目的 利用串联质谱标签（tandem mass tags，TMT）定量蛋白质组学技术研究痹祺胶囊治疗类风湿性关节炎（rheumatoid arthritis，RA）的作用机制。方法 建立II型胶原蛋白诱导的RA大鼠模型，并给予痹祺胶囊进行干预，取对照组（C）、模型组（M）、痹祺胶囊0.4 g/kg给药组（BQ）的左膝关节组织，采用TMT定量蛋白质组学技术分析鉴定各组大鼠膝关节组织中的蛋白，以表达差异倍数≥1.2或差异倍数≤0.83且P＜0.05筛选M vs C、BQ vs M、BQ vs C差异表达蛋白，并分析经痹祺胶囊干预后有回调趋势的差异表达蛋白。最后通过GeneCards、OMIN数据库检索RA疾病靶点，与回调蛋白进行交集分析得到与RA疾病相关的回调蛋白，对其进行基因本体（gene ontology，GO）功能和京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析，明晰痹祺胶囊对RA的作用机制。结果 经痹祺胶囊干预后有121个差异蛋白具有回调趋势，其中38个与RA疾病相关，如β2整合素（integrin subunit beta 2，ITGB2）、钙调蛋白2（calmodulin 2，CALM2）、钙调磷酸酶B亚基（calcineurin subunit B type 1，PPP3R1）、前列腺素合成酶（prostacyclin synthase，PTGIS）等。生物信息学分析发现这些回调蛋白参与了与RA相关的多条信号通路，如ITGB2参与调节RA及白细胞跨内皮细胞迁移信号通路；CALM2、PPP3R1通过调节钙信号通路，进而调节T细胞、B细胞受体信号通路、T细胞分化及破骨细胞分化等信号通路；PTGIS参与花生四烯酸代谢途径等。结论 痹祺胶囊可能通过干预ITGB2、PPP3R1、CALM2、PTGIS等蛋白的表达，调节了与RA相关的多条信号通路，发挥免疫抑制、抗炎、活血、抑制骨破坏和骨吸收、抑制血管翳生成等多方面药理作用，从而达到治疗RA的效果。

Objective To study the mechanism of Biqi Capsule (痹祺胶囊) in the treatment of rheumatoid arthritis (RA) by tandem mass tags (TMT) quantitative proteomics technology. Methods The rat model of RA induced by type Ⅱ collagen was established, and Biqi Capsule were given for intervention. The left knee joint tissues of control group (C), model group (M) and Biqi Capsule 0.4 g/kg administration group (BQ) were taken. TMT quantitative proteomics technology was used to analyze and identify the proteins in the knee joint tissues of rats in each group. The differentially expressed proteins of M vs C, BQ vs M, BQ vs C were screened by expression difference multiples ≥ 1.2 or difference multiples ≤ 0.83 and P < 0.05, and the differentially expressed proteins with a callback trend after Biqi Capsule intervention were analyzed. Finally, RA disease targets were retrieved through GeneCards and OMIN databases, and the callback proteins related to RA disease were obtained by intersection analysis. Gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were performed to clarify the mechanism of Biqi Capsule on RA. Results After the intervention of Biqi Capsule, a total of 121 differential proteins had a callback trend, of which 38 were related to RA diseases, such as integrin subunit beta 2 (ITGB2), calmodulin 2 (CALM2), calcineurin subunit B type 1 (PPP3R1), prostacyclin synthase (PTGIS), etc. Bioinformatics analysis showed that these callback proteins were involved in multiple signaling pathways related to RA, such as ITGB2 was involved in the regulation of RA and leukocyte transendothelial cell migration signaling pathways; CALM2 and PPP3R1 regulated T cell, B cell receptor signaling pathway, T cell differentiation and osteoclast differentiation by regulating calcium signaling pathway; PTGIS was involved in arachidonic acid metabolic pathway. Conclusion Biqi Capsule may regulate multiple signaling pathways related to RA by interfering with the expressions of ITGB2, PPP3R1, CALM2, PTGIS and other proteins, and exert immunosuppression, anti-inflammation, blood circulation, inhibition of bone destruction and bone resorption, inhibition of pannus formation and other pharmacological effects, so as to achieve the effect of treating RA.

R285.5

国家自然科学基金重点项目（U21A20406）