异基因脐血干细胞穴位移植对类风湿关节炎大鼠外周血CD_4~+CD_(25)~+调节性T细胞的影响

目的:观察异基因脐血干细胞(h UCBSCs)穴位移植对类风湿关节炎大鼠外周血CD4+CD25+调节性T细胞(CD4+CD25+Treg)的干预作用及相关机制。方法:先进行脐血CD34+细胞分选,将100只大鼠随机分为正常对照组、模型组对照组、h UCBSC尾静脉注射治疗组、h UCBSCs外关穴注射治疗组、甲氨喋呤(MTX)灌胃治疗组,每组20只。采用邓安梅法建立胶原诱导性关节炎(CIA)大鼠模型。于大鼠免疫后第31天(除正常组及模型组用等体积蒸馏水)向外关穴、尾静脉注射0.5 m L氯化钠注射液和0.5m L h UCBSCs悬液(含干细胞2×107/m L),4周后采用流式细胞仪,对外周血CD4+CD25+细胞的百分含量及其在CD4+T淋巴细胞中所占的比例进行检测。结果:h UCBSCs外关穴注射治疗组外周血CD4+CD25+细胞的百分含量及其在CD4+T淋巴细胞中所占的比例与甲氨喋呤组比较明显增高,差异有统计学意义(P<0.01),h UCBSC尾静脉注射治疗组与甲氨喋呤组比较,无显著性差异(P>0.05)。结论:h UCBSCs外关穴穴位注射治疗可能通过上调CD4+CD25+调节性T细胞水平,抑制关节炎症和关节骨破坏吸收,有助于类风湿关节炎的治疗。

Effects of Points Transplantation on Allogeneic Cord Blood Stem Cell in Rheumatoid Arthritis in Peripheral Blood CD_4~+CD_(25)~+ Regulatory T Cells

Objective: To observe allogeneic cord blood stem cells( h UCBSCs) points transplantation of peripheral blood of rheumatoid arthritis in rats CD4+CD25+regulatory T cells( CD4+CD25+Treg) and the intervention mechanisms. Methods: First,for cord blood CD34+cell sorting,100 rats were randomly divided into normal control group,model group,h UCBSC tail vein injection treatment group,h UCBSCs Waiguan injection treatment group,methotrexate( MTX) gavage treatment groups,20 in each group. DENG Anmei's method was established by using collagen-induced arthritis( CIA) rat model. On day 31 after immunization( except the normal group and model group with an equal volume of distilled water),0. 5 m L sodium chloride injection and 0.5m L h UCBSCs suspension( containing stem cells 2 × 107/ m L) was intravenously injected into Waiguan point. 4 weeks later,flow cytometry was used for peripheral blood CD4+CD25+cells and the percentage of CD4+T lymphocytes in the proportion of testing.Results: The h UCBSCs Waiguan injection therapy of peripheral blood CD4+CD25+cells and the percentage of CD4+T lymphocytes in the proportion was significantly higher compared with that of the methotrexate group and the difference was statistically significant( P < 0. 01). The h UCBSC tail vein injection treatment group had no significant difference when compared with the methotrexate group( P > 0. 05). Conclusion: The h UCBSCs Waiguan point injection therapy by up-regulating the CD4+CD25+regulatory T cells and inhibiting arthritis and joint destruction of bone absorption to help the treatment of rheumatoid arthritis.