背景:雷公藤甲素(TP),从草药雷公藤中提取的天然产物,表现出有效的免疫抑制活性。然而,其在类风湿性关节炎中的作用机制尚不完全清楚.
方法:用牛Ⅱ型胶原免疫Sprague-Dawley大鼠建立胶原诱导关节炎(CIA)模型,TP作为治疗。TP的治疗效果根据爪肿胀进行评估,组织病理学,血清炎症因子水平。从大鼠血清中分离的外泌体通过透射电子显微镜表征,动态光散射,和蛋白质印迹分析。通过直接DIA定量蛋白质组学分析来分析外泌体的蛋白质组学概况。基因本体论和京都百科全书的基因和基因组数据库被用于富集分析相关的分子功能,生物过程,和信号通路。蛋白质印迹分析用于分析差异表达的蛋白质。
结果:TP治疗改善了CIA大鼠的关节炎表型,关节炎评分降低,爪子肿胀,病理损伤严重程度评分,和血清炎性细胞因子水平。蛋白质组分析表明,TP处理显著抑制补体和凝血级联反应,白细胞介素-17信号通路,和胆固醇代谢,在CIA大鼠中重新激活。重要的是,在CIA组中,脂质运载蛋白2(LCN2)和髓过氧化物酶(MPO)水平显着上调,但在TP给药后受到抑制。此外,在滑膜组织中,CIA组LCN2和MPO表达水平也升高,但TP治疗后降低。
结论:我们的研究结果表明,TP缓解了A,可能通过外泌体LCN2和MPO蛋白的调节。
BACKGROUND: Triptolide (TP), a natural product derived from the herbal medicine Tripterygium wilfordii, exhibits potent immunosuppressive activity. However, the mechanisms underlying its effects in rheumatoid arthritis remain incompletely understood.
METHODS: Collagen-induced arthritis (CIA) model was induced in Sprague-Dawley rats by immunization with bovine type II collagen, and TP was administrated as treatment. The therapeutic effect of TP was evaluated based on paw swelling, histopathology, and serum levels of inflammatory factors. Exosomes isolated from rat serum were characterized by transmission electron microscopy, dynamic light scattering, and western blot analysis. Proteomic profiling of exosomes was analyzed by direct DIA quantitative proteomics analysis. Gene ontology and the Kyoto Encyclopedia of Genes and Genomes databases were employed for enrichment analysis related to molecular function, biological processes, and signaling pathways. Western blot analysis was used to analyze differentially expressed proteins.
RESULTS: TP treatment ameliorated arthritic phenotypes in CIA rats as evidenced by reduced arthritis score, paw swelling, pathological injury severity scores, and serum levels of inflammatory cytokines. The proteomic analysis revealed that TP treatment significantly inhibited complement and coagulation cascades, interleukin-17 signaling pathway, and cholesterol metabolism, which were reactivated in CIA rats. Importantly, lipocalin 2 (LCN2) and myeloperoxidase (MPO) levels were markedly upregulated in the CIA group but suppressed upon TP administration. Furthermore, in synovial tissues, LCN2 and MPO expression levels were also elevated in the CIA group but decreased following TP treatment.
CONCLUSIONS: Our findings demonstrate that TP alleviates CIA, possibly through modulation of exosomal LCN2 and MPO proteins.