类风湿性关节炎滑膜成纤维细胞骨保护素和骨形态发生蛋白-2的表达及干预

目的 探讨甲氨蝶呤（MTX）及白细胞介素-6受体（IL-6R）抗体对类风湿关节炎（RA）滑膜成纤维细胞增殖的干预及对骨保护素（OPG）、骨形态发生蛋白-2（BMP-2）的影响.方法 获取RA患者滑膜组织,消化并传代培养成纤维细胞.CCK-8法检测IL-6R抗体、甲氨蝶呤（MTX）对RA滑膜成纤维细胞活性影响;荧光定量（qRT）-PCR检测OPG和BMP-2表达.结果 与空白组相比,MTX组、IL-6R抗体组成纤维细胞的活性受到抑制（F=29.30,34.22,P＜0.05）,MTX联合IL-6R抗体组成纤维细胞的活性显著受到抑制（F=52.04,P ＜0.01）.qRT-PCR显示与空白组相比,IL-6R抗体组、MTX联合IL-6R抗体组OPG、BMP-2的表达均上升（P＜0.05）.与MTK组相比,IL-6R抗体组OPG、BMP-2的表达增加（P＜0.05）.结论 IL-6R抗体单独或联合MTK使用均能明显抑制RA滑膜成纤维细胞的活性,与MTX相比,IL-6R抗体能升高OPG、BMP-2的表达.本研究为应用IL-6R抗体预防和治疗RA关节破坏提供实验依据.

Expressing of OPG and BMP-2 in rheumatoid arthritis synovial fibroblasts

Objective To investigate the influence of MTX and IL-6R antibody on the expression of osteoprotegerin（OPG） and bone morphogenetic protein-2 （BMP-2） in rheumatoid arthritis（RA） synovial fibroblasts.Methods Synovial tissues were obtained from laparoscopic surgery.Fibroblasts were digested and subcultured from the tissues.Fibroblasts were divided into 4 groups,including control group,methotrexate （MTX） treated group,IL-6R antibody treated group,group treated with both IL-6R antibody and MTX.The inhibitor effect of IL-6R antibody and MTX in RA synovial fibroblasts was measured by cell counting kit-8 （CCK-8）.qRT-PCR was applied to detect the expression of OPG and BMP-2.Results The results of CCK-8 test showed that fibroblasts proliferations in MTX treated group,IL-6R antibody treated group was fairly lower than that of the control group（F =29.30,34.22 respectivly,P ＜ 0.05）,and group treated with both IL-6R antibody and MTX was significantly inhibited（F =52.04,P ＜ 0.01）.Results of qRT-PCR indicated that compared to control group,the expression of OPG、BMP-2 in IL-6R antibody treated group and group treated with both IL-6R antibody and MTX were increased.Compared to MTX treated group,the expression of OPG、BMP-2 in IL-6R antibody treated group were increased（P ＜ 0.05）.Conclusion IL-6R antibody alone or in both with MTX could significantly inhibit the proliferating activities of RA synovial fibroblast,and compared to MTX,IL-6R antibody could up regulated the expression of OPG、BMP-2.The reseach might provide basis of cellular and molecular biology for treatment of RA.