基于NLRP3通路研究miR-155对THP-1巨噬细胞炎症反应的调控作用  

Regulation of MiR-155 on THP-1 Macrophage Inflammatory Response Based on NLRP3 Pathway

作　　者：宁晓县 申元英 郭乐 Ning Xiaoxian;Shen Yuanying;Guo Le(Pre-clinical College,Dali University,Dali,Yunnan 671000,China)

机构地区：[1]大理大学基础医学院,云南大理671000

出　　处：《大理大学学报》2022年第10期32-38,共7页Journal of Dali University

基　　金：国家自然科学基金项目(81960371);云南省基础研究计划项目(202001AU070014)。

摘　　要：目的:基于核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体通路探讨miR-155对脂多糖(LPS)诱导的THP-1巨噬细胞炎症反应的调控作用。方法:构建LPS诱导的THP-1巨噬细胞炎症模型,将细胞分为空白对照组、LPS组、miR-155过表达组和miR-155阴性对照组,实时荧光定量聚合酶链反应检测白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)和NLRP3的表达变化;酶联免疫吸附试验测定IL-1β和TNF-α的分泌水平;蛋白质印迹法检测NLRP3和含半胱氨酸的天冬氨酸蛋白水解酶-1(Caspase-1)的表达。结果:LPS诱导细胞中炎症因子、NLRP3和miR-155表达;过表达miR-155,进一步促进炎症因子NLRP3和Caspase-1蛋白的表达。结论:miR-155通过激活其潜在作用靶点NLRP3促进LPS诱导的THP-1巨噬细胞炎症反应。Objective:To investigate the regulatory effect of miR-155 on lipopolysaccharide(LPS)-induced inflammatory response of THP-1 macrophages based on the nucleotide binding oligomerized domain-like receptor 3(NLRP3)inflammasome pathway.Method:The LPS-induced inflammatory model of THP-1 macrophages was constructed,the cells were divided into blank control group,LPS group,miR-155 overexpression group and miR-155 negative control group,and real-time quantitative polymerase chain reaction detected the expression changes of interleukin 1β(IL-1β),tumor necrosis factorα(TNF-α)and NLRP3.Enzyme-linked immunosorbent assay measures the secretion levels of IL-1βand TNF-α;Western blot detection of NLRP3 and cysteine-containing aspartate proteolytic enzyme-1(Caspase-1).Results:LPS induced the expression of inflammatory cytokines,NLRP3 and miR-155 in cells;overexpression of miR-155 further promoted the expression of inflammatory cytokines,NLRP3 and Caspase-1 proteins.Conclusion:MiR-155 promotes the LPS induced THP-1 macrophage inflammatory response by activating its potential target NLRP3.

关 键 词：NLRP3通路 MIR-155 THP-1细胞 炎症 

分 类 号：R392.12[医药卫生—免疫学]