miR-150-5p抑制TLR-5/NF-κB p65信号通路对大脑中动脉阻塞模型大鼠的神经保护效应  

Mi R-150-5p protects rats from middle cerebral artery occlusion by inhibiting the Toll-like receptor-5/nuclear factor-kappa B p65 signaling pathway

作　　者：谢媛媛[1] 张延军[1] 张晓曼[1] Xie Yuanyuan;Zhang Yanjun;Zhang Xiaoman(Department of Neurology,Zhengzhou First People’s Hospital,Zhengzhou 450004,Henan Province,China)

机构地区：[1]郑州市第一人民医院神经内科

出　　处：《中国组织工程研究》2020年第2期223-229,共7页Chinese Journal of Tissue Engineering Research

基　　金：河南省医学科技攻关计划项目(2018020725),项目负责人:张晓曼~~

摘　　要：背景:缺血性脑梗死的病变组织中发生炎症反应,且miR-150-5p表达明显下降,miR-150-5p是否经Toll样受体5/核因子κB途径抑制炎症因子释放并减轻缺血性脑梗死组织损伤尚不清楚。目的:探讨微小RNA-150-5p(miR-150-5p)在大鼠缺血性脑梗死中的作用及初步机制。方法:①构建大脑中动脉阻塞模型大鼠,建模后将大鼠分为5组:对照组、miR-150-5p agomir组、agomir control、miR-150-5p antagomir组和antagomir control组;②对照组大鼠给予生理盐水脑室注射,后4组大鼠脑室分别给予miR150-5p agomir(miR150-5p激动剂)、agomir阴性对照、miR150-5p antagomir(miR150-5p抑制剂)和antagomir阴性对照;③干预7 d后进行神经功能缺损程度(mNNS)评分,磁共振检测法测量脑梗死体积,苏木精-伊红染色观察脑组织病理学变化,qRT-qPCR法、ELISA法和免疫印迹法检测分别检测脑组织中miR-150-5p、白细胞介素6、肿瘤坏死因子α、Toll样受体5和核因子κB p65表达情况,通过检索生物信息学网站Targetscan预测miR-150-5p与Toll样受体5的结合位点,荧光素酶试验验证二者的靶向关系。结果与结论:①与对照组比较,miR-150-5p agomir组大鼠神经功能损伤评分、脑组织中白细胞介素6、肿瘤坏死因子α水平及Toll样受体5、核因子κB p65蛋白表达明显降低(P<0.05);miR-150-5p antagomir组生理评分及生化指标均明显升高(P<0.05);②苏木精-伊红染色显示,对照组神经细胞排列紊乱、轮廓模糊不清,神经细胞明显变性坏死;上述病理变化在miR-150-5p agomir组明显减轻,而在miR-150-5p antagomir组中明显加重。而agomir control组和antagomir control组与对照组各指标比较差异均无显著差异(P>0.05);③TargerScan网站预测结果和荧光素酶报告基因分析显示,miR-150-5p与Toll样受体5存在靶向结合位点;④结果证实,miR-150-5p可抑制缺血所致脑损伤过程中Toll样受体5/核因子κB p65炎性信号通路,降低炎性反应,从而减轻BACKGROUND:There is an inflammatory response in the lesion tissue of ischemic cerebral infarction,and the expression of miR-150-5p is significantly decreased.Whether miR-150-5p inhibits the release of inflammatory factors and alleviates the injury of ischemic cerebral infarction tissue through the Toll-like receptor-5/nuclear factor-κB pathway remains unclear.OBJECTIVE:To investigate the role and preliminary mechanism of miR-150-5p in ischemic cerebral infarction in rats.METHODS:(1)The rat models of middle cerebral artery occlusion were constructed and the rat models were divided into five groups:control,miR-150-5p agomir,agomir control,miR-150-5p antagomir and antagomir control groups.(2)The rats in the control group was given the intracerebroventricular injection of normal saline,and the rats in the latter four groups were given the intracerebroventricular injection of miR-150-5p agomir(miR-150-5p agonist),agomir negative control,miR150-5p antagomir(miR150-5p inhibitor)and antagomir negative control,respectively.(3)After 7 days,the brain was graded by modified neurological severity score,the cerebral infarct volume was measured by MRI,and the histopathological changes were observed by hematoxylin-eosin staining.The expression levels of miR-150-5p,interleukin-6,tumor necrosis factor-α,Toll-like receptor-5 and nuclear factor-κB p65 in brain tissues were detected by qRT-PCR,ELISA and western blot assay,respectively.The target relationship between miR150-5p and Toll-like receptor-5 was verified by luciferase assay by retrieving the bioinformatics website Targetscan to predict the binding sites of miR-150-5p and Toll-like receptor-5.RESULTS AND CONCLUSION:(1)Compared with the control group,the modified neurological severity score,and levels of interleukin-6,tumor necrosis factor-α,Toll-like receptor-5 and nuclear factor-κB p65 proteins were significantly decreased in the miR-150-5p agomir group(P<0.05).The physiological score and biochemical indexes in the miR-150-5p antagomir group were significantly increased

关 键 词：miR-150-5p Toll样受体5/核因子κB p65通路 炎症 缺血性脑梗死 神经功能 脑梗死体积 生物信息学 组织工程 

分 类 号：R446[医药卫生—诊断学] R363[医药卫生—临床医学]