NF-κB信号通路在小鼠脑缺血再灌注细胞凋亡中的作用  

The effect of NF-κB signaling pathway on apoptosis in cerebral ischemia reperfusion injury in mice

作　　者：朱晓瓞 余资江[1] 孙宝飞[1] 余彦[1] 李玉美[1] 罗时鹏[1] 肖朝伦[2] 贺菊芳[1] 

机构地区：[1]贵州医科大学基础医学院人体解剖学教研室,贵州贵阳550025 [2]贵州医科大学基础医学院人体解剖学实验中心,贵州贵阳550025

出　　处：《中风与神经疾病杂志》2016年第6期484-488,共5页Journal of Apoplexy and Nervous Diseases

基　　金：国家自然基金(No.81060108);贵州省科技厅2012年社会发展攻关项目[黔科合SY字(2012)3153号]

摘　　要：目的初探核因子-κB(nuclear factor-kappa B,NF-κB)信号通路在小鼠脑缺血再灌注损伤(cerebral ischemia reperfusion injury,CIRI)细胞凋亡中的作用。方法通过夹闭小鼠双侧颈总动脉建立动物模型。模型组分为3 h、6 h、12 h、1 d、3 d、7 d、14 d、21 d,共8组。TTC染色确定脑缺血损伤区域,TUNEL法检测不同时间缺血区细胞凋亡数,原位杂交法和Western blotting检测NF-κB信号通路靶基因/蛋白Bcl-2和C-myc mRNA及蛋白表达。PDTC抑制NF-κB信号通路后,TUNEL法和原位杂交检测建模后1 d、7 d、14 d神经细胞凋亡数及Bcl-2和C-myc mRNA变化情况。结果各模型组海马CA3区凋亡细胞数量、Bcl-2 mRNA和C-myc mRNA阳性细胞数及二者蛋白表达量高于正常组和假手术组(P<0.05);PDTC抑制NF-κB信号通路后,各抑制组与对应时间点模型组相比,Bcl-2 mRNA阳性细胞数减少,TUNEL阳性细胞数和C-myc mRNA阳性细胞数增加(P<0.05)。结论 CIRI早期即出现有凋亡细胞数的增加,并在其后较长一段时间内细胞凋亡过程仍存在着持续性的进展;NF-κB信号通路在CIRI病程进展中对神经细胞凋亡起抑制作用,其机制可能通过Bcl-2诱导和C-myc抑制共同发挥调节作用。Objective To investigate the possible effect of NF-κB signaling pathway on apoptosis in cerebral ischemia reperfusion injury（ CIRI） in mice. Methods CIRI models were copied by stopping the blood of common carotid arteries in both sides at the same time. Mice models were divided into 8 groups according to the reperfusion time： 3 h,6 h,12 h,1 d,3 d,7 d,14 d and 21 d. The accurate injured region was confirmed by TTC staining. The number of apoptotic cells was detected by TUNEL and the changes of the target gene / protein Bcl-2 and C-myc in NF-κB signaling pathway were detected by insitu hybridization（ ISH） detection and western blotting. And the TUNEL and ISH detection were used again for the groups in which the NF-κB signaling pathway was controlled by using PDTC. The groups were also divided into 1 d,7 d and 14 d group.Results The number of apoptotic cells,expression of mRNA and protein of Bcl-2 and C-myc significantly increased compared with the normal group and sham-operated group（ P 〈0. 05）. The number of Bcl-2 mRNA positive cells significantly decreased compared with the model group at the same reperfusion time when PDTC was used to restrain NF-κB,while the number of TUNEL and C-myc mRNA positive cells increased（ P 〈0. 05）. Conclusion Apoptosis happens in the early of CIRI and is in a sustainable progress for a longer time. NF-κB signaling pathway may play a suppressive role in the regulation of apoptosis in cerebral ischemia reperfusion injury by controling both of Bcl-2 up-regulation and C-myc down-regulation.

关 键 词：脑缺血再灌注 细胞凋亡 NF-ΚB信号通路 BCL-2 C-MYC 小鼠 

分 类 号：R743.3[医药卫生—神经病学与精神病学]