苏萸痛风方通过ROS/TXNIP/NLRP3信号通路抗痛风性关节炎的作用机制

Mechanism of Suyu tongfeng prescription in the treatment of gouty arthritis through ROS/TXNIP/NLRP3 signaling pathway

目的 研究苏萸痛风方抗痛风性关节炎的作用机制。方法 将雄性SD大鼠按体质量随机分为正常对照组、模型组、秋水仙碱片组(阳性对照药,0.3 mg/kg)和苏萸痛风方高、中、低剂量组(5、2.5、1.25 g/kg),每组10只。给药组大鼠每日灌胃相应药物1次(10 mL/kg),连续给药7 d。正常对照组和模型组大鼠同法灌胃等体积水。第6天给药后1 h,除正常对照组外,其余各组大鼠于关节腔注入尿酸钠复制痛风性关节炎模型。检测大鼠造模后2、6、24 h的关节肿胀度和炎症指数评分。末次给药后1 h,检测大鼠血清中氧化应激相关指标[超氧化物歧化酶(SOD)、黄嘌呤氧化酶(XOD)、丙二醛(MDA)]活性及炎症因子[白细胞介素1β(IL-1β)、IL-18、肿瘤坏死因子α(TNF-α)]水平,观察各组大鼠踝关节组织病理学变化,检测大鼠踝关节组织中硫氧还蛋白互作蛋白(TXNIP)、NOD样受体热蛋白结构域相关蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)的表达水平。结果 经苏萸痛风方干预后,大鼠关节肿胀度减轻、炎症指数评分降低、滑膜组织水肿改善、炎症细胞数量减少。苏萸痛风方高剂量组大鼠血清中SOD活性显著升高(P<0.01),XOD、MDA活性以及IL-1β、IL-18、TNF-α水平均显著降低(P<0.01),踝关节组织中ROS水平和TXNIP、NLRP3、ASC蛋白表达水平均显著降低(P<0.05或P<0.01)。苏萸痛风方中、低剂量组上述部分指标活性/水平也发生显著逆转(P<0.05或P<0.01)。结论 苏萸痛风方可通过ROS/TXNIP/NLRP3信号通路,抑制NLRP3炎性小体激活,进而发挥抗痛风性关节炎的作用。

OBJECTIVE To study the mechanism of Suyu tongfeng prescription against gouty arthritis.METHODS Male SD rats were randomly divided into normal control group,model group,colchicine tablets group(positive control drug,0.3 mg/kg)and high-dose,medium-dose and low-dose groups(5,2.5,1.25 g/kg)of Suyu tongfeng prescription according to body mass,with10 rats in each group.The rats in the administration group were orally given the corresponding drugs once a day(10 mL/kg)for 7consecutive days.Rats in the normal control group and model group were given equal volume of water intragastrically.On the 6th day,1 h after administration,except for normal control group,the rats in other groups were injected with sodium urate into the joint to replicate the gouty arthritis model.The degree of joint swelling and the score of inflammatory indexes were measured at 2,6 and 24 h after the establishment of the model.One hour after the last administration,the activity of oxidative stress related indicators [superoxide dismutase(SOD),xanthine oxidase(XOD),malondialdehyde(MDA)] and the levels of inflammatory factors [interleukin 1β(IL-1β),IL-18,tumor necrosis factor α(TNF-α)] were detected in the serum.The histopathological changes of ankle joints in each group were observed;the expressions level of thioredoxin interacting protein(TXNIP),NOD like receptor thermoprotein domain related protein 3(NLRP3),apoptosis related spot like protein(ASC) in the ankle joint of rats were detected.RESULTS After intervened with Suyu tongfeng prescription,the swelling degree of joints,the score of inflammation indexes,the edema of synovial tissue and the amount of inflammatory cells were reduced.The activity of SOD in Suyu tongfeng prescription high-dose group was increased significantly(P<0.01),while the activities of XOD and MDA as well as the levels of IL-1β,IL-18 and TNF-α were all decreased significantly(P<0.01).The level of ROS and the protein expressions of TXNIP,NLRP3 and ASC in ankle joint were all decreased significantly(P<0.05 or P<0.01).The activities/levels of the above indexes were also significantly reversed in the middle-dose and low-dose groups of Suyu tongfeng prescription(P<0.05 or P<0.01).CONCLUSIONS Suyu tongfeng prescription can inhibit the activation of NLRP3 inflammasome through ROS/TXNIP/NLRP3 signaling pathway,and then play the role of anti-gouty arthritis.