参附注射液对肾母细胞瘤细胞增殖、凋亡的影响及机制探讨

Effects of Shenfu injection on proliferation and apoptosis of Wilm's tumor cells

目的观察参附注射液对肾母细胞瘤细胞增殖及凋亡的影响,并探讨其机制。方法取适量对数生长期肾母细胞瘤细胞,分为低、中、高浓度组及对照组。低、中、高浓度组分别加入0.4、0.8、1.2 mg/m L参附注射液2m L,对照组加入等量无菌DMSO培养液。干预24 h时采用MTT法检测各组细胞增殖抑制率,采用流式细胞仪测算各组细胞凋亡率,采用Western blot法检测各组细胞p53、B细胞淋巴瘤因子l-2(bcl-2)蛋白。结果干预24 h时低浓度组、中浓度组、高浓度组、对照组的细胞增殖抑制率分别为22.83%±5.18%、35.76%±6.01%、54.51%±9.07%、6.39%±2.81%,组间比较,P均<0.05。干预24 h时低浓度组、终浓度组、高浓度组、对照组的细胞凋亡率分别为14.59%±1.24%、18.27%±1.73%、21.85%±1.62%、3.92%±1.36%,组间比较,P均<0.05。各组细胞p53的相对表达量为高浓度组<中浓度组<低浓度组<对照组,P均<0.05;bcl-2的相对表达量为高浓度组>中浓度组>低浓度组>对照组,P均<0.05。结论参附注射液可抑制肾母细胞瘤细胞增殖,促进细胞凋亡;其机制可能与降低p53表达、升高bcl-2表达有关。

Objective To investigate the effects and related mechanism of Shenfu injection on the proliferation and apoptosis of Wiling tumor cells in vitro. Methods Wilm＇s tumor ceils were divided into the control group, low-dose, medi- um-dose and high-dose groups. The low-dose, medium-dose and high-dose groups were treated with 2 ml, of 0.4 mg/mL, 0.8 mg/mL and 1.2 mg/mL Shenfu injection, and the control group was added with DMSO. After 24-hour intervention, the proliferation inhibition rate was detected by MTT, the rate of apoptosis and the expression of p53 and Bcl-2 protein was detected by flow cytometry and Western blotting, respectively. Results After 24-hour intervention, the proliferation inhibi- tion rates of the low-dose group, medium-dose group, high-dose group and the control group were respectively 22.83% ± 5.18% , 35.76% ±6.01%, 54.51% ±9.07% and 6.39% ±2.81%, all P〈0.05. After 24-hour intervention, the ap- optosis rates of the low-dose group, medium-dose group, high-dose group and the control group were 14.59% ± 1.24%, 18.27% ± 1.73%, 21.85% ± 1.62% and 3.92% ± 1.36%, respectively, all P 〈 0.05. The relative expression of p53 was high-dose group 〈 medium-dose group 〈 low-dose group 〈 control group, all P 〈 0.05 ; and the relative expression of Bcl-2 was high-dose group 〉 medium-dose group 〉 low-dose group 〉 control group, all P 〈 0.05. Conclusiion Shenfu in- jection can inhibit the proliferation of Wiling tumor ceils and induce its apoptosis, and the mechanism may be related with the down-regulated expression of p53 and up-regulated expression of Bcl-2.