维生素E对神经细胞氧化应激损伤的保护作用

The Protective Effect of VitE on PC12 Cell Injury Induced by Oxidative Stress

目的探讨维生素E对FeSO4和H2O2诱导PC12细胞氧化应激损伤模型的保护作用及其可能的机制。方法采用FeSO4和H2O2作用产生自由基的方法诱导建立PC12细胞氧化应激损伤模型。MTT比色法测定细胞活性,硫代巴比妥酸法测定MDA含量,黄嘌呤氧化酶法测定SOD活力,免疫细胞化学法测定α7烟碱型乙酰胆碱受体(nAchR)阳性表达及dot blot法测定α7nAchR亚单位水平。结果与FeSO4和H2O2损伤组(0.136±0.015)进行比较,维生素E组(0.178±0.025)和EGCG组(0.181±0.029)均能明显提高PC12细胞的活细胞数量(P<0.01),提高SOD活力(15.29±0.79)U.mL-1对(17.37±0.76)U.mL-1、(17.90±0.44)U.mL-1,P<0.01,有效降低MDA的含量(4.51±0.20)mmol.L-1对(3.39±0.11)mmol.L-1、(3.20±0.15)mmol.L-1,P<0.01,明显上调α7nAChR的表达(0.0910±0.0272)对(0.1895±0.0392)、(0.1956±0.0322),P<0.01。结论维生素E对FeSO4和H2O2诱导建立PC12细胞氧化损伤模型具有明显的保护作用,其作用机制可能与维生素E清除自由基,抑制脂质过氧化过程和保护α7nAchR有关。

Objective To explore the protective effect of Vitamin E on PC12 cell injury induced by H2O2 / Fe2+ and its underlying mechanisms.Methods PC12 cells were treated with H2O2 / Fe2+ to reproduce the cell injury model.Cell survival was assessed by Thiazzolyl blue(MTT) assay.The content of maleic diadehyde(MDA) and super oxide dismutase(SOD) activity were determined by thiobarbituric acid and xanthine oxidation enzyme respectively.The protein expression of α7 nicotinic acetylcholine receptor(nAchR) was examined by using immunocytochemical detecting kit,BCA protein kit and Dot blot assay.Results As compared with the group of FR(15.29±0.79) U·mL-1,the group of VitE(17.37±0.76) U·mL-1 and EGCG(17.90±0.44) U·mL-1 enhanced significantly the content of SOD and the PC12 cell survivability(P<0.01).In addition,α7nAChR in PC12 cell was significantly up-regulated.The expressions of α7nAChR(0.1956±0.0322)were significantly higher than those in the group of FR(0.0910±0.0272)(P<0.01).MDA in group VitE and group EGCG were significantly reduced from(4.51±0.20)mmol·L-1 to(3.39±0.11)mmol·L-1and(3.20±0.15)mmol·L-1(P<0.01).Conclusion Vitamin E can prevent against intoxication of PC12 induced by H2O2 / Fe2+.The mechanism maybe involve in free radical scavenging activity,lipid per oxidation process being inhabited and α7nAchR up-regulated.