基于TLR4/MyD88/NF-κB信号轴探讨闹羊花毒素Ⅲ对类风湿关节炎的影响  

Effects of RhodotoxinⅢon rheumatoid arthritis based on TLR4/MyD88/NF-κB signal axis

作　　者：刘笑蓉[1] 李硕夫[2] 刘湘丹[1] 王志辉[1] 曾娟 周日宝[1] LIU Xiaorong;LI Shuofu;LIU Xiangdan;WANG Zhihui;ZENG Juan;ZHOU Ribao(Hunan University of Chinese Medicine,Changsha,Hunan 410208,China;The First Hospital of HunanUniversity of Chinese Medicine,Changsha,Hunan 410007,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]湖南中医药大学第一附属医院,湖南长沙410007

出　　处：《湖南中医药大学学报》2023年第6期992-998,共7页Journal of Hunan University of Chinese Medicine

基　　金：湖南省自然科学基金项目(2022JJ80086);湖南省卫生健康委员会计划研究项目(D202302078705);湖南省大学生创新创业训练计划项目(2022-5313);湖南省教育厅科学研究项目(19C1384);湖南省中医药管理局科研计划项目(2021161);湖南中医药大学中药学一级学科开放基金项目(2020ZYX01);湖南中医药大学青苗计划(校行人字2017:25);湖南省一流学科中药学(校行科字2018:3);2020年湖南省一流本科专业建设点(湘教通2020:248号);2020年国家级一流本科专业建设点(教高厅函2021:7号):中药学专业。

摘　　要：目的本研究旨在探讨闹羊花毒素Ⅲ调控TLR4、MyD88和NF-κB的机制,以及闹羊花毒素Ⅲ对类风湿关节炎(rheumatoid arthritis,RA)的影响。方法RA大鼠模型通过Ⅱ型胶原诱导构建。随机将Wistar大鼠分成6组(n=8):假手术组(Sham组)、RA模型组(Model组)、阳性药物雷公藤组[TWG组,50 mg·(kg·d)^(-1)]、闹羊花毒素Ⅲ低剂量组[R-ⅢLo组,0.06 mg·(kg·d)^(-1)]、中剂量[R-ⅢMi组,0.12 mg·(kg·d)^(-1)]和高剂量组[R-ⅢHi组,0.24 mg·(kg·d)^(-1)]。测定各组大鼠的关节炎指数(arthritis index,AI);采用HE染色、ELISA、Western blot和RT-qPCR检测闹羊花毒素Ⅲ对TLR4/MyD88/NF-κB信号轴及RA的影响。结果与Model组比较,闹羊花毒素Ⅲ处理显著降低了RA大鼠的AI评分(P<0.05)。闹羊花毒素Ⅲ对滑膜组织的恶性增生和炎症细胞浸润有明显的抑制作用。与Model组比,闹羊花毒素Ⅲ各剂量组的TNF-α、IL-1β、IL-6、IL-17、VEGF、MMP-2、MMP-9、TLR4、MyD88、p-NF-κB/NF-κB因子的水平降低且呈浓度依赖性(P<0.05)。与TWG组比,R-ⅢLo和R-ⅢMi组的。TNF-α、IL-1β、IL-6、IL-17、VEGF、MMP-2、MMP-9、TLR4、MyD88、p-NF-κB/NF-κB因子的水平升高(P<0.05),而R-ⅢHi组无显著差异(P>0.05)。结论闹羊花毒素Ⅲ可以通过下调TLR4、MyD88、NF-κB因子的表达水平来改善RA。本研究为利用闹羊花毒素Ⅲ治疗RA提供了一定的实验基础。Objective To investigate the effects of rhodojaponinⅢon rheumatoid arthritis(RA)by regulating toll-likereceptor 4(TLR4),myeloid differentiation factor 88(MyD88),and nuclear factor-kappa-B(NF-κB).Methods TypeⅡcollagen was used to construct the RA rat model.Wistar rats were randomly divided into 6 groups,with 8 rats in each group:normal feeding group(sham group),RA model group(Model group),positive drug tripterygium wilfordii group[TWG group,50 mg·(kg·d)^(-1)],low-dose RhodojaponinⅢgroup[R-ⅢLo group,0.06 mg·(kg·d)^(-1)],medium-dose group[R-ⅢMi group,0.12 mg·(kg·d)^(-1)],and high-dose group[RhodojaponinⅢHi group,0.24 mg·(kg·d)^(-1)].The arthritis index(AI)of the rats was scored.Meanwhile,HE ELISA,Western blot,and real-time quantitative PCR(RT-qPCR)were used to detect the effects of RhodojaponinⅢon TLR4/MyD88/NF-κB signal axis and RA.Results RhodojaponinⅢsignificantly reduced the AI scores of RA rats compared with model group(P<0.05)and it effectively inhibited the development of malignant hyperplasia and inflammatory cell infiltration into synovial tissue.Meanwhile,compared with model group,the expresson levels of the tumor necrosis facor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-17(IL-17),vascular endothelial growth factor(VEGF),matrix metallopeptidase-2(MMP-2),matrix metallopeptidase-9(MMP-9),TLR4,MyD88,and phosphor-nuclear factor-kappa-B(p-NF-κB)/NF-κB were reduced in R-ⅢLo,Mi,and Hi groups(P<0.05).Compared with the TWG group,the levels of TNF-α,IL-1β,IL-6,IL-17,VEGF,MMP-2,MMP-9,TLR4,MyD88,p-NF-κB/NF-κB factors increased in the R-ⅢLo and R-ⅢMi groups(P<0.05),while no significant differences were observed in the R-ⅢHi group(P>0.05).Conclusion RhodojaponinⅢcan improve RA by down-regulating the expression levels of TLR4,MyD88,and NF-κB.This paper offered a solid experimental foundation for the use of RhodojaponinⅢin the treatment of RA.

关 键 词：闹羊花毒素Ⅲ TLR4/MyD88/NF-κB信号轴 类风湿关节炎 羊踯躅 炎症 血管生成 

分 类 号：R285.5[医药卫生—中药学]