体外人脐带间充质干细胞通过促分泌前列腺素E2肝细胞生长因子吲哚胺2，3-双加氧酶抑制干燥综合征CD4＋T细胞的活化增殖  

Mesenchymal stem cells inhibit the activation and proliferation of CD4＋ T cells in SjiSgren syndrome by promoting prostaglandin E2, hepatocyte growth factor and indoleamine 2, 3-dioxygenase secretion

作　　者：路臻豪 黄家树[2] 阮光峰[2] 房星星[2] 龚邦东[2] 汤建平[2] 

机构地区：[1]同济大学附属杨浦医院风湿免疫科,上海200090 [2]同济大学附属同济医院风湿免疫科

出　　处：《中华风湿病学杂志》2018年第3期181-185,F0003,共6页Chinese Journal of Rheumatology

基　　金：国家自然科学基金（81273295,81302562,81671598）;国家教育部留学回国人员科研启动基金（20101174）;上海市卫生局基金（2011276）;中华国际医学交流基金会项目（Z-2014.06.2-1620）

摘　　要：目的探讨体外人脐带MSCs促分泌的前列腺素E2（PGE2）、肝细胞生长因子（HGF）以及吲哚胺2，3-双加氧酶（IDO）在pSS中对外周血活化CD4＋T细胞的抑制作用。方法原代分离脐带MSCs，并经流式细胞术鉴定。荧光激活细胞分选术（FACS）分选pSS患者外周血CD4＋T细胞，分为3组：分别为活化CD4＋T细胞组（CD3、CD28抗体共刺激72h）、MSCs共培养组（活化CD4＋T细胞与MSCs共培养72h）和IFN-γ预刺激后MSCs共培养组（活化CD4＋T细胞与IFN-γ预刺激后的MSCs共培养72h），活化CD4＋T细胞组为对照。共培养72h后，收集悬浮的CD4＋T细胞，进行细胞计数，ELISA法检测共培养上清中细胞因子PGE2、HGF和IDO的表达水平。组间均数比较用单因素方差分析，多重比较采用LSD法。结果pSS患者活化CD4＋T细胞组、MSCs共培养组和IFN-γ预刺激后MSCs共培养组的培养上清中PGE2浓度[分别为（111±4）pg／ml、（2814±6）pg／ml和（2716±8）pg／ml，F=167292．12，P〈0．01]；HGF浓度[分别为（597±9）pg／ml、（383±9）pg／ml和（727±12）pg／ml，F=878．61，P〈0．01]；IDO浓度[分别为（143±4）pg／ml、（835±5）pg／ml和（588±3）pg/ml，F=21104．41，P〈0．01]。相对于活化CD4＋T细胞组，MSCs共培养组及IFN-γ预刺激后MSCs共培养组的培养上清中PGE2浓度升高（t=509．88，P〈0．01；t=491．48，P〈0．01），IDO浓度也升高（t=202．69，P〈0．01；t=130．39，P〈0．01），同时活化的CD4＋T细胞增殖受到抑制（t=-16．20，P〈0．01；t=-31．48，P〈0．01）；相对于MSCs共培养组，IFN-γ预刺激后MSCs共培养组的培养上清中PGE2、IDO浓度均有降低（t=-18．40，P〈0．01；t=-72．30，P〈0．01），而HGF浓度升高（t=41．51，P〈0．01），同时活化的CD4＋T细胞增殖受到进一步抑制（t=-15．28，P〈0．01）。结论MSCs在体外可以抑制DSS活化的CD4＋T细胞增殖，其对CD4＋T细胞的抑制�Objective To investigate the suppressive effect of prostaglandin E2 （PGE2）, hepatocyte growth factor （HGF） and indoleamine 2, 3-dioxygenase （IDO） whose secretion was promoted by human um- bilical cord mesenchymal stem eells（MSCs） on the activated peripheral blood CD4＋ T cells in primary Sjtigren syndrome （pSS） in vitro. Methods Primary cultured umbilical cord MSCs were identified by flow cytometry, and peripheral blood CD4＋ T cells were sorted in pSS patients. CD4＋ T cells were cultured with CD3, CD28 antibody for 72 h to be the activated（control group）; the activated CD4＋ T cells were co-cultured with MSCs for 72 h（MSCs group） or MSCs were pre-stimulated with interferon-γ（IFN-γ）, then the activated CD4＋ T cells were co-cultured with pre-stimulated MSCs for 72 h （pre-stimulated group）. The suspension of CD4＋ T cells were collected and counted. PGE2, HGF and IDO in the supematants were detected by ELISA. Mean in groups were compared using ANOVA, and multiple comparisons were used with LSD method. Results The concentrations of PGE2 in the supernatant of the control group, MSCs group and pre-stimulated group were （111±4） pg/ml, （2 814±6） pg/ml and （2 716±8） pg/ml （F=167 292.12, P〈0.01） respectively. The concentrations of HGF in the above groups were （597±9） pg/ml, （383±9） pg/ml and （727±12） pg/ml（F=878.61, P〈0.01） respectively. The concentrations of IDO in the above groups were （143±4） pg/ml, （835±5） pg/ml and （588±3） pg/ml （F=21 104.41, P〈0.01） respectively. Compared with the control group, levels of PGE2 significantly increased in the MSCs group and the pre-stimulated group that CD4＋ T cells were co-cultured with MSCs （t=509.88, P〈0.01 and t=491.48, P〈0.01）, and levels of IDO also significantly increased （t=202.69, P〈0.01 and t=130.39, P〈0.01）, while the activation and proliferation of CD4＋ T ceils were inhibited （t=-16.20, P〈0.01 and t=-31.48, P〈0.01）. Compared with MSCs gro

关 键 词：干燥综合征 间质干细胞 脐带 地诺前列酮 肝细胞生长因子 吲哚胺-毗咯2 3-双加氧酶 CD4阳性T淋巴细胞 

分 类 号：R593.2[医药卫生—内科学]