Liu G Y, Bian S, Li F, Li X F, Fan K, An H Z, Jia X X
Department of Rheumatism, the Second Hospital of Shanxi Medical University, Taiyuan 030001, China.
Zhonghua Yi Xue Za Zhi. 2017 Mar 7;97(9):698-702. doi: 10.3760/cma.j.issn.0376-2491.2017.09.014.
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. 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). After MSC transplantation, arthritis index, X-ray, HE staining of CIA rat showed that joint damage significantly reduced compared with 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, <0.05), (292.35±31.23 vs 257.27±13.99 ng/ml, <0.05) and CIA experiment group (125.79±9.12 vs 97.94±9.50 ng/ml, <0.05), (292.35±31.23 vs 262.16±22.02 ng/ml, <0.05) with statistically significant difference (<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. 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.
研究同种异体骨髓间充质干细胞移植对胶原诱导性关节炎大鼠血清及滑膜中白细胞介素-22(IL-22)、基质金属蛋白酶-3(MMP-3)表达的影响。通过两次注射Ⅱ型胶原建立胶原诱导性关节炎(CIA)大鼠模型。48只大鼠随机分为3组:对照组、CIA对照组、CIA实验组。采用骨髓法联合贴壁培养法培养大鼠骨髓间充质干细胞。鉴定后,将剩余细胞注入CIA实验组。分别采用酶联免疫吸附测定(ELISA)和免疫组织化学法检测大鼠血清及踝关节滑膜中IL-22和MMP-3的表达。分离培养滑膜细胞,用不同浓度的IL-22处理。在刺激前后通过蛋白质印迹法和实时定量聚合酶链反应(RT-qPCR)检测MMP-3蛋白和mRNA。MSC移植后,CIA大鼠的关节炎指数、X线、HE染色显示关节损伤较对照组明显减轻。ELISA结果显示,CIA对照组中MMP-3和IL-22的表达高于对照组(125.79±9.12 vs 102.00±7.63 ng/ml,<0.05),(292.35±31.23 vs 257.27±13.99 ng/ml,<0.05)以及CIA实验组(125.79±9.12 vs 97.94±9.50 ng/ml,<0.05),(292.35±31.23 vs 262.16±22.02 ng/ml,<0.05),差异有统计学意义(<0.05)。对照组和CIA实验组之间无显著差异。免疫组织化学结果与ELISA相似。蛋白质印迹法和RT-qPCR显示,IL-22刺激后MMP-3蛋白和mRNA表达呈剂量依赖性增加。IL-22和MMP-3在类风湿关节炎发病机制中起重要作用。IL-22可调节MMP-3的表达,骨髓间充质干细胞可通过降低IL-22的表达来降低类风湿关节炎治疗中MMP-3的表达。