补阳还五汤对脑缺血再灌注后星形胶质细胞connexin43影响机制的研究  

Mechanisms of Buyang Huanwu Decoction Influencing Connexin43 after Focal Ischemia

作　　者：郭建超[1] 苏宁[1] 刘微[1] 吴绍锋 周乐全[1] 关莉[1] 张洋[1] 李小英[1] 

机构地区：[1]广州中医药大学基础医学院,广东广州510006

出　　处：《辽宁中医杂志》2017年第11期2418-2420,I0006,共4页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81673772);广东省高等学校优秀青年教师培养项目(Yq2013041);高等学校博士学科点专项科研基金(20124425120011)

摘　　要：目的:探讨补阳还五汤对大鼠脑缺血再灌注后星形胶质细胞缝隙连接蛋白43(connexin43,Cx43)的影响机制。方法:运用线栓法建立大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)模型,补阳还五汤治疗1 d后ELISA法检测细胞因子[白细胞介素-1 beta,(IL-1β)和白细胞介素-10,(IL-10)]的表达,通过使用NF-κB激动剂增强炎症反应后,比较海马CA1区Cx43的表达,同时,使用HE染色观察细胞形态学改变。在补阳还五汤治疗7 d后,免疫组织化学染色法观察b FGF的表达情况,联合使用b FGF中和抗体后比较各组海马CA1区Cx43的表达同时HE染色观察细胞形态学改变。结果:同模型组相比,补阳还五汤组脑缺血再灌注1天后,IL-1β减少IL-10增加(P<0.05);联合使用NF-κB激动剂后补阳还五汤对海马CA1区Cx43的下调作用被抑制(P<0.05),HE染色显示联合组较补阳还五汤组神经元损伤加重。脑缺血7 d时补阳还五汤增强b FGF和Cx43的表达;联合使用b FGF中和抗体后海马CA1区Cx43的表达较补阳还五汤组降低(P<0.05),HE染色显示同补阳还五汤组比,联合组神经元状态更差。结论:补阳还五汤在脑缺血后1 d可能是通过抑制炎症下调Cx43发挥保护作用,而在脑缺血后7 d可能通过上调b FGF增强Cx43促进修复。Objective： To explore the mechanisms of Buyang Huanwu Decoction（ BYHWD） influencing connexin 43（ Cx43） expression after focal cerebral ischemia in rats. Methods： Middle cerebral artery occlusion（ MCAO） and reperfusion model was established by Longa method. One day after treatment with BYHWD,ELISA was used to detect levels of inflammatory cytokines（ interleukin-1,IL-1β,interleukin-10,IL-10）. Cx43 expression was compared by immunohistochemical staining after treatment with NF-κB agonist,and HE staining was used to observe cell morphological changes. Seven days after treatment with BYHWD,immunohistochemical staining was used to investigate the expression of b FGF and Cx43,and HE staining was used to observe cell morphology changes. Results： Compared with ischemia group,IL-1β was decreased whereas IL-10 was increased by BYHWD on day 1. The downregulation of Cx43 expression in hippocampus CA1 by BYHWD was inhibited by combination treatment with NF-κB agonist on day 1（ P〈0. 05） and neuronal damage was aggravated in combination group. On day 7 afer focal ischemia,b FGF and Cx43 expressions were higher in BYHWD group than that in ischemia group（ P〈0. 05）. Cx43 expression was significantly decreased after combination of b FGF neutralizing antibody with BYHWD（ P〈0. 05）,meanwhile neuronal damage was aggravated in combination group. Conclusion： The effects of BYHWD on Cx43 may be regulated by NFκB or b FGF and related with its curative effects of alleviating injury and facilitating functional restoration after focal cerebral ischemia and reperfusion.

关 键 词：补阳还五汤 脑缺血再灌注 星形胶质细胞 连接蛋白43 BFGF 

分 类 号：R743.33[医药卫生—神经病学与精神病学]