固脾消积饮诱导人肝癌HepG2细胞凋亡的分子机制

Molecular mechanisms of Gupi Xiaoji decoction inducing apoptosis of human hepatoma HepG2 cells

目的:探讨固脾消积饮对人肝癌细胞HepG2凋亡的分子机制。方法:将HepG2细胞分为4组:对照组(Control)、空白血清组(Blank)、固脾消积饮血清组(GPXJY)和顺铂组(Positive),每组设置8个复孔。固脾消积饮含药血清和顺铂干预24 h后,检测细胞活性、活细胞数量、细胞凋亡状态、细胞周期以及线粒体膜电位状况,检测细胞的脂质过氧化(MDA)水平、糖酵解速率和凋亡Bax、Bcl-2、Caspase-3蛋白的表达,检测细胞上清液三酰甘油(TG)、胆固醇(TC)、丙酮酸和葡萄糖的含量。结果:与Control组比较,GPXJY组(细胞的)抑制率增加、细胞数量减少、凋亡阳性细胞数增多(P<0.01),G1期细胞数目显著增加(P<0.05),细胞线粒体膜电位降低(P<0.01),糖酵解功能显著抑制,细胞中MDA水平升高,细胞Bax、Caspase-3的表达升高、Bcl-2的表达下降(P<0.05,P<0.01),细胞上清液中TC、TG、葡萄糖含量显著减少、丙酮酸含量显著增加(P<0.05,P<0.01)。结论:固脾消积饮可诱导HepG2细胞发生凋亡,可能对能量代谢发挥作用。

Objective:To investigate the molecular mechanisms of Gupi Xiaoji decoction on apoptosis of human hepatoma cells HepG2.Methods:HepG2 cells were divided into 4 groups:control group(Control),blank serum group(Blank),Gupi Xiaoji Yin serum group(GPXJY)and cisplatin group(Positive).Eight duplicate holes were set in each group.After treated with Gupi Xiaoji Decoction-containing serum or cisplatin for 24 hours,the cell viability,the number of viable cells,the state of apoptosis,the cell cycle and the mitochondrial membrane potential were detected,and the level of lipid peroxidation(MDA)and glycolysis rate of the cells were detected.The expressions of apoptotic Bax,Bcl-2,and Caspase-3 proteins,and the contents of triacylglycerol(TG),cholesterol(TC),pyruvate and glucose in the cell supernatant were detected.Results:Compared with the control group,in the GPXJY group,the inhibition rate was increased(P<0.05),the number of cells was decreased,the number of apoptosis-positive cells was increased(P<0.01),the number of cells in the G1 phase was increased significantly(P<0.05),and the cell membrane potential was decreased(P<0.05,P<0.01),the glycolytic function was inhibited significantly,the MDA level was increased,the expressions of Bax and Caspase-3 in the GPXJY group were increased,and the expression of Bcl-2 was decreased(P<0.05,P<0.01).In cell supernatant,the TC,TG and glucose contents were decreased significantly,and the pyruvate content was increased significantly(P<0.05,P<0.01).Conclusion:Gupi Xiaoji Decoction can induce apoptosis of HepG2 cells and may play a role in energy metabolism.