雷公藤甲素通过lncRNA GAS5/miR-21/TLR4信号轴抑制细胞焦亡改善佐剂型关节炎大鼠炎症  

Triptolide ameliorates inflammation in AA rats by inhibiting pyroptosis through regulation of the lncRNA GAS5/miR-21/TLR4 signaling axis

作　　者：王啊敏 曹云祥[2] 刘健[2] 朱南飞 王慧 WANG A’min;CAO Yunxiang;LIU Jian;ZHU Nanfei;WANG Hui(First Clinical College,Anhui University of Chinese Medicine,Hefei 230038,China;Department of Rheumatology,First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031,China)

机构地区：[1]安徽中医药大学第一临床医学院,合肥230038 [2]安徽中医药大学第一附属医院风湿病科,合肥230031

出　　处：《免疫学杂志》2024年第5期418-424,共7页Immunological Journal

基　　金：国家自然科学基金面上项目(82074090);国家中医药管理局高水平中医药重点学科建设项目(zyyzdxk-2023100);安徽省中医药领军人才项目(中医药发展秘[2018]23号);安徽省自然科学基金(2208085MH267);第七批全国老中医药专家学术经验继承项目(国中医药人教函[2022]76号);安徽省高校自然科学研究重点项目(2023AH050732)。

摘　　要：目的探讨雷公藤甲素(TP)通过lncRNA GAS5/miR-21/TLR4信号轴抑制细胞焦亡改善AA大鼠炎症的机制。方法将大鼠分为4组,每组6只:正常组(NC),模型组(MC),雷公藤甲素组(TP)和甲氨蝶呤组(MTX)。建立AA大鼠模型,观察大鼠一般状态、右足跖肿胀度和关节炎指数;HE染色检测各组大鼠关节滑膜组织病理形态;ELISA法检测IL-1β、IL-6、IL-18、TNF-α炎症因子水平;荧光定量PCR检测Caspase-1、IL-1β、NLRP3、GSDMD、GAS5、TLR4、NF-κB、miR-21 mRNA表达水平;Western blot检测Caspase-1、NLRP3、GSDMD、NF-κB、Caspase-4、TLR4蛋白的表达情况。结果与NC组比较,MC组大鼠肿胀度、关节炎指数显著升高(P<0.05),炎性细胞浸润增加;与MC组比较,TP组和MTX组IL-1β、IL-6、IL-18、TNF-α炎症因子水平下降(P<0.05),Caspase-1、IL-1β、NLRP3、GSDMD、TLR4、NF-κB、miR-21 mRNA表达水平减少,GAS5 mRNA表达增加(P<0.05),Caspase-1、NLRP3、GSDMD、NF-κB、Caspase-4、TLR4蛋白水平下降(P<0.05);与MTX组比较,TP组IL-1β、IL-6、IL-18、TNF-α炎症因子水平下降(P<0.05),Caspase-1、IL-1β、NLRP3、GSDMD、TLR4、NF-κB、miR-21 mRNA表达水平减少(P<0.05)。结论雷公藤甲素可能通过lncRNA GAS5/miR-21/TLR4信号轴,抑制细胞焦亡,抑制促炎因子产生,改善AA大鼠炎症。The aim of this study is to investigate the mechanism by which triptolide(TP)ameliorates inflammation in adjuvant arthritis(AA)rats by inhibiting pyroptosis through the lncRNA GAS5/miR-21/TLR4 signaling axis.Rats were divided into 4 groups:normal control group(NC),model control group(MC),triptolide group(TP)and methotrexate group(MTX),with 6 rats in each group.The AA rat model was established,and the general state,right foot-plantar swelling and arthritis index were observed;HE staining was used to detect the histopathological morphology of synovial joints of rats in each group;ELISA was used to detect the levels of inflammatory factors IL-1β,IL-6,IL-18 and TNF-α;fluorescence quantitative PCR was used to detect the mRNA expression levels of Caspase-1,IL-1β,NLRP3,GSDMD,GAS5,TLR4,NF-κB and miR-21;and Western blot was used to detect the expression levels of Caspase-1,NLRP3,GSDMD,NF-κB,Caspase-4 and TLR4 proteins.Compared with the NC group,rats in the MC group showed significantly higher levels of swelling,arthritis index(P<0.05)and inflammatory cell infiltration;compared with the MC group,the levels of IL-1β,IL-6,IL-18 and TNF-αwere decreased in the TP and MTX groups(P<0.05),and the levels of Caspase-1,IL-1β,NLRP3,GSDMD,TLR4,NF-κB,miR-21 mRNA expression decreased,GAS5 mRNA expression increased(P<0.05),and the protein levels of Caspase-1,NLRP3,GSDMD,NF-κB,Caspase-4 and TLR4 decreased(P<0.05);compared with the MTX group,the TP group showed lower levels of IL-1β,IL-6,IL-18,TNF-α(P<0.05),and lower expression levels of Caspase-1,IL-1β,NLRP3,GSDMD,TLR4,NF-κB and miR-21 mRNA(P<0.05).In conclusion,triptolide may inhibit the production of pro-inflammatory factors and inhibit pyroptosis through the lncRNA GAS5/miR-21/TLR4 signaling axis,thus ameliorate inflammation in AA rats.

关 键 词：雷公藤甲素 lncRNA GAS5/miR-21/TLR4信号轴 细胞焦亡 免疫炎症 佐剂型关节炎 

分 类 号：R593.22[医药卫生—内科学]