miR-204-5p靶向TRIB3对脂多糖诱导的肺微血管内皮细胞损伤的影响  

The effect of miR-204-5p on the injury of pulmonary microvascular endothelial cells induced by lipopolysaccharide by targeting TRIB3

作　　者：沈九 吴宏峰 刘子航 张明[1] SHEN Jiu;WU Hong-feng;LIU Zi-hang(Department of Emergency,Nanjing Jiangning Hospital,Nanjing 211100,China;Department of Basic Medicine,Kangda College,Nanjing Medical University,Lianyungang 222000,China;Emergency Care Unit,Nanjing Jiangning Hospital,Nanjing 211100,China)

机构地区：[1]南京市江宁医院急诊科,江苏南京211100 [2]南京医科大学康达学院基础医学部,江苏连云港222000 [3]南京市江宁医院急诊监护室,江苏南京211100

出　　处：《中国实验诊断学》2023年第4期477-482,共6页Chinese Journal of Laboratory Diagnosis

摘　　要：目的探讨miR-204-5p对脂多糖(LPS)诱导的肺微血管内皮细胞损伤的影响及其可能作用机制。方法采用LPS诱导大鼠肺微血管内皮细胞(PMVEC)建立细胞损伤模型,实验分组:NC组、LPS组、LPS+miR-con组、LPS+miR-204-5p组、LPS+si-con组、LPS+si-TRIB3组、LPS+miR-204-5p+pcDNA组、LPS+miR-204-5p+pcDNA-TRIB3组;MTT法检测细胞活力;qRT-PCR、Western blot检测miR-204-5p、TRIB3表达;流式细胞术检测细胞凋亡;ELISA检测TNF-α、IL-6水平;双荧光素酶实验检测miR-204-5p与TRIB3的靶向关系。结果与NC组比较,LPS组细胞活力、miR-204-5p表达量降低(1.00±0.10比0.43±0.04),细胞凋亡率、TRIB3 mRNA(1.00±0.09 vs 2.13±0.18)和蛋白(0.40±0.04 vs 0.83±0.08)水平、TNF-α、IL-6水平升高(P<0.05);过表达miR-204-5p或敲低TRIB3可升高细胞活力,降低细胞凋亡率和TNF-α、IL-6水平(P<0.05);TRIB3是miR-204-5p的靶基因,上调TRIB3可减弱过表达miR-204-5p对细胞增殖、凋亡和炎症反应的影响。结论miR-204-5p过表达可靶向负调控TRIB3表达促进细胞增殖,抑制细胞凋亡和炎症反应,从而减轻LPS诱导的肺微血管内皮细胞损伤。Objective To explore the effect of miR-204-5p on lipopolysaccharide(LPS)-induced pulmonary microvascular endothelial cell damage and its possible mechanism.Methods LPS was used to induce PMVEC of rat to establish a cell injury model.Experimental groups:NC group,LPS group,LPS+miR-con group,LPS+miR-204-5p group,LPS+si-con group,LPS+si-TRIB3 group,LPS+miR-204-5p+pcDNA group,LPS+miR-204-5p+pcDNA-TRIB3 group.MTT method was used to detect cell viability.qRT-PCR and Western blot were performed to examine of miR-204-5p and TRIB3 expression.Flow cytometry was used to detect the cell apoptosis.ELISA was used to detect the levels of TNF-αand IL-6.Dual luciferase assay was used to detect the targeting relationship between miR-204-5p and TRIB3.Results Compared with the NC group,cell viability and of miR-204-5p expression(1.00±0.10 vs 0.43±0.04)in the LPS group was decreased,cell apoptosis,TRIB3 mRNA(1.00±0.09 vs 2.13±0.18)and protein(0.40±0.04 vs 0.83±0.08)levels,and the levels of TNF-αand IL-6 were increased(P<0.05).Overexpression of miR-204-5p or knockdown of TRIB3 increased cell viability,decreased cell apoptosis rate,TNF-αand IL-6 levels(P<0.05).TRIB3 is the target gene of miR-204-5p.Upregulation of TRIB3 attenuates the effects of overexpression of miR-204-5p on cell proliferation,apoptosis and inflammation.Conclusion Overexpression of miR-204-5p could target negative regulation of TRIB3 expression to promote cell proliferation,inhibit cell apoptosis and inflammation response,thereby reducing LPSinduced pulmonary microvascular endothelial cell damage.

关 键 词：大鼠肺微血管内皮细胞 脂多糖 miR-204-5p TRIB3 炎症 

分 类 号：R459.7[医药卫生—急诊医学]