白藜芦醇对暂时性氧糖剥夺致神经元损伤的保护作用

Protective effects of resveratrol against neuronal injury induced by instant OGD in vitro

目的:探讨白藜芦醇对暂时性氧糖剥夺致神经元损伤的保护作用及机制。方法:以小鼠大脑皮层原代神经元氧糖剥夺4h"再灌注"20h模型为对照。白藜芦醇浓度分别为10、25、50、100μmol/L,治疗时间从缺氧开始,直至试验结束。台盼蓝染色法和LDH漏出率评估细胞活力。采用免疫印迹法分析基质金属蛋白酶-9(MMP-9)蛋白表达,采用RT-PCR检测MMP-9 mRNA水平。结果:台盼蓝染色和LDH漏出率结果显示,50μmol/L白藜芦醇即可对离体神经元缺血再灌注损伤具有良好保护作用,浓度高达100μmol/L亦没有发现明显的副作用。免疫印迹和半定量RT-PCR结果显示,与损伤组比较,50和100μmol/L两种浓度的白藜芦醇可降低MMP-9蛋白的表达及MMP-9 mRNA的水平(P<0.01)。结论:白藜芦醇可抑制神经元MMP-9的表达,从而发挥其对暂时性氧糖剥夺致神经元损伤的保护作用。

Objective To study the protective effects and mechanism of resveratrol against neuronal injury induced by instant oxygen glucose deprivation （OGD） in vitro. Methods Primary mouse cerebral cortex neuronal ischemia reperfusion model induced by OGD 4 h ＂reperfusion＂ 20 h in vitro was used as vehicle. For treatment, the resveratrol was added to cultures to give the desired final concentrations （10, 25, 50 and 100 μmol/L）. The duration of treatment is from OGD to the end of the experiment. Cell viability was evaluated by Typan Blue staining and LDH leakage rate. Western Blot and RT-PCR were used to study the expression and the mRNA level of matrix metallopmteinase 9 （MMP-9）. Results The results of Typan Blue staining and LDH leakage rate indicated that treatment with 50 μmol/L resveratrol showed better therapeutic outcome, and no harmful effects were found with 100 μmol/L resveratrol treatment. The results of Western Blot and semi-quantitative RT-PCR showed that better inhibition against MMP-9 were found at 50 and 100 μmol/L of resveratrol （P 〈 0.01 ）. Conclusion The neuroprotection of resveratrol against neuronal injury induced by instant OGD may be achieved by inhibition against MMP-9.