摘要
目的探究巨噬细胞极化对周细胞-肌成纤维细胞转分化及移植肾间质纤维化形成中的作用。方法分别收集5例2010~2015年在南京医科大学第一附属医院确诊慢性移植物失功(CGD)受者的移植肾组织和正常肾组织(对照组), 采用常规染色和免疫荧光染色的方法检测M1型和M2型巨噬细胞在肾组织中的表达与分布, 并用聚合酶链式反应(PCR)法检测样本中CD68、CD206和iNOS mRNA;采用转化生长因子-β1(TGF-β1)体外刺激小鼠血管周细胞系, 采用免疫印迹、细胞荧光的方法检测周细胞中α-平滑肌肌动蛋白(α-SMA)和血小板衍生生长因子受体-β(PDGFR-β)的表达;分别构建血管周细胞与M1型或M2型巨噬细胞联合培养模型, 采用免疫印迹、细胞荧光、PCR等方法检测周细胞中α-SMA和PDGFR-β的表达。结果在CGD受者的移植肾组织中观察到显著的CD68^(+)iNOS^(+)的M1型巨噬细胞浸润, 而CD68^(+)CD206^(+)细胞未显著浸润, 且CD68、iNOS、CD206 mRNA在CGD组中的表达均高于对照组(P<0.05);在CGD移植肾组织中, α-SMA和PDGFR-β的蛋白表达升高(P<0.05), 且α-SMA和PDGFR-β双染的细胞浸润于CGD受者移植肾间质中。体外实验中, TGF-β1可刺激小鼠周细胞中α-SMA和PDGFR-β升高(P<0.05), 且呈现时间依赖性;免疫印迹和细胞荧光中均可见M1型巨噬细胞可在体外促进小鼠周细胞中周细胞-肌成纤维细胞转分化相关指标升高, 而M2型巨噬细胞无法促进周细胞发生周细胞-肌成纤维细胞转分化过程。结论 M1型巨噬细胞极化可能通过促进周细胞-肌成纤维细胞转分化过程, 促进移植肾间质纤维化的形成。
Objective To explore the role of macrophage polarization on pericyte-to-myofibroblast transition and renal allograft fibrosis after kidney transplantation(KT).Methods Allograft tissues were harvestedfrom recipients with chronic allograft dysfunction(CGD)and normal kidney tissues.The expression and distribution of M1/M2 macrophages in kidney tissues were detected by routine and immunofluorescent staining;mRNA of CD68,CD206 and iNOS detected by polymerase chain reaction(PCR);Murine vascular pericytes subjected to TGF-β1 in vitro and the expressions ofα-SMA and PDGFR-βin perivascular cells detected by immunoblotting and cellular fluorescence;The co-culturing models of vascular pericytes and M1/M2 macrophages were constructed.The expressions ofα-SMA and PDGFR-βin pericytes were detected by immunoblotting,cellular fluorescence and PCR.Results A marked infiltration of CD68^(+)iNOS^(+)M1 macrophages was present in allograft tissues of recipients with CGD while no obvious infiltration of CD68^(+)CD206^(+)was observed.The mRNA levels of CD68,iNOS and CD206 were significantly higher in CGD group than those in control group(P<0.05);In CGD allograft tissues,protein expressions ofα-SMA and PDGFR-βspiked markedly(P<0.05)while cells with double staining ofα-SMA and PDGFR-βwere markedly infiltrated in interstitial area of CGD allograft.TGF-β1 could induce a marked elevation of PMT-related markers in a time-dependent manner(P<0.05);Immunoblotting and cellukar fluorescence indicated that M1 macrophages could promote the elevations ofα-SMA and PDGFR-βin pericytes in vitro while M2 macrophages showed no effect on pericyte-to-myofibroblast transition in pericytes.Conclusions M1 macrophage polarization may promote the formation of renal allograft interstitial fibrosis through promoting PMT.
作者
王子杰
桂泽平
郑明
杭周
韩志坚
陶俊
居小兵
谭若芸
顾民
Wang Zijie;Gui Zeping;Zheng Ming;Hang Zhou;Han Zhijian;Tao Jun;Ju Xiaobin;Tan Ruoyun;Gu Min(Department of Urology,the First Affiliated Hospital of Nanjing Medical University,Jiangsu 210029,China;Department of Urology,the Second Affiliated Hospital of Nanjing Medical University,Nanjing,Jiangsu 210003,China)
出处
《中华器官移植杂志》
CAS
2022年第6期346-351,共6页
Chinese Journal of Organ Transplantation
基金
国家自然科学基金项目(81900684)
江苏省自然科学基金青年基金项目(BK20191063)。
关键词
肾移植
巨噬细胞
慢性移植物失功
Kidney transplantation
Macrophage
Chronic graft dysfunction
