体外缺血再灌注脑损伤中NF-κB的表达及白藜芦醇苷的保护作用徐静1.DOC

体外缺血再灌注脑损伤中NF-κB的表达及白藜芦醇苷的保护作用 徐静徐静（1981），女（满族），辽宁省人，重庆医科大学附属儿童医院急诊医学专业在读硕士，email:xujingwws@126.com*国家自然科学基金资助项目 *重庆市自然科学基金资助项目（2005BB5228）王兴勇 杨廷芳 胡语航 熊燚 周海银 徐静（1981），女（满族），辽宁省人，重庆医科大学附属儿童医院急诊医学专业在读硕士，email:xujingwws@126.com *国家自然科学基金资助项目 *重庆市自然科学基金资助项目（2005BB5228） 重庆医科大学附属儿童医院（40014） 摘要：目的：探讨体外缺血再灌注（ischemia /reperfusion ,I/R）脑损伤中NF-κB的表达及白藜芦醇苷（polydatin, PD）的保护作用。方法：将体外培养7d的大鼠大脑皮层神经元随机分为3组：对照组、模型组和PD组。AO/EB及DAPI染色观察细胞凋亡的形态学变化，同时测定再灌注0h、6h、12h、24h、48h时细胞凋亡率、Bcl-2/Bax、NF-κBp65的动态变化。结果：模型组可见凋亡细胞的特征性变化；与对照组相比，模型组细胞凋亡率、NF-κBp65的表达量随再灌注时间的延长而明显增加（P＜0.05），24h达高峰，随后渐降低，Bcl-2/Bax的高峰点位于6h；PD组凋亡率及NF-κBp65的表达量较模型组均明显降低（P＜0.05），Bcl-2/Bax则明显升高（P＜0.05）。结论：NF-κBp65活化参与了体外I/R脑损伤的病理过程，PD可能通过抑制NF-κBp65的表达、上调Bcl-2/Bax，阻止神经元凋亡，从而减轻I/R脑损伤。 关键词：白藜芦醇苷、缺血再灌注、NF-κBp65、Bcl-2/Bax、凋亡 Expression of NF-ΚB and protective effect of polydatin in vitro following cerebral ischemia /reperfusion injury XU Jing, WANG Xing-yong, YANG Ting-fang, HU Yu-hang, XIONG Yi, ZHOU Hai-yin, HEI Rong． Children’s Hospital，Chongqing Medical University 400014．China Objective: To observe the expression of NF-κB and the influences of polydatin (PD) in vitro following cerebral ischemia /reperfusion (I/R) injury. Methods:Cortical neurons cultured 7 days in vitro from newborn rat were randomly divided into contraol group, I/R model group and PD group. Then mMorphological features of apoptosis were observed by fluorescence microscope. Apoptosis,Bcl-2/Bax, the expression of NF-κB p65 were determined on 0h,6h,12h,24h,48h after reperfusion. Results: Morphological diversify of apoptotic neurons can be observed by fluorescence microscope (AO/EB and DAPI) in I/R model group and PD group. Compared with contral control group ,the number of apoptotic cells and NF-κBp65 increased significantly（P＜0.05）with time after reperfusion and reached its peaked at 24h and then decreased. Bcl-2/Bax increased with to peak at 6h. In PD groups, the number of apoptotic cells and NF-κBp65 decreased obviously（P＜0.05） obviously co