金雀异黄素对类风湿关节炎患者成纤维样滑膜细胞骨保护素／细胞核因子κB受体活化因子／细胞核因子κB受体活化因子配体通路的影响

目的研究金雀异黄素（genistein，Gen）对人类风湿关节炎（rheumatoid arthritis，RA）成纤维样滑膜细胞（fibroblast—like synoviocytes，FLS）骨保护素（osteoprotegerin，OPG）／细胞核因子KB受体活化因子（receptor—activator of nuclear factor kappa bata，RANK）／细胞核因子KB受体活化因子配体（receptor—activator of nuclear factor kappa bata ligand，RANKL）通路的影响及机制。方法培养人RA—FLS，并分为对照组（RA—FLS细胞）、TNF—α组（RA—FLS细胞＋TNF-α 10ng／ml）、TNF—α＋Gen组（RA—FLS细胞＋TNF—α 10ng／ml＋Gen 50μM）、Gen组（RA—FLS细胞＋Gen 50μM）。应用免疫印迹检测Gen作用RA—FLS的RANK、RANKL、OPG、雌激素受体（estrogen receptora，ERa）表达情况，免疫荧光检测ERα细胞内分布情况。结果Gen组、TNF-α组及TNF—α＋Gen组细胞内RANK蛋白表达量较对照组无明显差异（P〉0．05），但OPG及RANKL蛋白表达明显高于对照组（均P〈0．05）。Gen组OPG／RANKL较对照组增高（P〈0．05）。Gen组、TNF-α组及TNF-α＋Gen组细胞内ERa蛋白表达量与对照组比较，差异无统计学意义（P〉0．05）。免疫荧光结果显示核内ERα表达量增加。结论Gen可能通过与雌激素受体结合，转入核内发挥免疫调节作用，从而调节0PG／RANKI／RANK通路，发挥抑制骨破坏的作用。Gen可以上调人RA—FLS中OPG及RANKL，且对OPG的上调作用更强。

Effect of Genistein on the Osteoprotegerin/Receptor-activator of Nuclear Factor Kappa Bata/Receptor-activator of Nuclear Factor Kappa Bata Ligand System of Fibroblast-like Synoviocytes

In order to investigate the effect of genistein （geM） on the secretion of receptor-activator of nuclear factor kappa bata ligand（ RANKL）/osteoprotegerin （OPG） secreted by fibroblast-like synoviocytes. Methods The expression of CD29, CD90 of human rheumatoid arthritis FLS was detected by Flow cytometry. The experience was divided into four groups： the control group, the TNF α treatment group, the TNF α ＋Gen group and the Gen group. Western blot was used to detect the expression of estrogen receptor α （ ER α）, receptor-activator of nuclear factor kappa bata （ RANK）, RANKL, OPG. Immunofluorescence was used to detect ER α. Results Western Blotting showed that Gen could significantly up-regulate the expression of OPG and RANKL relative to the control group （ P 〈 0. 05 ） ; but had no effect on RANK and ER α. The statistical analysis showed that the ratio of OPG/RANKL in the Gen group was increased. Compared with the control group, the results from the immunofluorescence also showed that most of the ER α were transferred into the nucleus. Conclusion Gen may influence ERα to regulate the OPG/RANKL/RANK pathway.