小鼠原发性胆汁性肝硬化与调节T细胞的相关性  

Relationship of Regulatory T Cells and Murine Primary Biliary Cirrhosis

作　　者：潘盈[1,2] 刘斌[1,3] 张奉春[1] 

机构地区：[1]中国医学科学院,北京协和医学院,北京协和医院风湿免疫科,北京100730 [2]西安交通大学第一附属医院风湿免疫科,西安710061 [3]青岛大学附属医院风湿科,山东青岛266000

出　　处：《中华临床免疫和变态反应杂志》2016年第2期131-136,F0003,共7页Chinese Journal of Allergy & Clinical Immunology

基　　金：国家十一五科技支撑计划项目(2008BAI59B03)

摘　　要：目的研究原发性胆汁性肝硬化(primary biliary cirrhosis,PBC)小鼠模型中调节T细胞(regulatory T cell,Treg)及其重要的调节因子转化生长因子β1(transforming growth factor-β1,TGF-β1)与发病的关系以及二者之间的关系。方法选取第8、16、24周PBC小鼠模型及正常对照小鼠(每组6只)-80℃冻存血清,使用酶联免疫吸附分析(enzyme-linked immunosorbent assay,ELISA)法测定血清TGF-β1含量;选取各组小鼠-80℃冻存肝脏标本,使用免疫印迹法半定量测定FOXP3含量;选取各组小鼠常温保存肝脏石蜡标本,使用免疫组化法标记FOXP3+Treg细胞,对汇管区淋巴细胞及FOXP3+Treg进行细胞计数。结果除16周PBC小鼠与24周PBC小鼠血清中TGF-β1含量存在统计学差异(0.174±0.084 vs.0.285±0.080,P=0.041)外,其他各组小鼠之间(包括8、16、24周PBC小鼠与正常对照比较)均无统计学差异;各组小鼠肝脏中均未测出FOXP3蛋白;根据PBC患者分期,PBC小鼠模型分期主要在1期和2期;各组PBC小鼠汇管区淋巴细胞数与同时期正常对照组相比,差异均具有显著统计意义(8周:394 vs.8,P=0.004,16周:392 vs.19,P=0.000,24周:171vs.18,P=0.000),且24周PBC小鼠汇管区淋巴细胞数显著低于16周PBC小鼠(171 vs.392,P=0.006);16、24周PBC小鼠Treg细胞数目显著高于同时期正常对照小鼠(11 vs.0,11 vs.0,P值分别为0.032,0.032)。结论 24周左右PBC小鼠肝脏病变进入纤维化阶段,同时期外周TGF-β1增高,肝脏组织Treg细胞数目增多;TGF-β1及Treg细胞与PBC肝纤维化有关,TGF-β1可能对Treg细胞的增生起促进作用。Objective To investigate whether Treg cells and TGF-β1 are involved in the pathogenesis of PBC in PBC mice model and investigate the relationship of Tregs and TGF-β1,which is one of most important regulatory cytokines. Methods PBC mice and normal control mice at 8w,16 w and 24 w were selected,6mice in each group. Serum were collected to evaluate the quantity of TGF-β1 with ELISA. Liver specimens were collected to evaluate the expression of FOXP3 with western blot and immunohistochemistry. Results The level of TGF-β1 in the serum of PBC mice was elevated than the control mice at 16 week and 24 week（ 0. 174 ± 0. 084 vs. 0. 285 ± 0. 080,P = 0. 041）. The expression of FOXP3 in liver couldn＇t be detected by western blot. According to the stages of PBC patients,PBC mice were all at stage Ⅰ and stage Ⅱ. Thenumber of lymphocytes in portal area in PBC mice was higher than controls（ 8 week： 394 vs. 8, P =0. 004,16 week： 392 vs. 19,P = 0,24 week： 171 vs. 18,P = 0）,while the lymphocytes in 24 week PBC mice were obviously fewer than 16 week PBC mice（ 171 vs. 392,P = 0. 006）. FOXP3＋Treg cells in16 week and 24 week PBC mice were obviously more than control mice（ 11 vs. 0,11 vs. 0,both P =0. 032）. Conclusions The infiltration of lymphocytes in the portal area is remarkable in PBC mice,and at about 24 week the inflammation is alleviated,which indicates the liver gets into the course of fibrosis. At the same period,TGF-β1 increased in the serum and Treg cells increase in the portal area of liver. TGF-β1 and Treg cells are involved in the fibrogenesis of primary biliary cirrhosis. TGF-β1 may accelerate the production of Treg cells.

关 键 词：肝硬化 胆汁性 模型 动物 T淋巴细胞调节性 转化生长因子Β 

分 类 号：R575.22[医药卫生—消化系统]