骨髓间充质干细胞对重症哮喘患儿外周血Th17/Treg的免疫调节作用  

Bone marrow-derived mesenchymal stem cells regulate the function of Th17 / Treg in peripheral blood of severe asthmatic children

作　　者：黄雪琼[1,2] 檀卫平[1] 吴葆菁[1] 蓝丹[1] 吴海飞[1] 麦贤弟[1] 

机构地区：[1]中山大学孙逸仙纪念医院儿科,广东广州510120 [2]中山大学附属第一医院儿童重症监护室,广东广州510080

出　　处：《中国病理生理杂志》2014年第9期1694-1697,1702,共5页Chinese Journal of Pathophysiology

基　　金：广东省自然科学基金资助项目(No.9451008901003013)

摘　　要：目的:探讨骨髓间充质干细胞(MSCs)在体外对重度哮喘患儿外周血辅助性T细胞17(Th17)和CD4+CD25+调节性T细胞(Treg)的免疫调节作用。方法:体外分离、培养和鉴定MSCs。MSCs经丝裂霉素处理后按不同比例(1∶1、1∶2、1∶10和1∶20)与哮喘患儿外周血T淋巴细胞(TLC)直接接触共培养,检测各组MSCs对TLC的增殖调节作用。选取上述1∶2比例共培养体系和单独TLC培养体系,ELISA法分别检测Th17的效应分子白细胞介素17(IL-17)和Treg效应分子转化生长因子β(TGF-β)水平,qRT-PCR法检测转录因子维甲酸相关孤儿核受体(RORC)及叉头框蛋白3(Foxp3)mRNA表达水平。结果:MSCs可显著抑制重度哮喘患儿TLC增殖,且随着MSCs数量的增加,抑制作用增强。MSCs+TLC共培养组Th17转录因子RORC mRNA和效应因子IL-17表达较TLC组下降,同时TGF-β表达增高,而Treg细胞调控基因Foxp3 mRNA表达无明显改变。结论:MSCs在体外可能通过抑制Th17分化及IL-17的分泌,同时上调TGF-β的表达,进而有效改善哮喘患儿的Th17/Treg失衡状态。AIM： To investigate the regulatory function of bone marrow-derived mesenchymal stem cells（ MSCs） on T helper 17 cells（ Th17） and regulatory T cells（ Treg） in peripheral blood of severe asthmatic children.METHODS： MSCs were isolated,cultured and identified in vitro. MSCs digested with mitomycin were cocultured with T lymphocytes（ TLC） at different ratios（ 1∶ 1,1∶ 2,1∶ 10 and 1∶ 20） from severe asthmatic children for 72 h. The proliferation of TLC was measured by CCK-8 method. In the coculture system of the 1∶ 2 ratio and the single TLC system,the supernatant levels of interleukin-17（ IL-17） and transforming growth factor-β（ TGF-β） were measured by ELISA. The mRNA expression of retinoic acid-related orphan nuclear receptor C（ RORC） and forkhead box protein 3（ Foxp3） in TLC was detected by qRT-PCR. RESULTS： After cocultured with MSCs,the proliferation of TLC decreased significantly in a dosedependent manner（ P〈 0. 05）. It also showed decreases in IL-17（ 3 799 ± 441 vs 4 890 ± 373,P〈 0. 05） and RORC mRNA level（ 1. 21 ± 0. 14 vs 3. 85 ± 0. 48,P 〈0. 05）,while an increase in TGF-β level（ 209 ± 32 vs 117 ± 26,P〈 0. 05） was observed. No influence on the mRNA expression of Foxp3 was found（ P〉 0. 05）. CONCLUSION： MSCs suppresses Th17 polarization of naive peripheral blood CD4^＋T cells and matures Th17 cells secreting IL-17,which may effectively revise Th17 /Treg imbalance of asthma.

关 键 词：间充质干细胞 哮喘 TH17细胞 调节性T细胞 

分 类 号：R725.6[医药卫生—儿科]