中国现代医学杂志2025，Vol.35Issue(6)：24-31,8.DOI:10.3969/j.issn.1005-8982.2025.06.005
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 lipopolysaccharide-induced injury in human umbilical vein endothelial cells
摘要
Abstract
Absract: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 cell scratch healing rate between the LPS group and the LPS+NC group was not statistically significant(P＞0.05),the cell scratch healing rate in the LPS+miR-132-3p mimic group was lower than that of the LPS group(P＜0.05),and the cell scratch healing rate in the LPS+miR-132-3p inhibitor group was higher than that in the LPS group(P＜0.05).LDH,TNF-α,IL-6,IL-1β,ROS,MDA and SOD levels in the LPS group were not different from those in the LPS+NC group(P＞0.05).The levels of LDH,TNF-α,IL-6,IL-1β,ROS,and MDA were higher,but the level of SOD was lower in the LPS+miR-132-3p mimic group than in the LPS group(P＜0.05).Compared with the LPS group,LDH,TNF-α,IL-6,IL-1β,ROS,and MDA levels were lower(P＜0.05),and the SOD levels were higher in the LPS+miR-132-3p inhibitor group(P＜0.05).The luciferase activities of Nrf2-WT in the control group and the negative control group were not statistically significant(P＜0.05).The miR-132-3p mimic inhibited the luciferase activity of Nrf2-WT(P＜0.05),and miR-132-3p inhibitor enhanced the luciferase activity of Nrf2-WT(P＜0.05).Comparison of the luciferase activities of Nrf2-MUT in each group showed no statistically significant difference(P＞0.05).Comparison of the relative mRNA expression of Nrf2 in the LPS group and the LPS+NC group showed no statistically significant difference(P＞0.05).The relative mRNA expression of Nrf2 in the LPS+miR-132-3p mimic group was lower compared with that in the LPS group(P＜0.05).The relative mRNA expression of Nrf2 in the LPS+miR-132-3p inhibitor group was higher compared with that in the LPS group(P＜0.05).Comparison of the relative protein expression of Nrf2 in the LPS group and the LPS+NC group showed no statistically significant difference(P＞0.05),the relative protein expression of Nrf2 in the LPS+miR-132-3p mimic group was lower compared with that in the LPS group(P＜0.05),and the relative protein expression of Nrf2 in the LPS+miR-132-3p inhibitor group was higher compared with that in the LPS group(P＜0.05).Conclusions The experiment determined that miR-132-3p aggravated LPS-induced endothelial cell injury by downregulating Nrf2 expression,indicating that miR-132-3p might be a novel potential target for sepsis treatment.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2Key words
sepsis/microRNA-132-3p/endothelial cells/lipopolysaccharide/nuclear factor erythroid 2-related factor 2分类
临床医学引用本文复制引用
马寒玉,赵宇浩,张铭,李真,王飞,陈书艳..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[J].中国现代医学杂志,2025,35(6):24-31,8.基金项目
国家自然科学基金(No:81974219) （No:81974219）
