发育型内皮基因座-1作用下氧化型低密度脂蛋白对巨噬细胞分泌TNF-α、MIP-1α及MIP-2的影响  

Effect of developmental endothelial locus-1 gene on the secretion of TNF-α,MIP-1αand MIP-2 by oxidized low-density lipoprotein-stimulated macrophages

作　　者：路艳林 李方琴 周明 彭进 万昌武[1] 梁静伟 陈灿 王杰[1] 夏冰[1] LU Yanlin;LI Fangqin;ZHOU Ming;PENG Jin;WAN Changwu;LIANG Jingwei;CHEN Can;WANG Jie;XIA Bing(School of Forensic Medicine,Guizhou Medical University,Guiyang 550004,Guizhou,China;Criminal Investigation Brigade,Xishui County Public Security Bureau,Zunyi 564600,Guizhou,China)

机构地区：[1]贵州医科大学法医学院,贵州贵阳550004 [2]贵州省习水县公安局刑事侦查大队,贵州遵义564600

出　　处：《贵州医科大学学报》2023年第8期888-894,共7页Journal of Guizhou Medical University

基　　金：国家自然科学基金(82060340);贵州省科技计划项目(黔科合基础-ZK〔2021〕一般487);贵州医科大学国家自然科学基金培育项目(20NSP083)。

摘　　要：目的探讨发育型内皮基因座-1(DEL-1)作用下氧化型低密度脂蛋白(OX-LDL)对巨噬细胞中肿瘤坏死因子α(TNF-α)、巨噬细胞炎性蛋白1α(MIP-1α)及巨噬细胞炎性蛋白2(MIP-2)表达的影响。方法体外培养人单核-巨噬细胞系THP-1细胞株至对数生长期,分为佛波脂(PMA)组(PMA诱导为贴壁的巨噬细胞)、OX-LDL组(OX-LDL诱导为泡沫细胞)、si-NC组(50 nmol/L siRNA-NC的小干扰转染细胞)及小干扰RNA-DEL-1(siRNA-DEL-1)组(50 nmol/L siRNA-DEL-1转染细胞),采用油红O染色鉴定泡沫细模型、并检测各组细胞中脂质积累,采用免疫荧光技术检测各组细胞中DEL-1的表达,采用实时聚合酶链反应(RT-PCR)及Western blot检测各组细胞中DEL-1、MIP-1α、MIP-2及TNF-α信使RNA(mRNA)和蛋白的表达;采用Pearson相关系数分析DEL-1蛋白表达与上述炎性因子的相关性。结果油红O染色结果显示,靶向敲低DEL-1泡沫细胞胞质内脂质积累减少;免疫荧光、Western blot及RT-PCR结果显示,OX-LDL组、siRNA-NC组细胞中DEL-1及下游炎性因子MIP-1α、MIP-2及TNF-α的蛋白及mRNA表达较PMA组上调(P<0.05),siRNADEL-1组细胞中上述因子表达较OX-LDL组及siRNANC组下调(P<0.05)。结论DEL-1介导了OX-LDL源性泡沫细胞分泌MIP-1α、MIP-2及TNF-α,可能参与了动脉粥样硬化(AS)的病变过程。Objective To investigate the effect of developmental endothelial locus-1 gene(DEL-1)on the expression of TNF-α,MIP-1α,and MIP-2,the inflammatory factors secreted by macrophages under the effect of OX-LDL.Methods The human monocyte-macrophage cell line THP-1 cell line was cultured in vitro to logarithmic growth phase and divided into the fobolipid(PMA)group(PMA induced as apposed macrophages),OX-LDL group(OX-LDL induced as foam cells),si-NC group(50 nmol/L siRNA-NC of small interfering transfected cells),and small interfering RNA-DEL-1(siRNA-DEL-1)group(50 nmol/L siRNA-DEL-1 transfected cells).Oil red O staining was used to identify the foam cell models,and detect the lipid accumulation of THP-1 cells in each group.The expression of DEL-1 of THP-1 cells in each group was detected by immunofluorescence technique;Real-time polymerase chain reaction(RT-PCR)and Western blotting were used to detect the expressions of DEL-1,MIP-1α,MIP-2,and TNF-αmessenger RNA(mRNA)and protein in each group.Pearson correlation coefficient method was used to analyze the correlation between DEL-1 protein table and the above-mentioned inflammatory factors.Results Oil Red O staining showed that intracytoplasmic lipid accumulation in foam cells was reduced with targeted knockdown of DEL-1;Immunofluorescence,Western blotting and RT-PCR showed that the protein and mRNA expression of DEL-1 and downstream inflammatory factors MIP-1α,MIP-2,and TNF-αwere up-regulated in THP-1 cells in OX-LDL and siRNA-NC groups compared with PMA group(P<0.05),while the expressions of these factors were down-regulated in THP-1 cells in the siRNADEL-1 group compared with OX-LDL and siRNANC groups(P<0.05).Conclusion DEL-1 may mediate the secretion of MIP-1α,MIP-2,and TNF-αby OX-LDL-derived foam cells and is likely involved in the pathological process of atherosclerosis(AS).

关 键 词：泡沫细胞 动脉粥样硬化 肿瘤坏死因子α 发育型内皮基因座-1基因 巨噬细胞炎性蛋白1Α 巨噬细胞炎性蛋白2 

分 类 号：R89[医药卫生—法医学]