AIM:To investigate the mechanism by which galangin,a polyphenolic compound derived from medicinal herbs,induces apoptosis of hepatocellular carcinoma(HCC) cells.METHODS:The 3-(4,5-Dimethyl-thiazol-2-yl)-2,5-diphenyl-t...AIM:To investigate the mechanism by which galangin,a polyphenolic compound derived from medicinal herbs,induces apoptosis of hepatocellular carcinoma(HCC) cells.METHODS:The 3-(4,5-Dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay was used to measure cell viability.Apoptosis was evaluated by in situ uptake of propidium iodide and Hoechst 33258 and was then detected by fluorescence microscopy.Protein expressions were detected by Western blotting.To confirm the apoptotic pathway mediated by galangin,cells were transfected by bcl-2 gene to overexpress Bcl-2 or siRNA to down-regulate Bcl-2 expression.RESULTS:Galangin(46.25-370.0 μmol/L) exerted an anti-proliferative effect,induced apoptosis,and decreased mitochondrial membrane potential in a dose and time-dependent manner.Treatment with galangin induced apoptosis by translocating the pro-apoptotic protein Bax to the mitochondria,which released apoptosis-inducing factor and cytochrome c into the cytosol.Overexpression of Bcl-2 attenuated galangin-induced HepG2 cell apoptosis,while decreasing Bcl-2 expression enhanced galangin-induced cell apoptosis.CONCLUSION:Our data suggests that galangin mediates apoptosis through a mitochondrial pathway,and may be a potential chemotherapeutic drug for the treatment of HCC.展开更多
Objective:To explore the cytotoxicity effects of Caspian cobra(Naja oxiana or N.oxiana) venom on hepatocytes and mitochondria obtained from the liver of HCC rats.Methods:In this study,HCC was induced by diethylnitrosa...Objective:To explore the cytotoxicity effects of Caspian cobra(Naja oxiana or N.oxiana) venom on hepatocytes and mitochondria obtained from the liver of HCC rats.Methods:In this study,HCC was induced by diethylnitrosamine(DEN),as an initiator,and 2-acetylaminofluorene(2-AAF),as a promoter.Rat liver hepatocytes and mitochondria for evaluation of the selective cytotoxic effect of N.oxiana venom were isolated and mitochondria and cellular parameters related to apoptosis signaling were then determined.Results:Our results showed a raise in mitochondrial reactive oxygen species(ROS)level,swelling in mitochondria,mitochondrial membrane potential(Djm) collapse and release of cytochrome c after exposure of mitochondria only isolated from the HCC group with the crude venom of the N.oxiana(12.5,25,and 50 mg/m L).This crude venom also induced caspase-3 activation(P < 0.001) in the hepatocytes obtained only from the HCC rat liver.Conclusions:Based on the over all results,we suggested that N.oxiana may be considered as a promising complementary therapeutic agent for the treatment of HCC.展开更多
目的:观察化瘀温胆汤对糖耐量低减大鼠肝细胞线粒体SOD及MDA水平的影响。方法:应用20周高脂高糖饮食诱导糖耐量低减大鼠模型,将模型鼠分为模型组、化瘀温胆汤组和二甲双胍组,并设立空白组对照;给药4周后,测定空腹血糖(FPG)和腹腔注射糖...目的:观察化瘀温胆汤对糖耐量低减大鼠肝细胞线粒体SOD及MDA水平的影响。方法:应用20周高脂高糖饮食诱导糖耐量低减大鼠模型,将模型鼠分为模型组、化瘀温胆汤组和二甲双胍组,并设立空白组对照;给药4周后,测定空腹血糖(FPG)和腹腔注射糖耐量实验2 h血糖(2 h PG)水平,提取肝细胞线粒体并测量其超氧化物歧化酶(SOD)及丙二醛(MDA)含量。结果:化瘀温胆汤组和二甲双胍组FPG、2 h PG及肝细胞线粒体MDA水平低于模型组,肝细胞线粒体SOD水平高于模型组(P<0.05)。结论:化瘀温胆汤可能通过抑制肝细胞线粒体氧化应激反应,发挥治疗糖耐量低减的作用。展开更多
Objective： To evaluate the effect of Bear Bile Powder （熊胆粉, BBP） on the growth and apoptosis of HepG2 human hepatocellular carcinoma cells, and investigate the possible molecular mechanisms mediating its anti-ca...Objective： To evaluate the effect of Bear Bile Powder （熊胆粉, BBP） on the growth and apoptosis of HepG2 human hepatocellular carcinoma cells, and investigate the possible molecular mechanisms mediating its anti-cancer activity. Methods： HepG2 cells were treated with 0.4-1.0 mg/mL of BBP for 24, 48 and 72 h. The viability of HePG2 cells was determined by MTT assay. Cellular morphology was observed via phase-contrast microscopy. Fluorescence-activated cell sorting analysis with Annexin-V/propidium idodide and 5,5＇,6,6＇-tetrachloro- 1 ,1＇,3,3＇-tetraethyl-benzimidazol-carbocyanine iodide （JC-1） staining was performed to determine cell apoptosis and the loss of mitochondrial membrane potential, respectively. Activation of caspase-9 and -3 was evaluated by a colorimetric assay. Results： The treatment with 0.4-1 mg/mL of BBP for 24, 48, or 72 h respectively reduced cell viability significantly by 7%-60%, 20%-90% or 25%-98%, compared with the untreated control cells （P〈0.01）. In addition, BBP treatment induced morphological changes in HepG2 cells. Furthermore, after treated with 0, 0.4, 0.6, 0.8 and 1.0 mg/mL of BBP, apoptosis cells （including early and late apoptotic cells） were 18.0% ± 1.3%, 34.9% ± 2.2%, 33.9% ± 2.8%, 37.4% ± 2.8% and 46.0% ± 2.5%, respectively （P〈0.05）; and the percentage of cells with reduced JC-1 red fluorescence were 6.6% ± 0.8%, 8.5% ± 0.8%, 13.5% ± 1.6%, 17.6%± 2.3% and 46.7% ± 3.6%, respectively （P〈0.01）. Finally, BBP treatment significantly and dose-dependently induced activation of both caspase-9 and caspase-3 in HepG2 cells （P〈0.05）. Conclusions： BBP could inhibit the growth of HepG2 hepatocellular cancer cells through mitochondrion-mediated apoptosis, which may, in part, explain its anti-cancer activity. BBP may be a potential novel therapeutic agent for the treatment of hepatocellular carcinoma.展开更多
基金Supported by The administration of traditional Chinese medicine of Guangdong province,China,No.1050047
文摘AIM:To investigate the mechanism by which galangin,a polyphenolic compound derived from medicinal herbs,induces apoptosis of hepatocellular carcinoma(HCC) cells.METHODS:The 3-(4,5-Dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay was used to measure cell viability.Apoptosis was evaluated by in situ uptake of propidium iodide and Hoechst 33258 and was then detected by fluorescence microscopy.Protein expressions were detected by Western blotting.To confirm the apoptotic pathway mediated by galangin,cells were transfected by bcl-2 gene to overexpress Bcl-2 or siRNA to down-regulate Bcl-2 expression.RESULTS:Galangin(46.25-370.0 μmol/L) exerted an anti-proliferative effect,induced apoptosis,and decreased mitochondrial membrane potential in a dose and time-dependent manner.Treatment with galangin induced apoptosis by translocating the pro-apoptotic protein Bax to the mitochondria,which released apoptosis-inducing factor and cytochrome c into the cytosol.Overexpression of Bcl-2 attenuated galangin-induced HepG2 cell apoptosis,while decreasing Bcl-2 expression enhanced galangin-induced cell apoptosis.CONCLUSION:Our data suggests that galangin mediates apoptosis through a mitochondrial pathway,and may be a potential chemotherapeutic drug for the treatment of HCC.
基金supported by Shahid Beheshti University of Medical Sciences,Deputy of Research(Grant No.1394-1-95-11835,2015)
文摘Objective:To explore the cytotoxicity effects of Caspian cobra(Naja oxiana or N.oxiana) venom on hepatocytes and mitochondria obtained from the liver of HCC rats.Methods:In this study,HCC was induced by diethylnitrosamine(DEN),as an initiator,and 2-acetylaminofluorene(2-AAF),as a promoter.Rat liver hepatocytes and mitochondria for evaluation of the selective cytotoxic effect of N.oxiana venom were isolated and mitochondria and cellular parameters related to apoptosis signaling were then determined.Results:Our results showed a raise in mitochondrial reactive oxygen species(ROS)level,swelling in mitochondria,mitochondrial membrane potential(Djm) collapse and release of cytochrome c after exposure of mitochondria only isolated from the HCC group with the crude venom of the N.oxiana(12.5,25,and 50 mg/m L).This crude venom also induced caspase-3 activation(P < 0.001) in the hepatocytes obtained only from the HCC rat liver.Conclusions:Based on the over all results,we suggested that N.oxiana may be considered as a promising complementary therapeutic agent for the treatment of HCC.
文摘目的:观察化瘀温胆汤对糖耐量低减大鼠肝细胞线粒体SOD及MDA水平的影响。方法:应用20周高脂高糖饮食诱导糖耐量低减大鼠模型,将模型鼠分为模型组、化瘀温胆汤组和二甲双胍组,并设立空白组对照;给药4周后,测定空腹血糖(FPG)和腹腔注射糖耐量实验2 h血糖(2 h PG)水平,提取肝细胞线粒体并测量其超氧化物歧化酶(SOD)及丙二醛(MDA)含量。结果:化瘀温胆汤组和二甲双胍组FPG、2 h PG及肝细胞线粒体MDA水平低于模型组,肝细胞线粒体SOD水平高于模型组(P<0.05)。结论:化瘀温胆汤可能通过抑制肝细胞线粒体氧化应激反应,发挥治疗糖耐量低减的作用。
基金Supported by Important Science&Technology Specific Projects of Fujian Province(No.2010YZ0001-1 and 2010Y2004)Developmental Fund of Chen Keji Integrative Medicine(No.CKJ2010019)
文摘Objective： To evaluate the effect of Bear Bile Powder （熊胆粉, BBP） on the growth and apoptosis of HepG2 human hepatocellular carcinoma cells, and investigate the possible molecular mechanisms mediating its anti-cancer activity. Methods： HepG2 cells were treated with 0.4-1.0 mg/mL of BBP for 24, 48 and 72 h. The viability of HePG2 cells was determined by MTT assay. Cellular morphology was observed via phase-contrast microscopy. Fluorescence-activated cell sorting analysis with Annexin-V/propidium idodide and 5,5＇,6,6＇-tetrachloro- 1 ,1＇,3,3＇-tetraethyl-benzimidazol-carbocyanine iodide （JC-1） staining was performed to determine cell apoptosis and the loss of mitochondrial membrane potential, respectively. Activation of caspase-9 and -3 was evaluated by a colorimetric assay. Results： The treatment with 0.4-1 mg/mL of BBP for 24, 48, or 72 h respectively reduced cell viability significantly by 7%-60%, 20%-90% or 25%-98%, compared with the untreated control cells （P〈0.01）. In addition, BBP treatment induced morphological changes in HepG2 cells. Furthermore, after treated with 0, 0.4, 0.6, 0.8 and 1.0 mg/mL of BBP, apoptosis cells （including early and late apoptotic cells） were 18.0% ± 1.3%, 34.9% ± 2.2%, 33.9% ± 2.8%, 37.4% ± 2.8% and 46.0% ± 2.5%, respectively （P〈0.05）; and the percentage of cells with reduced JC-1 red fluorescence were 6.6% ± 0.8%, 8.5% ± 0.8%, 13.5% ± 1.6%, 17.6%± 2.3% and 46.7% ± 3.6%, respectively （P〈0.01）. Finally, BBP treatment significantly and dose-dependently induced activation of both caspase-9 and caspase-3 in HepG2 cells （P〈0.05）. Conclusions： BBP could inhibit the growth of HepG2 hepatocellular cancer cells through mitochondrion-mediated apoptosis, which may, in part, explain its anti-cancer activity. BBP may be a potential novel therapeutic agent for the treatment of hepatocellular carcinoma.
