文献类型：期刊文献

中文题名：miR-129-5p靶向HMGB1减轻大鼠脑缺血再灌注损伤诱导的炎症和脑组织损伤

英文题名：MiR-129-5p targets HMGB1 to alleviate inflammation and brain tissue damage induced by cerebral ischemia-reperfusion injury in rats

作者：胡广泽[1];袁珊[1];何哲[1];许梅梅[1];杨欣原[1];孟平平[1];高蕊[1,2]

机构：[1]石河子大学医学院/新疆地方与民族高发病教育部重点实验室,新疆石河子832000;[2]广东海洋大学滨海农业学院,广东湛江524088

年份：2025

卷号：43

期号：4

起止页码：405

中文期刊名：石河子大学学报(自然科学版)

外文期刊名：Journal of Shihezi University(Natural Science)

收录：北大核心2023、、北大核心

基金：国家自然科学基金项目(32260142);新疆生产建设兵团区域创新引导计划项目(2021BB002)。

语种：中文

中文关键词：miR-129-5p;HMGB1;MCAO;炎症

外文关键词：miR-129-5p;HMGB1;MCAO;inflammation

中文摘要：目的通过前期课题组测序基础,筛选MCAO大鼠皮层组织中变化较显著的miR-129-5p及其可能的靶基因(high-mobility group box 1,HMGB1),探究二者靶向调控关系,明确miR-129-5p—HMGB1调控网络在缺血性卒中对炎症水平影响及脑损伤的作用。方法线栓法构建大鼠中动脉闭塞模型,通过行为学评分、全脑TTC染色及炎症相关因子(IL-1β、IL-6、TNF-α)指标评定缺血再灌注模型,应用qRT-PCR及Western Blot验证miR-129-5p与HMGB1的表达相关性。预测miR-129-5p与HMGB1的结合位点并比对相关物种保守性,双荧光素酶报告实验验证二者结合位点特异性。利用侧脑室注射过表达miR-129-5p agomir,在体水平验证miR-129-5p对HMGB1表达的负向调控及对MCAO大鼠脑组织炎症水平和梗死的影响。结果TTC染色显示MCAO组大鼠梗死面积大于sham组大鼠(P<0.001),longa评分显示MCAO组大鼠有明显神经行为缺损(P<0.05)。炎症因子IL-1β、IL-6、TNF-α显著升高(P<0.001),表明大鼠MCAO模型构建成功。大鼠脑缺血再灌注损伤后,miR-129-5p表达降低(P<0.001)、HMGB1表达升高(P<0.001),二者表达呈相反趋势。利用RNA hybrid生信软件分析预测miR-129-5p可靶向结合HMGB1的3′UTR区,且结合区域在大鼠、人、猪等物种间高度保守。通过双荧光素酶报告实验发现miR-129-5p agomir与HMGB13′UTR-WT共转染相较于HMGB13′UTR-野生型与agomir-NC共转染组荧光活性更高(P<0.001)。MCAO模型大鼠大脑过表达miR-129-5p显著下调HMGB1的表达(P<0.001),显著抑制炎症因子IL-1β、IL-6、TNF-α的上调(P<0.001),梗死面积显著缩小(P<0.001)。结论miR-129-5p通过靶向HMGB1来参与其转录后的负向调控,过表达miR-129-5p可下调HMGB1的表达从而抑制脑缺血再灌注损伤诱导的炎症,减轻脑损伤程度。

外文摘要：Objective Based on the previous sequencing work of our research group,to screen for the significantly altered miR-129-5p in the cortical tissue of middle cerebral artery occlusion(MCAO)rats and its potential target gene HMGB1,and to explore their targeted regulatory relationship.The aim is to clarify the role of the miR-129-5p-HMGB1 regulatory network in the impact on the inflammatory level and brain injury in ischemic stroke.Methods The rat middle artery occlusion model was established by suture method.The ischemia-reperfusion model was evaluated by behavioral scores,whole brain TTC staining and inflammation-related factors(IL-1β,IL-6,TNF-α).The expression correlation of miR-129-5p and HMGB1 was verified by qRT-PCR and Western Blot.The binding sites of miR-129-5p and HMGB1 were predicted and compared with the conservation of related species.The dual luciferase reporter experiment verified the specificity of the binding sites.The negative regulation of miR-129-5p on HMGB1 expression and its effect on the inflammation level and infarction of brain tissue in MCAO rats were verified in vivo by intracerebroventricular injection of overexpressed miR-129-5p agomir.Results TTC staining showed that the infarction area of MCAO rats was larger than that of sham rats(P<0.001).Longa score showed that MCAO rats had obvious neurobehavioral deficits(P<0.05).The inflammatory factors IL-1β,IL-6,and TNF-αincreased significantly(P<0.001),indicating that the rat MCAO model was successfully established.After cerebral ischemia-reperfusion injury in rats,the expression of miR-129-5p decreased(P<0.001)and the expression of HMGB1 increased(P<0.001),and the two expressions showed opposite trends.RNA hybrid bioinformatics software analysis predicted that miR-129-5p could target and bind to the 3′UTR region of HMGB1,and the binding region was highly conserved among species such as rats,humans,and pigs.The dual luciferase reporter assay showed that the fluorescence activity of miR-129-5p agomir co-transfection with HMGB13′UTR-WT was higher than that of HMGB13′UTR-wild type and agomir-NC co-transfection(P<0.001).Overexpression of miR-129-5p in the brain of MCAO model rats significantly downregulated the expression of HMGB1(P<0.001),significantly inhibited the upregulation of inflammatory factors IL-1β,IL-6,and TNF-α(P<0.001),and significantly reduced the infarct area(P<0.001).Conclusion MiR-129-5p can target HMGB1 and participate in its negative post-transcriptional regulation.Overexpression of miR-129-5p can down-regulate the expression of HMGB1 to inhibit the inflammation induced by cerebral ischemia-reperfusion injury and reduce the degree of brain injury.