贝参茱萸方通过线粒体途径诱导人肝癌HepG2细胞凋亡的分子机制  

Molecular mechanism research on Beishen Zhuyu Formula inducing hepatocellular carcinoma HepG2 cells through mitochondria apoptosis pathway

作　　者：晁旭[1,2] 张艳芳[3] 李宏[2] 史海龙[3] 黄峰[2] 王国全[3] 王瑞莉[3] CHAO Xu;ZHANG Yan-fang;LI Hong;SHI Hai-long;HUANG Feng;WANG Guo-quan;WANG Rui-li(Shaanxi University of Chinese Medicine,Xianyang 712046,China;The Second Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712000,China)

机构地区：[1]陕西中医药大学基础医学院,咸阳712046 [2]陕西中医药大学第二附属医院,咸阳712000 [3]陕西中医药大学,咸阳712046

出　　处：《中华中医药杂志》2020年第7期3353-3357,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81774132);陕西省中医药管理局科研项目(No.2019-zz-JC012);陕西中医药大学学科创新团队项目(No.2019-YS05)。

摘　　要：目的:探讨贝参茱萸方对人肝癌HepG2细胞增殖和凋亡的影响及其分子机制。方法:人肝癌HepG2细胞经贝参茱萸方含药血清处理后,用倒置显微镜观察肿瘤细胞的形态,MTT检测法检测细胞的生长抑制率,流式细胞术检测细胞的凋亡率和周期变化。采用罗丹明123染色法检测细胞的线粒体膜电位变化,Hoechst33258染色法检测细胞核变化,Western Blot检测细胞凋亡相关蛋白的表达。结果:贝参茱萸方含药血清处理人肝癌HepG2细胞后,细胞数量明显减少,细胞呈现典型的凋亡细胞形态;随着浓度升高,细胞生长抑制率显著增大(P<0.01),G1期细胞数明显增加,而G2/M期细胞数量减少,细胞线粒体膜电位显著降低(P<0.05)。贝参茱萸方可上调Bax、Caspase-3、Caspase-9和Cytochrome-c的表达,下调Bcl-2的表达(P<0.05)。结论:贝参茱萸方可阻滞细胞周期于G1期从而抑制肝癌HepG2细胞的增殖,并通过线粒体途径诱导细胞发生凋亡。Objective:To explore the apoptosis and mechanism of Beishen Zhuyu Formula(BSZY)on the hepatocellular carcinoma HepG2 cells.Methods:The cellular morphology of HepG2 cells treated by BSZY were observed by inverted microscope.MTT assay was used to detect the inhibitory effect of BSZY on cell proliferation.Fluorescence staining and flow cytometry were employed to analyze the effect of BSZY on apoptosis of HepG2 cells.Rho123 staining method was performed to detect the changes of mitochondrial membrane potential,Hoechst33258 staining was used to detect nuclear changes,and Western Blot was carried out to evaluate the expression of proteins related to apoptosis.Results:The number of hepatocellular carcinoma HepG2 cells treatment with serum contained BSZY were markedly reduced and cellular morphology significantly altered.MTT assay indicated that the inhibition rate increased with the increase of concentration of serum contained BSZY(P<0.01).The number of cells in G1 phase significantly increased and G2/M phase decreased after treatment with BSZY.Mitochondrial membrane potential(MMP)test showed that MMP of HepG2 cells were significantly decreased after treatment with serum contained BSZY(P<0.05).Western Blot results showed that expressions of Bax,Caspase-3,Caspase-9 and Cytochrome-c expression were significantly increased,while Bcl-2 expression decreased in the HepG2 cells treated serum contained BSZY(P<0.05).Conclusion:BSZY can significantly inhibit the proliferation of hepatocellular carcinoma HepG2 cells through arrest the cell cycle in G1 phase,and could induce apoptosis through the mitochondrial pathway.

关 键 词：贝参茱萸方 HEPG2细胞 线粒体途径 凋亡 

分 类 号：R285[医药卫生—中药学]