背景:急性痛风性关节炎(AGA)在中医(TCM)理论中被归类为“关节炎”。Shirebi颗粒(SGs),源自经典处方四妙丸,在临床上改善AGA方面发挥了令人满意的治疗效果。然而,SGs对抗AGA的潜在机制仍未阐明。
方法:与AGA相关的生物过程,通过生物信息学从GEO数据库中挖掘信号通路和生物标志物基因。使用UPLC-Q-TOF-MS/MS系统地识别SGs成分。基于生物标志物基因和化学成分建立了相关网络,从中选择用于进一步研究的信号通路。最后,我们使用在踝关节注射尿酸单钠(MSU)的SD大鼠建立了AGA模型,以进行实验验证。行为测试的组合,H&E,SafraninO-FastGreen,西方印迹,采用免疫荧光技术揭示SGs对AGA的作用机制。
结果:AGA的恶化与免疫和炎症之间的失衡显着相关,中性粒细胞趋化性和炎症因子激活。HDAC5,PRKCB,NFκB1,MPO,PRKCA,PIK3CA被确定为SGs针对AGA的候选目标,与中性粒细胞胞外陷阱(NETs)信号通路有关。动物实验证明SGs能有效修复软骨损伤,阻止TLR4激活,抑制NETs指标和炎症因子的表达。此外,SGs显著缓解关节红肿,改善关节功能障碍,抑制AGA大鼠的炎症浸润。
结论:我们的数据表明,SGs可以通过抑制NETs促进的免疫和炎症之间的失衡来有效缓解AGA的疾病严重程度。
BACKGROUND: Acute gouty arthritis (AGA) is classified as \'arthritis\' in traditional Chinese medicine (TCM) theory. Shirebi granules (SGs), derived from the classic prescription SiMiaoWan, exerts satisfying therapeutic efficacy in ameliorating AGA clinically. However, the underlying mechanisms of SGs against AGA remain unclarified.
METHODS: AGA-related biological processes, signal pathways and biomarker genes were mined from the GEO database through bioinformatics. SGs components were systematically recognized using the UPLC-Q-TOF-MS/MS. A correlation network was established based on the biomarker genes and the chemical components, from which the signal pathway used for further study was selected. Finally, we established an AGA model using SD rats injected with monosodium urate (MSU) in the ankle joint for experimental validation. A combination of behavioral tests, H&E, safranin O- fast green, western blotting, and immunofluorescence were employed to reveal the mechanism of action of SGs on AGA.
RESULTS: The deterioration of AGA was significantly related to the imbalance between immunity and inflammation, neutrophil chemotaxis and inflammatory factor activation. HDAC5, PRKCB, NFκB1, MPO, PRKCA, PIK3CA were identified to be the candidate targets of SGs against AGA, associated with neutrophil extracellular traps (NETs) signal pathway. Animal experiments demonstrated that SGs effectively repaired cartilage damage, blocked TLR4 activation, and inhibited the expression of NETs indicators and inflammatory factors. In addition, SGs prominently alleviated joint redness and swelling, improved joint dysfunction, inhibited inflammatory infiltration of AGA rats.
CONCLUSIONS: Our data reveal that SGs may effectively alleviate the disease severity of AGA by suppressing NETs-promoted imbalance between immunity and inflammation.