骨髓间充质干细胞移植治疗胶原诱导性关节炎的疗效和机制

目的 探讨骨髓间充质干细胞(MSCs)治疗胶原诱导型关节炎(CIA)疗效及可能治疗机制.方法 分离培养C57BL/6小鼠MSCs.健康对照组DBA-1小鼠3只(第0天和第21天尾静脉输注生理盐水0.2 m1)15只牛Ⅱ型胶原诱导的CIA小鼠随机分为CIA对照组(第0大和第21天尾静脉输注生理盐水0.2 ml)、预防组(第0天输注MSCs1×106细胞数/200 μl)和治疗组(第21天输注MSCsl×106细胞数/200 μl).另设健康对照组(不诱导CIA,第0天和第21天尾静脉输注生理盐水0.2 ml).疗效观察包括关节炎指数(AI)评分,关节病理分析,酶联免疫吸附试验(ELISA)测血清肿瘤坏死因子(TNF)-α,白细胞介素(IL)-1β水平,流式细胞术检测脾脏和淋巴结CD4+CD25+Foxp3+T淋巴细胞百分率等.结果 ①预防组和治疗组AI、关节病理评分明显下降(P<0.05);②健康对照组、预防组和治疗组血清TNF-α和IL-1β水平降低(P<0.05),与AI呈显著正相关(P<0.05),TNF-α与关节病理评分呈显著正相关(r=0.61,P<0.01);③治疗组小鼠脾脏和淋巴结中调节性T细胞明显升高(P<0.05).结论 MSCs治疗CIA有效,抑制TNF-α和IL-1β和上调调节性T细胞可能是MSCs治疗CIA有效的机制.

Effect and mechanism of bone marrow-derived mesenchymal stem cells transplantation on collagen-induced arthritis

Objective To investigate the effect of bone marrow-derived mesenchymal stem cells (MSCs) on collagen-induced arthritis (CIA) and the possible mechanisms. Methods MSCs from C57BL/6 mice bone marrow were isolated and expanded. Three DBA-I mice were as normal control group (Sodium Chloride injected on day 0 and 21) after primary immunization. After the bovine collagen Ⅱ induced-CIA models were established, fifteen male mice were randomly divided into 3 groups according to treatment regi-mens: CIA control group (0.2 ml Sodium Chloride injected through caudal vein on day 0 and 21), prevention group (1×10+6 cells MSC on day 0) and treatment group. (1×10+6 cells MSC on day 21 after primary immun-ization). The observed parameters included arthritis index (Al), articular pathology changes, levels of serum TNF-α, IL-1β detected hy ELISA. In addition, the percentage of CD4+CD25+Foxp3+Treg cells(Tregs) in CD4+T cells of spleen or lymphoid node from mice were also analyzed by flow cytometry (FCM). Results ①The Al and articular pathological score of the prevention group and treatment group were lower than the CIA control group (P<0.05). ②The levels of serum TNF-α and IL-1β of normal control, prevention group and treatment group were significantly decreased compared with CIA control group (P<0.05), and positively correlated with Al or articular pathological score (P<0.05). ③The percentage of Tregs from spleen and lymph nodes was significantly increased in MSCs treated mice (P<0.05). Conclusion MSCs transplantation is effective for CIA and suppression of TNF-α, IL-1β and up-regulation of Tregs by MSCs may be the possible mechanism in CIA.