IL-37通过促进巨噬细胞M2型极化在缺氧/复氧诱导的肝细胞损伤中的保护作用  

IL-37 protects hepatocyte injury against hypoxia/reoxygenation by promoting polarization of M2-type macrophages

作　　者：朱迪[1] 李婷婷[1] 陈青松[1] 牟童[1] 汤成泳[2] ZHU Di LI Ting-ting CHEN Qing-song MOU Tong TANG Cheng-yong(Department of Hepatobiliary Surgery Department of Pharmacy,the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China)

机构地区：[1]重庆医科大学附属第一医院肝胆外科,重庆400016 [2]重庆医科大学附属第一医院药学部,重庆400016

出　　处：《复旦学报（医学版）》2017年第4期453-460,共8页Fudan University Journal of Medical Sciences

基　　金：重庆市科委基金(CSTC2015shmszx120019)~~

摘　　要：目的探讨IL-37通过促进巨噬细胞M2型极化在缺氧/复氧(hypoxia/reoxygenation,H/R)诱导的肝细胞损伤中的保护作用及其相关分子机制。方法实时荧光定量PCR(qRT-PCR)和Western blot检测在不同极化类型的人单核巨噬细胞THP-1中IL-37 mRNA和蛋白质水平。通过慢病毒转染构建稳定过表达IL-37的细胞株,qRT-PCR检测CD206、CD86、ARG1和iNOS mRNA水平,流式细胞术检测CD163、CD86蛋白质水平。通过Transwell法将THP-1细胞与人肝细胞L02细胞共培养并构建H/R模型,CCK-8法及流式细胞术检测L02细胞存活率及凋亡率,ELISA检测细胞培养液中谷丙转氨酶(alanine transaminase,ALT)和谷草转氨酶(aspartate aminotransferase,AST)含量,HE染色观察细胞形态变化。Western blot检测THP-1细胞中STAT6及其磷酸化水平。结果 M2型巨噬细胞中IL-37 mRNA及蛋白质水平上调。IL-37促进巨噬细胞M2型极化。IL-37诱导的M2型巨噬细胞与L02细胞共培养,在H/R状态下显著提高肝细胞存活率(P=0.015),并降低细胞凋亡率和ALT、AST水平(P<0.001),减轻细胞损伤。Western blot提示过表达IL-37的THP-1细胞中STAT6蛋白质磷酸化水平上调(P<0.01)。结论 IL-37能促进巨噬细胞M2型极化,并对H/R诱导的肝细胞损伤有保护作用,其机制可能与STAT6信号通路有关。Objective To investigate the protective effect of IL-37 on hepatocyte injury against hypoxia/reoxygenation （H/R） by promoting polarization of M2-type macrophages and its molecular mechanisms. Methods Real-time fluorescent quantitative PCR （qRT-PCR） and Western blot were used to detect the levels of IL-37 mRNA and protein in human monocyte-macrophage THP-1 cells with different polarizations. The lentivirus with IL-37 gene was infected into THP-1 cells. The levels of CD206, CD86, ARG1 and iNOS mRNA was detected by qRT-PCR. The levels of CD163 and CD86 protein was detected by flow cytometric analysis. THP-1 cells and L02 cells were co-cultured by Transwell and treated with H/R. The survival rate and apoptotic rate of L02 cells were detected. The levels of alanine transaminase （ALT） and aspartate aminotransferase （AST） in culture medium were measured. The levels of STAT6 and its phosphorylation in THP-1 cells were detected by Western blot. Results The levels of IL-37 mRNA and protein were up-regulated in M2-type macrophages. IL-37 promoted the polarization of M2-type macropahges. M2-type macrophages induced by IL-37 were co-cultured with L02 cells, the survival rate was significantly increased by H/R treatment （P=0.015）, while the apoptotic rate, ALT level and AST level were significantly decreased （P〈0.001）. The level of phosphorylated STAT6 in THP-1 cells overexpressing IL-37 was up-regulated （P〈0.01）. Conclusions IL-37 can induce polarization of M2-type macrophages and protect hepatocyte injury against H/R. Its mechanism may be related to STAT6 signal pathways.

关 键 词：IL-37 巨噬细胞 缺氧/复氧 肝细胞损伤 

分 类 号：R575[医药卫生—消化系统] R364.4[医药卫生—内科学]