrease mortality rate associated with BLM instillation and it managed to improve lung architecture and lung fibrosis by decreasing hydroxyproline content and transforming growth factor-β1 (TGF-β1) protein expression. Chrysin showed anti-inflammatory effect displayed by the decrease in inflammatory cells infiltrates, the decline in permeability of the alveolar/capillary barrier and the reduction in lactate dehydrogenase (LDH) activity. Chrysin demonstrated potent antioxidant effect by decreasing lipid peroxidation, increasing antioxidant defense mechanisms by increasing superoxide dismutase (SOD) activity and reduced glutathione (GSH) content. Additionally, the effect of chrysin on nitric oxide (NOx) content was assessed, where chrysin decreased NOx, increased the protein expression of endothelial nitric oxide synthase (eNOS), and decreased inducible nitric oxide synthase (iNOS) protein expression. Chrysin also succeeded in decreasing thioredoxin-interacting protein (TXNIP), the negative regulator of thioredoxin system, showing potent antioxidant effect. Finally, both tissueand bronchoalveolar lavage fluid contents of hypoxia inducible factor one alpha (HIF1α) were decreased by chrysin indicating that chrysin decreased local tissue hypoxia. In conclusion, this study exposed a possible proof that chrysin could mitigate pulmonary fibrosis induced by BLM through its anti-inflammatory, antioxidant, antifibrotic effects and its effect in alleviating hypoxia.


greenmedinfo--2020-10-14--Chrysin inhibits NF-κB-dependent CCL5 transcription in the atopic dermatitis-like inflammatory micro

Chrysin inhibits NF-κB-dependent CCL5 transcription in the atopic dermatitis-like inflammatory microenvironment.

Chrysin ( 5,7-dihydroxyflavone ) is a natural polyphenolic compound that induces an anti-inflammatory response .
In this study , we @ @ @ @ @ the chrysin-induced suppression of C-C motif chemokine ligand 5 ( ) gene expression in atopic @ @ @ @ @ inflammatory microenvironment .
We showed that chrysin inhibitedexpression at the transcriptional level through the suppression @ @ @ @ @ ( NF-κB ) in the inflammatory environment .
Chrysin could bind to the ATP-binding pocket @ @ @ @ @ ( IκB ) kinase ( IKK ) and , subsequently , prevent IκB degradation and @ @ @ @ @ efficacy of chrysin in targeting IKK was evaluated in 2,4-dinitrochlorobenzene-induced skin lesions in BALB/c mice @ @ @ @ @ chrysin preventedexpression by targeting IKK to reduce the infiltration of mast cells to the inflammatory @ @ @ @ @ attenuate the inflammatory responses .
These findings suggested that chrysin might be useful as a @ @ @ @ @ synthesis of small-molecule IKK-targeting drugs for the treatment of chronic inflammatory diseases , such as @ @

Chrysin (5,7-dihydroxyflavone) is a natural polyphenolic compound that induces an anti-inflammatory response. In this study, we investigated the molecular mechanism underlying the chrysin-induced suppression of C-C motif chemokine ligand 5 () gene expression in atopic dermatitis (AD)-like inflammatory microenvironment. We showed that chrysin inhibitedexpression at the transcriptional level through the suppression of nuclear factor kappa B (NF-κB) in the inflammatory environment. Chrysin could bind to the ATP-binding pocket of the inhibitor of κB (IκB) kinase (IKK) and, subsequently, prevent IκB degradation and NF-κB activation. The clinical efficacy of chrysin in targeting IKK was evaluated in 2,4-dinitrochlorobenzene-induced skin lesions in BALB/c mice. Our results suggested that chrysin preventedexpression by targeting IKK to reduce the infiltration of mast cells to the inflammatory sites and at least partially attenuate the inflammatory responses. These findings suggested that chrysin might be useful as a platform for the design and synthesis of small-molecule IKK-targeting drugs for the treatment of chronic inflammatory diseases, such as AD.


greenmedinfo--2020-10-14--Chrysin reduces anxiety-like behavior through actions on GABA A receptors during metestrus-diestrus.

Low concentrations of ovarian hormones , among other factors , are associated with greater vulnerability to negative effects of environmental stressors and may trigger anxiety symptoms in females .
The flavonoid chrysin ( 5,7-dihydroxyflavone ) exerts anxiolytic-like effects in male and ovariectomized female rats , but it is unknown if chrysin could reduce anxiety-like behavior that naturally occurs through the ovarian cycle phases .
The present study evaluated the effect of chrysin on anxiety-like behavior associated with the ovarian cycle phases in rats and the participation ofγ-aminobutyric acid-A ( GABA ) receptors in these actions .
The acute effects of @ @ @ @ @ @ @ in female cycling Wistar rats in the elevated plus maze , locomotor activity test , and light/dark test .
Diazepam ( 2 mg/kg ) was used as reference anxiolytic drug .
The participation of GABAreceptor in the anxiolytic actions of chrysin was explored by pretreating the rats with the noncompetitive GABAchloride ion channel antagonist picrotoxin ( 1 mg/kg ) .
Chrysin and diazepam prevented anxiety-like behavior that was associated with the metestrus-diestrus phase in both the elevated plus maze and light/dark test , and these effects were reversed by picrotoxin , with no @ @ @ @ @ @ @ No significant motor effects of chrysin were detected in either behavioral test during proestrus-estrus or metestrus-diestrus phases , whereas diazepam produced motor hypoactivity in the locomotor activity test during proestrus-estrus phase .
These results indicate that the flavonoid chrysin prevents anxiety-like behavior that naturally occurs during metestrus-diestrus in two unconditioned models that are used to evaluate anxiety-like behavior , and these effects were mediated by actions on GABAreceptors .

Low concentrations of ovarian hormones, among other factors, are associated with greater vulnerability to negative effects of environmental stressors and may trigger anxiety symptoms in females. The flavonoid chrysin (5,7-dihydroxyflavone) exerts anxiolytic-like effects in male and ovariectomized female rats, but it is unknown if chrysin could reduce anxiety-like behavior that naturally occurs through the ovarian cycle phases. The present study evaluated the effect of chrysin on anxiety-like behavior associated with the ovarian cycle phases in rats and the participation ofγ-aminobutyric acid-A (GABA) receptors in these actions. The acute effects of chrysin (2 mg/kg) were investigated in female cycling Wistar rats in the elevated plus maze, locomotor activity test, and light/dark test. Diazepam (2 mg/kg) was used as reference anxiolytic drug. The participation of GABAreceptor in the anxiolytic actions of chrysin was explored by pretreating the rats with the noncompetitive GABAchloride ion channel antagonist picrotoxin (1 mg/kg). Chrysin and diazepam prevented anxiety-like behavior that was associated with the metestrus-diestrus phase in both the elevated plus maze and light/dark test, and these effects were reversed by picrotoxin, with no significant changes in spontaneous locomotor activity. No significant motor effects of chrysin were detected in either behavioral test during proestrus-estrus or metestrus-diestrus phases, whereas diazepam produced motor hypoactivity in the locomotor activity test during proestrus-estrus phase. These results indicate that the flavonoid chrysin prevents anxiety-like behavior that naturally occurs during metestrus-diestrus in two unconditioned models that are used to evaluate anxiety-like behavior, and these effects were mediated by actions on GABAreceptors.


greenmedinfo--2020-10-14--Chrysin-loaded PLGA attenuates OVA-induced allergic asthma by modulating TLR/NF-κB/NLRP3 axis.

Asthma , one of the significant public health problems , is triggered by certain inflammatory processes in the airways that @ @ @ @ @ current therapies .
Though pieces of evidence on allergic asthma mitigation by the anti-inflammatory bioflavonoid @ @ @ @ @ accumulating , poor bioavailability , and low solubility curtail drug development .
To overcome these @ @ @ @ @ ( CHR-NP ) was formulated , and its salutary effect in preclinical murine allergic asthma @ @ @ @ @ was evaluated .
The spherical nanosized particles showed slow , sustained release in vitro .
@ @ @ @ @ the serum IgE , ovalbumin ( OVA ) -induced lung histological alteration , as well @ @ @ @ @ ) cytokines in the bronchoalveolar lavage fluid ( BALF ) .
It also suppressed the @ @ @ @ @ their upstream TLR/NF-κB/NLRP3 pathway activation in lung superior to CHR and almost identical to dexamethasone @ @ @ @ @ this study suggests the potentiality of CHR-NP in ameliorating allergic asthma progression .

Asthma, one of the significant public health problems, is triggered by certain inflammatory processes in the airways that are not addressed propitiously by current therapies. Though pieces of evidence on allergic asthma mitigation by the anti-inflammatory bioflavonoid chrysin (CHR) are accumulating, poor bioavailability, and low solubility curtail drug development. To overcome these shortcomings, CHR loaded nanoparticle (CHR-NP) was formulated, and its salutary effect in preclinical murine allergic asthma model via the peroral route was evaluated. The spherical nanosized particles showed slow, sustained release in vitro. Moreover, CHR-NP dramatically reduced the serum IgE, ovalbumin (OVA)-induced lung histological alteration, as well as Th2 (T-helper 2) cytokines in the bronchoalveolar lavage fluid (BALF). It also suppressed the elevated serum pro-inflammatory cytokines and their upstream TLR/NF-κB/NLRP3 pathway activation in lung superior to CHR and almost identical to dexamethasone (DEX). Thus this study suggests the potentiality of CHR-NP in ameliorating allergic asthma progression.


greenmedinfo--2020-10-14--Chrysin attenuates the NLRP3 inflammasome cascade to reduce synovitis and pain in knee osteoarthriti

Chrysin attenuates the NLRP3 inflammasome cascade to reduce synovitis and pain in knee osteoarthritis.

Purpose : Our recent reports have revealed that inhibiting NLRP3 activation reduces synovial inflammation and fibrosis in knee osteoarthritis ( KOA ) .
Synovial inflammation is involved the entire process of KOA and promotes the progression of KOA .
Natural flavonoid Chrysin from Scutellariae Radix , a traditional Chinese medicine , exhibits multifarious biological activities and potentially has protective activity against osteoarthritis .
However , the mechanism of Chrysin in the treatment of synovial inflammation remains elusive .
The purpose of our research was to explore the anti-inflammatory effects of Chrysin on KOA , which was induced by monoiodoacetic acid @ @ @ @ @ @ @ the NLRP3 inflammasome in the hopes of identifying an effective drug to treat KOA .
Methods : The MIA-induced KOA model was used to evaluate the cold pain threshold and paw withdrawal threshold ( PWT ) of joints after MIA ( 40 mg/mL ) injection into the knee joints .
Microscopically , we used LPS ( 5 ug/mL ) and ATP ( 4 mmol/L ) to stimulate fibroblast-like synovial cells ( FLSs ) to explore the underlying mechanisms and effects of Chrysin .
Two staining methods , H & E and Sirius Red @ @ @ @ @ @ @ in synovial membranes .
Cellular signal transduction was determined by qRT-PCR and WB .
Cytokine expression ( inflammatory cytokines and pain-related cytokines ) was detected by ELISA .
The degree of chronic inflammatory pain was evaluated by c-Fos immunofluorescence .
Results : The results showed that Chrysin not only attenuated synovial inflammation but also reduced the secretion of pain-related factors and increased the PWT and cold pain threshold in rats .
Chrysin also inhibited NLRP3 inflammasome activation and increased IL-1β levels to alleviate the synovitis .
Conclusion : Chrysin can relieve knee synovial @ @ @ @ @ @ @ rats , which may be related to the ability of Chrysin to inhibit NLRP3 inflammasome activation .
Therefore , Chrysin may be developed as a new drug for the treatment of KOA .

Purpose: Our recent reports have revealed that inhibiting NLRP3 activation reduces synovial inflammation and fibrosis in knee osteoarthritis (KOA). Synovial inflammation is involved the entire process of KOA and promotes the progression of KOA. Natural flavonoid Chrysin from Scutellariae Radix, a traditional Chinese medicine, exhibits multifarious biological activities and potentially has protective activity against osteoarthritis. However, the mechanism of Chrysin in the treatment of synovial inflammation remains elusive. The purpose of our research was to explore the anti-inflammatory effects of Chrysin on KOA, which was induced by monoiodoacetic acid (MIA) in rats by targeting the NLRP3 inflammasome in the hopes of identifying an effective drug to treat KOA.
Methods: The MIA-induced KOA model was used to evaluate the cold pain threshold and paw withdrawal threshold (PWT) of joints after MIA (40 mg/mL) injection into the knee joints. Microscopically, we used LPS (5 ug/mL) and ATP (4 mmol/L) to stimulate fibroblast-like synovial cells (FLSs) to explore the underlying mechanisms and effects of Chrysin. Two staining methods, H&E and Sirius Red, were applied to assess histopathological changes in synovial membranes. Cellular signal transduction was determined by qRT-PCR and WB. Cytokine expression (inflammatory cytokines and pain-related cytokines) was detected by ELISA. The degree of chronic inflammatory pain was evaluated by c-Fos immunofluorescence.
Results: The results showed that Chrysin not only attenuated synovial inflammation but also reduced the secretion of pain-related factors and increased the PWT and cold pain threshold in rats. Chrysin also inhibited NLRP3 inflammasome activation and increased IL-1β levels to alleviate the synovitis.
Conclusion: Chrysin can relieve knee synovial inflammation and improve pain behavior in KOA rats, which may be related to the ability of Chrysin to inhibit NLRP3 inflammasome activation. Therefore, Chrysin may be developed as a new drug for the treatment of KOA.


greenmedinfo--2020-10-14--Chrysin iInhibits melanoma tumor metastasis.

Melanoma , which features high metastasis and high lethality , is one of the toughest tumors to treat .
Chrysin @ @ @ @ @ various plants , has shown a great inhibitory effect on melanoma proliferation .
Here , @ @ @ @ @ effect of chrysin on melanomaand.
, chrysin effectively inhibited ankios resistance from 5μM cell migration @ @ @ @ @ , and tube formation capacity of melanoma cells from 20 μM .
We discovered that @ @ @ @ @ transition via regulating FOXM1/β-catenin signaling , as the expression of key regulatory factors was downregulated @ @ @ @ @ overexpression of FOXM1 will attenuate the antimetastasis effect of chrysin .
We also tested chrysin @ @ @ @ @ metastasis , where we found fewer tumors were formed in the lungs of chrysin-treated mice @ @ @ @ @ expression of FOXM1 was also downregulated by chrysin .
Collectively , our findings suggested the @ @ @ @ @ lower the metastatic rate of melanoma through regulating FOXM1/β-catenin signaling , indicating the application potential @ @ @ @ @ .

Melanoma, which features high metastasis and high lethality, is one of the toughest tumors to treat. Chrysin, which is rich in various plants, has shown a great inhibitory effect on melanoma proliferation. Here, we evaluated the metastasis suppressive effect of chrysin on melanomaand., chrysin effectively inhibited ankios resistance from 5μM cell migration, invasion from 10 μM, and tube formation capacity of melanoma cells from 20 μM. We discovered that chrysin interfered with the mesenchymal-epithelial transition via regulating FOXM1/β-catenin signaling, as the expression of key regulatory factors was downregulated by chrysin treatment, and overexpression of FOXM1 will attenuate the antimetastasis effect of chrysin. We also tested chrysin on lung colonization in melanoma metastasis, where we found fewer tumors were formed in the lungs of chrysin-treated mice. In addition, the expression of FOXM1 was also downregulated by chrysin. Collectively, our findings suggested the ability of chrysin treatment to lower the metastatic rate of melanoma through regulating FOXM1/β-catenin signaling, indicating the application potential of chrysin for melanoma therapy.


greenmedinfo--2020-10-14--Investigation of the protective role of chrysin within the framework of oxidative and inflammatory m

Investigation of the protective role of chrysin within the framework of oxidative and inflammatory markers in experimental testicular ischaemia/reperfusion injury.

This study was performed to evaluate the effect of chrysin on testicular torsion and detorsion damage in rats in terms of biochemistry , histopathology and immunohistochemistry .
The study was performed on Wistar albino rats between 250 g and 300 g. A total of 40 rats were used .
Five groups were created with eight rats in each group .
Group 1 was the control group , and no torsion procedure was performed .
In Group 2 , 2 hr of torsion and 2 hr of detorsion were applied .
In Group 3 , 2 hr of torsion and 24 @ @ @ @ @ @ @ Group 4 , 2 hr of torsion , 2 hr of detorsion and 50 mg/kg intraperitoneal chrysin were applied .
In Group 5 , 2 hr of torsion , 24 hr of detorsion and 50 mg/kg of chrysin were applied .
In the torsion/detorsion groups , the study determined decreases in glutathione and testosterone levels , increases in tumour necrosis factor-α , interleukin-4 , interleukin-6 and interleukin-10 levels , and increases in expression levels of caspase-3 and caspase-8 .
Chrysin application reduced malondialdehyde , tumour necrosis factor-α , caspase-3 and caspase-8 expression levels @ @ @ @ @ @ @ be used to reduce damage in cases of testicular ischaemia/reperfusion .
For more reliable results , further clinical trials are recommended .

This study was performed to evaluate the effect of chrysin on testicular torsion and detorsion damage in rats in terms of biochemistry, histopathology and immunohistochemistry. The study was performed on Wistar albino rats between 250 g and 300 g. A total of 40 rats were used. Five groups were created with eight rats in each group. Group 1 was the control group, and no torsion procedure was performed. In Group 2, 2 hr of torsion and 2 hr of detorsion were applied. In Group 3, 2 hr of torsion and 24 hr of detorsion were applied. In Group 4, 2 hr of torsion, 2 hr of detorsion and 50 mg/kg intraperitoneal chrysin were applied. In Group 5, 2 hr of torsion, 24 hr of detorsion and 50 mg/kg of chrysin were applied. In the torsion/detorsion groups, the study determined decreases in glutathione and testosterone levels,increases in tumour necrosis factor-α, interleukin-4, interleukin-6 and interleukin-10 levels, and increases in expression levels of caspase-3 and caspase-8. Chrysin application reduced malondialdehyde, tumour necrosis factor-α, caspase-3 and caspase-8 expression levels. We can say that chrysin can be used to reduce damage in cases of testicular ischaemia/reperfusion. For more reliable results, further clinical trials are recommended.


greenmedinfo--2020-10-15--Anti-cancer effect of gingerol in cancer prevention and treatment.

Cancer is one of the most lethal diseases in the world .
Because of high death rate associated with cancer and the side effects of chemotherapy and radiation therapy , patients required alternative strategies for its treatment .
Ginger ( Zingiber officinale ) has an enormous medicinal properties and health beneficial effects .
In this review , we discuss the basic mechanism by which gingerol ( a active component of ginger ) modulate a variety of cell signaling pathways linked to cancer , including nuclear factors ( NF-κB ) , signal transducer and activator of transcription 3 ( STAT3 ) @ @ @ @ @ @ @ β- catenin , growth factors receptors ( EGFR , VEGFR ) ; mitogen activated protein kinases ( MAPK ) and proinflammatory mediators ( TNF-α and COX-2 ) .
Both in vitro and in vivo studies support the role of gingerolin cancer .
The efficacy of gingerol by clinical trials has also been reported .
Importantly , natural agents are already in clinical trials against various kinds of cancer .
Effort has been made through this comprehensive review to highlight the recent developments and milestones achieved in cancer therapies via studies based on different @ @ @ @ @

Cancer is one of the most lethal diseases in the world. Because of high death rate associated with cancer and the side effects of chemotherapy and radiation therapy, patients required alternative strategies for its treatment. Ginger (Zingiber officinale) has an enormous medicinal properties and health beneficial effects. In this review, we discuss the basic mechanism by which gingerol (a active component of ginger) modulate a variety of cell signaling pathways linked to cancer, including nuclear factors (NF-κB), signal transducer and activator of transcription 3 (STAT3), activator protein1 (AP-1), β- catenin, growth factors receptors (EGFR, VEGFR); mitogen activated protein kinases (MAPK) and proinflammatory mediators (TNF-α and COX-2). Both in vitro and in vivo studies support the role of gingerolin cancer. The efficacy of gingerol by clinical trials has also been reported. Importantly, natural agents are already in clinical trials against various kinds of cancer. Effort has been made through this comprehensive review to highlight the recent developments and milestones achieved in cancer therapies via studies based on different cell lines using gingerol.


greenmedinfo--2020-10-15--6-gingerol pre-treatment can alleviate hypoxia/reoxygenation-induced H9C2 myocardial injury.

Objetive : To observe the effect of 6-gingerol ( 6-G ) pretreatment on hypoxia/reoxygenation ( H/R ) induced injury in H9C2 myocardial cell and investigate its related mechanism .
Methods : The H/Rmodel of cardiomyocytes was prepared by conventional methods .
In detail , H9C2 cells were added with the nitrogen-saturated hypoxic liquid , and placed in an incubator , mixed with gas ( 1 % O , 5 % CO , 94 % N ) applying for 15 min .
After culturing for 3 h , the cells were taken out and placed in an incubator ( 37℃ , @ @ @ @ @ @ @ Before establishing the cell model , the cells were pretreated with 6-G , and the cell viability was measured by MTT method to observe the protective effect of different concentrations of 6-G on H/R-induced cell damage .
The 6-G mass concentration for pretreatment that led to the highest cell viability was used for follow-up experiments .
DCFH-DA fluorescent probe was used to detect the effect of 6-G pretreatment on H9C2 oxidative stress level , and the intracellular oxidative stress was observed with fluorescence microscope and flow cytometry .
Western blot method was used @ @ @ @ @ @ @ factor-α ( TNF-α ) , interleukin-6 ( IL-6 ) and interleukin-1β ( IL-1β ) in H/R-induced cell inflammatory responses .
Results : Compared with the H/R group , the cell viability of the 6-G+H/R group began to increase when the concentration of 6-G promoted to50μg/mL .
The cell viability was the highest after pretreated with 200 μg/mL 6-G .
Therefore , 200 μg/mL was considered as the best 6-G intervention concentration for subsequent experiment .
The content of reactive oxygen species ( ROS ) in the 200 μg/mL 6-G group had no significant @ @ @ @ @ @ @ > 0.05 ) , and the ROS fluorescence peak did not migrate significantly .
However the ROS content in the H/R group increased significantly compared with the control ( < 0.05 ) , and the ROS fluorescence peak shifted to the right .
Compared with the H/R group , the ROS content of the 6-G+H/R group decreased ( < 0.05 ) , and the ROS fluorescence peak shifted to the left .
Compared with the control group , the expressions of TNF-α , IL-6 , IL-1β in the 6-G group had no significant @ @ @ @ @ @ @ expressions of TNF-α , IL-6 , IL-1β in the H/R group increased ( < 0.05 ) .
Compared with H/R group , the expressions of TNF-α , IL-6 and IL-1β in 6-G+H/R group decreased ( < 0.05 ) .
Conclusion : 6-G pretreatment can alleviate H/R-induced H9C2 myocardial injury , which may be related to the inhibition of oxidative stress and inflammatory responses .

Objetive: To observe the effect of 6-gingerol (6-G) pretreatment on hypoxia/reoxygenation (H/R) induced injury in H9C2 myocardial cell and investigate its related mechanism.
Methods: The H/Rmodel of cardiomyocytes was prepared by conventional methods. In detail, H9C2 cells were added with the nitrogen-saturated hypoxic liquid, and placed in an incubator, mixed with gas (1% O, 5% CO, 94% N) applying for 15 min. After culturing for 3 h, the cells were taken out and placed in an incubator (37℃, 5% CO) for 1 h. Before establishing the cell model, the cells were pretreated with 6-G, and the cell viability was measured by MTT method to observe the protective effect of different concentrations of 6-G on H/R-induced cell damage. The 6-G mass concentration for pretreatment that led to the highest cell viability was used for follow-up experiments. DCFH-DA fluorescent probe was used to detect the effect of 6-G pretreatment on H9C2 oxidative stress level, and the intracellular oxidative stress was observed with fluorescence microscope and flow cytometry. Western blot method was used to detect the expression of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) in H/R-induced cell inflammatory responses.
Results: Compared with the H/R group, the cell viability of the 6-G+H/R group began to increase when the concentration of 6-G promoted to50μg/mL. The cell viability was the highest after pretreated with 200 μg/mL 6-G. Therefore, 200 μg/mL was considered as the best 6-G intervention concentration for subsequent experiment. The content of reactive oxygen species (ROS) in the 200 μg/mL 6-G group had no significant changes compared with the control group (>0.05), and the ROS fluorescence peak did not migrate significantly. However the ROS content in the H/R group increased significantly compared with the control (<0.05), and the ROS fluorescence peak shifted to the right. Compared with the H/R group, the ROS content of the 6-G+H/R group decreased (<0.05), and the ROS fluorescence peak shifted to the left. Compared with the control group, the expressions of TNF-α, IL-6, IL-1β in the 6-G group had no significant changes (>0.05); the expressions of TNF-α, IL-6, IL-1β in the H/R group increased (<0.05). Compared with H/R group, the expressions of TNF-α, IL-6 and IL-1β in 6-G+H/R group decreased (<0.05).
Conclusion: 6-G pretreatment can alleviate H/R-induced H9C2 myocardial injury, which may be related to the inhibition of oxidative stress and inflammatory responses.


greenmedinfo--2020-10-15--Shikonin induces cell death by inhibiting glycolysis in human testicular cancer I-10 and seminoma TC

Shikonin induces cell death by inhibiting glycolysis in human testicular cancer I-10 and seminoma TCAM-2 cells.

OBJECTIVE : To investigate the pattern of shikonin-induced cell death in testicular cancer cell I-10 and seminoma TCAM-2 cells and explore the possible mechanism in light of mitochondrial function and glycolysis .
METHODS : I-10 cells treated with 0 , 1.2 , 1.4 and 1.6μmol/L shikonin and TCAM-2 cells treated with 0 , 0.5 , 1 and 1.5 μmol/L shikonin were examined for mitochondrial membrane potential and production of reactive oxygen species ( ROS ) using JC-1 kit and ROS kit , respectively .
The levels of intracellular lactic acid in the cells were detected using a lactic acid kit @ @ @ @ @ @ @ the proliferation of the cells was assessed with MTT assay .
The death patterns of the cells were observed by transmission electron microscopy , and annexin V-FITC/PI double staining was used to detect cell apoptosis .
Western blotting was used to detect the relative expression levels of the apoptotic proteins Bax , Bcl-2 , and cleaved caspase-3 , the autophagy- related protein LC3B and glycolysis- related proteins PKM2 , GLUT1 and HK2 .
RESULTS : MTT assay showed that shikonin significantly inhibited the proliferation of I-10 and TCAM-2 cells in a time- and @ @ @ @ @ @ @ The ICvalues of shikonin in I-10 cells at 24 , 48 , and 72 h were 1.8 , 1.36 and 1.16μmol/L , as compared with 2.37 , 0.8 and 0.41 μmol/L in TCAM-2 cells , respectively .
Shikonin treatment significantly reduced mitochondrial membrane potential , increased ROS levels and lower the level of lactic acid in both I-10 and TCAM-2 cells ( < 0.05 ) .
Transmission electron microscopy and annexin V-FITC/PI double staining demonstrated that shikonin induced apoptosis and excessive autophagy in I-10 and TCAM-2 cells ( < 0.05 ) .
In @ @ @ @ @ @ @ treatment significantly down- regulated the expressions of Bax , Bcl-2 , cleaved caspase-3 , PKM2 , GLUT1 and HK2 , and up-regulated the expression of autophagy-related protein LC3B ( < 0.05 ) .
CONCLUSIONS : Shikonin can inhibit the proliferation , induce apoptosis and increase autophagy in both I-10 and TCAM-2 cells probably by affecting energy metabolism of the cells .

OBJECTIVE: To investigate the pattern of shikonin-induced cell death in testicular cancer cell I-10 and seminoma TCAM-2 cells and explore the possible mechanism in light of mitochondrial function and glycolysis.
METHODS: I-10 cells treated with 0, 1.2, 1.4 and 1.6μmol/L shikonin and TCAM-2 cells treated with 0, 0.5, 1 and 1.5 μmol/L shikonin were examined for mitochondrial membrane potential and production of reactive oxygen species (ROS) using JC-1 kit and ROS kit, respectively. The levels of intracellular lactic acid in the cells were detected using a lactic acid kit. The inhibitory effect of shikonin on the proliferation of the cells was assessed with MTT assay. The death patterns of the cells were observed by transmission electron microscopy, and annexin V-FITC/PI double staining was used to detect cell apoptosis. Western blotting was used to detect the relative expression levels of the apoptotic proteins Bax, Bcl-2, and cleaved caspase-3, the autophagy- related protein LC3B and glycolysis- related proteins PKM2, GLUT1 and HK2.
RESULTS: MTT assay showed that shikonin significantly inhibited the proliferation of I-10 and TCAM-2 cells in a time- and dose-dependent manner (<0.05). The ICvalues of shikonin in I-10 cells at 24, 48, and 72 h were 1.8, 1.36 and 1.16μmol/L, as compared with 2.37, 0.8 and 0.41 μmol/L in TCAM-2 cells, respectively. Shikonin treatment significantly reduced mitochondrial membrane potential, increased ROS levels and lower the level of lactic acid in both I-10 and TCAM-2 cells (<0.05). Transmission electron microscopy and annexin V-FITC/PI double staining demonstrated that shikonin induced apoptosis and excessive autophagy in I-10 and TCAM-2 cells (<0.05). In both I-10 and TCAM cells, shikonin treatment significantly down- regulated the expressions of Bax, Bcl-2, cleaved caspase-3, PKM2, GLUT1 and HK2, and up-regulated the expression of autophagy-related protein LC3B (<0.05).
CONCLUSIONS: Shikonin can inhibit the proliferation, induce apoptosis and increase autophagy in both I-10 and TCAM-2 cells probably by affecting energy metabolism of the cells.


greenmedinfo--2020-10-15--Citral interacts synergistically with norfloxacin against methicillin resistant Staphylococcus aureu

Citral interacts synergistically with norfloxacin against methicillin resistant Staphylococcus aureus.

BACKGROUND : Staphylococcus aureus ( SA ) , is a major human pathogen causing wide range of clinical infections , which has been further complicated by drug resistance like methicillin resistant S. aureus ( MRSA ) , vancomycin intermediate S. aureus ( VISA ) /vancomycin resistant S. aureus ( VRSA ) , etc .
The present study was aimed at determining anti-staphylococcal potential of citral against drug resistant clinical isolates alone and in combination with antibiotics .
PURPOSE : To assess the potential of citral in combination with norfloxacin in treating drug resistant infections of SA .
STUDY DESIGN : @ @ @ @ @ @ @ of citral and norfloxacin against drug resistant SA strains was evaluated .
Further the efficacy and possible mechanism of action of the combination was also evaluated using in vitro and in vivo assays .
METHOD : The anti-staphylococcal activity of each of the monoterpene and the antibiotic was determined in terms of MIC and the effective concentration of both compounds in combination was obtained by checkerboard assay .
In vivo efficacy and oral acute toxicity was evaluated in Swiss albino mice model .
To understand the mechanism of action , time-kill curve , @ @ @ @ @ @ @ salt tolerance and ethidium bromide efflux assays were performed .
RESULTS : Citral was found effective against clinical isolates of SA with MIC values ranging from 75 to 150 µg mlexhibiting bacteriostatic activity .
Citral interacted synergistically , reducing MIC of norfloxacin up to 32-folds with FICI ≤ 0.50 .
Citral did not affect cell wall , but could damage cell membrane , inhibit efflux pump and affect the membrane potential .
Citral could reduce the staphylococcal load of spleen and liver tissues in a dose-dependent manner which was further reduced when used @ @ @ @ @ @ @ not exhibit any mortality or morbidity up to 500 mg kgbody weight and found to prolong the post-antibiotic effect of norfloxacin .
CONCLUSION : Based on these observations , citral could be a lead candidate phytomolecule for further developing it into an anti-staphylococcal agent .
The observations of combination study will help in reducing the burden of antibiotics leading to delayed resistance development .

BACKGROUND: Staphylococcus aureus (SA), is a major human pathogen causing wide range of clinical infections, which has been further complicated by drug resistance like methicillin resistant S. aureus (MRSA), vancomycin intermediate S. aureus (VISA)/vancomycin resistant S. aureus (VRSA), etc. The present study was aimed at determining anti-staphylococcal potential of citral against drug resistant clinical isolates alone and in combination with antibiotics.
PURPOSE: To assess the potential of citral in combination with norfloxacin in treating drug resistant infections of SA.
STUDY DESIGN: In the present study, synergistic interaction of citral and norfloxacin against drug resistant SA strains was evaluated. Further the efficacy and possible mechanism of action of the combination was also evaluated using in vitro and in vivo assays.
METHOD: The anti-staphylococcal activity of each of the monoterpene and the antibiotic was determined in terms of MIC and the effective concentration of both compounds in combination was obtained by checkerboard assay. In vivo efficacy and oral acute toxicity was evaluated in Swiss albino mice model. To understand the mechanism of action, time-kill curve, bacteriolysis, leakage, membrane depolarization, salt tolerance and ethidium bromide efflux assays were performed.
RESULTS: Citral was found effective against clinical isolates of SA with MIC values ranging from 75 to 150 µg mlexhibiting bacteriostatic activity. Citral interacted synergistically, reducing MIC of norfloxacin up to 32-folds with FICI ≤ 0.50. Citral did not affect cell wall, but could damage cell membrane, inhibit efflux pump and affect the membrane potential. Citral could reduce the staphylococcal load of spleen and liver tissues in a dose-dependent manner which was further reduced when used in combination with norfloxacin. Citral did not exhibit any mortality or morbidity up to 500 mg kgbody weight and found to prolong the post-antibiotic effect of norfloxacin.
CONCLUSION: Based on these observations, citral could be a lead candidate phytomolecule for further developing it into an anti-staphylococcal agent. The observations of combination study will help in reducing the burden of antibiotics leading to delayed resistance development.


greenmedinfo--2020-10-15--Citral attenuated intestinal inflammation induced by Cronobacter sakazakii in newborn mice.

Necrotizing enterocolitis ( NEC ) is a serious inflammatory intestinal disorder with a high mortality rate , which occurs most commonly in newborn infants.
, a common contaminant in infant formula , is associated with NEC .
However , its role in NEC pathogenesis is unknown , and there are still no effective treatments for NEC .
Currently , natural bioactive products have been investigated for their beneficial effects in preventing microbial infection .
In this study , a neonatal mouse intestinal inflammation model was used to examine the protective effects of citral ( a natural bioactive product ) on-induced @ @ @ @ @ @ @ shown that citral reduced the number ofcells in ileal tissues , and mice treated with citral had a significantly higher body weight than-infected mice .
Citral treatment also ameliorated serious ileal tissue damages , including epithelial sloughing , villous rupture , and enterocyte apoptosis.infection upregulated the messenger RNA transcription levels of several inflammation-associated genes , increased production of IL-6 and TNF-α , and activated the NF-κB and MAPK signaling pathways in ileal tissues .
Citral treatment mitigated these inflammatory responses .
The apoptotic index and activities of caspase 3 , 8 , and @ @ @ @ @ @ @ but citral inhibited both enterocyte apoptosis and activations of these caspase .
These findings suggest that citral has protective effects on-induced intestinal inflammation in newborn mice , and it may play a future role in the management ofassociated infections and diseases .

Necrotizing enterocolitis (NEC) is a serious inflammatory intestinal disorder with a high mortality rate, which occurs most commonly in newborn infants., a common contaminant in infant formula, is associated with NEC. However, its role in NEC pathogenesis is unknown, and there are still no effective treatments for NEC. Currently, natural bioactive products have been investigated for their beneficial effects in preventing microbial infection. In this study, a neonatal mouse intestinal inflammation model was used to examine the protective effects of citral (a natural bioactive product) on-induced intestinal inflammation and damages. It was shown that citral reduced the number ofcells in ileal tissues, and mice treated with citral had a significantly higher body weight than-infected mice. Citral treatment also ameliorated serious ileal tissue damages, including epithelial sloughing, villous rupture, and enterocyte apoptosis.infection upregulated the messenger RNA transcription levels of several inflammation-associated genes, increased production of IL-6 and TNF-α, and activated the NF-κB and MAPK signaling pathways in ileal tissues. Citral treatment mitigated these inflammatory responses. The apoptotic index and activities of caspase 3, 8, and 9 increased in murine ileum afterinfection, but citral inhibited both enterocyte apoptosis and activations of these caspase. These findings suggest that citral has protective effects on-induced intestinal inflammation in newborn mice, and it may play a future role in the management ofassociated infections and diseases.


greenmedinfo--2020-10-15--Cytotoxicity of citral against melanoma cells.

Citral is a natural compound that has shown cytotoxic and antiproliferative effects on breast and hematopoietic cancer cells ; however , there are few studies on melanoma cells .
Oxidative stress is known to be involved in all stages of melanoma development and is able to modulate intracellular pathways related to cellular proliferation and death .
In this study , we hypothesize that citral exerts its cytotoxic effect on melanoma cells by the modulation of cellular oxidative status and/or intracellular signaling .
To test this hypothesis , we investigated the antiproliferative and cytotoxic effects of citral on B16F10 murine melanoma @ @ @ @ @ @ @ stress , DNA damage , cell death , and important signaling pathways , as these pathways , namely , extracellular signal-regulated kinases 1/2 ( ERK1/2 ) , AKT , and phosphatidylinositol-3 kinase , are involved in cell proliferation and differentiation .
The p53 and nuclear factor kappa B were also investigated due to their ability to respond to intracellular stress .
We observed that citral exerted antiproliferative and cytotoxic effects in B16F10 ; induced oxidative stress , DNA lesions , and p53 nuclear translocation ; and reduced nitric oxide levels and nuclear factor @ @ @ @ @ @ @ .
To investigate citral specificity , we used non-neoplastic human and murine cells , HaCaT ( human skin keratinocytes ) and NIH-3T3 cells ( murine fibroblasts ) , and observed that although citral effects were not specific for cancer cells , non-neoplastic cells were more resistant to citral than B16F10 .
These findings highlight the potential clinical utility of citral in melanoma , with a mechanism of action involving the oxidative stress generation , nitric oxide depletion , and interference in signaling pathways related to cell proliferation .

Citral is a natural compound that has shown cytotoxic and antiproliferative effects on breast and hematopoietic cancer cells; however, there are few studies on melanoma cells. Oxidative stress is known to be involved in all stages of melanoma development and is able to modulate intracellular pathways related to cellular proliferation and death. In this study, we hypothesize that citral exerts its cytotoxic effect on melanoma cells by the modulation of cellular oxidative status and/or intracellular signaling. To test this hypothesis, we investigated the antiproliferative and cytotoxic effects of citral on B16F10 murine melanoma cells evaluating its effects on cellular oxidative stress, DNA damage, cell death, and important signaling pathways, as these pathways, namely, extracellular signal-regulated kinases 1/2 (ERK1/2), AKT, and phosphatidylinositol-3 kinase, are involved in cell proliferation and differentiation. The p53 and nuclear factor kappa B were also investigated due to their ability to respond to intracellular stress. We observed that citral exerted antiproliferative and cytotoxic effects in B16F10; induced oxidative stress, DNA lesions, and p53 nuclear translocation; and reduced nitric oxide levels and nuclear factor kappa B, ERK1/2, and AKT. To investigate citral specificity, we used non-neoplastic human and murine cells, HaCaT (human skin keratinocytes) and NIH-3T3 cells (murine fibroblasts), and observed that although citral effects were not specific for cancer cells, non-neoplastic cells were more resistant to citral than B16F10. These findings highlight the potential clinical utility of citral in melanoma, with a mechanism of action involving the oxidative stress generation, nitric oxide depletion, and interference in signaling pathways related to cell proliferation.


greenmedinfo--2020-10-15--Citral reduces breast tumor growth by inhibiting the cancer stem cell marker ALDH1A3.

Breast cancer stem cells ( CSCs ) can be identified by increased Aldefluor fluorescence caused by increased expression of aldehyde dehydrogenase 1A3 ( ALDH1A3 ) , as well as ALDH1A1 and ALDH2 .
In addition to being a CSC marker , ALDH1A3 regulates gene expression via retinoic acid ( RA ) signaling and plays a key role in the progression and chemotherapy resistance of cancer .
Therefore , ALDH1A3 represents a druggable anti-cancer target of interest .
Since to date , there are no characterized ALDH1A3 isoform inhibitors , drugs that were previously described as inhibiting the activity of other @ @ @ @ @ @ @ .
Twelve drugs ( 3-hydroxy-dl-kynurenine , benomyl , citral , chloral hydrate , cyanamide , daidzin , DEAB , disulfiram , gossypol , kynurenic acid , molinate , and pargyline ) were compared for their efficacy in inducing apoptosis and reducing ALDH1A3 , ALDH1A1 and ALDH2-associated Aldefluor fluorescence in breast cancer cells .
Citral was identified as the best inhibitor of ALDH1A3 , reducing the Aldefluor fluorescence in breast cancer cell lines and in a patient-derived tumor xenograft .
Nanoparticle encapsulated citral specifically reduced the enhanced tumor growth of MDA-MB-231 cells overexpressing ALDH1A3 @ @ @ @ @ @ @ citral-mediated tumor growth inhibition , we performed cell proliferation , clonogenic , and gene expression assays .
Citral reduced ALDH1A3-mediated colony formation and expression of ALDH1A3-inducible genes .
In conclusion , citralis an effective ALDH1A3 inhibitor and is able to block ALDH1A3-mediated breast tumor growth , potentially via blocking its colony forming and gene expression regulation activity .
The promise of ALDH1A3 inhibitors as adjuvant therapies for patients with tumors that have a large population of high-ALDH1A3 CSCs is discussed .

Breast cancer stem cells (CSCs) can be identified by increased Aldefluor fluorescence caused by increased expression of aldehyde dehydrogenase 1A3 (ALDH1A3), as well as ALDH1A1 and ALDH2. In addition to being a CSC marker, ALDH1A3 regulates gene expression via retinoic acid (RA) signaling and plays a key role in the progression and chemotherapy resistance of cancer. Therefore, ALDH1A3 represents a druggable anti-cancer target of interest. Since to date, there are no characterized ALDH1A3 isoform inhibitors, drugs that were previously described as inhibiting the activity of other ALDH isoforms were tested for anti-ALDH1A3 activity. Twelve drugs (3-hydroxy-dl-kynurenine, benomyl, citral, chloral hydrate, cyanamide, daidzin, DEAB, disulfiram, gossypol, kynurenic acid, molinate, and pargyline) were compared for their efficacy in inducing apoptosis and reducing ALDH1A3, ALDH1A1 and ALDH2-associated Aldefluor fluorescence in breast cancer cells. Citral was identified as the best inhibitor of ALDH1A3, reducing the Aldefluor fluorescence in breast cancer cell lines and in a patient-derived tumor xenograft. Nanoparticle encapsulated citral specifically reduced the enhanced tumor growth of MDA-MB-231 cells overexpressing ALDH1A3. To determine the potential mechanisms of citral-mediated tumor growth inhibition, we performed cell proliferation, clonogenic, and gene expression assays. Citral reduced ALDH1A3-mediated colony formation and expression of ALDH1A3-inducible genes. In conclusion, citralis an effective ALDH1A3 inhibitor and is able to block ALDH1A3-mediated breast tumor growth, potentially via blocking its colony forming and gene expression regulation activity. The promise of ALDH1A3 inhibitors as adjuvant therapies for patients with tumors that have a large population of high-ALDH1A3 CSCs is discussed.


greenmedinfo--2020-10-15--In vivo anti-tumor effects of citral on 4T1 breast cancer cells.

Breast cancer is the most commonly diagnosed cancer and the leading cause of cancer death among females globally .
The tumorigenic activities of cancer cells such as aldehyde dehydrogenase ( ALDH ) activity and differentiation have contributed to relapse and eventual mortality in breast cancer .
Thus , current drug discovery research is focused on targeting breast cancer cells with ALDH activity and their capacity to form secondary tumors .
Citral ( 3,7-dimethyl-2,6-octadienal ) , from lemon grass ( ) , has been previously reported to have a cytotoxic effect on breast cancer cells .
Hence , this study was @ @ @ @ @ @ @ of citral in targeting ALDH activity of breast cancer cells .
BALB/c mice were challenged with 4T1 breast cancer cells followed by daily oral feeding of 50 mg/kg citral or distilled water for two weeks .
The population of ALDHtumor cells and their capacity to form secondary tumors in both untreated and citral treated 4T1 challenged mice were assessed by Aldefluor assay and tumor growth upon cell reimplantation in normal mice , respectively .
Citral treatment reduced the size and number of cells with ALDHactivity of the tumors in 4T1-challenged BALB/c mice .
@ @ @ @ @ @ @ with smaller tumor size and delayed tumorigenicity after reimplantation of the primary tumor cells into normal mice .
These findings support the antitumor effect of citral in targeting ALDHcells and tumor recurrence in breast cancer cells .

Breast cancer is the most commonly diagnosed cancer and the leading cause of cancer death among females globally. The tumorigenic activities of cancer cells such as aldehyde dehydrogenase (ALDH) activity and differentiation have contributed to relapse and eventual mortality in breast cancer. Thus, current drug discovery research is focused on targeting breast cancer cells with ALDH activity and their capacity to form secondary tumors. Citral (3,7-dimethyl-2,6-octadienal), from lemon grass (), has been previously reported to have a cytotoxic effect on breast cancer cells. Hence, this study was conducted to evaluate the in vivo effect of citral in targeting ALDH activity of breast cancer cells. BALB/c mice were challenged with 4T1 breast cancer cells followed by daily oral feeding of 50 mg/kg citral or distilled water for two weeks. The population of ALDHtumor cells and their capacity to form secondary tumors in both untreated and citral treated 4T1 challenged mice were assessed by Aldefluor assay and tumor growth upon cell reimplantation in normal mice, respectively. Citral treatment reduced the size and number of cells with ALDHactivity of the tumors in 4T1-challenged BALB/c mice. Moreover, citral-treated mice were also observed with smaller tumor size and delayed tumorigenicity after reimplantation of the primary tumor cells into normal mice. These findings support the antitumor effect of citral in targeting ALDHcells and tumor recurrence in breast cancer cells.


greenmedinfo--2020-10-15--Effect and mechanism of citral against methicillin-resistant Staphylococcus aureus in vivo.

BACKGROUND : Citral is an active component of many plant extracts , and it is a safe additive used in food and cosmetics .
A previous study showed that citral has a good antibacterial effect against methicillin-resistant Staphylococcus aureus ( MRSA ) in vitro , but its in vivo anti-infective activity has not been studied .
Anti-MRSA activity and the preliminary mechanism of citral against MRSA were investigated in MRSA-infected KM mice .
The EDwas calculated using Karber 's method .
Groups were selected for inflammatory and oxidative stress level tests , and lung and liver tissues were counterstained with @ @ @ @ @ @ @ Cytokines and oxidative factors were evaluated using the ELISA method ( one-way ANOVA computed using SPSS 19.0. )
.
RESULTS : With the increase in the concentration of citral , the survival rate of MRSA-infected mice increased accordingly .
The EDvalues of citral for intramuscular injection and intragastric administration were 0.09 and 0.26 g kgrespectively .
Citral significantly reduced cytokines ( IL-1β , IL-6 , TNF-α ) and oxidative factors ( malondialdehyde and hydroxyl radicals ) of MRSA-infected mice , whereas it increased gluthtione and superoxide dismutase levels .
Citral can reduce the @ @ @ @ @ @ @ CONCLUSIONS : Citral exerted a dose-dependent anti-MRSA effect and ameliorated MRSA-induced abnormal changes in inflammation and oxidative stress .
This indicates that citral has the potential for development as a new anti-MRSA drug.© 2019 Society of Chemical Industry .

BACKGROUND: Citral is an active component of many plant extracts, and it is a safe additive used in food and cosmetics. A previous study showed that citral has a good antibacterial effect against methicillin-resistant Staphylococcus aureus (MRSA) in vitro, but its in vivo anti-infective activity has not been studied. Anti-MRSA activity and the preliminary mechanism of citral against MRSA were investigated in MRSA-infected KM mice. The EDwas calculated using Karber's method. Groups were selected for inflammatory and oxidative stress level tests, and lung and liver tissues were counterstained with HE for detection of pathological changes. Cytokines and oxidative factors were evaluated using the ELISA method (one-way ANOVA computed using SPSS 19.0.).
RESULTS: With the increase in the concentration of citral, the survival rate of MRSA-infected mice increased accordingly. The EDvalues of citral for intramuscular injection and intragastric administration were 0.09 and 0.26 g kgrespectively. Citral significantly reduced cytokines (IL-1β, IL-6, TNF-α) and oxidative factors (malondialdehyde and hydroxyl radicals) of MRSA-infected mice, whereas it increased gluthtione and superoxide dismutase levels. Citral can reduce the lung inflammatory infiltrates infected by MRSA.
CONCLUSIONS: Citral exerted a dose-dependent anti-MRSA effect and ameliorated MRSA-induced abnormal changes in inflammation and oxidative stress. This indicates that citral has the potential for development as a new anti-MRSA drug.© 2019 Society of Chemical Industry.


greenmedinfo--2020-10-15--Antipyretic effects of citral and possible mechanisms of action.

Citral is a mixture of the two monoterpenoid isomers ( neral and geranial ) widely used as a health-promoting food additive safe for human and animal ( approved by the US Food and Drug Administration ) .
In vitro studies have reported on the capability of citral to reduce inflammation .
Here , we report antipyretic effects of citral in vivo using the most well-accepted model of sickness syndrome , i.e.
, systemic administration of lipopolysaccharide ( LPS ) to rats .
Citral given by gavage caused no change in control euthermic rats ( treated with saline ) but blunted @ @ @ @ @ @ @ the sickness syndrome [ fever ( hallmark of infection ) , plasma cytokines ( IL-1β , IL-6 , and TNF-α ) release , and prostaglandin E ( PGE ) synthesis ( both peripherally and hypothalamic ) ] .
Moreover , LPS caused a sharp increase in plasma corticosterone levels that was unaltered by citral .
These data are consistent with the notion that citral has a corticosterone-independent potent antipyretic effect , acting on the peripheral febrigenic signaling ( plasma levels of IL-1β , IL-6 , TNF-α , and PGE ) , eventually down-modulating @ @ @

Citral is a mixture of the two monoterpenoid isomers (neral and geranial) widely used as a health-promoting food additive safe for human and animal (approved by the US Food and Drug Administration). In vitro studies have reported on the capability of citral to reduce inflammation. Here, we report antipyretic effects of citral in vivo using the most well-accepted model of sickness syndrome, i.e., systemic administration of lipopolysaccharide ( LPS ) to rats. Citral given by gavage caused no change in control euthermic rats (treated with saline) but blunted most of the assessed parameters related to the sickness syndrome [fever (hallmark of infection), plasma cytokines (IL-1β, IL-6, and TNF-α) release, and prostaglandin E(PGE) synthesis (both peripherally and hypothalamic)]. Moreover, LPS caused a sharp increase in plasma corticosterone levels that was unaltered by citral. These data are consistent with the notion that citral has a corticosterone-independent potent antipyretic effect, acting on the peripheral febrigenic signaling (plasma levels of IL-1β, IL-6, TNF-α, and PGE), eventually down-modulating hypothalamic PGEproduction.


greenmedinfo--2020-10-15--The results suggest that citral could be a potential new agent which can eliminate drug-resistant br

The results suggest that citral could be a potential new agent which can eliminate drug-resistant breast cancer cells.

BACKGROUND : Breast cancer remains a leading cause of death in women worldwide .
Although breast cancer therapies have greatly advanced in recent years , many patients still develop tumour recurrence and metastasis , and eventually succumb to the disease due to chemoresistance .
Citral has been reported to show cytotoxic effect on various cancer cell lines .
However , the potential of citral to specifically target the drug resistant breast cancer cells has not yet been tested , which was the focus of our current study .
METHODS : The cytotoxic activity of citral was first tested on MDA-MB-231 @ @ @ @ @ @ @ Subsequently , spheroids of MDA-MB-231 breast cancer cells were developed and treated with citral at different concentrations .
Doxorubicin , cisplatin and tamoxifen were used as positive controls to evaluate the drug resistance phenotype of MDA-MB-231 spheroids .
In addition , apoptosis study was performed using AnnexinV/7AAD flowcytometry .
Aldefluor assay was also carried out to examine whether citral could inhibit the ALDH-positive population , while the potential mechanism of the effect of citral was carried out by using quantitative real time- PCR followed by western blotting analysis .
RESULTS : Citral was @ @ @ @ @ @ @ MDA-MB-231 spheroids when compared to a monolayer culture of MDA-MB-231 cells at a lower ICvalue .
To confirm the inhibition of spheroid self-renewal capacity , the primary spheroids were then cultured to additional passages in the absence of citral .
A significant reduction in the number of secondary spheroids were formed , suggesting the reduction of self-renewal capacity of these aldehyde dehydrogenase positive ( ALDH ) drug resistant spheroids .
Moreover , the AnnexinV/7AAD results demonstrated that citral induced both early and late apoptotic changes in a dose-dependent manner compared to the vehicle @ @ @ @ @ @ @ showed lower cell renewal capacity compared to the vehicle control spheroids in the mammosphere formation assay .
Gene expression studies using quantitative real time PCR and Western blotting assays showed that citral was able to suppress the self-renewal capacity of spheroids and downregulate the Wnt/β-catenin pathway .
CONCLUSION : The results suggest that citral could be a potential new agent which can eliminate drug-resistant breast cancer cells in a spheroid model via inducing apoptosis .

BACKGROUND: Breast cancer remains a leading cause of death in women worldwide. Although breast cancer therapies have greatly advanced in recent years, many patients still develop tumour recurrence and metastasis, and eventually succumb to the disease due to chemoresistance. Citral has been reported to show cytotoxic effect on various cancer cell lines. However, the potential of citral to specifically target the drug resistant breast cancer cells has not yet been tested, which was the focus of our current study.
METHODS: The cytotoxic activity of citral was first tested on MDA-MB-231 cells in vitro by MTT assay. Subsequently, spheroids of MDA-MB-231 breast cancer cells were developed and treated with citral at different concentrations. Doxorubicin, cisplatin and tamoxifen were used as positive controls to evaluate the drug resistance phenotype of MDA-MB-231 spheroids. In addition, apoptosis study was performed using AnnexinV/7AAD flowcytometry. Aldefluor assay was also carried out to examine whether citral could inhibit the ALDH-positive population, while the potential mechanism of the effect of citral was carried out by using quantitative real time- PCR followed by western blotting analysis.
RESULTS: Citral was able to inhibit the growth of the MDA-MB-231 spheroids when compared to a monolayer culture of MDA-MB-231 cells at a lower ICvalue. To confirm the inhibition of spheroid self-renewal capacity, the primary spheroids were then cultured to additional passages in the absence of citral. A significant reduction in the number of secondary spheroids were formed, suggesting the reduction of self-renewal capacity of these aldehyde dehydrogenase positive (ALDH) drug resistant spheroids. Moreover, the AnnexinV/7AAD results demonstrated that citral induced both early and late apoptotic changes in a dose-dependent manner compared to the vehicle control. Furthermore, citral treated spheroids showed lower cell renewal capacity compared to the vehicle control spheroids in the mammosphere formation assay. Gene expression studies using quantitative real time PCR and Western blotting assays showed that citral was able to suppress the self-renewal capacity of spheroids and downregulate the Wnt/β-catenin pathway.
CONCLUSION: The results suggest that citral could be a potential new agent which can eliminate drug-resistant breast cancer cells in a spheroid model via inducing apoptosis.


greenmedinfo--2020-10-15--Apple polyphenol extract alleviates lipid accumulation in free-fatty-acid-exposed HepG2 cells.

Defective degradation of intracellular lipids induced by autophagy is causally linked to the development of non-alcoholic fatty liver disease ( @ @ @ @ @ that can restore autophagy could therefore have the potentials for clinical applications for this public @ @ @ @ @ we investigated the effects of apple polyphenol extract ( APE ) on fatty acid-induced lipids @ @ @ @ @ APE treatment alleviated palmitic acid and oleic acid-induced intracellular lipid accumulation , concomitant with the @ @ @ @ @ acidification , inhibited lipid synthesis and slight promotion of fatty acid oxidation .
Mechanistically , @ @ @ @ @ SIRT1 , activated LKB1/AMPK pathway and inhibited mTOR signaling .
Over-expressed or knock-down SIRT1 positively @ @ @ @ @ .
SIRT1 and ATG7 knock-down impaired APE induction of improved lipid accumulation , increased intracellular @ @ @ @ @ APE induction of autophagy to ameliorate fatty acid-induced lipid deposition is SIRT1 dependent , APE @ @ @ @ @ hepatosteatosis .

Defective degradation of intracellular lipids induced by autophagy is causally linked to the development of non-alcoholic fatty liver disease (NAFLD). Natural agents that can restore autophagy could therefore have the potentials for clinical applications for this public health issue. Herein, we investigated the effects of apple polyphenol extract (APE) on fatty acid-induced lipids depositions in HepG2 cells. APE treatment alleviated palmitic acid and oleic acid-induced intracellular lipid accumulation, concomitant with the increased autophagy, restored lysosomal acidification, inhibited lipid synthesis and slight promotion of fatty acid oxidation. Mechanistically, APE up-regulated the expression of SIRT1, activated LKB1/AMPK pathway and inhibited mTOR signaling. Over-expressed or knock-down SIRT1 positively regulated AMPK and ATG7 expressions. SIRT1 and ATG7 knock-down impaired APE induction of improved lipid accumulation, increased intracellular TG content. Thus, APE induction of autophagy to ameliorate fatty acid-induced lipid deposition is SIRT1 dependent, APE conserved preventive potentials for clinical hepatosteatosis.


greenmedinfo--2020-10-15--Annurca apple polyphenol extract promotes mesenchymal-to-epithelial transition and inhibits migratio

Annurca apple polyphenol extract promotes mesenchymal-to-epithelial transition and inhibits migration in triple-negative breast cancer cells.

Aberrant activation of epithelial-to-mesenchymal transition has been shown to correlate with triple-negative breast cancer ( TNBC ) progression and metastasis .
Thus , the induction of the reverse process might offer promising opportunities to restrain TNBC metastatic spreading and related mortality .
Recently , the Annurca apple polyphenol extract ( APE ) has been highlighted as a multi-faceted agent that selectively kills TNBC cells by ROS generation and sustained JNK activation .
Here , by qualitatively and quantitatively monitoring the real-time movements of live cells we provided the first evidence that APE inhibited the migration of MDA-MB-231 and MDA-MB-468 TNBC @ @ @ @ @ @ @ In MDA-MB-231 cells APE decreased SMAD-2/3 and p-SMAD-2/3 levels , increased E-cadherin/N-cadherin protein ratio , induced the switch from N-cadherin to E-cadherin expression and greatly reduced vimentin levels .
Confocal and scanning electron microscopy imaging of APE-treated MDA-MB-231 cells evidenced a significant cytoskeletal vimentin and filamentous actin reorganization and revealed considerable changes in cell morphology highlighting an evident transition from the mesenchymal to epithelial phenotype with decreased migratory features .
Notably , all these events were reverted by N-acetyl-L-cysteine and JNK inhibitor SP600125 furnishing evidence that APE exerted its effects through the activation @ @ @ @ @ @ @ highlighted APE as a potential preventing agent for TNBC metastasis .

Aberrant activation of epithelial-to-mesenchymal transition has been shown to correlate with triple-negative breast cancer (TNBC) progression and metastasis. Thus, the induction of the reverse process might offer promising opportunities to restrain TNBC metastatic spreading and related mortality. Recently, the Annurca apple polyphenol extract (APE) has been highlighted as a multi-faceted agent that selectively kills TNBC cells by ROS generation and sustained JNK activation. Here, by qualitatively and quantitatively monitoring the real-time movements of live cells we provided the first evidence that APE inhibited the migration of MDA-MB-231 and MDA-MB-468 TNBC cells and downregulated metalloproteinase-2 and metalloproteinase-9. In MDA-MB-231 cells APE decreased SMAD-2/3 and p-SMAD-2/3 levels, increased E-cadherin/N-cadherin protein ratio, induced the switch from N-cadherin to E-cadherin expression and greatly reduced vimentin levels. Confocal and scanning electron microscopy imaging of APE-treated MDA-MB-231 cells evidenced a significant cytoskeletal vimentin and filamentous actin reorganization and revealed considerable changes in cell morphology highlighting an evident transition from the mesenchymal to epithelial phenotype with decreased migratory features. Notably, all these events were reverted by N-acetyl-L-cysteine and JNK inhibitor SP600125 furnishing evidence that APE exerted its effects through the activation of ROS/JNK signaling. The overall data highlighted APE as a potential preventing agent for TNBC metastasis.