抗白细胞介素-6抗体对类风湿关节炎大鼠的治疗作用

The therapeutic effect of anti-interleukin-6 on the rheumatoid arthritis rats

目的 探讨抗白细胞介素（IL）-6抗体对类风湿关节炎大鼠的治疗作用及其作用机制.方法 构建大鼠类风湿性关节炎模型（CIA模型）,给予抗IL-6中和性抗体（anti-IL-6）药物治疗,通过检测相关指标,证实anti-IL-6对CIA模型大鼠的治疗效果,包括临床评分、足趾肿胀程度;分离的大鼠滑膜组织,采用定量聚合酶链反应（PCR）方法检测关节滑膜组织凋亡相关基因[主要包括B细胞淋巴瘤/白血病-2（bcl-2）、bcl-2相关X蛋白（bax）]与凋亡相关基因（Nfkb1、Ras等）的mRNA表达,并采用Western blot方法检测细胞功能表型基因[凋亡相关膜联蛋白V（Annexin V）,增殖相关Ki-67和增殖细胞核抗原（PCNA）]的蛋白表达,初步阐明anti-IL-6的治疗作用机制.结果 与模型组比较,anti-IL-6治疗组大鼠关节炎评分和足趾肿胀程度降低,CIA大鼠滑膜组织的IL-6受体IL-6R mRNA表达水平明显高于正常组,滑膜组织中细胞促凋亡基因bax的mRNA表达水平显著增强（CIA组：0.43 ±0.08,CIA＋ anti-IL-6组：0.89 ±0.05;P＜0.01）,抗凋亡基因bcl-2则降低（CIA组：3.21 ±1.22,CIA＋ anti-IL-6组：0.98 ±0.07;P＜0.01）,细胞凋亡标志Annexin V蛋白表达增强.同时,anti-IL-6治疗组大鼠滑膜组织中细胞增殖相关基因Nfkb1（CIA组：3.23±0.65,CIA＋ anti-IL-6组：1.23 ±0.31;P ＜0.01）和Ras的mRNA（CIA组：6.87±1.31,CIA＋ anti-IL-6组：3.21±1.35;P＜0.01）表达水平显著降低,细胞增殖标志Ki-67和PCNA蛋白表达水平下调.结论 anti-IL-6能特异性调节滑膜组织凋亡基因和增殖基因,有效改善Ⅱ型胶原酶诱导的大鼠关节炎模型.

Objective To investigate the therapeutic effect of anti-interleukin （IL）-6 on the rheumatoid arthritis and the mechanism.Methods The rat rheumatoid arthritis （CIA） model was constructed and the neutralized antibody against IL-6 （anti-IL-6） was administrated.The efficacy of anti-IL-6 on CIA was proven and the mRNA expression of proliferation and apoptosis genes in synovial joint was detected.The protein expression of genes indicating the cell function phenotype was also checked.Results The results showed that anti-IL-6 could alleviate the arthritis score and paw swelling in rat CIA model.Anti-IL-6 increased the expression of B cell lymphoma/leukemia-2 associated X protein （for bax,CIA group：0.43 ± 0.08,and CIA ＋ anti-IL-6 group：0.89 ± 0.05;P ＜ 0.01）,annexin V and decreased B cell lymphoma/leukemia-2 （for bcl-2,CIA group：3.21 ± 1.22,and CIA ＋ anti-IL-6 group：0.98 ± 0.07;P ＜ 0.01）.Meanwhile,anti-IL-6 reduced the expression of Nfkb1 （CIA group：3.23 ±0.65,and CIA ＋ anti-IL-6 group：1.23 ±0.31;P ＜0.01） and Ras （CIA group：6.87 ± 1.31,and CIA ＋ anti-IL-6 group：3.21 ± 1.35;P ＜ 0.01）,and the proliferation markers Ki-67 and proliferating cell nuclear antigen （PCNA） were also downregulated.Conclusion Anti-IL-6 could ameliorate the symptoms of CIA in rats via specifically mediating the expression of proliferation genes and apoptosis genes in the synovial joint.Conclusion Anti-IL-6 could ameliorate the symptom of CIA in rat via specifically mediating the expression of proliferation genes and apoptosis genes in the synovial joint.