Li Xin, Mao Xia, Jiang Hong, Xia Cong, Fu Lu, Gao Wenjing, Chen Wenjia, Li Weijie, Wang Ping, Zhang Yanqiong, Xu Haiyu
State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, People's Republic of China.
Chin Med. 2024 Aug 9;19(1):105. doi: 10.1186/s13020-024-00962-6.
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.
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.
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.
Our data reveal that SGs may effectively alleviate the disease severity of AGA by suppressing NETs-promoted imbalance between immunity and inflammation.
急性痛风性关节炎（AGA）在中医理论中被归类为“痹症”。四妙颗粒（SGs）源自经典方剂四妙丸，在临床上对AGA的改善具有令人满意的治疗效果。然而，SGs治疗AGA的潜在机制仍不明确。
通过生物信息学从基因表达综合数据库（GEO数据库）中挖掘与AGA相关的生物学过程、信号通路和生物标志物基因。使用超高效液相色谱-四极杆飞行时间串联质谱（UPLC-Q-TOF-MS/MS）系统识别SGs的成分。基于生物标志物基因和化学成分建立相关网络，从中选择用于进一步研究的信号通路。最后，我们通过向SD大鼠踝关节注射尿酸钠（MSU）建立AGA模型进行实验验证。采用行为学测试、苏木精-伊红染色（H&E）、番红O-固绿染色、蛋白质免疫印迹法和免疫荧光法相结合的方法揭示SGs对AGA的作用机制。
AGA的恶化与免疫和炎症失衡、中性粒细胞趋化和炎症因子激活显著相关。组蛋白去乙酰化酶5（HDAC5）、蛋白激酶Cβ（PRKCB）、核因子κB1（NFκB1）、髓过氧化物酶（MPO）、蛋白激酶Cα（PRKCA）、磷脂酰肌醇-3激酶催化亚基α（PIK3CA）被确定为SGs治疗AGA的候选靶点，与中性粒细胞胞外陷阱（NETs）信号通路相关。动物实验表明，SGs能有效修复软骨损伤，阻断Toll样受体4（TLR4）激活，并抑制NETs指标和炎症因子的表达。此外，SGs显著减轻关节红肿，改善关节功能障碍，抑制AGA大鼠的炎症浸润。
我们的数据表明，SGs可能通过抑制NETs促进的免疫和炎症失衡来有效减轻AGA的疾病严重程度。
