CAJ | 학술논문

目的探讨白细胞介素6受体抗体(IL-6R-Ab)对类风湿关节炎(RA)滑膜成纤维细胞增殖与核因子-κB受体激活因子配体(RANKL)表达的影响.方法从膝关节镜术中获取RA患者滑膜组织,培养成纤维细胞.实验分为4组:甲氨蝶呤(MTX)组、IL-6R-Ab组、IL-6R-Ab联合MTX组、空白对照组.分别采用CCK-8检测IL-6R-Ab、MTX对RA滑膜成纤维细胞增殖活力影响；荧光定量(FQ)-PCR检测各组培养体系中的滑膜成纤维细胞RANKL mRNA的表达.结果 CCK-8检测结果显示:与空白对照组比较,MTX组、IL-6R-Ab组及联合组成纤维细胞的生长受到抑制(F=54.64,P＜0.05)；IL-6R-Ab联合MTX组RA滑膜成纤维细胞的生长显著受到抑制(P＜0.01)；但MTX组与IL-6R-Ab组未见统计学差异.ELISA检测结果提示:与MTX组相比,IL-6R-Ab组的RANKL表达量降低(P＜0.05),IL-6R-Ab联合MTX组的RANKL表达量明显降低(P＜0.01).FQ-PCR检测显示,MTX组、IL-6R-Ab组、联合组均能抑制RANKL mRNA的表达(F =32.17,P＜0.05)；与MTX组或IL-6R-Ab组相比,IL-6R-Ab联合MTX组RANKL mRNA的表达也受到抑制(P＜0.05).结论 IL-6R-Ab单独或联合MTX使用均能抑制RA滑膜成纤维细胞的增殖活力,显著抑制RANKL的表达.本研究为IL-6R-Ab防治RA滑膜成纤维细胞所造成的关节破坏提供细胞与分子生物学方面的理论依据.
목적 탐토백세포개소6수체항체(IL-6R-Ab)대류풍습관절염(RA)활막성섬유세포증식여핵인자-κB수체격활인자배체(RANKL)표체적영향.방법 종슬관절경술중획취RA환자활막조직,배양성섬유세포.실험분위4조:갑안접령(MTX)조、IL-6R-Ab조、IL-6R-Ab연합MTX조、공백대조조.분별채용CCK-8검측IL-6R-Ab、MTX대RA활막성섬유세포증식활력영향；형광정량(FQ)-PCR검측각조배양체계중적활막성섬유세포RANKL mRNA적표체.결과 CCK-8검측결과현시:여공백대조조비교,MTX조、IL-6R-Ab조급연합조성섬유세포적생장수도억제(F=54.64,P＜0.05)；IL-6R-Ab연합MTX조RA활막성섬유세포적생장현저수도억제(P＜0.01)；단MTX조여IL-6R-Ab조미견통계학차이.ELISA검측결과제시:여MTX조상비,IL-6R-Ab조적RANKL표체량강저(P＜0.05),IL-6R-Ab연합MTX조적RANKL표체량명현강저(P＜0.01).FQ-PCR검측현시,MTX조、IL-6R-Ab조、연합조균능억제RANKL mRNA적표체(F =32.17,P＜0.05)；여MTX조혹IL-6R-Ab조상비,IL-6R-Ab연합MTX조RANKL mRNA적표체야수도억제(P＜0.05).결론 IL-6R-Ab단독혹연합MTX사용균능억제RA활막성섬유세포적증식활력,현저억제RANKL적표체.본연구위IL-6R-Ab방치RA활막성섬유세포소조성적관절파배제공세포여분자생물학방면적이론의거.
Objective To investigate the influence of interleukin-6 receptor (IL-6R-Ab) antibody on proliferation and expression of nuclear factor-κB-activating factor receptor ligand (RANKL) in rheumatoid arthritis(RA) synovial fibroblasts.Methods Synovial tissue obtained from laparoscopic surgery,and then digested and subcultured.The experiment was divided into 4 groups according to different cultural conditions,methotrexate (MTX) group,IL-6R a(n)tibody (IL-6R-Ab) group,IL-6R antibody combined MTX (combination )group,and the control group respectively.Cells growth inhibition and medicine toxicity of IL-6R antibody,MTX in RA synovial fibroblasts were measured by cell counting kit-8 (CCK-8).FQ-PCR was applied to detect the expression of RANKL mRNA.Results The results of CCK-8 test showed that the cells proliferation in the presence of MTX,1L-6R-Ab and combination group were fairly lower than that of in the control group (F =54.64,P ＜0.05),and IL-6R antibody combined MTX group was significantly inhibited (P ＜ 0.01);comparing to the MTX group,there was no significant difference in IL-6R antibody group.ELISA tests results indicated that compared to the MTX group,expression of RANKL in IL-6R-Ab and IL-6R-Ab combined MTX group all decreased (F =32.17,P ＜ 0.05).Further more,the expression in IL-6R antibody combined MTX group is the lowest (P ＜ 0.01).Results of FQ-PCR indicated that compared to the control cells,the expression of RANKL mRNA in other three groups were inhibited (P ＜ 0.05).Compared to the MTX group or IL-6R-Ab group,the expression of RANKL mRNA was inhibited in combination group obviously (P ＜ 0.05).Conclusion IL-6R-Ab alone or combinate to MTX could significantly inhibit the proliferating activities of RA synovial fibroblasts and significantly down regulated the expression of RANKL,which might provide theoretical basis of cellular and molecular biology for treatment of RA.