MSCs联合雷帕霉素对异基因鼠脾淋巴细胞的免疫调节作用

山东大学学报(医学版)

MSCs联合雷帕霉素对异基因鼠脾
淋巴细胞的免疫调节作用

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

(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)

收稿日期:1900-01-01 修回日期:1900-01-01 发布日期:2009-04-16
通讯作者: 李晶

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

YIN Yujun¹, LI Jing², TANG Yu², YOU Haiyan³, ZHU Wei⁴,　XU Wengrong⁴, JIAO Zhijun³

（1. Department of Dermatology, 2. Department of Rheumatology, 3. Department Laboratory, Affiliated Hospital of Jiangsu University,
Zhenjiang 212001, Jiangsu, China； 4. School of Medical Technology, Jiangsu University, Zhenjiang 212013, Jiangsu, China）

Received:1900-01-01 Revised:1900-01-01 Published:2009-04-16
Contact: LI Jing

摘要/Abstract

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 Mouse 1X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。

关键词: 间充质干细胞, 雷帕霉素, 淋巴细胞, 免疫调节

Abstract: To investigate the immunoregulatory effects of mesenchymal stem cells(MSCs) combined with rapamycin on T and B cells in vitro and its potential mechanism. MethodsMSCs 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 lymphocytes were isolated by EZSep(tm) Mouse 1X. Under ConA or LPS stimulation, lymphocytes were treated with MSCs and/or rapamycin. 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. ResultsBoth MSCs and MSCs combined with rapamycin could significantly inhibit proliferation of lymphocytes and the apoptosis of T cells. However, the inhibitory 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. ConclusionMSCs 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，apoptosis and mRNA expressions of IFNγ and IL10.

Key words: Mesenchymal stem cells, Rapamycin, Lymphocytes, Immunoregulatory

中图分类号:

R392

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
. MSCs联合雷帕霉素对异基因鼠脾
淋巴细胞的免疫调节作用[J]. 山东大学学报(医学版), 2009, 47(04): 58-61.

YIN Yujun¹, LI Jing², TANG Yu², YOU Haiyan³, ZHU Wei⁴,　XU Wengrong⁴, JIAO Zhijun³
. Mesenchymal stem cells combined with rapamycin immunoregulates
the spleniclymphocytes in allogenic mice[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2009, 47(04): 58-61.

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