半乳糖凝集素1在脐带来源间充质干细胞调控类风湿关节炎患者T细胞功能中的作用  

Role of galectin-1 in regulation of umbilical cord mesenchymal stem cells on T cells of rheumatoid arthritis

作　　者：孙琳[1] 安卓[1] 李常虹[1] 刘蕊[1] 赵金霞[1] 刘湘源[1] SUN Lin AN Zhuo LI Chang-hong LIU Rui ZHAO Jin-xia LIU Xiang-yuan(Department of Rheumatology and Immunology, Peking University Third Hospital, Beijing 100191, China)

机构地区：[1]北京大学第三医院风湿免疫科,北京100191

出　　处：《北京大学学报（医学版）》2016年第6期964-969,共6页Journal of Peking University:Health Sciences

基　　金：国家自然科学基金(81273293)资助~~

摘　　要：目的:明确半乳糖凝集素1(galectin-1)在脐带间充质干细胞(umbilical cord mesenchymal stem cells,UCMSCs)调控类风湿关节炎(rheumatoid arthritis,RA)患者T细胞功能中的作用。方法:应用慢病毒构建galectin-1低表达的UC-MSCs,即UC-MSCs(Gal-1-),采用直接接触培养方式分别将UC-MSCs、UC-MSCs(Gal-1-)与RA患者的CD4^+T细胞共培养。设立阴性对照组(CD4^+T)、阳性对照组[CD4^+T培养过程中加入植物血凝素(phytohaemagg lutinin,PHA)2 mg/L]、UC-MSCs-CD4^+T共培养组、UC-MSCs(对照shRNA)-CD4^+T共培养组及UC-MSCs(Gal-1-)-CD4^+T共培养组。MTS法检测CD4^+T细胞增殖,ELISA方法检测共培养上清液中肿瘤坏死因子α(tumor necrosis factorsα,TNF-α)水平,流式细胞术检测Th1/Th2/Th17细胞亚型。结果:UC-MSCs和UC-MSCs(对照shRNA)与CD4^+T细胞共培养组可显著抑制PHA诱导的CD4^+T细胞增殖及TNF-α表达,而UC-MSCs(Gal-1-)组对此无明显抑制作用。与阴性对照组(8.51%±2.04%)相比,UC-MSCs及UC-MSCs(对照shRNA)组可显著抑制Th1细胞分化(分别为4.83%±1.37%和5.13%±0.87%,P值分别为0.012和0.018),而UC-MSCs(Gal-1-)组对此无明显抑制作用(6.41%±0.96%,P=0.101)。与阴性对照组相比,UC-MSCs及UC-MSCs(对照shRNA)组可上调Th2并下调Th17比例,但差异并无统计学意义,UC-MSCs(Gal-1-)组的这一作用不明显。结论:UC-MSCs可抑制RA患者CD4^+T细胞的增殖、活化并可降低Th1细胞比例,但敲低galectin-1分子的表达后该作用减弱,提示galectin-1参与了UC-MSCs抑制RA患者CD4^+T细胞功能的过程。Objective： The therapeutic potential of umbilical cord mesenchymal stem cells（ UC-MSCs）in rheumatoid arthritis（ RA） has attracted more and more attention,because of it can suppress the various inflammatory effects of T cells. Galectin-1 is highly expressed in UC-MSCs,as the first lectin mediating the immunomodulatory effect of MSCs. Our study will investigate the effects of galectin-1 in regulation of UC-MSCs on rheumatoid arthritis T cells. Methods： Lentivirus transfected shRNA technique was used to knock down the expression of galectin-1 in UC-MSCs to construct UC-MSCs（ Gal-1^-）. The effects of UC-MSCs and UC-MSCs（ Gal-1^-） on CD4^＋T cells in RA patients were investigated by contact system,including negative control group（ CD4^＋T cells）,positive control group [CD4^＋T-phytohemagg lutinin（ PHA） ],UC-MSCs-CD4^＋T cells co-culture group,UC-MSCs（ control shRNA）-CD4^＋T cells co-culture group,and UC-MSCs（ Gal-1^-）-CD4^＋T cells co-culture group. The proliferation of CD4^＋T cells was detected by MTS assay. The level of tumor necrosis factors α（ TNF-α） in cells supernatant was detected by enzyme linked immunosorbent assay（ ELISA）. The effect of UC-MSCs on helper T cell（ Th）subset was detected by flow cytometry. Results： In vitro,UC-MSCs were capable of inhibiting PHA induced proliferation of CD4^＋T cells from RA patients,but UC-MSCs（ Gal-1^-） did not show the significant inhibitory effect. Galectin-1 affect the TNF-α level of CD4^＋T cells regulated by UC-MSCs. UCMSCs and UC-MSCs（ control shRNA） significantly inhibited the expression of TNF-α in PHA-induced CD4^＋T cells. However,UC-MSCs（ Gal-1^-） had no significant inhibitory effect. Furthermore,the Th1 cells were also significantly suppressed by UC-MSCs and UC-MSCs（ control shRNA）（ 4. 83% ± 1. 37%and 5. 13% ± 0. 87%,P = 0. 012 and P = 0. 018）. These was no significant difference in the proportionof the Th1 cells between the control group and UC-MSCs（ Gal-1^-） group（ 8. 51

关 键 词：关节炎 类风湿 间质干细胞 半乳糖凝集素类 

分 类 号：R593.22[医药卫生—内科学]