祛寒逐风颗粒对骨性关节炎大鼠炎症反应的影响

Efficacy of Quhan Zhufeng Granule on Inflammatory Response in Rats with Osteoarthritis

目的:观察祛寒逐风颗粒对膝骨性关节炎(KOA)大鼠炎症反应和NF-κB介导NLRP3炎性小体活化的影响。方法:建立骨性关节炎大鼠模型,随机分为假手术组、模型组、祛寒逐风颗粒组(3,6,9g/kg)和阳性药组,测量大鼠机械性痛阈值和膝关节直径,采用酶联免疫吸附实验测量大鼠血清中白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α和IL-6的含量变化,采用RT-PCR法测量大鼠组织中IL-1β、TNF-α和IL-6的mRNA含量,采用苏木精-伊红染色和阿尔新蓝染色观察组织的病理学变化,采用蛋白免疫印迹法检测大鼠软骨组织中p-p65、p65、p-IκBα、NLRP3、Caspase-1p20、IL-1βp17的蛋白水平。结果:与模型组比较,祛寒逐风颗粒组和阳性药组大鼠的机械性痛阈值增大,膝关节直径变小,血清中IL-1β、TNF-α和IL-6含量减少,组织中IL-1β、TNF-α和IL-6mRNA水平下调,软骨损伤和纤维化缓解,软骨组织中p-p65、p65、p-IκBα、NLRP3、Caspase-1p20、IL-1βp17蛋白表达下调。结论:祛寒逐风颗粒可能通过抑制NF-κB介导的NLRP3炎性小体活化显着改善KOA大鼠的炎症反应并缓解疼痛,对骨关节炎发挥治疗作用。

Objective:To observe the efficacy of Quhan Zhufeng granules on inflammatory response and NF-κB-mediated activation of NLRP3inflammasomes in rats with knee osteoarthritis.Methods:Rat models with osteoarthritis were established and randomly divided into sham operation group,model group,Quhan Zhufeng granule(3,6,9g/kg)group and positive drug group.Mouse mechanical pain threshold and knee joint diameter was analyzed.The enzyme-linked immunosorbent assay was used to measure the level changes of interleukin(IL)-1β,tumor necrosis factor(TNF)-αand IL-6in rat serum.The RT-PCR was used to measure the levels of IL-1β,TNF-αand IL-6mRNA in rat tissues.Hematoxylin-eosin staining and Alcian blue staining were used to observe the pathological changes of the tissues.The protein levels of p-p65,p65,p-IκBα,NLRP3,Caspase-1p20,and IL-1βp17in rat cartilage tissues were detected by Western Blot.Results:Compared with the model group,the mechanical pain threshold was increased,the diameter of the knee joint was decreased in the Quhan Zhufeng granule group and the positive drug group,the contents of IL-1β,TNF-αand IL-6in serum were decreased,the levels of IL-1β,TNF-αand IL-6mRNA in tissues were down-regulated,the cartilage damage and fibrosis were relieved,and the protein expression of p-p65,p65,p-IκBα,NLRP3,Caspase-1p20,IL-1βp17in cartilage tissue of the rats in the Quhan Zhufeng granule group and the positive drug group was down-regulated.Conclusion:Quhan Zhufeng granules may significantly improve the inflammatory response and relieve pain of KOA rats by inhibiting the activation of NLRP3inflammatory bodies mediated by NF-κB,and play a therapeutic efficacy on osteoarthritis.