糖肾安对高糖环境下人肾系膜细胞Nrf2/ARE通路的影响

Effect of Tangshen'an on Nrf2/ARE Pathway in Human Renal Mesangial Cells Under High Glucose

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 explore whether the renal protective effect of Tangshen＇an is related to the activation of Nrf2/ARE pathway,the role of anti-oxidative stress and reduction of inflammatory response. Method：Male Wistar rats were randomly divided into 6 groups： normal group,model group,Tangshen＇an low,middle and high dose groups,and irbesartan group. Their drug-containing serum was prepared from each group. The serum of normal group was added into mesangial cells cultured in normal concentration of glucose,and the serum from other5 groups was respectively added into the mesangial cells cultured in high concentration of glucose. The mRNA and protein expression levels of nuclear factor E2-related factor 2（ Nrf2） and its downstream factorγ-glutamylcysteinesynthetase（ γ-GCS） were detected by Real-time polymerase chain reaction（ Real-time PCR）and Western blot respectively. The levels of interleukin-6（ IL-6） and tumor necrosis factor-α（ TNF-α） in the supernatant of each group were detected by ELISA. Result： As compared with the model group,the mRNA and protein expression levels of Nrf2 and γ-GCS were significantly up-regulated（ P〈0. 01）,and the contents of IL-6 and TNF-α were decreased（ P〈0. 01） in various treatment groups. Conclusion： Tangshen＇an can activate Nrf2/ARE pathway,decrease oxidative stress,reduce inflammation,and eventually achieve the role of renal protection.