HIE幼龄大鼠神经元自噬诱导线粒体功能障碍的研究  

Autophagy induces mitochondrial dysfunction in neurons from neonatal rats with hypoxic ischemic encephalopathy

作　　者：李鹏[1] 李晓华[1] 苏秦[1] 白静[2] 郝雷[2] LI Peng;LI Xiao-hua;SU Qin;BAI Jing;HAO Lei(Departmengt of Pediatrics,Affiliated Hospital of Inner Mongolian Medical University,Hohhot 010050,China;Drpartment of Pathophysiology,Inner Mongolian Medical University,Hohhot 010059,China)

机构地区：[1]内蒙古医科大学附属医院儿科,内蒙古呼和浩特010050 [2]内蒙古医科大学病理生理学教研室,内蒙古呼和浩特010059

出　　处：《中国病理生理杂志》2020年第2期366-370,共5页Chinese Journal of Pathophysiology

基　　金：内蒙古自然科学基金资助项目[No.2016MS(LH)0815;No.2017MS0899]

摘　　要：目的:探讨细胞自噬对缺氧缺血性脑病(HIE)幼龄大鼠神经元线粒体功能的影响。方法:随机选取10日龄SPF级SD大鼠30只,分为假手术(sham)组和HIE组,后者结扎单侧颈总动脉复制缺血缺氧模型。取脑组织行镜下病理观察,免疫组化分析活化型caspase-3和LC3B-II蛋白表达;体外实验中观察缺氧诱导的原代大鼠神经元的自噬过程,Western blot检测相关蛋白,并对大鼠神经元线粒体功能进行测试。结果:(1)与sham组相比,HIE组大鼠出现脑萎缩和脑室增宽;HIE组免疫组化显示活化型caspase-3和LC3B-II蛋白表达上调(P<0.01);(2)体外细胞实验发现,缺氧可诱导大鼠神经元出现自噬和凋亡;(3)与sham组相比,单纯缺氧的神经元内活性氧簇增加,线粒体超氧化物上调,线粒体跨膜电位降低(P<0.01)。结论:在HIE大鼠模型中,缺氧诱导的神经元线粒体功能障碍,可能与缺氧时神经元出现的自噬和凋亡有关。这一结果将为临床上开发细胞自噬因子类药物治疗HIE提供了新的思路。AIM:To explore the influence of autophagy on the induction of mitochondrial dysfunction in the neurons in a neonatal rat hypoxic ischemic encephalopathy(HIE)model.METHODS:Ten-day-old rat pups(n=30)were randomly divided into sham group and model group.The rats in the latter group were subject to hypoxia-ischemia treatment via unilateral common carotid artery ligation.The rats were sacrificed for brain pathological examination,and the protein levels of cleaved caspase-3 and LC3B-Ⅱwere detected by immunohistochemical analysis.For the in vitro experiments,the autophagy of primarily cultured rat neurons was observed after hypoxia,and Western blot and mitochondrial function testing were also performed.RESULTS:Compare with sham group,the hypoxia-ischemia treatment caused atrophy and apoptosis of neurons,and ventricular area enlargement of rat brains.Immunohistochemical results demonstrated significantly higher levels of apoptosis-and autophagy-associated proteins,such as cleaved caspase-3 and LC3B-II(P<0.01).In vitro experiments demonstrated that hypoxia induced autophagy and apoptosis in the neurons.Compared with sham group,there were higher levels of reactive oxygen species and mitochondrial superoxide,and lower mitochondrial membrane potential in the model group(P<0.01).CONCLUSION:In neonatal HIE rat model,the hypoxia-induced mitochondrial dysfunction is related to apoptosis and autophagy.It will provide a new idea for administration of autopahgy inducer agents in treatment of HIE.

关 键 词：细胞凋亡 自噬 线粒体功能障碍 缺氧缺血性脑病 神经元 

分 类 号：R722[医药卫生—儿科] R363.2[医药卫生—临床医学]