复方木鸡颗粒联合顺铂对人肝癌HepG2细胞增殖及凋亡的机制研究

Effects of Compound Muji Granules combined with cisplatin on proliferation and apoptosis of human hepatoma HepG2 and its potential mechanism

目的探讨复方木鸡颗粒联合顺铂对人肝癌HepG2细胞增殖、凋亡的影响及其潜在的作用机制。方法体外培养人肝癌HepG2细胞,取对数生长期细胞,将其消化后随机分为对照组,复方木鸡颗粒高、低(10、5 mg/mL)剂量组,顺铂组(1 mg/mL)和复方木鸡颗粒(5 mg/mL)联合顺铂(1 mg/mL)组,采用CCK-8法检测细胞增殖情况,采用Hoehcst染色法检测细胞凋亡情况,采用免疫荧光联合激光共聚焦分别检测各组细胞中凋亡相关蛋白B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、蛋白天冬氨酸蛋白水解酶-9(Caspase-9)、细胞色素C氧化酶(Cyt-C)表达水平。结果与对照组比较,复方木鸡颗粒10、5 mg/mL组及顺铂组、联合组的细胞活力显著降低(P<0.01),细胞凋亡显著增加,细胞内Bcl-2蛋白表达显著下调(P<0.05、0.01),而Bax、Caspase-9、Cyt-C蛋白显著上调(P<0.05、0.01),且联合组在抑制人肝癌HepG2细胞增殖、促进其凋亡、调控凋亡相关蛋白表达的作用效果最佳。结论复方木鸡颗粒可显著抑制HepG2细胞增殖并促进其凋亡,其与顺铂联合起到协同增效的效果,其作用机制可能与下调人肝癌HepG2细胞内抑凋亡蛋白Bcl-2,上调促凋亡蛋白Bax、Caspase-9、Cyt-C表达水平有关。

Objective To discuss the effects of Compound Muji Granules combined with cisplatin on proliferation and apoptosis of human hepatoma HepG2 and its potential mechanism.Methods HepG2 cells were cultured in vitro,and then were divided into the control group,Compound Muji Granules high and low(10 and 5 mg/mL)groups,cisplatin group(1 mg/mL),Compound Muji Granules(5 mg/mL)+cisplatin(1 mg/mL)group.The cell proliferation inhibition was detected by CCK-8.Hoehcst staining was used to detect the apoptosis of HepG2 cells,and the immunofluorescent staining was used to detect the protein expressions of Bcl-2,Bax,Caspase 9,and Cyt C in HepG2 cells.Results Compared with the control group,the cell viability of Compound Muji Granules 10 and 5 mg/mL groups,cisplatin group,and Compound Muji Granules+cisplatin group was significantly reduced(P<0.01),while the apoptosis of HepG2 cells was increased,and the relative expression of Bcl-2 protein was decreased,but the relative expression of Bax,Caspase 9,and Cyt C protein was significantly increased(P<0.05,0.01).And Compound Muji Granules+cisplatin group had the best effect on inhibiting the proliferation,promoting the apoptosis and regulating the expression of apoptosis-related proteins of HepG2 cells.Conclusion Compound Muji Granules inhibits the proliferation and promotes the apoptosis of HepG2 cells,and the effect enhanced while combined with cisplatin by reducing Bcl-2 and increasing Bax,Caspase-9,and Cyt C.