漆黄素调控焦亡相关蛋白对人类风湿关节炎滑膜成纤维细胞增殖、迁移及周期阻滞影响  

Effects of butin on regulation of pyroptosis related proteins on proliferation,migration and cycle arrest of human rheumatoid arthritis synovial fibroblast

作　　者：李浩 姚血明 姚晓玲 娄华勇 潘卫东 马武开 LI Hao;YAO Xue-ming;YAO Xiao-ling;LOU Hua-yong;PAN Wei-dong;MA Wu-kai(Dept of Rheumatology and Immunology,the Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guizhou University of Traditional Chinese Medicine,Guiyang 550001,China;the Second Clinical Medical College,Guizhou University of Traditional Chinese Medicine,Guiyang 550001,China;Natural Products Research Center of Guizhou Province,Guiyang 550014,China)

机构地区：[1]贵州中医药大学第二附属医院风湿免疫科,贵州贵阳550001 [2]贵州中医药大学第二临床医学院,贵州贵阳550001 [3]贵州省天然产物研究中心,贵州贵阳550014

出　　处：《中国药理学通报》2024年第10期1937-1944,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 82274678,32060100)。

摘　　要：目的 探讨漆黄素对人类风湿关节炎滑膜成纤维细胞(human fibroblast-like synoviocytes of rheumatoid arthritis, HFLS-RA)增殖、迁移、周期阻滞及焦亡相关炎症因子调控机制。方法 采用细胞迁移及侵袭实验研究细胞增殖、迁移及侵袭;流式细胞术检测细胞周期;ELISA法、RT-qPCR法和Western blot法检测焦亡相关炎症因子IL-1β、IL-18、caspase-1和caspase-3的表达。结果 迁移与侵袭实验表明,漆黄素低、中、高剂量组细胞增殖率较空白对照组降低(P<0.05)。细胞周期分析显示,在G_(0)/G_(1)期,漆黄素中、高剂量组DNA表达量升高(P<0.05),G_(2)、S期,漆黄素中、高剂量组DNA表达量降低(P<0.05)。ELISA、RT-qPCR和Western blot检测结果显示,与IL-1β+caspase-3抑制剂组比较,漆黄素低、中、高剂量组IL-1β、IL-18、caspase-1、caspase-3表达降低(P<0.05)。结论 漆黄素能够抑制HFLS-RA增殖,在G_(1)期发挥明显阻滞作用,其通过调控caspase-1/IL-1β焦亡通路,抑制caspase-1活化,从而减少通路下游IL-1β、IL-18等炎症因子的生成与释放,这可能是漆黄素治疗RA潜在的作用机制之一。Aim To investigate the regulatory mechanism of butin on the proliferation,migration,cycle blockage and pyroptosis related inflammatory factors in human fibroblast-like synoviocytes of rheumatoid arthritis(HFLS-RA).Methods Cell proliferation,migration and invasion were studied using cell migration and invasion assays.Cell cycle was detected by flow cytometry,and the expression of the pyroptosis-associated inflammatory factors IL-1β,IL-18,caspase-1 and caspase-3 was detected by ELISA,RT-qPCR and Western blot.Results Migration and invasion experiments showed that the cell proliferation rate of the butin group was lower than that of the blank control group(P<0.05).Cell cycle analysis demonstrated that in the G_(0)/G_(1) phase,the DNA expression was elevated in the medium and high-dose groups of butin(P<0.05),while in the G_(2) and S phases,the DNA expression was reduced in the medium and high-dose groups of butin(P<0.05).The results of ELISA,RT-qPCR and Western blot assay revealed that the expression of IL-1β,IL-18,caspase-1,and caspase-3 decreased in the butin group compared with the IL-1β+caspase-3 inhibitor group(P<0.05).Conclusions Butin inhibits HFLS-RA proliferation by inhibiting the synthesis of inflammatory vesicles by caspase-1 in the pyroptosis pathway,thereby reducing the production and release of inflammatory factors such as IL-1βand IL-18 downstream of the pathway,and also inhibits HFLS-RA proliferation by exerting a significant blocking effect in the G1 phase,which may be one of the potential mechanisms of butin in the treatment of RA.

关 键 词：漆黄素 类风湿关节炎 细胞周期 炎性细胞因子 焦亡机制 民族医药 

分 类 号：R284.1[医药卫生—中药学] R329.28[医药卫生—中医学] R392.12R593.22