MSCs联合雷帕霉素对异基因鼠脾淋巴细胞的免疫调节作用  

Mesenchymal stem cells combined with rapamycin immuno-regulates the spleniclymphocytes in allogenic mice

作　　者：殷玉俊[1] 李晶[2] 汤郁[2] 尤海燕[3] 朱伟[4] 许文荣[4] 焦志军[3] 

机构地区：[1]江苏大学附属医院皮肤科,江苏镇江212001 [2]江苏大学附属医院风湿科,江苏镇江212001 [3]江苏大学附属医院检验科,江苏镇江212001 [4]江苏大学医学技术学院,江苏镇江212013

出　　处：《山东大学学报（医学版）》2009年第4期58-61,64,共5页Journal of Shandong University:Health Sciences

基　　金：江苏省科技厅社会发展项目(BS2005043)

摘　　要：目的探讨体外骨髓间充质干细胞(MSCs)联合雷帕霉素对BALB/c鼠T、B淋巴细胞的免疫调节作用及可能机制。方法从5～6周龄BALB/c鼠骨髓中分离培养MSCs并鉴定其纯度。用EZ-SepTM Mouse1X分离异体BALB/c鼠脾脏淋巴细胞,分别在刀豆蛋白A(ConA)、脂多糖(LPS)刺激下,用MSCs和/或雷帕霉素处理,MTT法检测淋巴细胞的增殖,流式细胞术检测T、B淋巴细胞CD69、CD28和CD86的表达和T淋巴细胞凋亡情况,Real-time PCR测定T淋巴细胞IL-10 mRNA、IFN-γ mRNA的表达水平。结果MSCs明显抑制T、B淋巴细胞增殖,联合雷帕霉素组抑制作用更加显著(P<0.01)。MSCs可抑制T淋巴细胞的凋亡,联合雷帕霉素组较单独MSCs组抑制作用更加明显(P<0.01)。MSCs联合雷帕霉素具有协同促进IFN-γ mRNA表达和协同抑制IL-10 mRNA表达的作用。单独MSCs或MSCs联合雷帕霉素对T、B淋巴细胞CD69、CD28和CD86的表达无显著影响。结论MSCs联合雷帕霉素对BALB/c鼠T、B淋巴细胞有免疫负调节作用,可能与协同抑制淋巴细胞增殖、T淋巴细胞凋亡和干预IFN-γ mRNA、IL-10 mRNA表达有关。Objective To investigate the immunoregulatory effects of mesenchymal stem cells（MSCs） combined with rapamycin on T and B cells in vitro and its potential mechanism. Methods MSCs were isolated and cultivated from the bone marrow of 5- 6 week-old BALB/c mice in vitro. The purity was detected through surface markers by flow cytometry（FCM）. Splenic lympho- cytes were isolated by EZ-Sep（tm） Mouse IX. Under ConA or LPS stimulation, lymphocytes were treated with MSCs and/or rapa- mycin. The proliferation was assessed by MTT coloremetry. FCM was used to analyze the apoptosis and surface markers-CD69, CD28 and CD86. mRNA expressions of cytokines were detected by real-time quantitative PCR. Results Both MSCs and MSCs combined with rapamycin could significantly inhibit proliferation of lymphocytes and the apoptosis of T cells. However, the inhibi- tory effect of the latter was more obvious than that of the former （P 〈 0.01 ）. Both MSCs and MSCs combined with rapamycin could significantly increase expression of interferon （IFN）-γ mRNA but decrease expression of interleukin （IL）-10. No differences of CD69, CD28 and CD86 expressions were observed among all groups. Conclusion MSCs combined with rapamycin could down-modulate the function of T and B cells, and the potential mechanism may be related to its effect on the proliferation, apopto- sis and mRNA expressions of IFN-γ and IL-10.

关 键 词：间充质干细胞 雷帕霉素 淋巴细胞 免疫调节 

分 类 号：R392[医药卫生—免疫学]