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目的:探讨糖肾安对肾脏的保护作用是否与激活核转录因子NF-E2相关因子(neclear factorerythroid 2-related factor 2,Nrf2)/抗氧化反应元件(antioxidant response element,ARE)通路,发挥抗氧化应激作用,减轻炎症反应有关。方法:健康雄性Wistar大鼠随机分6组,正常组、模型组、糖肾安低、中、高剂量组、厄贝沙坦组,制备含药血清;将正常组含药血清加入正常糖浓度培养的肾系膜细胞中培养,其他5组加到高糖培养的肾系膜细胞中培养;实时荧光定量聚合酶链式反应(Real-time PCR)与蛋白免疫印迹法(Western blot)检测Nrf2和谷氨酰半胱氨酸合成酶(γ-glutamylcysteinesynthetase,γ-GCS)的mRNA及蛋白表达水平;酶联免疫吸附法(ELISA)检测各组细胞上清液中白细胞介素-6(interleukin-1,IL-6)及肿瘤坏死因子-α(tumor necrosis factor-α)的含量。结果:与模型组比较,各给药组Nrf2,γ-GCS mRNA及蛋白表达显著上调(P<0.01),IL-6和TNF-α的含量下降(P<0.01)。结论:糖肾安能通过激活Nrf2/ARE通路,发挥抗氧化应激作用,减轻炎症反应,最终实现肾脏保护作用。
OBJECTIVE: To investigate whether the protective effect of Tang Shen An on kidneys is related to the activation of the Nrf2 / ARE (antioxidant factor (NF)) -related pathway (NF-E2) Stress, reduce inflammation related. Methods: Healthy male Wistar rats were randomly divided into 6 groups, normal group, model group, Tangshenan low, medium and high dose group, irbesartan group, preparation of serum containing; normal group serum containing normal glucose concentration Cultured in the mesangial cells, the other five groups were cultured in high glucose cultured mesangial cells; Real-time PCR and Western blot were used to detect the expression of Nrf2 And γ-glutamylcysteine synthetase (γ-GCS) mRNA and protein expression were detected by enzyme-linked immunosorbent assay (ELISA). The levels of interleukin- 1, IL-6) and tumor necrosis factor-α. Results: Compared with the model group, the mRNA and protein expressions of Nrf2 and γ-GCS in each group were significantly increased (P <0.01) and the contents of IL-6 and TNF-α were decreased (P <0.01). Conclusion: Tang Shen An can activate the Nrf2 / ARE pathway to exert anti-oxidative stress and reduce inflammation, and eventually achieve renal protection.