扶正消瘤颗粒含药血清对肝癌HepG2细胞体外增殖和凋亡相关蛋白的影响

Effect of Fuzheng Xiaoliu Granule Drug-containing Serum on Proliferation and Apoptosis-related Proteins of Hepatocarcinoma HepG2 Cells in Vitro

目的:观察扶正消瘤颗粒含药血清对肝癌HepG2细胞增殖及凋亡相关蛋白的影响。方法:60只雄性SD大鼠,随机分为6个组,每组10只,分别为扶正消瘤颗粒低剂量组、扶正消瘤颗粒中剂量组、扶正消瘤颗粒高剂量组、5-氟尿嘧啶(5-fluorouracil,5-FU)组、联合组、对照组。中药各剂量组给予不同剂量扶正消瘤颗粒灌胃;5-FU组给予5-FU腹腔灌注;联合组给予中剂量的扶正消瘤颗粒灌胃,同时给予5-FU腹腔灌注;对照组给予生理盐水灌胃。采用四甲基偶氮唑盐比色法检测肝癌HepG2细胞增殖;Annexin V-FITC/PI双染流式细胞法检测肝癌HepG2细胞凋亡;Western-Blot法观察各组肝癌HepG2细胞P53蛋白、Bcl-2蛋白和Survivin蛋白的表达。结果:扶正消瘤颗粒含药血清可抑制肝癌HepG2细胞的增殖,并呈时间、剂量依赖性;其中联合组对肝癌HepG2细胞的抑制率最高,与对照组比较,差异有统计学意义(P<0.05);不同浓度扶正消瘤颗粒含药血清均可诱导肝癌HepG2细胞的凋亡,联合组凋亡率最高,与对照组比较,差异有统计学意义(P<0.05);与对照组比较,各扶正消瘤颗粒含药血清组均可下调Bcl-2和Survivin蛋白的表达,增强P53蛋白的表达。结论:扶正消瘤颗粒含药血清可诱导肝癌HepG2细胞凋亡,其作用可能是通过抑制Bcl-2和Survivin蛋白的表达,增强P53蛋白的表达实现的。

Objective： To observe the effect of Fuzheng Xiaoliu Granule drug-containing serum on the proliferation and apoptosis-related proteins of HepG2 cells. Methods： Sixty male Sprague-Dawley rats were randomly divided into 6 groups with 10 rats in each group： Fuzheng Xiaoliu Granule low dose group,Fuzheng Xiaoliu Granule medium dose group,Fuzheng Xiaoliu Granule high dose group,5-fluorouracil(5-fluorouracil,5-FU） group,combination group,and control group. Each dose group of traditional Chinese medicine was given different doses of Fuzheng Xiaoliu Granule to gavage; 5-FU group was given 5-FU intraperitoneal perfusion; the combination group was given moderate dose of Fuzheng Xiaoliu Granule by intragastric administration and 5-FU intraperitoneal perfusion; the control group was given physiological salt water gavage. The proliferation of hepatocellular carcinoma HepG2 cells was detected by MTT assay; the apoptosis of HepG2 cells was detected by Annexin V-FITC/PI double staining flow cytometry; the P53 protein and Bcl were observed by Hepatoma HepG2 cells by Western-Blot.-2 protein and Survivin protein expression. Results：The Fuzheng Xiaoliu Granule drug-containing serum could inhibit the proliferation of hepatocellular carcinoma HepG2 cells in a time-and dose-dependent manner. Among them,the combined group had the highest inhibition rate of hepatocellular carcinoma HepG2 cells,and the difference was statistically significant compared with the control group(P 〈 0. 05）. Different concentrations of Fuzheng Xiaoliu Granule drug-containing serum can induce HepG2 cell apoptosis and realize the highest rate of apoptosis in the combined group. Compared with that of the control group,the difference was statistically significant(P 〈 0. 05）. Compared with the control group,the expression of Bcl-2 and Survivin protein in each Fuzheng Xiaoliu Granule drug-containing serum group was down-regulated and the expression of P53 protein was enhanced. Conclusion： Fuzheng Xiaoliu Granule drug-containing serum can induce HepG2 cell apoptosis,which may be achieved by inhibiting the expression of Bcl-2 and Survivin protein and enhancing the expression of P53 protein.