M1型巨噬细胞极化在内皮细胞转分化及慢性移植肾失功中的作用  

The role of M1 polarized-macrophage in endothelial-to-myofibroblast transition and chronic allograft dysfunction

作　　者：张翔[1] 王子杰[1] 郑明 韩前广 黄正楷[1] 易小敏 陶俊[1] 韩志坚[1] 谭若芸 居小兵[1] 顾民[1] ZHANG Xiang;WANG Zijie;ZHENG Ming;HAN Qianguang;HUANG Zhengkai;YI Xiaomin;TAO Jun;HAN Zhijian;TAN Ruoyun;JU Xiaobing;GU Min(Department of Urology,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China)

机构地区：[1]南京医科大学第一附属医院泌尿外科,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2021年第9期1296-1303,1309,共9页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金(81900684,81870512);江苏省自然科学基金青年基金(BK20191063)。

摘　　要：目的:探究M1型巨噬细胞极化在内皮细胞向间充质细胞转分化(endothelium-to-myofibroblast transition,EndMT)及慢性肾移植失功(chronic allograft dysfunction,CAD)中的作用。方法:从GEO公共数据库中下载GSE21374转录组测序数据,并使用Cibersort软件分析CAD患者移植肾中的免疫细胞的浸润状态。收集本中心CAD患者的移植肾切除标本,使用免疫荧光、聚合酶链式反应(polymerase chain reaction,PCR)和蛋白质印迹法(Western blot,WB)等方法观察M1型巨噬细胞在移植肾组织中的浸润情况。体外脂多糖(lipopolysaccharide,LPS)和干扰素-γ(interferon-γ,IFN-γ)诱导Raw264.7巨噬细胞向M1极化。Transwell小室构建M1型巨噬细胞和主动脉内皮细胞的共培养模型。使用PCR、WB和细胞免疫荧光观察共培养模型中内皮细胞的CD31与α-SMA的表达情况。结果:基于GEO公共数据库,发现单核-巨噬细胞明显浸润于CAD移植肾组织中(P<0.05);在本中心的CAD患者移植肾标本中,CD68(+)iNOS(+)M1型巨噬细胞亦明显浸润于肾小球及间质中,且M1型巨噬细胞标志物诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)的mRNA表达量明显增加(P<0.05)。在细胞共培养模型中发现,M1型巨噬细胞可明显促进内皮细胞发生EndMT过程。结论:M1型巨噬细胞明显浸润于CAD患者的移植肾组织中,并可能通过诱导内皮细胞发生EndMT促进CAD的进展。Objective:This study aims to explore the role of M1-polarized macrophages in the pathogenesis of endothelial-to-mesenchymal transition(EndMT)and chronic allograft dysfunction(CAD).Methods:The GSE21374 transcriptome array from GEO public database was downloaded,and cibersort software was used to analyze the immune cell infiltration status in allograft tissues of CAD patients.Then the allograft tissues were collected from patients diagnosed with CAD in our center.Immunofluorescence,polymerase chain reaction(PCR)and Western blot(WB)were used to observe infiltration of M1-polarized macrophages in the renal allografts.Finally,in vitro lipopolysaccharide(LPS)and interferon-γ(IFN-γ)were used to induce the polarization of Raw264.7 macrophage cell line into M1 macrophages.A transwell chamber was used to establish a co-culture system for the M1 macrophages and Balb/c mouse-derived aortic endothelial cells.PCR and WB assays,as well as cell immunofluorescence,were performed to examine the expression of CD31 andα-SMA in the mouse-derived aortic endothelial cells.Results:Based on the public database,monocytes and macrophages were observed to be highly expressed in allograft tissues from CAD patients(P<0.05).Similarly,significant infiltration of CD68(+)iNOS(+)M1 macrophages in glomerulus and interstitial area of allograft with CAD was observed,and PCR results showed that iNOS(inducible nitric oxide synthase),the surface marker of M1 macrophages was increased significantly compared to the normal kidney tissues(P<0.05).Co-cultured with M1 macrophages,the EndMT of endothelial cells was promoted.Conclusion:We have observed significant M1 macrophages infiltration in the allograft tissues diagnosed with CAD,which may induce the occurrence of EndMT and CAD progression in renal transplant recipients.

关 键 词：巨噬细胞极化 内皮细胞向间充质细胞转分化 慢性移植肾失功 肾脏移植 

分 类 号：R692.5[医药卫生—泌尿科学]