复元醒脑汤对MCAO模型大鼠血脑屏障的保护作用及机制研究  

Study on the protective effect and mechanism of Fuyuan Xingnao Decoction on blood-brain barrier in MCAO model rats

作　　者：孙玉婷 张文 蒲玉婷 吴霖光缙 陈采玉 徐湘茹 费月蓉 方邦江[1,2] SUN Yu-ting;ZHANG Wen;PU Yu-ting;WU Lin-guangjin;CHEN Cai-yu;XU Xiang-ru;FEI Yue-rong;FANG Bang-jiang(Longhua Hospital of Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China;Institute of Acute and Critical Care,Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)

机构地区：[1]上海中医药大学附属龙华医院,上海200032 [2]上海中医药大学急危重症研究所,上海200032

出　　处：《时珍国医国药》2024年第4期783-787,共5页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金面上项目(81973811);华东片区及市级中医专科专病联盟建设[ZY(2021-2023)-0302]。

摘　　要：目的探究复元醒脑汤对急性缺血性脑梗死模型大鼠血脑屏障的影响及机制研究。方法取40只SD大鼠随机分为假手术组、模型组、复元醒脑汤组和尼莫地平组,除假手术组外,其余各组建立大鼠大脑中动脉栓塞制备缺血性脑梗死模型。假手术组和模型组给予0.9%氯化钠溶液、复元醒脑汤组和尼莫地平组给予相应的等效剂量药物,每日灌胃2次,连续灌胃3d。采用Bederson神经功能评分;HE染色观察各组大鼠脑组织病理变化;采用IHC观察ZO-1和Claudin-5的表达;Western blot检测大鼠脑组织MMP-2、MMP-9、ZO-1和Claudin-5的蛋白表达情况;RT-qPCR检测大鼠脑组织MMP-2、MMP-9、ZO-1和Claudin-5基因表达水平。结果与假手术组比较,模型组大鼠神经功能缺损评分升高;与假手术组相比,模型组大鼠脑组织MMP-2和MMP-9蛋白和mRNA表达升高(P<0.05,P<0.01),ZO-1和Claudin-5蛋白mRNA表达降低(P<0.05,P<0.01);与模型组相比,复元醒脑汤组的大鼠脑组织缺血半暗带MMP-2和MMP-9蛋白mRNA表达显著降低(P<0.05,P<0.01);ZO-1和Claudin-5蛋白mRNA表达显著升高(P<0.05,P<0.01)。结论复元醒脑汤保护血脑屏障通透性,可能与抑制MMP-2和MMP-9的表达,抑制紧密连接蛋白中ZO-1和claudin-5降解及丢失,从而发挥保护血脑屏障结构功能的作用。Objective To investigate the effects of Fuyuan Xingnao Decoction on the blood-brain barrier in acute ischemic cerebral infarction model rats and its mechanism.Methods 40 SD rats were randomly divided into the sham operation group,model group,Fuyuan Xingnao Decoction group,and Nimodipine group.Middle cerebral artery embolization was established in the other groups to prepare the ischemic cerebral infarction model except for the sham operation group.The sham operation group and model group were given 0.9%sodium chloride solution,Fuyuan Xingnao decoction group and Nimodipine group were given the corresponding equivalent dose of drugs,and gavage twice a day,continuous gavage for 3 days.Bederson neural function score was used.The pathological changes in brain tissue were observed by hematoxylin-eosin staining.The expressions of ZO-1 and Claudin-5 were observed by immunohistochemistry.The protein expression of MMP-2,MMP-9,ZO-1,and Claudin-5 in rat brains was detected by Western blot.The expression levels of MMP-2,MMP-9,ZO-1 and Claudin-5 genes in rat brain were detected by RT-qPCR.Results Compared with sham operation group,the neurological impairment of rats in model group was higher.Compared with sham operation group,the protein and mRNA expressions of MMP-2 and MMP-9 in brain tissue of model group were increased(P<0.05,P<0.01),while the protein mRNA expressions of ZO-1 and Claudin-5 were decreased(P<0.05,P<0.01).Compared with model group,the mRNA expressions of MMP-2 and MMP-9 protein in ischemic penumbra of brain tissue in Fuyuan Xingnao Decotion group were significantly decreased(P<0.05,P<0.01).The mRNA expressions of ZO-1 and Claudin-5 protein were significantly increased(P<0.05,P<0.01).Conclusion Fuyuan Xingnao Decoction can protect the permeability of the blood-brain barrier,and may inhibit the expression of MMP-2 and MMP-9,and inhibit the degradation and loss of ZO-1 and claudin-5 in tight junction protein,so as to protect the structure and function of the blood-brain barrier.

关 键 词：复元醒脑汤 脑梗死 血脑屏障 金属基质蛋白酶 紧密连接蛋白 

分 类 号：R285.5[医药卫生—中药学]