摘要
目的:探讨萝卜硫素(SFN)对肾小管上皮细胞(HK-2)的氧化应激及核转录因子E2相关因子2(Nrf2)/血红素加氧酶(HO)-1信号通路的影响。方法:体外培养HK-2细胞,将细胞分为空白对照组、H_2O_2处理组、H_2O_2+10μmol·L^(-1)SFN组、H_2O_2+20μmol·L^(-1)SFN组和H_2O_2+40μmol·L^(-1)SFN组;测定HK-2细胞中丙二醛(MDA)、一氧化氮(NO)、谷胱甘肽(GSH)的含量及超氧化物歧化酶(SOD)的活性;RT-PCR和Western Blot法分别检测Nrf2和HO-1 mRNA及蛋白表达水平。结果:与空白对照组相比,H_2O_2处理组中MDA、NO含量明显升高,GSH水平和SOD酶活性显著降低(P<0.05);经高、中、低3种浓度的萝卜硫素处理后MDA、NO含量显著降低,GSH水平显著升高,SOD酶活性也显著升高,Nrf2和HO-1mRNA及蛋白表达明显上调(P<0.05)。结论:萝卜硫素有抗HK-2细胞氧化应激损伤的作用,其作用机制可能与激活Nrf2/HO-1信号通路有关。
Objective: To investigate the effects of sulforaphane (SFN) on oxidative stress and Nrt2 ( Nuclear factor E2-related factor 2)/HO-1 (heine oxygenase-1 ) signaling pathway in renal tubular epithelial cells (HK-2). Methods: HK-2 ceils were cultured in vitro and divided into five groups, including the control group, H2O2 group, H2O2 +10 μmol·L^-1 SFN group, H2O2 +20μmol·L^-1 SFN group and H2O2 + 40μmol·L^-1 SFN group. The levels of MDA, NO, GSH and the activity of SOD were measured. The expression levels of Nrf2, HO-1 mRNAR and proteins were analyzed using RT-PCR and Western Blot methods. Resell: The levels of MDA and NO in H2O2group significantly increased when compared with those in the control group, while the levels of GSH and the activity of SOD significantly decreased( P 〈0.05). The levels of MDA and NO in the cells after the treatment of sulforaphane at different concentrations significantly reduced when compared with those in H2O2group, while the levels of GSH and the aotivity of SOD significantly increased( P 〈0.05). The results of RT-PCR and Western Blot analysis showed that the expression levels of Nrf2, HO-1 mBNAR and proteins in the ceils after the treatment of stdforaphane at different concentrations were significantly up-regulated when compared with those in H2O2 group ( P 〈 0.05). Conclusion: Sulforaphane can strengthen the antagonism effects on oxidative stress injury in HK-2 cells and the underlying mechanism might he associated with activating Nrf2/HO-1 signal transductiou pathway.
出处
《中国药师》
CAS
2017年第5期809-812,共4页
China Pharmacist