海南医学院学报2023，Vol.29Issue(23)：1786-1793,8.DOI:10.13210/j.cnki.jhmu.20230928.001
基于NLRP3炎症小体探讨三石汤治疗痛风性关节炎的机制研究
Mechanism of Sanshi decoction in the treatment of gouty arthritis by NLRP3 inflammasome
摘要
Abstract
Objective:To observe the effect of Sanshi decoction on P2X7R/PKR pathway-mediated activation of macro-phage NLRP3 inflammasome in order to elucidate the molecular mechanism of Sanshi decoction in the treatment of gouty arthritis.Methods:THP-1 macrophages were divided into control group,model group,low dose group,medium dose group,high dose group of Sanshi decoction and inhibitor group.The remaining groups were induced with monosodium urate crystals to establish a gouty arthritis cell model except the control group.Flow cytometry was used to detect macrophage ROS levels in each group,ELI-SA to detect MDA levels and SOD and GSH-PX activities in each group,and Western blot to detect P2X7R/PKR pathway and NLRP3 inflammasome-associated protein expression.We also used CCK-8 and flow cytometry to measure MH7A activity and apoptotic levels.Results:Compared with the control group,the ROS level,the content of MDA,the activities of SOD and GSH-PX were significantly increased,and the expression levels of NLRP3,full-length IL-1β,pro-IL-1β,full-length IL-18,pro-IL-18,full-length caspase-1,GSDMD-NT,P2X7R and p-PKR protein expression were significantly upregulated,and GSDMD-FL protein expression was significantly downregulated in the model group,and that the differences between them were statistically significant(P<0.05 and P<0.01).Compared with the model group,Sanshi decoction could reduce macrophage ROS levels,MDA content,SOD and GSH-PX activities,and downregulate macrophage NLRP3,mature IL-1β,pro IL-1β,mature IL-18,pro IL-18,mature caspase-1,GSDMD-NT,P2X7R and p-PKR protein expression,and upregulate GSDMD-FL protein expression,with statistically significant differences(P<0.05 and P<0.01).In addition,MH7A activity was downregulated,and apoptosis level was upregulated in the model group in comparison with the control group,and differences were all significantly dif-ferent(P<0.05).As compared to the model group,Sanshi decoction could significantly increase the activity of MH7A and inhibit the level of apoptosis,and that the differences between them were statistically significant(P<0.05 and P<0.01).Conclusion:Sanshi decoction can achieve the therapeutic effect of gouty arthritis by inhibiting P2X7R/PKR pathway activation,thus reducing the activation level of NLRP3.关键词
痛风性关节炎/三石汤/NLRP3炎症小体/P2X7R/PKR信号通路/巨噬细胞Key words
Gouty arthritis/Sanshi decoction/NLRP3 inflammasome/P2X7R/PKR signaling pathway/Macrophages分类
医药卫生引用本文复制引用
朴勇洙,齐明明,聂双莲,潘国雄,张皓,王欣波..基于NLRP3炎症小体探讨三石汤治疗痛风性关节炎的机制研究[J].海南医学院学报,2023,29(23):1786-1793,8.基金项目
This study was supported by Heilongjiang Province Traditional Chinese Medicine Research Project(ZHY19-006) 黑龙江省中医药科研项目(ZHY19-006) （ZHY19-006）
