益肾清络活血方对CFA大鼠OPG/RANKL通路的影响及机制  

Effectof Yishen-Qingluo-Huoxue prescription on OPG/RANKL pathway in CFA rats and its mechanism

作　　者：李丹凤 王雯 许萍 陈修平 李艳[2] LI Dan-feng;WANG Wen;XU Ping;CHEN Xiu-ping;LI Yan(Graduate School of Wannan Medical College,Yijishan Hospital,Wannan Medical College,Wuhu,Anhui 241002,China;Dept of Traditional Chinese Medicine,Yijishan Hospital,Wannan Medical College,Wuhu,Anhui 241002,China)

机构地区：[1]皖南医学院研究生学院,安徽芜湖241002 [2]皖南医学院弋矶山医院中医科,国医大师李济仁工作室,安徽芜湖241002

出　　处：《中国药理学通报》2022年第11期1747-1752,共6页Chinese Pharmacological Bulletin

基　　金：国家中医药管理局重点学科中医痹病学建设项目(国中医药人教发[2012]32号);中部地区名老中医学术观点、特色诊疗方法和重大疾病防治经验研究(No.2018YFC1704103)。

摘　　要：目的探讨新安方益肾清络活血方(Yisheng-Qingluo-Huoxie prescription,YSF)对完全弗氏佐剂(complete freund's adjuvant,CFA)诱导的类风湿关节炎大鼠中OPG/RANKL通路的影响及机制。方法取雄性SD大鼠32只,随机分为正常(Normal)组、模型(Adjuvantinduced arthritis,AIA)组、清络饮(Qing-luo-yin,QLY)组和YSF组,每组8只。除正常组外,其余均诱导为AIA大鼠,模型成功后首日灌胃给QLY和YSF中药,连续30 d。观察大鼠体质量、AI评分;HE染色法观察大鼠踝关节组织形态学变化;免疫组化法观察MMP3与p-p65的表达;酶联免疫吸附法检测大鼠血清TNF-α、IL-1β、OPG、RANKL水平;Western blot法检测大鼠滑膜OPG、RANKL蛋白的表达;RT-PCR检测滑膜中OPG、RANKLmRAN的表达。结果与AIA组相比,经过YSF给药治疗后,大鼠体质量渐渐恢复正常,炎症评分降低,HE染色显示骨质保存完好,未见滑膜浸润;MMP3、p-p65出现低表达;血清中TNF-α、IL-1β、RANKL水平明显下调,OPG、OPG/RANKL水平明显上升;Western blot和RT-PCR均显示大鼠滑膜中OPG、OPG/RANKL出现高表达,而RANKL则相反。结论YSF有可能通过抑制炎症反应,降低炎症评分及血清中TNF-α、IL-1β的表达,升高OPG/RANKL的比值来调节关节骨破坏的环境,进而缓解RA病情。Aim To investigate the effect of Xinan Fang Yisheng-Qingluo-Huoxie prescription(YSF)on OPG/RANKL pathway in rats with rheumatoid arthritis induced by complete Freund′s adjuvant(CFA)and its mechanism.Methods Thirty-two male SD rats were randomly divided into normal group,AIA group,QLY group and YSF group,with 8 rats in each group.AIA rats were induced except for the normal group.QLY and YSF were administered intragastrically on the first day after the model was successful for 30 days.The body weight and AI score of rats were observed.HE staining was used to observe the histomorphological changes of the ankle joint of rats.The expression of MMP3 and P-P65 was observed by immunohistochemistry.Serum levels of TNF-α,IL-1β,OPG and RANKL were detected by ELISA.The expression of OPG and RANKL proteins in synovium of rats was detected by Western blot and RT-PCR.Results Compared with AIA model,the body weight of the rats gradually returned to normal,and inflammation score decreased.HE staining showed that the bone was well preserved,no synovial infiltration was observed after YSF administration,and MMP3 and P-P65 showed low expression.The levels of TNF-α,IL-1βand RANKL in serum were significantly down-regulated,while the levels of OPG and OPG/RANKL were significantly up-regulated.In addition,Western blot and RT-PCR showed that OPG and OPG/RANKL were highly expressed in synovium of rats,while RANKL was on the contrary.Conclusions YSF may decrease inflammatory score and expression of TNF-αand IL-1βin serum by inhibiting inflammatory response.What′s more,the increase of OPG/RANKL ratio can regulate the environment of joint bone destruction,thus alleviating RA.

关 键 词：益肾清络活血方 清络饮 佐剂性关节炎 OPG/RANKL 骨保护 炎症 新安方 

分 类 号：R-332[医药卫生] R287R322.72R364.5R392.12R593.22