白藜芦醇在缺糖缺氧/再灌注损伤不同时间窗对大鼠皮质神经元的保护作用  

Effects of resveratrol on oxygen and glucose deprivation/reperfusion injury of rat primary cortical neurons at different time windows

作　　者：黄家贵[1,2] 张黎黎[1] 沈长波[1] 刘舒[1] 徐兰[1] 杨琴[1] 

机构地区：[1]重庆医科大学附属第一医院神经内科,重庆400016 [2]宜宾市第二人民医院神经内科,四川宜宾644000

出　　处：《中国药理学通报》2013年第9期1230-1234,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金面上项目(No 81071119);重庆市卫生局科研基金资助项目(No 2009-2-359)

摘　　要：目的研究白藜芦醇在缺糖缺氧/再灌注损伤不同时间窗内对大鼠皮质神经元的保护作用及机制。方法大鼠皮质神经元缺糖缺氧处理150 min,随即恢复正常培养24 h。实验分为正常组,模型组,白藜芦醇预处理组、造模时处理组、造模后处理组和全程处理组。倒置显微镜下观察细胞形态。化学比色法测定LDH活性和GSH含量。MTT法检测细胞活力。TUNEL检测细胞凋亡。免疫印迹法检测Bcl-2、Caspase-3蛋白表达。结果在白藜芦醇全程处理组中,各浓度白藜芦醇均可降低培养上清液LDH活性,升高细胞内GSH含量(P<0.05),其中以40μmol·L-1白藜芦醇作用最佳。在缺糖缺氧再灌注损伤的不同时间内给予白藜芦醇,均较模型组增强神经元活力,减少神经元凋亡,上调Bcl-2蛋白、下调Caspase-3蛋白的表达(P<0.05),其中全程处理组作用最佳,其次为预处理组。结论白藜芦醇对缺糖缺氧/再灌注损伤的神经元有剂量依赖性保护作用,而且全程处理组作用最佳,预处理组次之。其保护作用机制可能至少部分是通过调节凋亡相关蛋白而实现。Aim To study the protective effect of res- veratrol on oxygen and glucose deprivation/reperfusion injury（OGD/Rep） of rat cortical neurons at different time windows and its possible mechanisms. Methods Rat cortical neurons were cultured with oxygen and glucose deprivation for 150 minutes and returned to normal culture for 24 hours. The cells were divided in- to normal group（Nor）, model group（Mod）, pre-treat- ment group （ Pre）, OGD-treatment group （ OGD）, post- treatment group （Post） and the whole processing group （WP）. The cell morphology was observed under invert microscope. The activity of L-lactate dehydrogenase （LDH） and the content of glutathione （GSH） were de- tected by chemical colorimetry. Cell viability was de- tected with MTT assay. Apoptosis of neurons was ana- lyzed by TUNEL. The expression levels of Bcl-2 and Caspase-3 protein were observed by Western blot. Re- suits Resveratrol of various concentrations could sig- nificantly reduce the activity of LDH for the cultured supernatant and elevate the content of intracellular GSH in the whole processing group（P 〈 0. 05 ）. More- over, 40μmol·L^-1 resveratrol had the best effect on LDH and GSH. Resveratrol significantly strengthened cell viability, reduced the numbers of apoptotic cells, increased the expression level of Bcl-2 protein, and de- creased the expression level of Casepase-3 protein com- pared with model group at any stage of OGD/Rep. Furthermore, there was the best effect in the whole processing group and better effect in the pre-treatment group. Conclusions Resveratrol has dose dependent protective effects on rat primary cortical neurons during OGD/Rep. Moreover, there is the best effect in the whole processing group and better effect in the pre- treatment group. Its protective mechanism may be at least partly associated with the regulation of apoptosis related proteins.

关 键 词：白藜芦醇 神经元 缺糖缺氧 再灌注损伤 细胞凋 亡 保护作用 时间窗 

分 类 号：R-332[医药卫生] R284.1