白藜芦醇通过Nrf2/ARE信号通路降低AA大鼠脾脏中氧化应激反应

Resveratrol reduces oxidative stress in spleen of adjuvant arthritis rats through Nrf2/ARE signaling pathway

目的探讨白藜芦醇(Res)通过核因子NF-E2相关因子/抗氧化反应元件(Nrf2/ARE)通路对佐剂性关节炎(AA)大鼠脾脏氧化应激调节作用及机制。方法 SD大鼠50只,随机均分为5组:对照组、模型组、Res低剂量组、Res高剂量组和阳性药组,左侧足趾皮下注射弗氏完全佐剂(FCA)建立AA大鼠模型。12 d后,给予相应药物灌胃并测量大鼠关节炎指数及足趾肿胀度。连续灌胃12 d后处死大鼠。HE染色观察脾脏病理学改变,脾脏组织匀浆检测抗氧化能力,免疫荧光及Western blot法检测脾脏组织Nrf2、Kelch样环氧氯丙烷相关蛋白1(Keap1)、锰超氧化物歧化酶(MnSOD)蛋白表达。结果 Res处理后关节炎评分降低;HE染色显示Res处理组脾脏损伤低于模型组;氧化应激指标检测显示,Res处理组较模型组氧化应激水平减轻;免疫荧光及Western blot结果表明Res降低Nrf2的表达,升高MnSOD水平,减轻氧化应激,抑制Nrf2过度激活。结论 Res能通过Nrf2/ARE通路降低Nrf2蛋白表达,增加AA大鼠脾脏抗氧化应激蛋白的表达,减轻氧化应激。

Objective To investigate the regulatory effects and mechanism of resveratrol(Res)on oxidative stress in spleen of adjuvant arthritis(AA)rats via Nrf2/ARE pathway.Methods 50 SD rats were randomly divided into 5 groups:control group,model group,Res low dose group,Res high dose group,positive drug group,Freund’s complete adjuvant was injected subcutaneously into left toes to induce AA model.After 12 days,the index of arthritis and the degree of toe swelling in rats were measured.After 12 days of continuous gastric perfusion,the rats were sacrificed to observe the pathological change of spleen,the spleen homogenate was used to detect the antioxidant ability,the expression of Nrf2,Keap1,MnSOD protein in spleen tissue were detected by immunofluorescence and Western blot.Results The arthritis score decreased after Res treatment,HE staining showed that pathological damage of spleen in the Res treated group was less than that in the model group,the oxidative stress in the Res treated group was less than that in the model group.The results of immunofluorescence and Western blot showed that Res inhibited the expression of Nrf2,increased the level of MnSOD,alleviated oxidative stress and inhibited the excessive activation of Nrf2.Conclusion Res can reduce the expression of Nrf2 protein through Nrf2/ARE pathway,enhance the expression of antioxidant stress protein in spleen tissue and alleviate oxidative stress.