首页|补阳还五汤减轻氧化应激保护脑缺血再灌注损伤大鼠血脑屏障的作用研究

补阳还五汤减轻氧化应激保护脑缺血再灌注损伤大鼠血脑屏障的作用研究

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目的 探究补阳还五汤(Buyang Huanwu Decoction，BYHWD)降低氧化应激水平保护脑缺血再灌注损伤(cerebral ischemia/reperfusion injury，CIRI)大鼠血脑屏障(blood-brain barrier，BBB)的机制。方法 采用线栓法建立大鼠大脑中动脉栓塞再灌注(middle cerebral artery occlusion/reperfusion，MCAO/R)模型，应用 PeriCam PSI 激光散斑血流成像系统检测脑血流量判定模型建立是否成功。通过Zea Longa评分评价大鼠神经功能缺损情况;HE染色观察大鼠脑组织病理学改变;干湿重法检测脑水肿程度;伊文思蓝(Evans blue，EB)染色检测BBB通透性;透射电镜观察BBB超微结构改变;试剂盒检测氧化应激相关指标活性氧(ROS)、丙二醛(MDA)的含量及总超氧化物歧化酶(SOD)活性;免疫组织化学染色及Western blot检测基质金属蛋白酶-9(MMP-9)的表达水平;免疫荧光双染及 W estern blot 检测紧密连接蛋白(tight junction protein，TJP)中 Occludin、ZO-1、Claudin-5 蛋白的表达水平。结果 BYHWD能够降低MCAO/R大鼠神经功能缺损评分，减轻脑组织病理学损伤，减轻血脑屏障结构破坏，延长紧密连接结构致密区，减轻缺血侧大脑脑水肿情况，降低BBB通透性。BYHWD可以降低ROS与MDA水平，提高SOD活性，降低MMP-9表达水平，提高Occludin、Claudin-5、ZO-1蛋白表达水平。结论 BYHWD可以升高BBB紧密连接蛋白的表达水平，降低血脑屏障通透性，保护血脑屏障超微结构，减轻脑水肿，其机制可能与BYHWD抗氧化应激，抑制MMP-9的激活相关。
Effect of Buyang Huanwu Decoction in reducing oxidative stress and protecting cerebral ischemia-reperfusion injury to rat blood-brain barrier
Objective To explore the mechanisms of Buyang Huanwu Decoction(BYHWD)in reducing oxidative stress levels to protect the blood-brain barrier(BBB)in cerebral ischemia/reperfusion injury(CIRI)rats.Methods A middle cerebral artery occlusion/reperfusion(MCAO/R)model in rats was established via wire embolization method.PeriCam PSI laser speckle flow imaging was applied to detect whether the model was successfully established.Neurological deficits in the rats were evaluated by Zea Longa score,and histopathological changes in the rat brain were observed by HE staining.The degree of brain edema was detected by the dry and wet weight method.BBB permeability was detected by Evans blue staining,and ultrastructural changes to the BBB were observed by transmission electron microscopy.The levels of ROS,MDA and SOD activities,which are related to oxidative stress,were detected using kits.The expression levels of matrix metalloproteinase-9(MMP-9)were detected by immunohistochemical staining and Western blot.The expression levels of Occludin,ZO-1,and Claudin-5 tight junction proteins were determined via immunofluorescence and Western blot.Results BYHWD reduced neurological deficit scores,alleviated brain histopathological damage,alleviated BBB structural disruption,prolonged the appearance of dense regions in the tight junction structure,attenuated edema of the brain on the ischemic side,and reduced BBB permeability in MCAO/R rats.BYHWD decreased the levels of ROS and MDA,increased the activity of SOD,decreased the expression levels of MMP-9,and increased the expression levels of Occludin,Claudin-5 and ZO-1.Conclusions BYHWD can increase BBB tight junction protein expression levels,reduce the permeability of the BBB,protect the ultrastructure of the BBB,and reduce brain edema,and its mechanisms may be related to its antioxidant activity and inhibition of MMP-9 activation.

Buyang Huanwu Decoctioncerebral ischemia-reperfusion injurytransmission electron microscopyoxidative stressblood-brain barriermatrix metalloproteinase-9

马娴、高萍、刘真一、辛紫媛、靳晓飞、周晓红、高维娟

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河北中医学院,石家庄 050000

河北省心脑血管病中医药防治研究重点实验室,石家庄 050000

承德医学院附属医院,河北承德 067000

补阳还五汤 脑缺血再灌注损伤 透射电镜 氧化应激 血脑屏障 基质金属蛋白酶-9

中央引导地方科技发展资金项目河北省科技研发平台与新型研发机构建设专项河北中医学院科技能力提升重点项目

206Z7706G20567626HKTZ2019018

2024

10.3969/j.issn.1671-7856.2024.03.010

中国比较医学杂志
中国实验动物学会，中国医学科学院医学实验动物研究所

中国比较医学杂志

CSTPCD北大核心
影响因子：0.473
ISSN：1671-7856
年,卷(期)：2024.34(3)
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