IL-10/TGF-β诱导的巨噬细胞对肾缺血再灌注损伤的作用研究  

Effects of interleukin （IL）-10/ transforming growth factor （TGF）-β-modified macrophages on renal ischemia reperfusion injury

作　　者：赵艳龙[1] 田普训[1] 郑瑾[1] 丁晨光[1] 高祎 薛武军[1] 丁小明[1] 刘静[3] 韩锋[1] 夏欣欣[4] Zhao Yanlong , Tian Puxun , Zheng Jin , Ding Chenguang , Gao Yi , Xue Wujun, Ding Xiaoming, Liu Jing, Han Feng, Xia Xinzcin.(Department of Kidney Transplantation, Hospital of Nephropathy , the First Affiliated Hospital of Xi＇an Jiaotong University, Xi＇an 710061, China)

机构地区：[1]西安交通大学第一附属医院肾脏病医院肾移植科,710061 [2]西安市第三医院肾脏内科 [3]西安交通大学第二附属医院肾脏内科 [4]西安交通大学第一附属医院中医科

出　　处：《中华器官移植杂志》2017年第12期734-740,共7页Chinese Journal of Organ Transplantation

基　　金：国家自然科学基金（81670681,81670682）;陕西省自然科学基础研究计划--重大基础研究项目（2017ZDJC-09）

摘　　要：目的分析白细胞介素10（IL-10）/转化生长因子-β（TGF-β）诱导的巨噬细胞对肾缺血再灌注损伤的影响。方法IL-10/TGF-β联合诱导骨髓源性巨噬细胞，获得M2c型巨噬细胞，尾静脉输注给肾缺血再灌注损伤（IRI）模型小鼠。肾IRI后3 d检测肾功能，HE染色分析肾组织病理变化，脱氧核糖核苷酸末端转移酶介导的缺口末端标记法检测肾小管细胞的凋亡，增殖细胞核抗原的免疫荧光染色分析小管细胞增值，聚合酶链反应及流式细胞仪分别分析肾组织炎症因子和调整性T淋巴细胞（Treg）表达，评估M2c巨噬细胞对肾缺血再灌注损伤的影响。结果成功获得成熟巨噬细胞（纯度〉95%，增殖能力〈1%）及M2c型巨噬细胞，与经典活化的M1型比较，M2c表达低水平的MHCⅡ（P〈0.01）、CD86（P〈0.01）及炎症因子TNF-α（P〈0.01）、IL-1β（P〈0.01），但却表达高水平的IL-10（P〈0.01）。M2c型巨噬细胞可以明显改善肾IRI小鼠肾功能（P〈0.01或P〈0.05）及病理损伤（P〈0.05）；并能抑制小管上皮细胞的凋亡（P〈0.01），促进肾小管上皮细胞的增殖（P〈0.05），减轻肾组织内炎症因子TNF-α（P〈0.05）、IL-1β（P〈0.01）和IL-6（P〈0.05）浸润，增加调节性T淋巴细胞表达（P〈0.01）。结论IL-10/TGF-β诱导的M2c型巨噬细胞通过抑制小管细胞凋亡、炎症细胞浸润及促进小管细胞增殖等对肾缺血再灌注损伤发挥保护作用。M2c巨噬细胞可能成为干预肾移植等过程导致的肾IRI的新策略。Objective To investigate the effects of IL-10/TGF-β-modified macrophages on renal ischemia reperfusion injury （IRI）. Methods Bone marrow-derived macrophages were modified ex vivo by IL-10/TGF-β to acquire M2c （a subset of activated macrophages）. M2c were transferred into treated C57BL/6 mice by a single tail-vein injection at 6 h after renal IRI. Mice were killed on the day 3 after renal IRI. Blood samples were collected to check renal function. Kidneys were harvested to determine tubular necrosis and apoptosis by H＆E staining and TUNEL assay. Immunofluorescence was performed to analyze the proliferating tubular cell nuclear antigen. Meanwhile, proinflammatory cytokines and regulatory T cells in renal tissues were analyzed with real-time PCR and flow cytometry. Results In comparison with M1, M2c expressed lower levels of MHC Ⅱ （P〈0. 01）, CD86 （P〈 0.01）, TNFα （P〈0.01） and IL-1β （P〈0.01） and higher level of IL-10 （P〈0.01）. M2c significantly attenuated renal functional decline （P〈 0. 01 or 0.05）, structural injury （P 〈 0. 05 ）, apoptosis of tubular cells （P〈0. 01） and inflammation factors infiltration （P〈0. 01 or 0. 05）. Whars more, the cells could promote tubular cells proliferation （P%0. 05） and regulatory T cells expression （P〈0. 01）. Conclusion Our results demonstrated that M2c macrophages effectively protect against renal IRI and may become a therapeutic strategy for renal IRI.

关 键 词：肾 缺血再灌注损伤 巨噬细胞 小鼠 

分 类 号：R699.2[医药卫生—泌尿科学]