当归拈痛汤防治类风湿性关节炎大鼠滑膜病变的蛋白质组学研究

Proteomic Study of Danggui Niantong Decoction on Synovium Pathological Changes in Rheumatoid Arthritis Rat Model

目的观察当归拈痛汤干预类风湿性关节炎(RA)大鼠滑膜病变的蛋白质改变,分析当归拈痛汤治疗RA可能的作用机制。方法将SD大鼠随机分为空白对照组、模型组、当归拈痛汤组3组,每组10只。双向电泳(2-DE)分离滑膜组织蛋白质,用PDQuest7.1.0软件包进行差异表达蛋白质组分析,基质辅助激光解吸附离子化飞行时间质谱(MALDI-TOF)鉴定蛋白质。选择2个差异蛋白用实时荧光定量PCR(Q-PCR)和蛋白质印迹法(Western-blot)进行验证。结果获得8个差异表达的蛋白:Myosin regulatory light chain 2,40S ribosomal protein SA,Mimecan,Heat shock protein beta-1,Vimentin,Peroxiredoxin-2,Interleukin-1 receptor antagonist protein,Lamin A。Q-PCR和Western-blot验证Hsp27 Peroxiredoxin-2表达水平变化与2-DE结果一致。结论当归拈痛汤可能通过以下机制抑制RA滑膜组织的炎症反应:(1)抑制组织局部的炎症反应信号通路;(2)保护细胞结构;(3)抑制氧化应激反应。

Objective To observe the effect of Danggui Niantong Decoction on the synovial proteome of rheumatoid arthritis （RA） rats by and to analyze the therapeutic mechanism of Danggui Niantong Decoction for RA. Methods Thirty rats were equally divided into blank control group, model group and Danggui Niantong Decoction group. Two-dimensional eleetrophoresis （2-DE） was used for the isolation of proteins. PDQuest 7.1.0 software was used to analyze differential expression of proteome. Matrix associated laser desorption ionization- time of the flight （MALDI-TOF） was used to identify proteins. Two differential proteins were chosen for confirmation by fluorescence quantitative PCR and Western blotting methods. Results Eight differently-expressed proteins were obtained, and they were： Myosin regulatory light chain 2, 40S ribosomal protein SA, Mimecan, Heat shock protein beta-l, Vimentin, Peroxiredoxin-2, Interleukin-I receptor antagonist protein and Lamin A. The change of Hsp27 Peroxiredoxin-2 expression confirmed by Q-PCR and western-blot was consistent with that detected by 2-DE. Conclusion The mechanism of Danggui Niantong Decoction in inhibiting the inflammation of RA synovial tissue maybe associated with inhibiting the signaling pathway of inflammatory response in local tissue, protecting cell structure, and inhibiting oxidative stress response.