黑升麻提取物对人肝癌细胞系HepG2增殖和凋亡的影响  

Effect of Black Cohosh Extracts on Proliferation and Apoptosis of Hepatocellular Carcinoma HepG2 Cells

作　　者：倪璠 柯阳[1] 邹仁超 艾润垚 马瑞成 周剑 郭志唐 王琳[1] NI Fan;KE Yang;ZOU Renchao;AI Runyao;MA Ruicheng;ZHOU Jian;GUO Zhitang;WANG Lin(Department of Hepatobiliary Surgery,the Second Affiliated Hospital of Kunming Medical University,Kunming 650101,China;Kunming Medical University,Kunming 650500,China)

机构地区：[1]昆明医科大学第二附属医院肝胆胰外科,昆明650101 [2]昆明医科大学,昆明650500

出　　处：《医学综述》2019年第5期1030-1035,共6页Medical Recapitulate

基　　金：国家自然科学基金(81660399;81860423);云南省创新团队计划(2015HC033);云南省外国专家局项目(H162039011);云南省卫生和计划生育委员会医学领军人才培养计划(L-201622);昆明医科大学重大科技成果培育项目(CGPY201607)

摘　　要：目的研究黑升麻提取物对人肝癌细胞系HepG2增殖和凋亡的影响。方法将肝癌HepG2细胞依据随机数字法分为黑升麻药物0、10、20、30、40、50μg/mL组,采用细胞计数试剂盒8检测实验组不同浓度(10、20、30、40、50μg/mL)黑升麻处理后HepG2细胞的增殖抑制率。流式细胞仪、Annexin V-异硫氰酸荧光素/碘化丙啶双染检测对照组(黑升麻药物浓度0μg/mL)及不同浓度(10、20、32μg/mL)黑升麻处理后实验组的HepG2凋亡细胞比率; Western blot检测凋亡相关胱天蛋白酶(caspase)-3、caspase-8、caspase-9蛋白的表达。结果 0、10、20、30、40、50μg/mL组抑制率分别为0%、(12. 51±9. 31)%、(27. 85±1. 51)%、(53. 20±5. 55)%、(67. 42±3. 54)%、(81. 43±0. 73)%,各组比较差异有统计学意义(P <0. 05),随着各组给药浓度的增加抑制率逐渐升高(P <0. 05)。黑升麻IC50为32. 010μg/mL。与0μg/mL组比较,10、20、32μg/mL组凋亡逐渐增加(P <0. 05),Cleaved caspase-3、Cleaved caspase-8蛋白表达上调(P <0. 05)。结论黑升麻提取物可有效抑制肝癌HepG2细胞系增殖,促进细胞凋亡,该作用至少部分依赖于外源性死亡受体途径。Objective To explore the effect of black cohosh extracts on the proliferation and apoptosis of hepatocellular carcinoma HepG2 cells. Methods The HepG2 cells were divided into 0,10,20,30,40,50 μg/mL group.The inhibitory rate of proliferation of HepG2 cells in the experimental group treated with different concentrations of black cohosh (10,20,30,40,50 μg/mL) were analyzed by cell counting Kit-8 assay.The ratio of apoptotic cells in the blank control group and experimental group(10,20,32 μg/mL) was detected by Annexin V-fluoresceine isothiocyanate/propidium iodide double staining and flow cytometry.Expressions of apoptosis-related caspase-3,caspase-8 and caspase-9 proteins were detected by Western blot. Results The inhibitory rates of 0,10,20,30,40,50 μg/mL groups were 0%,(12.51±9.31)%,(27.85±1.51)%,(53.20±5.55)%,(67.42±3.54)% and (81.43±0.73)%,there was statistical significance in comparison among groups( P <0.05),and the inhibition rate gradually increased with the increase of drug concentration in each group( P <0.05).The IC 50 of black cohosh was 32.010 μg/mL. Compared with 0 μg/mL group,10,20,32 μg/mL group apoptosis increased gradually( P <0.05),expression of Cleaved caspase-3,Cleaved caspase-8 protein increased( P <0.05). Conclusion Black cohosh extract can effectively inhibit the proliferation of HepG2 cell lines and promote apoptosis,which is at least partially dependent on the exogenous death receptor pathway.

关 键 词：肝细胞癌 黑升麻 增殖 凋亡 胱天蛋白酶3 胱天蛋白酶8 

分 类 号：R735.7[医药卫生—肿瘤]