山茱萸-白芍酒炙配伍前后通过调节OPG/RANKL/RANK信号通路抗类风湿性关节炎药效机制研究  

Anti-rheumatoid arthritis efficacy and mechanism of Corni Fructus-Paeoniae Radix Alba herbal pair with wine before and after processing based on OPG/RANKL/RANK pathway

作　　者：郑寅涛 曹岗[1] 吴鑫 ZHENG Yin-tao;CAO Gang;WU Xin(School of Pharmaceutical Sciences,Zhejiang Chinese Medical University,Hangzhou 310053,China)

机构地区：[1]浙江中医药大学药学院,浙江杭州310053

出　　处：《中草药》2022年第10期3084-3092,共9页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金项目(81773900)。

摘　　要：目的探讨山茱萸-白芍酒炙配伍前后对类风湿性关节炎小鼠关节评分、足趾红肿度、胫骨关节与踝关节损伤度及膝关节滑膜组织中骨保护素(osteoprotegerin,OPG)/核因子-κB(nuclear factor-κB,NF-κB)受体激活因子配体(receptor activator of NF-κB ligand,RANKL)/NF-κB受体激活因子(receptor activator of NF-κB,RANK)信号通路的影响。方法尾根部sc牛II型胶原和完全弗氏佐剂的混合乳状液建立类风湿性关节炎(rheumatoid arthritis,RA)小鼠模型。将DBA/1雄性小鼠按照体质量分布随机分为9组,即对照组、模型组、来氟米特阳性药组、生山茱萸组、酒山茱萸组、生白芍组、酒白芍组、生山茱萸-生白芍配伍组、酒山茱萸-酒白芍配伍组,每组8只。除对照组外,首次免疫造模7 d后进行第2次加强免疫,第30天造模完成后,各给药组第31天起每天ig相应剂量的75%乙醇提取物无菌水溶液,阳性药组给予来氟米特无菌水溶液(6 mg/kg),对照组、模型组小鼠ig相应体积的无菌水,连续给药30 d。显微镜下观察小鼠踝关节HE病理变化,断层扫描(Micro-CT)观察小鼠胫骨关节及踝关节损伤程度,RT-qPCR法测定膝关节滑膜组织中OPG、RANKL、肿瘤坏死因子受体相关因子6(tumor necrosis factor receptor-associated factor 6,TRAF6)、酪氨酸蛋白激酶(sarcoma,Src)、核因子-p65(nuclear factor-p65,P65)基因表达水平及Western blotting法测定OPG、RANKL、RANK、TRAF6、Src、磷酸化NF-κB抑制蛋白α(phosphorylation inhibitoryκBα,p-IκBα)、磷酸化P65(p-P65)蛋白的表达。结果与对照组相比,模型组小鼠踝关节组织有明显的滑膜成纤维细胞及破骨细胞等炎性浸润,足趾关节及踝关节发生变形,OPG蛋白和基因表达水平明显降低(P<0.001),RANKL、TRAF6、Src、P65基因表达水平及RANKL、RANK、TRAF6、Src、p-IκBα/IκBα、p-P65/P65蛋白表达水平显著升高(P<0.001);与模型组相比,山茱萸和白芍酒炙配伍前后各组小鼠踝关�Objective To investigate the effects of Shanzhuyu(Corni Fructus)-Baishao(Paeoniae Radix Alba)herbal pair with wine before and after processing on joint scores,toe swelling degree,tibial articular and ankle joint injury degree,and osteoprotegerin(OPG)/receptor activator of nuclear factor-κB ligand(RANKL)/receptor activator of NF-κB(RANK)pathway of knee synovial tissuein rheumatoid arthritis mice.Methods The emulsion of bovine type II collagen and complete Freund’s adjuvant were injected subcutaneously into the tail root of mice to establish the rheumatoid arthritis model.According to the weight distribution,the DBA/1 male mice were randomly divided into nine groups(eight per group).Namely,the control group,the model group,the leflunomide positive drug group,the raw Corni Fructus group,the wine-processed Corni Fructus group,the raw Paeoniae Radix Alba group,the wine-processed Paeoniae Radix Alba group,the raw Corni Fructus and Paeoniae Radix Alba compatibility group,and the wine-processed Corni Fructus and Paeoniae Radix Alba compatibility group,respectively.The second booster immunization was performed 7 d after the first immunization and then the rheumatoid arthritis model was established on the 30th d.Furthermore,each administration group was given a corresponding dose of 75%ethanol extract sterile aqueous solution,the positive drug group was given a sterile aqueous solution of leflunomide(6 mg/kg),and mice in the control group and model group were given a corresponding volume of sterile water for the following 30 consecutive days.Finally,the pathological changes of HE in the mouse ankle joint were observed under microscope,and the injury degree of the mouse tibial articular and ankle joint were observed by Micro-CT tomography,respectively.Moreover,the OPG,RANKL,tumor necrosis factor receptor-associated factor 6(TRAF6),sarcoma(Src),nuclear factor-p65(P65)genes and the OPG,RANKL,RANK,TRAF6,Src,phosphorylation inhibitoryκBα(p-IκBα),p-P65 protein in the synovial tissue of the knee joint were determined by R

关 键 词：山茱萸 白芍 酒炙 配伍 类风湿性关节炎 OPG/RANKL/RANK信号通路 

分 类 号：R285[医药卫生—中药学]