Wenshuang Wang, Shanshan Zhai, Wen Yang, He Gao, Nianwei Chang, Man Zhang, Yuanyuan Hou, Gang Bai
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引用次数: 0
Abstract
Background: Inflammation plays a significant role in initiating and sustaining rheumatoid arthritis (RA). Acacetin, a natural flavonoid compound, exhibits excellent anti-inflammatory effects specifically for RA. However, its relevant targets and molecular mechanisms remain to be elucidated.
Purpose: This study aims to investigate the mechanism of acacetin in the therapeutic efficacy of acacetin in RA and search for new therapeutic options for RA treatment.
Methods: A collagen-induced RA mouse model was established to evaluate the therapeutic effect of acacetin. Acacetin functional probes were synthesized to capture potential target proteins in RAW264.7 cells. Various small molecule-protein interaction methods were conducted to verify the binding of acacetin to target protein. Molecular docking and site directed mutagenesis tests were performed to analyze the specific binding sites. Co-immunoprecipitation, immunofluorescence assay and western blot were engineered to explore the effect of acacetin on COX-2 degradation by targeting HSP90.
Results: Acacetin specifically binds to the ATP domain of HSP90, to facilitate the dissociation between HSP90 and COX-2, inducing the ubiquitin-degradation of COX-2 in macrophages. Acacetin suppressed the production of pro-inflammatory cytokines, as well as inflammatory related pathways, exerting excellent anti-inflammatory effects in RA.
Conclusions: This research proved that acacetin, a novel HSP90 ATPase inhibitor, inhibits the functional folding of the client protein COX-2, promoting its ubiquitin degradation for anti-inflammation. Targeting HSP90 is a viable strategy to inhibit inflammation, affording a distinct way to managing joint inflammation and pains associated with RA.
背景：炎症在类风湿性关节炎（RA）的发病和持续发展中起着重要作用。作为一种天然黄酮类化合物，乙酰丙酮（Acacetin）对类风湿性关节炎（RA）具有卓越的抗炎作用。目的：本研究旨在探讨阿卡西汀对 RA 的疗效机制，并寻找治疗 RA 的新疗法：方法：建立胶原蛋白诱导的RA小鼠模型，评估阿卡匹林的治疗效果。方法：建立了胶原蛋白诱导的 RA 小鼠模型，以评估阿卡西汀的治疗效果。合成了阿卡西汀功能探针，以捕获 RAW264.7 细胞中的潜在靶蛋白。采用多种小分子与蛋白质相互作用的方法来验证阿卡西汀与靶蛋白的结合。分子对接和定点突变试验分析了特定的结合位点。通过免疫共沉淀、免疫荧光检测和 Western 印迹技术，探讨了阿卡西汀通过靶向 HSP90 对 COX-2 降解的影响：结果：Acacetin能特异性地与HSP90的ATP结构域结合，从而促进HSP90与COX-2的解离，诱导巨噬细胞中COX-2的泛素降解。阿卡西汀可抑制促炎细胞因子以及炎症相关通路的产生，对RA具有良好的抗炎作用：这项研究证明，新型 HSP90 ATP 酶抑制剂 Acacetin 可抑制客户蛋白 COX-2 的功能性折叠，促进其泛素降解以达到抗炎目的。以HSP90为靶点是一种抑制炎症的可行策略，为控制与RA相关的关节炎症和疼痛提供了一种独特的方法。
期刊介绍：
Phytomedicine is a therapy-oriented journal that publishes innovative studies on the efficacy, safety, quality, and mechanisms of action of specified plant extracts, phytopharmaceuticals, and their isolated constituents. This includes clinical, pharmacological, pharmacokinetic, and toxicological studies of herbal medicinal products, preparations, and purified compounds with defined and consistent quality, ensuring reproducible pharmacological activity. Founded in 1994, Phytomedicine aims to focus and stimulate research in this field and establish internationally accepted scientific standards for pharmacological studies, proof of clinical efficacy, and safety of phytomedicines.
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