异基因骨髓间充质干细胞移植对小鼠胶原性关节炎CD4＋CD25＋调节性T细胞表达的影响

摘要

Objective To study the influence of allogenetic mesenchymal stem cells (MSCs) transplantation on the expression of collagen induced arthritis CD4 + CD25 + regulatory T cell in mice .Methods A total of 40 C57BL/6 (H-2b) mice were selected and randomly divided into normal control group ,model group ,MSCs transplant treatment group and methotrexate treatment of positive control group .Ten mice in each group .Except for mice in the normal control group ,the others were treated with Freund's complete adjuvant + Collagen Ⅱ to induce C57BL/6(H-2b) mice that to make mouse model .Bone marrow mononuclear cells were isolated ,then the MSCs were identified and screened by FAcscaliburTM flow cytometry analyzer .MSCs was transplanted by tail vein injection with the number of cells as 2 × 106 .All the mice in four groups were put to death after transplanting 42 days ,and observed the symptoms and the degree of swelling of the joints .Using flow cytometry to detect the contents of CD4 + CD25 + ,and collected data and made statistical analysis .Results There was no significant difference on the swelling degree of joints between the MSCs transplant treatment group and the normal control group (P> 0 .05) .Compared with methotrexate treatment of positive control group ,there was significant difference on CD4 + CD25 + regulatory T cell expression in the MSCs transplant treatment group (P< 0 .05) .But there was no significant difference compared with that of the normal con -trol group(P> 0 .05) .Conclusion MSCs transplantion could significantly improve the swelling degree of joints ,and significantly increase the expression of CD4 + CD25 + .Therefore ,MSCs could be used as a new source of replacement cells for transplantation therapy of rheumatoid arthritis .%目的：探讨异基因骨髓间充质干细胞（MSCs）移植对小鼠胶原性关节炎 CD4＋、CD25＋调节性 T 细胞表达的影响。方法选取40只 C57BL/6（H-2b）小鼠，随机分为正常对照组、Ⅱ型胶原性关节炎（CIA）模型组、MSCs 移植治疗组、甲氨蝶呤治疗阳性对照组，每组10只小鼠。除正常对照组外，其余各组弗氏完全佐剂＋Ⅱ型胶原诱导小鼠建立 CIA 小鼠模型。分离骨髓单个核细胞后，进行 MSCs 细胞分选及鉴定。美国 BD 公司 FAcscal-iburTM 型流式细胞分析仪对小鼠骨髓 MSCs 进行分选和鉴定。 MSCs 移植采用尾静脉注射，所含细胞数为2×106。移植后第42天全部处死动物观察不同分组小鼠的关节症状及肿胀度，采用流式细胞术检测 CD4＋ CD25＋浓度，收集数据并做统计分析。结果 MSCs 移植治疗组小鼠关节的肿胀度，与正常对照组比较，差异无统计学意义（P＞0．05）。 MSCs 移植组 CD4＋ CD25＋调节性 T 细胞的表达，与甲氨蝶呤治疗阳性对照组比较，差异有统计学意义（P＜0．05）；与正常对照组比较，差异无统计学意义（P＞0．05）。结论 MSCs 移植治疗小鼠 CIA ，能显著改善关节的肿胀度，并明显上调 CD4＋ CD25＋表达，故 MSCs 可作为一种新的替代细胞来源用于类风湿性关节炎的移植治疗。