背景:滑膜炎性增生是类风湿关节炎(RA)进展中导致关节进一步损伤的关键病理过程。Kadsuraheteroclita(Roxb)Craib,在中国土家族民族医学中也被称为雪通,用于其药用特性,包括促进血液循环,驱散“风邪”,并减轻“潮湿的邪恶”。它已用于治疗关节痛和RA,在土家族民族医学实践中。雪通素(XTS),雪通的主要组成部分,已显示抑制RA成纤维细胞样滑膜细胞(RAFLS)细胞的增殖。然而,XTS在RA治疗中的分子机制需要进一步研究.
目的:观察XTS对类风湿关节炎滑膜炎性增生的治疗作用,重点研究其潜在的分子机制,涉及janus激酶2(JAK2)/转录激活因子3(STAT3)和核因子κB(NF-κB)信号通路。
方法:使用蛋白质-蛋白质相互作用(PPI)和分子对接来寻找XTS治疗RA的主要靶标。在脂多糖(LPS)诱导的RAFLS和RAW264.7细胞的体外模型中,使用酶联免疫吸附试验(ELISA)分析炎症细胞因子的水平,以及JAK2、STAT3和NF-κB信号通路的表达,以及环氧合酶-2(COX-2),通过蛋白质印迹试验进行分析。溶血试验用于证明XTS的生物安全性。用40只雄性大鼠建立佐剂性关节炎(AIA)模型,并给予不同剂量的XTS,然后是自动血常规,ELISA测定,苏木精和伊红(H&E)染色,和放射学分析无XTS对血液细胞因子的影响,组织学变化,AIA大鼠后爪骨破坏的改善。通过免疫荧光分析炎性细胞因子的蛋白质水平,免疫组织化学,或者westernblot.最后,H&E染色检测XTS对心脏的损伤,肝脏,脾,脾肺,和AIA大鼠的肾脏。
结果:我们的结果表明,XTS通过调节JAK2/STAT3和NF-κB信号通路,有效抑制LPS诱导的RAFLS和RAW264.7细胞炎症反应。此外,在AIA大鼠模型中施用XTS显著改善了爪肿胀。组织学分析显示XTS还通过调节JAK2/STAT3和NF-κB信号通路抑制了爪组织中的炎症反应。重要的是,在治疗过程中,XTS表现出优异的安全性,因为它没有引起大鼠血常规参数的任何异常或引起器官损伤。
结论:我们的研究结果强调XTS是抑制RA滑膜增生的一种有前途的天然药物。XTS作为开发针对RA滑膜增生的新型治疗策略的前所未有的天然药物具有巨大潜力。
BACKGROUND: Synovial inflammatory hyperplasia is the key pathological process that leads to further joint damage in rheumatoid arthritis (RA) progress. Kadsura heteroclita (Roxb) Craib, also called Xuetong in Chinese Tujia ethnomedicine, is utilized for its medicinal properties, including promoting blood circulation, dispelling \"wind evil\", and relieving \"damp evil\". It has been used in the treatment of arthralgia and RA, within Tujia ethnomedicinal practices. Xuetongsu (XTS), the main component of Xuetong, has been shown to inhibit the proliferation of RA fibroblast-like synovial cells (RAFLS) cells. However, the molecular mechanism of XTS in RA treatment requires further investigation.
OBJECTIVE: To observe the therapeutic effect of XTS on synovial inflammatory hyperplasia in rheumatoid arthritis, focusing on its underlying molecular mechanisms involving the janus kinase 2 (JAK2)/transducer/activator of transcription 3 (STAT3) and nuclear factor kappa B (NF-κB) signaling pathways.
METHODS: Protein-protein interaction (PPI) and molecular docking were used to find the main targets of XTS treatment for RA. In lipopolysaccharide (LPS)-induced RAFLS and RAW264.7 cells in vitro models, the levels of inflammatory cytokines were analyzed using enzyme linked immunosorbent assay (ELISA), and the expression of JAK2, STAT3, and NF-κB signaling pathways, as well as cyclooxygenase-2 (COX-2), were analyzed through western blotting test. A hemolysis assay was used to certify the biosecurity of XTS. A model of adjuvant arthritis (AIA) was established in 40 male rats, and different doses of XTS were administered, followed by an automatic blood routine, ELISA assay, hematoxylin and eosin (H&E) staining, and radiological analysis of the effect of no XTS on blood cytokines, histological changes, and improvement of posterior paw bone destruction in AIA rats. The protein levels of inflammatory cytokines were analyzed by immunofluorescence, immunohistochemistry, or western blot. Finally, H&E staining was used to detect the damage of XTS on the heart, liver, spleen, lung, and kidney of AIA rats.
RESULTS: Our results demonstrate that XTS effectively inhibited LPS-induced inflammatory responses in RAFLS and RAW264.7 cells by modulating the JAK2/STAT3 and NF-κB signaling pathways. Moreover, XTS administration in the AIA rats model significantly ameliorated paw swelling. Histological analysis revealed that XTS also suppressed the inflammatory response in paw tissue by modulating the JAK2/STAT3 and NF-κB signaling pathways. Importantly, during the treatment, XTS exhibited excellent safety profiles, as it did not induce any abnormalities in blood routine parameters or cause organ damage in the rats.
CONCLUSIONS: Our findings highlight XTS as a promising natural agent for inhibiting synovial hyperplasia in RA. XTS holds great potential as an unprecedented natural agent for developing novel therapeutic strategies to target synovial hyperplasia in RA.