葛根素对痛风性关节炎小鼠炎性相关细胞因子表达的影响

Effect of puerarin on the expression of inflammatory cytokines in gouty arthritis mice

目的探究葛根素对痛风性关节炎(GA)小鼠炎性相关细胞因子表达的影响。方法 30只小鼠随机分为sham组、model组及puerarin组,每组10只,微晶尿酸钠注射用于构建小鼠GA模型,造模后puerarin组小鼠每日给予100 mg/kg葛根素注射治疗5 d。观察并记录各组小鼠关节肿胀度变化及后肢功能障碍指数;ELISA检测血清UA、NO含量及关节组织液中IL-1β、IL-10、TNF-α等炎症因子表达水平;血液分析仪用于检测小鼠全血中白细胞、中性粒细胞及淋巴细胞数量变化;HE染色观察各组小鼠后肢足掌关节滑膜组织病理变化;Western blot及qRT-PCR用于检测p-AMPKα、Arginase-1、NOS2、NLRP3蛋白及mRNA表达水平。结果与model组相比,葛根素能有效减少puerarin组小鼠关节肿胀度、后肢功能障碍指数并缓解关节滑膜组织病理变化。ELISA结果显示,puerarin组小鼠血清UA含量较model组显著降低,NO表达增加,关节组织液中IL-1β、TNF-α含量减少,IL-10表达量增加;全血分析结果显示puerarin组小鼠血液白细胞、中性粒细胞及淋巴细胞数量较Model组显著减少;Western blot结果显示,与model组相比,puerarin组小鼠关节组织中p-AMPKα和Arginase-1蛋白表达量增加,NOS2、NLRP3蛋白表达量降低,同时qRT-PCR结果显示Arginase-1、NOS2、NLRP3 mRNA表达趋势与蛋白表达变化趋势一致。结论葛根素可能通过影响炎性细胞因子表达水平发挥对GA小鼠的治疗作用。

To investigate the effect of puerarin on the expression of inflammatory cytokines in gouty arthritis(GA) mice, total of 30 mice were recruited and randomly divided into sham group, model group and puerarin group,with 10 mice in each group. Monosodium urate was introduced to construct the GA model in mice, and the mice in Puerarin group were given 100 mg/kg puerarin injection for 5 days after modeling. The joint swelling degree,dysfunction index and the pathological changes of synovial tissues were measured. The changes of UA and NO contents in serum, IL-1β, IL-10 and TNF-α contents in synovial joint fluid, and p-AMPKα, Arginase-1, NOS2,NLRP3 proteins and mRNAs expression levels were detected by various methods. Data showed that puerarin effectively reduced the joint swelling degree and hindlimb dysfunction index in puerarin group and alleviated the pathological changes of joint synovial tissues. ELISA showed that when compared with the model group, the serum UA content in puerarin group was significantly lower, the content of NO was increased, the content of IL-1β, TNF-αin joint tissue fluid were decreased, and the expression of IL-10 was increased. The number of WBC, neutrophils and lymphocytes in the puerarin group were significantly lower than those in model group. Western blot showed that the expression of p-AMPKα and Arginase-1 proteins in joint tissues were increased in puerarin group while the expression of NOS2 and NLRP3 protein were decreased, as compared with the model group;qRT-PCR showed that the expression of Arginase-1, NOS2 and NLRP3 mRNA were consistent with the trend of protein expression. In conclusion, puerarin may exerts a therapeutic effect on GA mice by regulating the expression levels of inflammatory cytokines.