淫羊藿苷对IL-1β刺激RA滑膜成纤维细胞增殖及TNF-α和RANKL分泌的影响  

Effects of Icariin on IL-1β-induced rheumatoid arthritis synovial fibroblasts proliferation and TNF-α and RANKL production

作　　者：刘益杰[1] 冯伟[1] 褚立希[1] 杨松滨[1] 朱鸿飞[1] 宁长青[1] 

机构地区：[1]上海中医药大学康复医学院,上海201203

出　　处：《上海中医药杂志》2012年第3期77-80,共4页Shanghai Journal of Traditional Chinese Medicine

基　　金：上海市自然科学基金资助项目(09ZR1432000);上海市浦江计划基金资助项目(10PJ1409300);博士点建设基金资助项目(2010);上海高校创新团队(第二期)建设项目(沪教委科[2009]6号;26号文件);上海市浦东新区卫生系统重点学科群建设项目(PWZxkq2011-01)

摘　　要：目的观察淫羊藿苷(Icariin,ICA)对IL-1β刺激类风湿性关节炎滑膜成纤维细胞(rheumatoid arthritis synovial fibroblasts,RASFs)增殖及培养上清TNF-α和RANKL含量的影响。方法体外分离培养RA患者来源滑膜成纤维细胞,取第二代细胞分别接种于96孔板和6孔板,除正常对照组(N组)外其余各组均以终浓度为5μg/L的IL-1β刺激24 h后加入不同药物干预,分别设置空白对照组(C组)、MTX组(50 mg/L)、淫羊藿高剂量组(IH组,50μmol/L)和淫羊藿低剂量组(IL组,5μmol/L);继续培养72 h后,以WST-1细胞增殖及细胞毒性检测试剂盒检测各组细胞增殖情况,以ELISA法检测各组培养上清中TNF-α和RANKL含量。结果 C组以IL-1β连续刺激RASFs 96 h后,细胞增殖率为17.9%,培养上清中TNF-α、RANKL含量分别为(361.542±7.178)ng/L和(428.468±11.118)ng/L,与N组[TNF-α、RANKL含量分别为(346.458±3.329)ng/L和(400.630±7.007)ng/L]比较,差异具有统计学意义(P<0.05)。以不同药物干预后,与C组比较,MTX组细胞增殖率为-34.70%(P<0.05);IH组培养上清中RANKL含量降低,为(375.766±7.806)ng/L(P<0.05);IL组细胞增殖率为-31.90%,培养上清中TNF-α、RANKL含量分别为(334.542±6.691)ng/L和(365.495±5.280)ng/L,差异具有统计学意义(P<0.05)。结论 IL-1β能促进RA滑膜成纤维细胞增殖和分泌细胞因子TNF-α、RANKL;淫羊藿苷能抑制IL-1β刺激RA滑膜成纤维细胞的增殖及TNF-α、RANKL的分泌。Objective To observe the effects of Icariin （ICA） on proliferation of rheumatoid arthritis synovial fibroblasts（RASFs） stimulated by IL-1β and the expression of TNF-α and RANKL. Methods RASFs were isolated form Synovial tissue of RA patients and cultured in vitro;second-generation RASFs were plated in 96 / 6-well, setting with normal group ,control group ,MTX group ,high-dose ICA group and low-dose ICA group ;all groups except the normal ,after stimulated by 5 ng/ml IL-1β for 24 hours, were continuously treated with different medicine for 72 hours. Then the RASFs proliferation was evaluated with WST-1 Cell Proliferation and Cytotoxicity Assay Kit, the expression of TNF-α and RANKL in supernatant of each group were detected by ELISA. Results Compared with normal group, Control group has a significantly higher absorbance values at 17.9% proliferation ratio（ P 〈 0.05 ）, also TNF-α and RANKL were higher expressed at the level of （361. 542 ± 7,178 ） ng/L and （428. 468±11.118 ） ng/L respectively （ P 〈 0.05 ）. Compared with control group after being intervened for 72 hours, MTX can inhibit the proliferation of RASFs （P 〈0.05）, and reduce the production of TNF-α and RANKL while without significant difference（P 〉0.05） ; RANKL levels in IH group was （375.766 ±7.806） ng/L（P〈0.05） ; low-dose ICA can inhibit RASFs proliferation with a proliferation ratio of -31.90%, and down-regulate the concentration of TNF-α at （334. 542 ± 6.691 ）ng/L or RANKL （365. 495 ± 5.280）ng/L（P〈 0.05）. Conclusion IL-1β can promote proliferation of RASFs and stimulate the production of cytokines through TNF-α and RANKL; icariin has the effect in inhibiting proliferation of IL-1β - stimulating RASFs and TNF-α, RANKL production.

关 键 词：淫羊藿苷 RA滑膜成纤维细胞 肿瘤坏死因子-Α NF-KB配体的受体或激活因子 白介素1-β 

分 类 号：R259[医药卫生—中西医结合]