【摘要】 目的研究叔丁基对苯二酚(tert-butylhydroquinone,tBHQ)对高糖培养的小鼠肾小球系膜细胞株(mouse mesang-ial cells,MMCs)核因子NF-E2相关因子(nuclear factor ery-throid 2-related factor 2,Nrf2)表达及氧化应激的影响,探讨tBHQ保护糖尿病肾脏的部分机制。方法 MMCs随机分为4组,正常糖对照组(C组)、高糖组(HG组)、高糖+溶剂对照组(HG+D组)、高糖+tBHQ预处理组(HG+t组),于高糖刺激48 h后收集细胞及上清液,2’7’-二氯荧光素探针(2’,7’-Dichlorofluorescin diacetate,2.7-DCFH-DA)检测细胞内活性氧族(reactive oxygen species,ROS);硫代巴比妥法检测细胞内丙二醛(malondialdehyd,MDA)浓度;流式细胞学检测细胞增殖;酶联免疫吸附试验(enzyme-linked immunosor-bent assay,ELISA)检测细胞上清液转化生长因子-β1(trans-forming growth factor-β1,TGF-β1)水平;Western blot及反转录-多聚酶链反应(reverse transcription polymerse chain reac-tion,RT-PCR)检测系膜细胞Nrf2、血红素氧化酶-1(hemeoxygenase-1,HO-1)、γ-谷氨酰半胱氨酸合成酶(γ-glutamyl-cysteine synthethase,γ-GCS)的表达水平。结果①高糖刺激使MMCs的ROS、MDA、细胞增殖及细胞上清液TGF-β1水平明显升高,而tBHQ预处理可以使以上指标明显下降。②tBHQ预处理可以增加高糖刺激下系膜细胞内Nrf2在细胞内和细胞核的表达以及细胞内HO-1、γ-GCS蛋白和mR-NA的表达水平。结论 tBHQ减轻了高糖诱导的肾小球系膜细胞的氧化应激损伤,其作用机制可能是通过激活Nrf2-ARE信号通路,进而增强抗氧化蛋白、HO-1、γ-GCS的表达而实现的。更多还原

【Abstract】 Aim To investigate the effects of tert-bu-tylhydroquinone(tBHQ) on activation of Nrf2 and its target antioxidant genes of mouse mesangial cells(MMCs)under high glucose stimulation.Methods MMCs were randomized and cultured in medium containing 5.5 mmol·L-1 of glucose as C group;30 mmol·L-1 of glucose as HG group;30 mmol·L-1 of glucose and 0.01% DMSO as HG+D group;30 mmol·L-1 of glucose and 40 μmol·L-1 tBHQ as HG+t group for 48 h,respectively.2’7’-Dichloro-fluorescein diacetate(2’7’-DCFH-DA) and malondialdehyde(MDA)were assessed as cellular ROS generation and lipid peroxidation.Proliferation activity of mesangial cells was determined by flow cytometry;the contents of transforming growth factor-β1(TGF-β1) in the supernatants were measured by enzyme linked immunosorben assay(ELISA).Nrf2(total and nucleus),Heme oxygenase-1(HO-1),γ-glutamylcysteine synthethase(γ-GCS) protein and mRNA were analysed by Western-blot and Reverse transcription polymerse chain reaction(RT-PCR).Results ]①In MMCs,high glucose induced ROS production,lipid peroxidation,cell proliferation and TGF-β1 secretion compared with those of control cells.All these parameters were markedly reduced in tBHQ pretrated cells.②The expressions of Nrf2,HO-1 and γ-GCS were up-regulated in MMCs pretreated with tBHQ and then cultured in HG than those of HG group.Conclusion Treatment with tBHQ protects against hyperglycemia-induced oxidative stress damages by enhancing Nrf2,and downstream antioxidant HO-1 and g-GCS expression in mouse mesangial cells.更多还原