生脉散对H_(2)O_(2)损伤PC12细胞保护作用的研究

Study on the Protective Effect of Shengmai-san on PC12 Cells Damaged by H_(2)O_(2)

目的探究生脉散(SMS)对PC12细胞增殖的影响和经H_(2)O_(2)损伤后对PC12细胞的保护作用。方法用H_(2)O_(2)处理小鼠PC12细胞,构建氧化应激损伤细胞模型;通过CCK8法检测SMS对PC12细胞增殖的影响以及SMS对H_(2)O_(2)诱导损伤后的PC12细胞存活率的影响;将PC12细胞分为空白对照组、H_(2)O_(2)损伤模型组、SMS低剂量组、SMS中剂量组、SMS高剂量组,对照组不做处理,其他组均采用H_(2)O_(2)诱导损伤,SMS低、中、高剂量组则分别加入浓度为0.1、1.0、5.0 g/ml的SMS,然后检测胞内外乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)的表达水平。通过蛋白质印迹法检测SMS对PC12细胞的核因子E2相关因子2(Nrf-2)蛋白表达水平的影响。结果SMS各含量组能促进正常PC12细胞的增殖、提高氧化损伤的PC12细胞的存活率;能使细胞外LDH活性减弱、MDA含量下降,SOD活力增强,差异有统计学意义(P<0.05);SMS可上调PC12损伤细胞中Nrf-2蛋白的表达,差异有统计学意义(P<0.05)。结论SMS能促进PC12细胞的增殖且对H_(2)O_(2)诱导的细胞损伤有保护作用,其机制可能与增强核内Nrf-2蛋白表达有关。

Objective To investigate the effect of Shengmai-san(SMS)on the proliferation of PC12 cells and the protective effect of SMS on PC12 cells after H_(2)O_(2) injury.Methods PC12 cells were treated with H_(2)O_(2) to construct a model of oxidative stress injury;the effect of SMS on the proliferation of PC12 cells and the effect of SMS on the survival rate of PC12 cells after H_(2)O_(2)-induced injury were detected by CCK8 method.PC12 cells were divided into blank control group,H_(2)O_(2) injury model group,SMS low dose group,SMS medium dose group and SMS high dose group,while the control group was not treated,and the other groups were induced by H_(2)O_(2) injury.SMS low,medium and high dose groups were added with SMS at concentrations of 0.1,1.0 and 5.0 g/ml,and then the expression levels of intracellular and extracellular lactate dehydrogenase(LDH),malondialdehyde(MDA)and superoxide dismutase(SOD)were detected.The effect of SMS on the expression of nuclear factor E2-related factor 2(Nrf-2)protein in PC12 cells was detected by Western blotting.Results Different concentrations of SMS groups could promote the proliferation of normal PC12 cells and increase the survival rate of oxidative damaged PC12 cells,meanwhile the activity of extracellular LDH was decreased,the content of MDA was decreased,and the activity of SOD was increased,the difference was statistically significant(P<0.05).SMS could up-regulate the expression of Nrf-2 protein in PC12 damaged cells,and the difference was statistically significant(P<0.05).Conclusion SMS can promote the proliferation of PC12 cells and protect against H_(2)O_(2)-induced cell injury,and the mechanism may be related to the enhancement of Nrf-2 protein expression in the nucleus.