白芍总苷调控NF-κB/NLRP3信号通路对急性痛风性关节炎大鼠的影响机制研究

Effect mechnism of total glucosides of paeoniae on NF-κB/NLRP3 signaling pathway in rats with acute gouty arthritis

目的:探究白芍总苷(TGP)通过调控核因子-κB(NF-κB)/含NLR家族Pyrin域蛋白3(NLRP3)信号通路对急性痛风性关节炎(AGA)大鼠炎症的影响。方法:将60只SD大鼠以每组10只随机分为对照组、AGA组、秋水仙碱组、TGP-L组、TGP-M组、TGP-H组。对照组和AGA组每天进行生理盐水灌胃,秋水仙碱组、TGP-L组、TGP-M组、TGP-H组每天分别灌胃相应药物,共7 d,除对照组外均于灌胃第5天时构建尿酸钠(MSU)诱导的AGA大鼠模型;观察各组大鼠一般情况;检测大鼠步态及关节炎症指数评价、关节肿胀程度;HE染色观察大鼠踝关节滑膜组织病理学变化;ELISA法检测大鼠关节液中炎症因子水平;Western blot检测大鼠踝关节滑膜组织NF-κB p65/NLRP3通路相关蛋白表达情况。结果:与对照组相比,AGA组大鼠关节肿胀、跛行、皮毛无光泽、精神倦怠;秋水仙碱组、TGP-L组、TGP-M组、TGP-H组大鼠相关症状均得到有效改善;与对照组相比,AGA组大鼠步态级别、关节炎症指数、关节肿胀程度、组织病理学程度、TNF-α、IL-6、IL-1β、p-NF-κB p65/NF-κB p65、NLRP3和Caspase-1水平显著升高,p-IκBα/IκBα水平显著降低(P<0.05);与AGA组相比,秋水仙碱组、TGP-L组、TGP-M组和TGP-H组大鼠步态级别、关节炎症指数、关节肿胀程度、组织病理学程度、TNF-α、IL-6、IL-1β、p-NF-κB p65/NF-κB p65、NLRP3和Caspase-1水平显著降低,p-IκBα/IκBα水平显著升高,存在TGP剂量依赖性(P<0.05);与秋水仙碱组相比,TGP-H组大鼠步态级别、关节炎症指数、关节肿胀程度、组织病理学程度、TNF-α、IL-6、IL-1β、p-NF-κB p65/NF-κB p65、p-IκBα/IκBα、NLRP3和Caspase-1水平差异无统计学意义(P>0.05)。结论:TGP对AGA大鼠炎症状态的改善可能与其抑制NF-κB/NLRP3信号通路有关。

Objective:To explore the effect of total glucosides of paeony(TGP)on inflammation in rats with acute gouty arthritis(AGA)by regulating nuclear factor-kappa B(NF-κB)/NLR family Pyrin domain containing protein 3(NLRP3)signaling pathway.Methods:Sixty SD rats were randomly divided into control group,AGA group,colchicine group,TGP-L group,TGP-M group and TGP-H group,with ten rats in each group.The control group and the AGA group were given normal saline gavage everyday,and the colchicine group,TGP-L group,TGP-M group and TGP-H group were given the corresponding drugs every day for 7 days,except for the control group,a rat model of AGA induced by sodium urate(MSU)was constructed on the 5th day after intragastric administration;the general conditions of rats in each group were observed;the gait and joint inflammation index evaluation,and joint swelling detection were implemented;the histopathological changes of the synovium of the ankle joint in rats were observed with HE staining;the levels of inflammatory factors in the synovial fluid of rats were detected with ELISA method;the expression of NF-κB p65/NLRP3 pathway related proteins in synovial tissue of rat ankle joint was detected with Western blot.Results:Compared with control group,rats in AGA group had swollen joints,lameness,dull fur,and mental fatigue;the related symptoms of rats in colchicine group,TGPL group,TGP-M group and TGP-H group were all effectively improved;compared with control group,the gait grade,joint inflammation index,joint swelling degree,histopathological degree,TNF-α,IL-6,IL-1β,p-NF-κB p65/NF-κB p65,NLRP3 and Caspase-1 levels were increased significantly in AGA group,the p-IκBα/IκBαlevel reduced significantly(P<0.05);compared with AGA group,the gait grade,joint inflammation index,joint swelling degree,histopathological degree,TNF-α,IL-6,IL-1β,p-NF-κB p65/NF-κB p65,NLRP3 and Caspase-1 levels were decreased significantly in colchicine group,TGP-L group,TGP-M group and TGP-H group,ence in the gait grade,joint inflammation index,joint swelling degree,histopathological degree,TNF-α,IL-6,IL-1β,p-NF-κB p65/NF-κB p65,p-IκBα/IκBα,NLRP3 and Caspase-1 levels between colchicine group and TGP-H group(P>0.05).Conclusion:The improvement of TGP on the inflammatory state of AGA rats may be related to its inhibition on NF-κB/NLRP3 signaling pathway.