miR-132靶向FoxO3a抑制细胞自噬在脑出血模型大鼠中的神经保护作用  

Neuroprotective effect of miR-132 in rats with intracerebral hemorrhage by targeting FOXO3a and inhibiting autophagy

作　　者：陆飞宇 李剑侠 黄先锋 王文宏 林小祥 Lu Feiyu;Li Jianxia;Huang Xianfeng;Wang Wenhong;Lin Xiaoxiang(Department of Neurosurgery,the Zhongda Hospital Southeast University,Nanjing 210043,China)

机构地区：[1]东南大学附属中大医院神经外科,南京210043

出　　处：《脑与神经疾病杂志》2021年第10期629-634,共6页Journal of Brain and Nervous Diseases

摘　　要：目的研究微小RNA(miR)-132靶向叉形头转录因子O亚型3a(Fox03a)抑制细胞自噬在脑出血模型大鼠中的神经保护作用。方法成年雄性SD大鼠随机分为对照组、模型组、miR-阴性对照(NC)组、miR-132组,后三组采用Ⅶ型胶原酶注入苍白球的方式建立脑出血模型,对照组和模型组给予生理盐水侧脑室注射,miR-NC组和miR-132组分别给予miR-NC及miR-132侧脑室注射。造模后第7d,比较各组大鼠神经功能评分的差异,苏木精-伊红染色观察血肿周围脑组织病理改变,PCR检测miR-132的表达水平,Western blot检测FoxO3a、自噬基因(Beclin-1、Atg12、LC3)的表达水平;培养PC12神经元细胞,转染miR-NC或miR-132后,采用双荧光素酶报告基因实验验证miR-132靶向FoxO3a。结果在动物实验中,模型组大鼠的神经功能评分增加,血肿周围脑组织中FoxO3a、Beclin-1、Atg12、LC3-Ⅱ/LC3-Ⅰ的表达增加,miR-132的表达减少(P<0.05);miR-132组大鼠的神经功能评分降低,血肿周围脑组织中FoxO3a、Beclin-1、Atg12、LC3-Ⅱ/LC3-Ⅰ的表达减少,miR-132的表达增加(P<0.05)。在细胞实验中,miR-132组细胞中FoxO3a的表达水平及野生型FoxO3a 3’UTR双荧光素酶报告基因的荧光活力均低于miRNC组(P<0.05)。结论miR-132在脑出血模型大鼠中起保护作用,靶向FoxO3a并抑制自噬是可能的分子机制。Objective To study the neuroprotective effect of microRNA(miR)-132 targeting forkhead transcription factor 03 A(FoxO3 a)in rats with intracerebral hemorrhage by targeting FOXO3 a and inhibiting autophagy.Methods Adult male SD rats were randomly divided into control group,model group,miR-negative control(NC)group and miR-132 group.Intracerebral hemorrhage model was established by injecting type VII collagenase into globus pallidus.Then normal saline was injected into lateral ventricle of control group and model group,miR-NC and miR-132 were injected into lateral ventricle of miR-NC group and miR-132 group.On the7 th day after modeling,the neurological function score were compared,pathological changes of brain tissue around hematoma were observed by HE staining,the expression of miR-132 was detected by PCR,the expression of FOXO3 a and autophagy gene(beclin-1,Atg12,LC3)were detected by western blot;PC12 neurons were cultured and transfected with miR-NC or miR-132,and double luciferase reporter gene experiment was used to verify miR-132 targeting FoxO3 a.Results In the animal experiment,the neurological function score increased,the expression of FoxO3 a,beclin-1,atg12 and LC3-II/LC3-I increased,and the expression of miR-132 decreased in the model group(P<0.05);the neurological function score decreased,and the expression of FoxO3 a,beclin-1,atg12 and LC3-II/LC3-I decreased,and the expression of miR-132 increased in the miR-132 group(P<0.05).In cell experiment,the expression level of FOXO3 a and the fluorescence activity of wild-type FOXO3 a 3’UTR double luciferase reporter gene in miR-132 group were lower than those in miR-NC group(P<0.05).Conclusion miR-132 plays a protective role in ICH rats,targeting FOXO3 a and inhibiting autophagy may be the related molecular mechanism.

关 键 词：脑出血 miR-132 叉形头转录因子O亚型3a 自噬 

分 类 号：R743.34[医药卫生—神经病学与精神病学]