摘要
目的探讨白藜芦醇预处理对缺糖缺氧再灌注所致大鼠原代皮质神经元损伤的保护作用及机制。方法 40μmol/L白藜芦醇预处理大鼠原代皮质神经元24 h,皮质神经元缺糖缺氧150 min,恢复正常培养24 h,构建缺糖缺氧再灌注损伤。实验分为正常组,模型组,白藜芦醇组。倒置显微镜下观察细胞形态。免疫荧光染色鉴定神经元。MTT法检测细胞活力。化学比色法测定培养上清液LDH活性和细胞内SOD活性。TUNEL检测细胞凋亡。免疫印迹法和免疫荧光法检测Bcl-2、Caspase-3、Nrf-2、NQO-1蛋白表达。结果细胞培养6 d,免疫荧光染色显示90%的细胞是神经元特异性烯醇化酶阳性。与模型组相比,白藜芦醇预处理能明显降低培养上清液乳酸脱氢酶活性,增强细胞内SOD活性和神经元细胞活力,减少细胞凋亡,上调Bcl-2、Nrf-2和NQO-1蛋白表达,下调Caspase-3蛋白表达;促进Nrf-2蛋白从细胞浆转移到细胞核。结论白藜芦醇对大鼠原代皮质神经元缺糖缺氧再灌注损伤有良好的保护作用,其机制可能是通过抗凋亡和抗氧化途径实现。
AIM To study the effect and mechanism of resveratrol on rat primary cortical neurons during oxygen and glucose deprivation / reperfusion injury（ OGD / Rep）. METHODS Rat primary cortical neurons were pretreated with 40 μmol / L resveratrol for 24 hours,then cultured with oxygen and glucose deprivation for 150 minutes and returned to normal culture for 24 hours. Cells were divided into three groups including the normal group,model group and resveratrol group. MTT assay detected cell viability. Chemical colorimetry detected SOD and LDH activity. TUNEL detected cell apoptosis. The expression levels of Bcl-2,Caspase-3,nuclear factor erythroid 2-related factor 2（ Nrf-2）,and NAD（ P） H quinone oxidoreductase-1（ NQO-1） proteins were detected by Western blot. RESULTS Compared with model group,resveratrol significantly reduced LDH activity,enhanced intracellular SOD activity and neuronal cell viability,decreased the numbers of apoptosis and the expression level of Caspase-3 protein,increased the expressions of Bcl-2,Nrf-2,and NQO-1 proteins,and promoted the Nrf-2 to enter into the nuclei from the cytosol. CONCLUSION Resveratrol has a good protective effect on rat primary cortical neurons during OGD / Rep. Its protective mechanism may be associated with antioxidation and anti-apoptosis.
出处
《中成药》
CAS
CSCD
北大核心
2014年第5期897-903,共7页
Chinese Traditional Patent Medicine
基金
国家自然科学基金面上项目(81071119)
重庆市卫生局科研基金(2009-2-359)
