miR-129-2靶向PIK3R1通过PI3K/AKT信号通路减轻肺内皮细胞损伤  

miR-129-2 targeting PIK3R1 alleviates pulmonary endothelial cell injury through PI3K/AKT signaling pathway

作　　者：袁江汉 郑喜盛 贾明雅[1] 冯永利[1] 李长力[1] 周发春[2] YUAN Jianghan;ZHENG Xisheng;JIA Mingya;FENG Yongli;LI Changli;ZHOU Fachun(Department of Critical Care Medicine,Nanyang Central Hospital,Nanyang 473000,China;Department of Critical Care Medicine,First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]南阳市中心医院重症医学科,473000 [2]重庆医科大学附属第一医院重症医学科,400016

出　　处：《免疫学杂志》2023年第7期586-593,共8页Immunological Journal

基　　金：重庆市自然科学基金面上项目(cstc2020jcyjmsxmX1096)。

摘　　要：目的探讨miR-129-2在肺血管内皮细胞损伤中的作用及其机制。方法SPF级小鼠40只随机分为对照组、脂多糖(LPS)组、空载组、过表达组,给予气管滴注LPS诱导急性肺损伤(ALI),尾静脉注射高表达慢病毒来过表达miR-129-2,观察ALI小鼠呼吸频率、体质量等一般情况,检测miR-129-2及炎症因子IL-6、IL-10、TNF-α表达水平和肺损伤程度。人肺微血管内皮细胞(HPMECs)分为Control组、LPS组、NC组和mimic组,LPS处理诱导细胞损伤,转染miR-129-2 mimic来上调miR-129-2的水平,划痕实验及Transwell实验检测细胞迁移能力,检测各组细胞中miR-129-2、IL-6、IL-10、TNF-α及PIK3R1 mRNA的表达水平。双荧光素酶实验验证miR-129-2与PIK3R1的靶向关系,Western blot检测miR-129-2对PIK3R1及PI3K/AKT信号通路蛋白表达的影响。结果在小鼠中,miR-129-2过表达能使小鼠进食增加、呼吸频率减慢、体质量增加、肺组织损伤减轻,IL-6、TNF-α表达降低,IL-10表达升高(P<0.05)。在HPMECs中,mimic转染使miR-129-2表达水平上调,炎症因子表达水平降低,细胞迁移能力增强,PIK3R1表达降低(P<0.05);双荧光素酶报告显示miR-129-2与PIK3R1存在结合位点,miR-129-2下调使PIK3R1表达增加,PI3K/AKT信号通路被激活。结论miR-129-2过表达能够减轻小鼠炎症反应和肺损伤,并能介导PIK3R1通过PI3K/AKT信号通路缓解肺内皮细胞损伤。This study was designed to investigate the role of miR-129-2 in pulmonary vascular endothelial cell injury and its mechanism.Forty SPF mice were randomly divided into control group,lipopolysaccharide(LPS)group,LV-NC group and overexpression group.Acute lung injury(ALI)was induced by intratracheal instillation of LPS.Lentivirus was injected into the tail vein to overexpress miR-129-2.The respiratory rate,body mass and other general conditions of ALI mice were observed;the expression levels of miR-129-2 and inflammatory factors IL-6,IL-10,TNF-αand the degree of lung injury were detected.Human pulmonary microvascular endothelial cells(HPMECs)were divided into control group,LPS group,NC group and mimic group.LPS was used to induce cell damage,and miR-129-2 mimic was transfected to increase the level of miR-129-2.Scratch test and Transwell test were used to detect cell migration.The expression levels of miR-129-2,IL-6,IL-10,TNF and PIK3R1 mRNA in cells of each group were detected;the targeting relationship between miR-129-2 and PIK3R1 was verified by double luciferase experiment;the effect of miR-129-2 on PIK3R1 and PI3K/AKT signaling pathway protein expression was detected by Western blotting.In mice,the overexpression of miR-129-2 can increase the food intake,slow down the respiratory rate,increase the body mass,reduce the lung tissue damage,reduce the expressions of IL-6 and TNF-α,and increase the expression of IL-10(P<0.05).In HPMECs,mimic transfection up-regulated the expression of miR-129-2,reduced the expression of inflammatory factors,enhanced the cell migration ability,and decreased the expression of PIK3R1(P<0.05).Dual luciferase reports showed that there was a binding site between miR-129-2 and PIK3R1.Downregulation of miR-129-2 increased the expression of PIK3R1 and activated the PI3K/AKT signaling pathway.Taken together,the overexpression of miR-129-2 can reduce the inflammatory response and lung injury in mice,and can mediate PIK3R1 to alleviate cell injury in pulmonary endothelial cells through the PI

关 键 词：miR-129-2 PIK3R1 PI3K/AKT 内皮细胞损伤 炎症因子 

分 类 号：R563.8[医药卫生—呼吸系统]