摘要目的 探讨异体骨髓间充质干细胞移植对胶原诱导关节炎大鼠血清及关节滑膜白细胞介素-22(IL-22)、基质金属蛋白酶-3(MMP-3)表达的影响.方法 2次注射Ⅱ型胶原建立胶原诱导关节炎(CIA)大鼠模型,48只大鼠随机分为3组:空白对照组、CIA对照组及CIA实验组;骨髓培养法结合贴壁法培养大鼠骨髓间充质干细胞(MSC),并鉴定,其余按1×107/kg注入实验组大鼠体内;酶联免疫吸附法(ELISA)、免疫组化分别检测各组大鼠血清、踝关节IL-22及MMP-3表达量;分离并培养各组大鼠膝关节滑膜细胞,并给予不同浓度IL-22刺激,蛋白质印迹及实时定量聚合酶链反应(RQ-PCR)分别检测IL-22刺激前后MMP-3蛋白及mRNA表达量.结果 MSC移植后,CIA大鼠关节炎指数、X线、HE染色均显示关节破坏较对照组明显减轻;ELISA结果显示,CIA对照组IL-22、MMP-3表达量最高,与正常对照组[(125.79±9.12)比(102.00±7.63)ng/ml,P<0.05]、[(292.35±31.23)比(257.27±13.99)ng/ml,P<0.05]及CIA实验组[(125.79±9.12)比(97.94±9.50)ng/ml,P<0.05]、[(292.35±31.23)比(262.16±22.02)ng/ml,P<0.05]相比差异有统计学意义(P<0.05),空白对照组与CIA实验组差异无统计学意义;免疫组化结果与ELISA结果一致;Western blot、RQ-PCR结果示,IL-22刺激后MMP-3蛋白及mRNA表达量增高,表达量与刺激量呈剂量依赖性.结论 IL-22、MMP-3参与了类风湿关节炎的发病;IL-22可调控MMP-3的表达;骨髓间充质干细胞通过降低IL-22从而降低MMP-3的表达治疗类风湿关节炎.

abstractsObjective To study the effect of allogeneic bone marrow mesenchymal stem cells transplantation on the expression of interleukin -22 (IL-22), matrix metalloproteinase -3 (MMP-3) in serum and synovial of rats with collagen induced arthritis .Methods Type Ⅱ collagen were injected twice to establish the collagen induced arthritis ( CIA ) rat model.Forty-eight rats were randomly divided into 3 groups:control group , CIA control group , CIA experiment group .Rat bone marrow mesenchymal stem cells were cultured by bone marrow method combined with adherent culture method .After identify, the remaining cells were injected in the CIA experimental group .Enzyme linked immunosorbent assay ( ELISA ) and immunohistochemistry were used to detect the expression of IL-22 and MMP-3 in serum and anklebone joint′s synovium of rats, respectively.Synovial cells were isolated and cultured , and were treated with different concentrations of IL-22.MMP-3 protein and mRNA were detected before and after stimulation by Western blot and real-time quantitative polymerase chain reaction (RT-qPCR).Results After MSC transplantation, arthritis index, X-ray, HE staining of CIA rat showed that joint damage significantly reduced compared with&nbsp;the control group.The ELISA results showed that the expression of MMP-3 and IL-22 in CIA control group was higher than those in the control group ( 125.79 ±9.12 vs 102.00 ±7.63 ng/ml, P<0.05 ) , (292.35 ±31.23 vs 257.27 ±13.99 ng/ml, P<0.05 ) and CIA experiment group ( 125.79 ±9.12 vs 97.94 ±9.50 ng/ml, P<0.05), (292.35 ±31.23 vs 262.16 ±22.02 ng/ml, P<0.05) with statistically significant difference ( P<0.05 ) .There was no significant difference between the control group and CIA experimental group.Immunohistochemical showed similar results with ELISA .Western blotting and RT-qPCR showed that MMP-3 protein and mRNA expression was increased after IL-22 stimulation in a dose-dependent manner .Conclusion IL-22 and MMP-3 play an important role in the pathogenesis of rheumatoid arthritis.IL-22 could regulate the expression of MMP-3, and bone marrow mesenchymal stem cells could reduce the expression of MMP-3 in the treatment of rheumatoid arthritis by reducing the expression of IL-22.