黄芪甲苷对局灶性脑缺血后适应中Wnt/β-catenin信号通路关键因子表达的影响  

Study of astragaloside-IV on the expression of key factors of Wnt/β-catenin signaling pathway in local cerebralischemic post-conditioning

作　　者：徐玥玮[1] 王丽[1] 高晓明[1] 谢若男[1] 王雷 杨满琴[1] XU Yuewei;WANG Li;GAO Xiaoming;XIE Ruonan;WANG Lei;YANG Manqin(Department of Pharmacy,the 2^(nd) Affiliated Hospital of Anhui University of Traditional Chinese Medicine,Hefei 230061,Anhui,China;School of Pharmacy,Anhui University of Traditional Chinese Medicine,Hefei 230061,Anhui,China)

机构地区：[1]安徽中医药大学第二附属医院药学部,安徽合肥230061 [2]安徽中医药大学药学院,安徽合肥230061

出　　处：《贵州医科大学学报》2022年第2期171-178,共8页Journal of Guizhou Medical University

基　　金：安徽省自然科学基金(1908085QH351);安徽中医药大学科研项目(2019zryb23)。

摘　　要：目的探讨黄芪甲苷对局灶性脑缺血后适应中Wnt/β-连环蛋白(Wnt/β-catenin)信号通路关键因子表达的影响。方法将SD大鼠随机分为假手术组(Sham组)、缺血再灌注模型组(I/R组)、缺血后适应组(IPC)、黄芪甲苷(低、中、高)+缺血后适应组(IPC+AS-IV10组、IPC+AS-IV20组、IPC+AS-IV50组)、尼莫地平+缺血后适应组(IPC+Nim组),共7组;采用线栓法制作大脑中动脉闭塞后缺血再灌注模型;对各组大鼠进行神经功能损伤评分,采用2,3,5-氯化三苯基四氮唑(TCC)染色检测各组大鼠脑梗死区面积,HE染色检测各组大鼠缺血侧脑组织的病理学变化,ELISA试剂盒检测各组大鼠血清中脑组织损伤标志物大鼠源高敏c反应蛋白(hs-CRP)和大鼠源S100钙结合蛋白B(S100-B)的表达,Real time PCR和Western blot检测大鼠脑组织中Wnt家族成员3A(Wnt3a)、β-连环蛋白(β-catenin)、脑源性神经营养因子(BDNF)mRNA和蛋白以及磷酸化β-catenin(p-β-catenin)蛋白的表达。结果神经损伤评分结果显示,各处理组较I/R组评分降低(P<0.05);梗死面积结果显示,IPC组、IPC+AS-IV20组、IPC+AS-IV50组、IPC+Nim组较I/R组减少,IPC+AS-IV50组、IPC+Nim组较IPC组减少(P<0.05);血清S-100B、hs-CRP含量结果显示,I/R组明显高于Sham组(P<0.05),IPC组、IPC+AS-IV10组、IPC+AS-IV20组、IPC+AS-IV50组、IPC+Nim组低于I/R组(P<0.05),IPC+AS-Ⅳ10组、IPC+AS-IV20组、IPC+AS-IV50组、IPC+Nim组低于IPC组(P<0.05);病理结果显示,各处理组较I/R组有不同程度减轻;Real time PCR和Western blot结果显示,与Sham组比较,I/R组大鼠脑组织中Wnt3a、β-catenin、BDNF mRNA和蛋白的表达降低(P<0.05),而p-β-catenin蛋白升高(P<0.05);与I/R组比较,IPC组及各处理组Wnt3a、β-catenin、BDNF mRNA和蛋白的表达增加(P<0.05),而p-β-catenin蛋白水平下降(P<0.05);与IPC组比较,IPC+AS-IV20组、IPC+AS-IV50组、IPC+Nim组大鼠脑组织中BDNF、Wnt 3 a、β-catenin mRNA升高(P<0.05),大鼠脑组织中Wnt 3 a、β-catenin mRNObjective To investigate the effect of astragaloside-IV on expression of key factors of Wnt/β-catenin signaling pathway in local cerebral-ischemic post-conditioning.Methods SD rats were randomly divided into 7 groups:Sham group,ischemic reperfusion model group(I/R group),ischemic post-conditioning group(IPC group),astragaloside(low,medium,high)+ischemic post-conditioning group(IPC+AS-IV10 group,IPC+AS-IV20 group,IPC+AS-IV50 group),nimodipine+ischemic post-conditioning group(IPC+Nim group).Suture method was adopted to create cerebral arterial occlusion then ischemic reperfusion model.Neurological Deficiency Score was adopted to evaluate all groups,2,3,5-TCC staining was used to detect cerebral infarct area;pathological changes of ischemic cerebral tissues were evaluated by HE staining in each group.ELISA kit was used to detect cerebral tissue damage markers:expression of hs-CRP and S100-B;Real time PCR and Western blot were used to detect expression of Wnt3a,β-catenin,BDNF mRNA and p-β-catenin.Results Neural injury scores indicated that treated groups showed lowered scores compared with I/R group(P<0.05);infarct area results indicated that IPC group,PC+AS-IV20 group,IPC+AS-IV50 group,IPC+N im group showed reduced area compared with I/R group;IPC+AS-IV50 group,IPC+Nim group showed reduced area compared with IPC group(P<0.05).Serum S-100B,hs-CRP content results indicated that I/R group was obviously higher than Sham group(P<0.05);IPC group,IPC+AS-IV10 group,IPC+AS-IV20 group,IPC+AS-IV50 group,IPC+Nim group were lower than I/R group(P<0.05);IPC+AS-IV10 group,IPC+AS-IV20 group,IPC+AS-IV50 group,IPC+Nim group were lower than PC group(P<0.05).Pathological results indicated that all groups showed various degree of improvement compared with I/R group;Real time PCR and Western blot results indicated that Wnt3a,β-catenin,BDNF mRNA and expression of protein in rat cerebral tissues of I/R group were lowered compared with Sham group(P<0.05),while p-β-catenin protein was improved(P<0.05);comparing with I/R group,Wnt3a,β-c

关 键 词：黄芪甲苷 大鼠 脑缺血再灌注损伤 WNT/Β-CATENIN通路 缺血后适应 神经保护 

分 类 号：R743[医药卫生—神经病学与精神病学]