黄芪甲苷通过PINK1/parkin通路介导的线粒体自噬途径减轻大鼠脑缺血再灌注损伤  

Astragaloside IV protects against rat cerebral ischemia-reperfusion injury via PINK1/parkin mitophagy-associated pathway

作　　者：马莉[1] 赵俊杰 王鹏 钱建华 李良勇[2] MA Li;ZHAO Junjie;WANG Peng;QIAN Jianhua;LI Liangyong(Clinical College of Integrated Traditional Chinese and Western Medicine,Anhui University of Traditional Chinese Medicine,Hefei 230001,China;Department of Neurology,the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine,Hefei 230031,China)

机构地区：[1]安徽中医药大学中西医结合学院,安徽合肥230001 [2]安徽中医药大学第一附属医院,安徽合肥230031

出　　处：《中国病理生理杂志》2025年第3期501-508,共8页Chinese Journal of Pathophysiology

基　　金：安徽省高等学校自然科学基金资助项目(No.2023AH050828,No.2024AH050955);安徽中医药大学人才支持计划项目(No.2023rcyb023)。

摘　　要：目的:探讨黄芪甲苷(astragaloside IV,AS-IV)通过PTEN诱导激酶1(PTEN-induced kinase 1,PINK1)/parkin通路介导的线粒体自噬途径,抑制氧化应激及减轻脑缺血再灌注损伤的作用机制。方法:构建大鼠大脑中动脉阻塞/再灌注(middle cerebral artery occlusion/reperfusion,MCAO/R)模型,将SD大鼠随机分为sham、MCAO/R、AS-IV及线粒体分裂抑制剂1(mitochondrial division inhibitor-1,Mdivi-1)组。AS-IV及Mdivi-1组腹腔注射AS-IV(20 mg/kg,每天1次,连续7 d),Mdivi-1组同时给予Mdivi-1腹腔注射(1.2 mg·kg^(-1)·d^(-1)),sham和MCAO/R组则给予等体积的蒸馏水。Zea-Longa法检测神经功能缺损评分,TTC染色检测脑梗死体积,HE染色观察脑组织病理形态学改变,透射电子显微镜观察线粒体自噬,酶标法测定丙二醛(malondialdehyde,MDA)含量和超氧化物歧化酶(superoxide dismutase,SOD)活性,流式细胞术检测活性氧(reactive oxygen species,ROS)含量,Western blot和RTqPCR检测PINK1、parkin、微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)蛋白和mRNA表达水平。结果:MCAO/R组大鼠经AS-IV治疗后,脑组织神经元及线粒体的损伤明显减轻,自噬小体增多(P<0.05),脑梗死体积显著减小,神经功能缺损情况亦明显减轻(P<0.05),PINK1、parkin及LC3蛋白和mRNA表达水平显著上调(P<0.05),ROS和MDA含量明显降低,SOD活性显著增加(P<0.05);Mdivi-1干预后则完全逆转了ASIV的作用,阻抑了PINK1/parkin通路的激活,LC3蛋白和mRNA表达水平下调,线粒体明显损伤,自噬小体减少,ROS和MDA含量增加,SOD活性下降,脑梗死体积增加,神经功能缺损加剧(P<0.05)。结论:AS-IV可能通过激活PINK1/parkin通路介导的线粒体自噬,抑制氧化应激反应,从而减轻大鼠脑缺血再灌注损伤。AIM:To clarify the molecular mechanism by which astragaloside IV(AS-IV)suppresses oxida-tive stress and alleviates cerebral ischemia-reperfusion injury(CIRI)via the PTEN-induced kinase 1(PINK1)/parkin mi-tophagy-associated pathway.METHODS:A middle cerebral artery occlusion/reperfusion(MCAO/R)model was estab-lished in Sprague-Dawley rats.The animals were allocated to sham,MCAO/R,AS-IV,and mitochondrial division inhibi-tor-1(Mdivi-1)treatment groups.The rats in AS-IV and Mdivi-1 groups were intraperitoneally injected once daily with AS-IV(20 mg/kg)for 7 d,while those in Midivi-1 group also received intraperitoneal injection of Mdivi-1(1.2 mg·kg^(-1)·d^(-1)).The rats in sham and MCAO/R groups were given equivalent volume of distilled water.Neurological deficits were as-sessed using Zea Longa scoring,infarcted area volumes were measured using TTC staining,and brain tissue pathology was examined using hematoxylin and eosin staining.The levels of malondialdehyde(MDA)and superoxide dismutase(SOD)were assessed by ELISA,while those of reactive oxygen species(ROS)were measured using flow cytometry.The expres-sion levels of PINK1,parkin and microtubule-associated protein 1 light chain 3(LC3)were quantified using Western blot and RT-qPCR.RESULTS:AS-IV administration significantly alleviated neuronal and mitochondrial damage in MCAO/R model rat brains(P<0.05),together with significant reductions in the cerebral infarct volume and neurological dysfunc-tion(P<0.05).Significant increases in PINK1,parkin and LC3 protein and mRNA levels were observed in response to AS-IV(P<0.05),SOD activity rose,and ROS and MDA levels declined significantly(P<0.05).The co-administration of Mdivi-1 abrogated the protective benefits of AS-IV,inhibited activation of the PINK1/parkin pathway,down-regulated LC3 at the mRNA and protein levels,and significantly increased mitochondrial damage.Mdivi-1 also markedly reduced autophagosome formation and SOD activity level,but increased both ROS and MDA levels,cerebral infarct volume,and the severity of neu

关 键 词：黄芪甲苷 线粒体自噬 氧化应激 脑缺血再灌注损伤 

分 类 号：R743[医药卫生—神经病学与精神病学] R285.5[医药卫生—临床医学] R363.2