雷公藤内酯酮联合甘草次酸对促进人肝癌HepG2细胞凋亡及相关蛋白表达的影响  

Enhanced Apoptosis of Triptonide Combined with Glycyrrhetinic Acid for Hepatocellular Carcinoma HepG2 Cells and Its Effect on Related Protein Expression

作　　者：戴雅彬 黄慧[1] 张月芬[1] 郭丹[1] 邓艳平[2] DAI Yabin;HUANG Hui;ZHANG Yuefen;GUO Dan;DENG Yanping(Department of Science and Technology Service Center,Fujian Health College,Fuzhou 350101,China;School of Pharmacy,Fujian Medical University,Fuzhou 350122,China)

机构地区：[1]福建卫生职业技术学院科技服务中心,福州350101 [2]福建医科大学药学院,福州350122

出　　处：《福建医科大学学报》2023年第1期1-8,共8页Journal of Fujian Medical University

基　　金：福建省科技创新联合资金项目(2019Y9010);福建省自然科学基金项目(2020J01630);福建卫生职业技术学院校级科研项目(MWY2021-1-04)。

摘　　要：目的研究雷公藤内酯酮(TPN)单独或联用甘草次酸(GA)对人肝癌HepG2细胞增殖、迁移、凋亡及相关信号通路Caspase-3、Bax和Bcl-2蛋白表达的影响。方法采用CCK-8法测定TPN单独或TPN(0.3、0.6、1.2μmol/L)联用GA(2、10、20μmol/L)对HepG2细胞24 h的增殖抑制作用,利用增效指数(q值)判定协同药效;Annexin V-FITC/PI染色法检测给药后细胞凋亡率变化;细胞划痕实验检测药物对细胞迁移能力的影响;Western-blot法检测Caspase-3、Bax和Bcl-2蛋白的表达水平。结果TPN、GA单独作用对HepG2细胞24 h的半数抑制浓度(IC 50)分别为(7.28±2.13)和(26.56±6.32)μmol/L。TPN(1.2μmol/L)和GA(2μmol/L)联用组的q值(1.505)最大,细胞抑制率为(62.28±6.13)%;该联用组对HepG2细胞的总凋亡率高于单独使用TPN(P<0.05),同时抑制HepG2细胞的迁移,使细胞内Caspase-3蛋白表达水平升高、Bcl-2蛋白表达水平降低(P<0.05)。结论TPN联用GA对HepG2细胞的增殖具有良好的协同抑制效果,可能与其通过下调Bcl-2基因表达与激活Caspase基因家族共同诱导肝癌细胞的凋亡有关,两者联用能增强抑制肝癌HepG2细胞的迁移。Objective The effects of triptonide(TPN)alone or its combination with glycyrrhetnic acid(GA)on proliferation,migration,and expression levels of Caspase-3,Bax,and Bcl-2,which are related to apoptosis in human hepatocellular carcinoma HepG2 cells,were studied.Method The proliferation inhibitory effects of TPN alone or TPN(0.3,0.6,1.2μmol/L)combined with GA(2,10,20μmol/L)on HepG2 cells were determined using Cell Counting Kit-8(CCK-8)method.The widths of the cell wounds were continuously recorded via microscopic photographs.The cell apoptosis rates of HepG2 cells were recorded using Annexin V-FITC/PI staining.Western-blot was used to determine the protein levels of Caspase-3,Bcl-2,and Bax.Results The median inhibitory concentrations(IC 50)of TPN or GA alone on HepG2 cells for 24 h were(7.28±2.13)and(26.56±6.32)μmol/L,respectively.The maximum synergy index q value of TPN(1.2μmol/L)combined with GA(2μmol/L)was 1.505,and the cell inhibition rate was(62.28±6.13)%.Compared with TPN alone,TPN(1.2μmol/L)combined with GA(2μmol/L)significantly enhanced the total apoptosis rate(P<0.05),inhibited the migration,increased the protein expression level of Caspase-3,and decreased the protein expression level of Bcl-2 in HepG2 cells(P<0.05).Conclusion TPN combined with GA had a good synergistic inhibitory effect on the proliferation of HepG2 cells,which might be related to the enhanced apoptosis that could up-regulate Caspase-3 and down-regulate Bcl-2 as well as reduce migration at the same time.

关 键 词：雷公藤内酯酮 甘草次酸 联合用药 肝癌 细胞凋亡 

分 类 号：R285[医药卫生—中药学]