片仔癀对MCAO大鼠血脑屏障通透性及AMPK/PKC通路的影响  

Effects of Pien Tze Huang on blood brain barrier permeability and AMPK/PKC pathway in MCAO rats

作　　者：张小琴[1] 林钰欣 韩美婷 黄莉莉 吴珍琴 黄鸣清[1] ZHANG Xiaoqin;LIN Yuxin;HAN Meiting;HUANG Lili;WU Zhenqin;HUANG Mingqing(College of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Department of Encephalopathy,Fuzhou Traditional Chinese Medicine Hospital,Fuzhou 350001,China)

机构地区：[1]福建中医药大学药学院,福建福州350122 [2]福建中医药大学附属福州中医院脑病科,福建福州350001

出　　处：《药学研究》2025年第2期113-117,共5页Journal of Pharmaceutical Research

基　　金：国家自然科学基金项目(No.81973437、82274080);福建省自然科学基金项目(No.2022J01869、2023J011565);福建中医药大学校管项目(No.X2021002)。

摘　　要：目的研究片仔癀对脑缺血/再灌注大鼠血脑屏障损伤及腺苷酸活化蛋白激酶(AMPK)/蛋白激酶C(PKC)通路的影响。方法将健康成年雄性SD大鼠随机分为假手术组、模型组和片仔癀干预组(180 mg·kg-1)。大鼠于造模前4 d开始灌胃给药,末次给药30 min后采用线栓法制作脑缺血/再灌注大鼠模型(MCAO)。Zea-Longa神经功能评分评价片仔癀对MCAO大鼠的保护效应;伊文思蓝染色法观察血脑屏障通透性变化;Western blotting法测定大鼠缺血侧脑组织闭合蛋白(Occludin)、紧密连接蛋白5(Claudin-5)、闭锁小带蛋白1(ZO-1)、基质金属蛋白酶9(MMP-9)、PKC、AMPK及其磷酸化水平(p-AMPK)蛋白表达的影响。结果与模型组比较,片仔癀能够减轻MCAO大鼠神经功能损伤(P<0.05),降低血脑屏障通透性(P<0.05),上调Occludin、Claudin-5、ZO-1、PKC蛋白表达(P<0.01),下调MMP-9、p-AMPK蛋白表达(P<0.01,P<0.05)。结论片仔癀能够上调紧密连接蛋白、PKC表达,降低MMP-9蛋白表达,抑制AMPK蛋白激活,修复血脑屏障,从而减轻脑损伤。Objective To investigate the effects of Pien Tze Huang(PTH)on blood-brain barrier damage and related regulatory proteins in rats with cerebral ischemia/reperfusion injury.Methods Healthy adult male Sprague-Dawley rats were randomly divided into sham operation group,model group and PTH group.Rats were orally administered PTH for 4 days before modeling,and the last dose was given 30 minutes before middle cerebral artery occlusion(MCAO)using the suture method.Zea-longa neurofunctional score was used to evaluate the protective effect of PTH on MCAO rats.Evans blue staining method was used to detect the changes in BBB permeability of cerebral ischemia.The expression of Occludin,Claudin-5,zonula occluden-1(ZO-1),matrix metalloproteinases-9(MMP-9),protein kinase C(PKC),adenine monophosphate activated protein kinase(AMPK)and its phosphorylated proteins(p-AMPK)in MCAO rats were determined by Western blotting.Results Compared with the model group,PTH significantly improved the neurological deficit(P<0.05),decreased BBB permeability(P<0.05),promoted Occludin,Claudin-5,ZO-1 and PKC protein expression(P<0.01),decreased MMP-9 protein expression(P<0.01),inhibited AMPK phosphorylation(P<0.05).Conclusion Pien Tze Huang could regulate the expression of tight junction protein and PKC protein,reduce the expression of MMP-9 protein,inhibit the activation of AMPK protein,repair the blood-brain barrier,and reduce brain injury.

关 键 词：片仔癀 脑缺血/再灌注 血脑屏障 紧密连接蛋白 腺苷酸活化蛋白激酶 蛋白激酶C 

分 类 号：R285.5[医药卫生—中药学]