MicroRNA-132-3p靶向Nrf2加重脂多糖诱导的人脐静脉内皮细胞损伤的机制研究  

MicroRNA-132-3p targets Nrf2 to exacerbate lipopolysaccharideinduced injury in human umbilical vein endothelial cells

作　　者：马寒玉 赵宇浩 张铭 李真 王飞[1] 陈书艳[1] Ma Han-yu;Zhao Yu-hao;Zhang Ming;Li Zhen;Wang Fei;Chen Shu-yan(Department of Geriatrics,Xinhua Hospital of Shanghai Jiao Tong University School of Medicine,Shanghai 200092,China)

机构地区：[1]上海交通大学医学院附属新华医院老年医学科,上海200092

出　　处：《中国现代医学杂志》2025年第6期24-31,共8页China Journal of Modern Medicine

基　　金：国家自然科学基金(No:81974219)。

摘　　要：目的探讨microRNA-132-3p(miR-132-3p)靶向Nrf2加重脂多糖(LPS)诱导的人脐静脉内皮细胞(HUVECs)损伤的机制。方法用LPS刺激HUVECs建立体外脓毒症细胞模型,引起内皮细胞损伤。采用CCK-8法测定细胞活力,EdU法检测细胞增殖能力。转染miR-132-3p模拟物/抑制剂后,检测细胞迁移能力、乳酸脱氢酶(LDH)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、IL-1β、活性氧(ROS)、超氧化物歧化酶(SOD)和丙二醛(MDA)水平。通过荧光素酶报告基因验证miR-132-3p与其靶基因的结合。结果对照组细胞活力、细胞阳性比高于LPS组(P<0.05)。LPS组LDH、TNF-α、IL-6、IL-1β、ROS、MDA水平均较对照组升高(P<0.05),SOD水平较对照组降低(P<0.05)。LPS组miR-132-3p mRNA相对表达量较对照组升高(P<0.05),Nrf2 mRNA相对表达量较对照组降低(P<0.05)。LPS组Nrf2蛋白相对表达量较对照组降低(P<0.05)。LPS组与LPS+阴性对照组细胞活力比较,差异无统计学意义(P>0.05),LPS+miR-132-3p模拟物组细胞活力较LPS组降低(P<0.05),LPS+miR-132-3p抑制剂组细胞活力较LPS组升高(P<0.05)。LPS组与LPS+阴性对照组迁移细胞数比较,差异无统计学意义(P>0.05),LPS+miR-132-3p模拟物组迁移细胞数较LPS组减少(P<0.05),LPS+miR-132-3p抑制剂组迁移细胞数较LPS组增多(P<0.05)。LPS组与LPS+阴性对照组细胞划痕愈合率比较,差异无统计学意义(P>0.05),LPS+miR-132-3p模拟物组细胞划痕愈合率较LPS组降低(P<0.05),LPS+miR-132-3p抑制剂组细胞划痕愈合率较LPS组升高(P<0.05)。LPS组与LPS+阴性对照组LDH、TNF-α、IL-6、IL-1β、ROS、MDA和SOD水平比较,差异无统计学意义(P>0.05);LPS+miR-132-3p模拟物组LDH、TNF-α、IL-6、IL-1β、ROS、MDA水平均较LPS组升高(P<0.05),SOD水平较LPS组降低(P<0.05);LPS+miR-132-3p抑制剂组LDH、TNF-α、IL-6、IL-1β、ROS、MDA水平均较LPS组降低(P<0.05),SOD水平较LPS组升高(P<0.05)。对照组与阴性对照组Nrf2-WT的荧光素酶活性比较,差�Objective To investigate the mechanism by which microRNA-132-3p(miR-132-3p)targets Nrf2 to exacerbate lipopolysaccharide(LPS)-induced injury in human umbilical vein endothelial cells(HUVECs).Methods HUVECs were stimulated with LPS to establish an in vitro sepsis cell model,inducing endothelial cell injury.Cell viability was measured using the CCK-8 assay,and cell proliferation was assessed using the EdU assay.After transfection with miR-132-3p mimics/inhibitors,cell migration ability and levels of lactate dehydrogenase(LDH),tumor necrosis factor-alpha(TNF-α),interleukin-6(IL-6),IL-1β,reactive oxygen species(ROS),superoxide dismutase(SOD),and malondialdehyde(MDA)were measured.The binding of miR-132-3p to its target genes was confirmed by luciferase reporter assay.Results The cell viability and percentage of positive cells in the control group were higher than those in the LPS group(P<0.05).The levels of LDH,TNF-α,IL-6,IL-1β,ROS,and MDA were higher and the level of SOD was lower in the LPS group than in the control group(P<0.05).The relative expression of miR-132-3p in the LPS group was higher than that in the control group(P<0.05),and relative mRNA expression of Nrf2 was lower in the LPS group than that in the control group(P<0.05).The relative protein expression of Nrf2 was lower in the LPS group than that in the control group(P<0.05).The difference in cell viability between the LPS group and the LPS+NC group was not statistically significant(P>0.05).The cell viability of the LPS+miR-132-3p mimic group was lower than that of the LPS group(P<0.05).The cell viability of the LPS+miR-132-3p inhibitor group was higher than that of the LPS group(P<0.05).Comparing the number of migrated cells in the LPS group and the LPS+NC group,the difference was not statistically significant(P>0.05).The number of migrated cells in the LPS+miR-132-3p mimic group was decreased(P<0.05),and the number of migrated cells in the LPS+miR-132-3p inhibitor group was increased(P<0.05)compared with that in LPS group.The difference in the cel

关 键 词：脓毒症 microRNA-132-3p 内皮细胞 脂多糖 核因子红细胞2相关因子2 

分 类 号：R631[医药卫生—外科学]