摘要
目的观察间充质干细胞（MSCs）对Ⅱ型胶原诱导的关节炎（CIA）大鼠Treg细胞数量和功能的影响。方法以Ⅱ型胶原蛋白免疫SD大鼠建立CIA动物模型；分离培养正常SD大鼠骨髓MSCs，尾静脉移注CIA大鼠；流式细胞仪检测大鼠脾脏CD4^＋CD25^＋T细胞比例变化；实时定量聚合酶链反应（Real—timePCR）检测脾脏叉状头／翅膀状螺旋转录因子（Foxp3）mRNA水平及白细胞介素（IL）-10、转化生长因子-β1（TGF—β1）的表达；免疫磁珠法分选各组大鼠脾脏CD4^＋CD25^＋T细胞，体外检测CD4^＋CD25^＋T细胞对CD4^＋CD25^-T细胞（Teff）增殖的抑制作用。结果MSCs移植改善了CIA大鼠的关节炎症状；与对照组比较，MSCs治疗组大鼠的脾脏内Treg细胞比例逐渐增加，到30d时，和正常对照组比较差异无统计学意义（P〉0．05）。Foxp3mRNA水平逐渐升高，同时IL-10、TGF-β 1mRNA水平亦逐渐增加；MSCs治疗组Treg细胞对Teff细胞增殖作用与CIA组比较显著增强（P〈0．05）。结论通过移植MSCs可提高CIA大鼠体内Treg细胞的数量和功能，通过免疫调节改善了CIA大鼠关节炎症的进程。
Objective To observe the immunologic effect of transplantation of mesenchymal stem cells （MSCs） onCD4^＋CD25^＋ Treg cells frequency and function in collagen-induced arthritis （CIA） rat model. Methods Rat CIA model was established by immunizing SD rats with type Ⅱ collagen. MSCs were purified and cultured from the bone marrow of SD rats, then transplanted to CIA model. The frequency Of CD4^＋CD25^＋ Treg cells in the spleen was measured by flow cytometry （FCM） respectively after the MSCs transplantation, and the expression of forkhead box P3 （Foxp3） , transforming growth factor-β1 （TGF-β1） and interleukin （IL）-10 mRNA was evaluated by real-time quantitative polymerase chain reaction （Real- time PCR）. CD4^＋CD25^＋ Treg cells were isolated from the spleen of rats by immunomagnetic beads and the inhibitory effect of CD4^＋CD25^＋ Treg cells on proliferation of CD4^＋CD25^- Teff cells was estimated by methyl thiazol tetrazolium （MTT） colorimetry. Results The MSCs transplantation relieved the arthritis symptom of the CIA rats. The frequency of CD4^＋CD25^＋ Treg ceils in the spleen of MSCs-treated CIA rats was increased significantly （P 〈 0.05 ） as compared with the CIA group, and the levels of Foxp3, IL-10 and TGF-β1 mRNA were also significantly increased （ P 〈 0. 05 ）. CD4^＋CD25^＋ Treg cells isolated from treatment group showed higher inhibition on the proliferation of CD4^＋CD25^- Teff ceils versus CD4^＋CD25^＋ Treg cells isolated from CIA rats. Conclusion Transplantation of MSCs is effective on preventing the development of CIA. The increased frequency and reinforced function of CD4^＋CD25^＋ Treg cells may be one of the mechanisms for MCSs transplantation treating CIA in rats.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2014年第5期1091-1093,共3页
Chinese Journal of Experimental Surgery
关键词
骨髓间充质干细胞
类风湿关节炎
调节性T细胞
Bone marrow mesenchymal stem cells
Rheumatoid arthritis
Regulatory T cells
