骨髓来源间充质干细胞通过JAK-STAT信号通路抑制树突状细胞功能的体外研究

Mesenchymal stem cells derived from bone marrow inhibit dendritic cell function via JAK-STAT signaling pathway

目的探讨白介素-10(interleukin-10,IL-10)及其下游的JAK/STAT信号转导通路在骨髓来源的间充质干细胞(mesenchymal stem cells,MSCs)对树突状细胞(dendritic cells,DCs)的免疫抑制机制中的作用。方法 SD大鼠骨髓来源的MSCs和DCs按1∶10比例共培养,在共培养体系中分别加入IL-10、IL-10中和抗体(neutralizing antibody,NA)和JAK抑制剂(AG490)干预。ELISA法检测培养细胞上清液IL-10和IL-12水平的变化;流式检测DCs表面分子标志物CD80、CD86、OX62及MHC-II表达情况;Western blot检测(P-)JAK1,(P-)STAT3蛋白的表达情况。结果成功在体外分离和培养MSCs和DCs并得到流式鉴定证实。MSCs与DCs培养后上清液分泌IL-10水平明显升高。MSCs处理后DCs没有典型的成熟DCs表面标志,CD80、CD86、MHC-II及OX62的表达下降,IL-12分泌的水平也降低。实验中还发现外源性IL-10加到共培养板IL-10水平增加而IL-12水平减少,DCs的表面标志物表达下降,而加入IL-10NA与IL-10恰恰表现了相反的作用效果。加入JAK抑制剂(AG490)到共培养板后,IL-10分泌和(P-)JAK1和(P-)STAT3表达明显下降。结论大鼠骨髓来源的MSCs能抑制DCs成熟和功能,可能是通过IL-10和JAK1/STAT3信号转导通路来发挥作用的。

To investigate the role of interleukin-10（IL-10） and its downstream JAK/STAT signaling pathway in the immunosuppressive effects of bone marrow-derived mesenchymal stem cells（MSCs） on dendritic cells（DCs）,SD rats bone marrow-derived MSCs and DCs were co-cultured by proportion of 1∶10, with or without intervention of IL-10, IL-10 neutralizing antibody（IL-10NA） or JAK inhibitor（AG490）. Then the IL-10 and IL-12 of the cell culture supernatants were detected by ELISA; surface molecular markers of DCs CD80, CD86, OX62 and MHC-II were observed by FACS; and（P-） JAK1,（P-） STAT3 protein expressions of DCs was tested by Western blot. Data showed that IL-10 was significantly increased when MSCs and DCs co-cultured. After MSCs intervention, DCs lost typical mature DCs surface markers expression and the CD80, CD86, OX62 and MHC-II expression also decreased,in addition, IL-12 secretion was reduced. It was found that exogenous IL-10 addition to the co-culture plate increased IL-10 secretion, while reduced IL-12 secretion and surface markers of DCs expression. IL-10NA showed exactly the opposite effect. When AG490 was added to the co-culture system, IL-10 secretion as well as（P-） JAK1and（P-） STAT3 expressions decreased significantly. In conclusion, rat bone marrow-derived MSCs may inhibit DCs maturation and function via IL-10 and JAK1/STAT3 signal transduction pathway.