Li Yang, Liu Jian, Sun Yue, Hu Yuedi, Cong Chengzhi, Chen Yiming, Fang Yanyan
Department of Rheumatology, The First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei, Anhui 230031, China; First Clinical Medical School, Anhui University of Chinese Medicine, Hefei, Anhui 230038, China.
Department of Rheumatology, The First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei, Anhui 230031, China; Institute of Rheumatology, Anhui Academy of Chinese Medicine, Hefei, Anhui 230031, China.
Int Immunopharmacol. 2025 Feb 20;148:114112. doi: 10.1016/j.intimp.2025.114112. Epub 2025 Jan 21.
Rheumatoid arthritis (RA) is a common chronic autoimmune disease. Neutrophils release and their extracellular traps (NETs) tend to result in synovial inflammation and cartilage damage. Huangqin Qingre Chubi Capsule (HQC) is an important herbal formulation for RA treatment and has been used for many years. This study applied an interdisciplinary and integrated strategy combining clinical data, integrative bioinformatics, molecular modeling, and both in vitro and in vivo experiments to elucidate the efficacy and potential anti-inflammatory mechanisms of HQC for RA. Retrospective clinical data mining demonstrated that patients with RA treated with HQC exhibited recovery of immuno-inflammatory markers and self-perception. By applying network pharmacological analysis, molecular docking, molecular dynamics simulation, and cellular thermal shift assays, we determined that HQC can directly bind to MAPK14 and target p38 MAPK signaling pathway and NETs formation. Treatment of an adjuvant-induced arthritis rat model combining damp-heat patterns using HQC demonstrated it's effectiveness in reducing the severity of inflammatory arthritis in a dose-dependent manner and downregulated phosphorylation of p38 MAPK and NETs release. In vitro co-culture system mimicking inflammatory microenvironment of RA in vivo revealed that crosstalk between neutrophils (PMNs) and fibroblast-like synoviocytes (FLSs) was manifested by activation of p38 MAPK signaling pathway, release of NETs, and amplification of inflammation, which was blocked by HQC. Mechanistic validation using the p38 MAPK agonist diprovocim demonstrated that HQC inhibited NET formation by modulating the p38 MAPK pathway (mainly by inhibiting its phosphorylation) to exert anti-inflammatory effects. In conclusion, our interdisciplinary and integrated study demonstrated that HQC can target the p38 MAPK signaling pathway to inhibit NET formation and inflammatory response, thereby blocking the crosstalk between PMNs and FLSs to ameliorating RA.
类风湿性关节炎（RA）是一种常见的慢性自身免疫性疾病。中性粒细胞的释放及其细胞外陷阱（NETs）往往会导致滑膜炎症和软骨损伤。黄芩清热除痹胶囊（HQC）是治疗RA的一种重要中药制剂，已应用多年。本研究采用跨学科综合策略，结合临床数据、整合生物信息学、分子建模以及体外和体内实验，以阐明HQC治疗RA的疗效和潜在抗炎机制。回顾性临床数据挖掘表明，接受HQC治疗的RA患者免疫炎症标志物和自我感觉有所恢复。通过网络药理学分析、分子对接、分子动力学模拟和细胞热位移分析，我们确定HQC可直接与MAPK14结合，靶向p38 MAPK信号通路和NETs形成。用HQC治疗佐剂诱导的湿热型关节炎大鼠模型，结果表明其能以剂量依赖的方式有效减轻炎症性关节炎的严重程度，并下调p38 MAPK的磷酸化和NETs的释放。体外共培养系统模拟体内RA的炎症微环境，结果显示中性粒细胞（PMNs）与成纤维细胞样滑膜细胞（FLSs）之间的相互作用表现为p38 MAPK信号通路的激活、NETs的释放和炎症的放大，而HQC可阻断这种相互作用。使用p38 MAPK激动剂双丙戊酸进行的机制验证表明，HQC通过调节p38 MAPK通路（主要是抑制其磷酸化）来抑制NET形成，从而发挥抗炎作用。总之，我们的跨学科综合研究表明，HQC可靶向p38 MAPK信号通路，抑制NET形成和炎症反应，从而阻断PMNs与FLSs之间的相互作用，改善RA。
