miR-210过表达对急性脑梗死大鼠脑损伤的影响及其机制  

Effect and its mechanism of miR⁃210 overexpression on cerebral injury in rats with acute cerebral infarction

作　　者：陈伟 任蓓 杨云鹏 蒋锋[3] 山媛[3] 王静 CHEN Wei;REN Bei;YANG Yunpeng;JIANG Feng;SHAN Yuan;WANG Jing(Neurological Intensive Care Center,Xi'an People's Hospital(Xi'an Fourth Hospital),Xi'an 710004,China;Department of Neurology,Xi'an People's Hospital(Xi'an Fourth Hospital);Department of Neurology,Shaanxi Provincial People's Hospital)

机构地区：[1]西安市人民医院(西安市第四医院)神经重症中心,西安710004 [2]西安市人民医院(西安市第四医院)神经内科 [3]陕西省人民医院神经内一科

出　　处：《山西医科大学学报》2024年第12期1495-1504,共10页Journal of Shanxi Medical University

基　　金：陕西省自然科学基础研究计划项目(2022JM-594,2022JQ-912)。

摘　　要：目的 探究miR-210在急性脑梗死(ACI)中的作用和分子调控机制。方法 采用改良线栓法制造大脑中动脉栓塞构建ACI模型大鼠,将ACI大鼠分为模型组、agomir-NC组和agomir-210组,另设置健康SD大鼠为假手术组。造模完成后72 h,采用Zea-Longa评分对大鼠进行神经功能评估,TTC染色检测脑梗死面积,HE染色检测脑基底动脉损伤,TUNEL染色检测脑基底动脉血管内皮细胞凋亡,ELISA法检测血清中MDA、SOD、GSH-Px和ROS水平。将人脑微细血管内皮细胞HBMEC-D3细胞分为常氧组、缺氧组、mimic-NC组(缺氧培养)和miR-210 mimic组(缺氧培养),采用CCK-8检测HBMEC-D3细胞增殖,流式细胞术检测HBMEC-D3细胞凋亡,DCFH-DA探针法检测HBMEC-D3细胞内ROS含量。采用RT-PCR检测大鼠脑组织和HBMECD3细胞中miR-210表达水平,Western blot检测大鼠脑组织和HBMEC-D3细胞中凋亡相关蛋白(Bax、cleaved Caspase-3和Bcl-2)和STAT3/PI3K/AKT信号通路相关蛋白(p-STAT3、STAT3、p-PI3K、PI3K、p-AKT和AKT)表达水平。结果 与假手术组比较,模型组大鼠脑血管病变严重,神经功能评分、脑梗死面积比和细胞凋亡率均升高(P<0.05)。与agomir-NC组比较,agomir-210组大鼠脑血管壁内皮状态正常,miR-210、SOD、GSH-Px、Bcl-2、p-STAT3/STAT3、p-AKT/AKT和p-PI3K/PI3K水平均升高(P<0.05),神经功能评分、脑梗死面积比和细胞凋亡率均降低(P<0.05),MDA、ROS、Bax和cleaved Caspase-3表达量均降低(P<0.05)。与常氧组比较,缺氧组细胞增殖水平以及miR-210、Bcl-2、p-STAT3/STAT3、p-AKT/AKT和p-PI3K/PI3K水平均降低(P<0.05),细胞凋亡率以及ROS、Bax和cleaved Caspase-3表达量升高(P<0.05)。与mimic-NC组比较,miR-210 mimic组细胞增殖水平以及miR-210、Bcl-2、p-STAT3/STAT3、p-AKT/AKT和p-PI3K/PI3K水平均升高(P<0.05),细胞凋亡率以及ROS、Bax和cleaved Caspase-3表达量降低(P<0.05)。结论 过表达miR-210可以抑制ACI大鼠脑损伤,其机制可能与抑制血管内皮细胞凋亡和ROS生成Objective To explore the role and molecular regulatory mechanism of miR‐210 in acute cerebral infarction(ACI).Methods Middle cerebral artery embolization was induced by modified thread embolization method to construct ACI model rats.ACI rats were di‐vided into model group,agomir‐NC group and agomir‐210 group,and healthy SD rats were chosen as sham operation group.At 72 h af‐ter the modeling,the neural function of rats was evaluated by Zea‐Longa scoring system.Cerebral infarction size was detected by TTC staining,the cerebral basilar artery injury was detected by HE staining,and the cerebral basilar artery endothelial cell apoptosis was detected by TUNEL staining.The levels of MDA,SOD,GSH‐Px and ROS in serum were detected by ELISA.Human brain microvascu‐lar endothelial cells D3(HBMEC‐D3)were divided into normoxia group,hypoxia group,mimic‐NC group(hypoxia culture)and miR‐210 mimic group(hypoxia culture).HBMEC‐D3 cell proliferation was detected by CCK‐8,HBMEC‐D3 cell apoptosis was detected by flow cytometry,and ROS content in HBMEC‐D3 cells was detected by DCFH‐DA probe.The expression of miR‐210 in rat brain tissue and HBMEC‐D3 cells was detected by RT‐PCR,and the protein expression levels of apoptosis‐related proteins(Bax,cleaved Caspase‐3 and Bcl‐2)and STAT3/PI3K/AKT signaling pathway proteins(p‐STAT3,STAT3,p‐PI3K,PI3K,p‐AKT and AKT)in rat brain tis‐sue and HBMEC‐D3 cells were analyzed by Western blot.Results Compared with sham group,the cerebral vascular lesions of rats were serious in model group,and the neural function score,the infarct area percentage and the apoptosis rate were increased(P<0.05).Compared with agomir‐NC group,the cerebral vascular lesions of rats were serious in agomir‐210 group,the levels of miR‐210,SOD,GSH‐Px,Bcl‐2,p‐STAT3/STAT3,p‐AKT/AKT and p‐PI3K/PI3K were increased(P<0.05),the neural function score,the in‐farct area percentage,the apoptosis rate were decreased(P<0.05),and the expression levels of MDA,ROS,Bax and cle

关 键 词：MIR-210 急性脑梗死(ACI) 脑损伤 脑血管内皮细胞 凋亡 活性氧(ROS) STAT3/PI3K/AKT信号通路 

分 类 号：R743.3[医药卫生—神经病学与精神病学]