基于NLRP3介导的细胞焦亡探讨痹通方含药血清对类风湿关节炎成纤维样滑膜细胞损伤的保护作用及机制  

Protective Effect and Mechanism of Serum Containing Bitong Decoction(痹通方)on Fibroblast-Like Synovial Cell Injury in Rheumatoid Arthritis Based on NLRP3 Mediated Pyroptosis

作　　者：齐庆[1,2] 孙蓬远[1,2] 许诺 关蕊[1] 郑建南 赵夜雨[2] 于静[2] 高明利[2] QI Qing;SUN Pengyuan;XU Nuo;GUAN Rui;ZHENG Jiannan;ZHAO Yeyu;YU Jing;GAO Mingli(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]辽宁中医药大学附属医院,辽宁沈阳110032

出　　处：《中华中医药学刊》2024年第10期44-47,I0006-I0010,共9页Chinese Archives of Traditional Chinese Medicine

基　　金：国家重点研发计划“中医药现代化研究”重点专项(2018YFC1705202);辽宁省教育厅项目(L201916)。

摘　　要：目的探讨痹通方含药血清对脂多糖(LPS)诱导的类风湿关节炎成纤维样滑膜细胞(MH7A)的细胞焦亡的保护作用机制及对NLRP3通路的影响。方法SD大鼠随机分成3组:空白对照组,痹通方低、高剂量组分别给予4.2 g/(kg·d)、16.8 g/(kg·d)灌胃;空白对照组给予等量蒸馏水灌胃。连续灌胃10 d后制备含药血清。将细胞分为5组:空白对照组,LPS处理组,痹通方含药血清低、高剂量组,MCC950组。收集上述各组细胞,CCK8检测细胞活力,RT-qPCR检测各组细胞肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-18、IL-1β、单核细胞趋化蛋白(MCP)-1、MIP-1α基因表达水平及Western blotting检测AIM2、ASC、Caspase-1、IL-18、IL-1β、cleaved-GSDMD、GSDMD、NLRP3蛋白表达。免疫荧光染色法检测NLRP3表达情况。结果与空白组比较,LPS处理组细胞活力下降(P<0.01),TNF-α、IL-18、IL-1β、MCP-1、MIP-1αmRNA表达升高(P<0.01),AIM2、ASC、Caspase-1、IL-18、IL-1β、cleaved-GSDMD、GSDMD、NLRP3蛋白表达升高(P<0.01);免疫荧光结果显示,与空白对照比较,模型组NLRP3荧光强度显著升高(P<0.01)。与LPS处理组相比,给予NLRP3抑制剂MCC950组和痹通方含药血清组均可改善上述指标变化。结论痹通方能有效抑制NLRP3炎症小体表达,减少MH7A细胞焦亡及改善炎症损伤,其机制可能与调控NLRP3通路相关。Objective To investigate the protective mechanism of Bitong Decoction(痹通方)containing serum on pyroptosis of fibroblast-like synoviocytes(MH7A)induced by lipopolysaccharide(LPS)and its effect on NOD-like receptor thermal protein domain associated protein 3(NLRP3)pathway in rheumatoid arthritis.Methods SD rats were randomly divided into 3 groups:control group,Bitong Decoction low and high dose groups[4.2 g/(kg·d)and 16.8 g/(kg·d)by gavage,respectively].The control group was given the same volume of distilled water by gavage.The drug-containing serum was prepared after 10 days of continuous intragastric administration.The cells were divided into 5 groups:control group,LPS treatment group,low and high dose serum containing Bitong Decoction group,MCC950 group.The cells in the above groups were collected,and the cell viability was detected by CCK8.The expression levels of tumor necrosis factor-α(TNF-α),interleukin-18(IL-18),interleukin-1β(IL-1β),monocyte chemotactic protein-1(MCP-1)and macrophage inflammatory protein 1α(MIP-1α)were detected by RT-qPCR.The protein expressions of AIM2,ASC,Caspase-1,IL-18,IL-1β,cleaved-GSDMD,GSDMD and NLRP3 were detected by Western blotting.The expression of NLRP3 was detected by immunofluorescence staining.Results Compared with that of the control group,the viability of cells in LPS treatment group was decreased(P<0.01),and the mRNA expressions of TNF-α,IL-18,IL-1β,MCP-1 and MIP-1A were increased(P<0.01).The protein expressions of AIM2,ASC,Caspase-1,IL-18,IL-1β,cleaved-GSDMD,GSDMD and NLRP3 were significantly increased(P<0.01).The results of immunofluorescence showed that the fluorescence intensity of NLRP3 in the model group was significantly higher than that in the blank control group(P<0.01).Compared with LPS treatment group,NLRP3 inhibitor MCC950 group and serum containing Bitong Decoction groups could improve the changes of above indexes.Conclusion Bitong Decoction can effectively inhibit the expression of NLRP3 inflammasome,reduce pyroptosis of MH7A cells and impr

关 键 词：类风湿关节炎 MH7A细胞 细胞焦亡 痹通方 NLRP3 

分 类 号：R289.5[医药卫生—方剂学]