摘要
目的研究紫檀茋对脑缺血再灌注大鼠的脑保护作用,并考察其对血管内皮间质过度转化与蛋白激酶B/糖原合成激酶-3β(protein kinase B/glycogen synthesizing kinase-3β,Akt/GSK-3β)通路的影响。方法建立脑缺血再灌注大鼠模型,给予紫檀茋和硫酸氢氯吡格雷片干预6 d后,检测大鼠神经功能缺损程度;采用ELISA法检测各组大鼠血清血小板活化因子(platelet activating factor,PAF)、血小板源性CD40配体(CD40L)和血小板P选择素(P-slelctin,CD62P)水平;采用试剂盒检测各组大鼠脑组织超氧化物歧化酶(superoxide dismutase,SOD)活性和丙二醛(malondialdehyde,MDA)水平;采用苏木素-伊红(HE)染色法观察各组大鼠脑皮质病理变化;采用免疫荧光染色法检测各组大鼠脑血管内皮和间质标志物血小板-内皮细胞黏附分子-1(platelet endothelial cell adhesion molecule-1,PECAM-1,又名CD31)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)表达;采用Western blotting法检测各组大鼠脑血管CD31、α-SMA和脑皮质Akt和GSK-3β蛋白表达情况;采用免疫组化法检测各组大鼠脑皮质Akt和GSK-3β蛋白表达。结果与假手术组比较,模型组大鼠神经功能缺损评分显著升高(P<0.05);血清PAF、CD40L和CD62P水平显著升高(P<0.05);脑组织SOD活性显著降低,MDA水平升高(P<0.05);脑皮质细胞形态破坏,呈空泡样坏死,细胞间隙增大,染色变浅,残存细胞体积缩小,细胞核固缩、深染,细胞边界不清;脑血管CD31表达减弱,α-SMA表达增强;脑血管CD31和脑皮质p-Akt、p-GSK-3β蛋白表达水平显著降低(P<0.05),脑血管α-SMA蛋白表达水平显著升高(P<0.05)。与模型组比较,各给药组大鼠神经功能缺损评分显著降低(P<0.05);血清PAF、CD40L和CD62P水平降低(P<0.05);脑组织SOD活性升高(P<0.05),MDA水平降低(P<0.05);脑皮质细胞形态、排列、细胞核清晰度以及间质染色均匀度等均改善;脑血管CD31表达增强,α-SMA表达减弱;脑血管CD31和脑皮
Objective To study the cerebral protective effect of pterostilbene on cerebral ischemia-reperfusion rats,and investigate its effect on vascular endothelial mesenchymal transition and protein kinase B/glycogen synthesizing kinase-3β(Akt/GSK-3β)pathway.Methods A rat model of cerebral ischemia-reperfusion was established,and after 6 d of intervention with pterostilbene and clopidogrel hydrogen sulfate tablets,degree of neurological deficit in rats was detected;Platelet activating factor(PAF),platelet-derived CD40 ligand(CD40L)and platelet P-selectin(CD62P)levels in serum were detected by ELISA;Kits were used to detect superoxide dismutase(SOD)activity and malondialdehyde(MDA)level in brain tissue of rats in each group;Hematoxylin-eosin(HE)staining was used to observe the pathological changes of cerebral cortex of rats in each group;Immunofluorescence staining was used to detect the cerebral vascular endothelial and interstitial markers platelet endothelial cell adhesion molecule-1(PECAM-1,also known as CD31)andα-smooth muscle actin(α-SMA)expressions of rats in each group;Western blotting was used to detect CD31 andα-SMA protein expressions in cerebrovascular,Akt and GSK-3βprotein expressions in cerebral cortex of rats;Akt and GSK-3βexpressions in cerebral cortex of rats in each group were detected by immunohistochemistry.Results Compared with sham group,neurological deficit score of rats in model group was significantly increased(P<0.05);Levels of PAF,CD40L and CD62P in serum were significantly increased(P<0.05);SOD activity in brain tissue was significantly decreased,and MDA level was increased(P<0.05);Cerebral cortical cells were morphologically damaged,showing vacuolar necrosis,intercellular space was increased,staining was lighted,residual cell volume was reduced,nuclei were pyknotic and hyperchromatic,and cell boundaries were unclear;CD31 expression in cerebral blood vessels was weakened,α-SMA expression was enhanced;CD31 protein expression in cerebrovascular,p-Akt and p-GSK-3βprotein expressions in ce
作者
汪宁
高军
白方会
刘圆方
温昌明
WANG Ning;GAO Jun;BAI Fang-hui;LIU Yuan-fang;WEN Chang-ming(Interventional Ward of Cerebrovascular Disease,Department of Neurology,Nanyang Central Hospital,Nanyang 473000,China;Department of Pediatric Surgery,Nanyang Central Hospital,Nanyang 473000,China)
出处
《中草药》
CAS
CSCD
北大核心
2022年第8期2399-2406,共8页
Chinese Traditional and Herbal Drugs
基金
河南省科技发展计划项目(192102310349)。
关键词
紫檀茋
脑缺血再灌注
血管内皮-间质过度转化
蛋白激酶B
糖原合成激酶-3β
pterostilbene
cerebral ischemia-reperfusion
endothelial-mesenchymal excessive transition
protein kinase B
glycogen synthesis kinase-3β
