中国病理生理杂志2025，Vol.41Issue(3)：481-491,11.DOI:10.3969/j.issn.1000-4718.2025.03.008
补阳还五汤通过抑制脑微血管内皮细胞自噬减轻大鼠脑缺血再灌注损伤
Buyang-Huanwu decoction attenuates rat cerebral ischemia-reperfusion injury by inhibiting autophagy of cerebral microvascular endothelial cells
摘要
Abstract
AIM:This study aims to investigate the protective effect of Buyang-Huanwu decoction(BYHWD)on cerebral ischemia-reperfusion injury(CIRI)in rats,focusing on its role in regulating the autophagy of cerebral micro-vascular endothelial cells(BMECs).METHODS:(1)We established a rat model of middle cerebral artery occlusion/re-perfusion(MCAO/R)and divided the subjects into four groups:sham group,model(MCAO/R)group,BYHWD group,and 3-n-butylphthalide(NBP)group.Neurological deficits were assessed using the Zea Longa score,while the volume of cerebral infarction was measured through 2,3,5-triphenyltetrazolium chloride(TTC)staining.Pathological damage in the ischemic penumbra was evaluated using HE staining,and blood-brain barrier(BBB)permeability was assessed by Evans blue(EB)staining.The ultrastructure of BMECs was analyzed by transmission electron microscopy,and the co-expres-sion and positive cell rate of microtubule-associated protein 1 light chain 3(LC3)in BMECs were determined through im-munofluorescence double staining.Additionally,the protein expression levels of ZO-1,claudin-5 and occludin in the cor-tical region of the ischemic penumbra in rats were examined using Western blot analysis.(2)A rat BMEC model of oxy-gen-glucose deprivation/reoxygenation(OGD/R)was also established.Rat BMECs were categorized into normal control(CON),OGD/R,dimethyl sulfoxide(DMSO),rapamycin and 3-methyladenine groups to observe autophagy levels by monodansylcadaverine(MDC)staining.Furthermore,rat BMECs were divided into CON,OGD/R,BYHWD-containing serum(BHDS)and NBP groups.The cell autophagy was assessed by MDC staining and Western blot,while cell viability was measured by CCK-8 assay.RESULTS:(1)The rats in MCAO/R group exhibited significantly higher neurological scores(P＜0.01)and increased cerebral infarction volumes(P＜0.01)compared with sham group.Severe damage in the ischemic penumbra was observed,characterized by disordered tissue structure,widened intercellular spaces,and compro-mised cellular integrity.The EB dye permeability was notably elevated(P＜0.01),and BMECs showed structural destruc-tion,including damaged cell membranes,swollen Golgi apparatus,dilated endoplasmic reticulum vesicles,and damaged mitochondria.The ratio of LC3+CD31+/CD31+and the protein levels of ZO-1,claudin-5 and occludin were significantly el-evated(P＜0.01).In contrast,the rats in BYHWD and NBP groups demonstrated lower neurological scores(P＜0.01)and reduced cerebral infarction volumes(P＜0.01).Furthermore,EB permeability decreased(P＜0.01),BMEC morphol-ogy improved,and the protein expression levels of ZO-1,claudin-5 and occludin increased(P＜0.05).(2)Rat BMECs in OGD/R group had a significantly elevated autophagy level compared with CON group(P＜0.01),with increased expres-sion of LC3 and beclin-1 proteins and decreased level of P62 protein(P＜0.05).Notably,the cells in BHDS and NBP groups displayed decreased autophagy level compared with OGD/R group,with increased cell viability(P＜0.01),re-duced LC3 and beclin-1 protein expression,and increased P62 protein expression(P＜0.05).CONCLUSION:Buyang-Huanwu decoction alleviates cerebral ischemia-reperfusion injury in rats by inhibiting the autophagy of cerebral microvas-cular endothelial cells.关键词
补阳还五汤/脑微血管内皮细胞/自噬/血脑屏障/脑缺血再灌注损伤Key words
Buyang-Huanwu decoction/brain microvascular endothelial cells/autophagy/blood-brain barrier/cerebral ischemia-reperfusion injury分类
中医学引用本文复制引用
李猛,左春月,靳晓飞,张天赐,周晓红,高维娟..补阳还五汤通过抑制脑微血管内皮细胞自噬减轻大鼠脑缺血再灌注损伤[J].中国病理生理杂志,2025,41(3):481-491,11.基金项目
国家自然科学基金资助项目(No.81873180) （No.81873180）
河北省自然科学基金资助项目(No.H2022423382) （No.H2022423382）
河北省中医药管理局科研计划项目(No.2021101) （No.2021101）
校级燕赵医学项目(No.YZZZ2023009) （No.YZZZ2023009）
青年教师科研能力提升项目(No.JCYJ2022009) （No.JCYJ2022009）
