叔丁基对苯二酚对高糖环境下大鼠视网膜Müller细胞的抗氧化应激作用及其机制  

Antioxidative effect and mechanism of tert-butyl hydroquinone on rat retinal Müller cells under high glucose condition

作　　者：何薇 田敏 余曦 曹阳 郑文丽 吕红彬 He Wei;Tian Min;Yu Xi;Cao Yang;Zheng Wenli;Lyu Hongbin(Department of Ophthalmology,the People's Hospital of Chengdu Wenjiang District,Chengdu 611130,China;Department of Ophthalmology,the Affiliated Hospital of Southwest Medical University,Luzhou 646099,China;Department of Ophthalmology,The Second People's Hospital of Luzhou;The Affiliated Hospital Eye Hospital of Panzhihua College,Panzhihua 617000,China)

机构地区：[1]成都市温江区人民医院眼科,611130 [2]西南医科大学附属医院眼科,泸州646099 [3]泸州市第二人民医院眼科,646100 [4]攀枝花学院附属医院眼科医院,617000

出　　处：《中华实验眼科杂志》2020年第1期16-22,共7页Chinese Journal Of Experimental Ophthalmology

基　　金：四川省科技支撑计划项目(2015SZ0086)。

摘　　要：目的研究叔丁基对苯二酚(tBHQ)对高糖环境下SD大鼠视网膜Müller细胞氧化应激的影响及其机制。方法体外培养SD大鼠视网膜Müller细胞。将实验分为正常对照组、高糖组和tBHQ干预组,采用Western blot法和实时荧光定量PCR测定各组Müller细胞核因子-E2相关因子(Nrf2)、血红素氧化酶1(HO-1)、缺氧诱导因子1α(HIF-1α)和血管内皮生长因子(VEGF)蛋白及其mRNA的相对表达量。结果体外培养Müller细胞的细胞体扁平不规则,细胞核呈卵圆形,细胞质丰富,相邻细胞交织形成网状;Western blot法检测结果显示,正常对照组、高糖组和tBHQ干预组Müller细胞中Nrf2、HO-1、HIF-1α和VEGF的蛋白相对表达量总体比较,差异均有统计学意义(F=73.831、148.618、152.269、91.217,均P<0.001);其中高糖组Nrf2、HO-1、HIF-1α和VEGF蛋白相对表达量分别是0.17±0.02、0.47±0.02、0.67±0.07和0.60±0.05,较正常对照组的0.06±0.01、0.19±0.03、0.06±0.00和0.07±0.02明显增加,差异均有统计学意义(t=4.114、9.275、16.479、13.353,均P<0.001);tBHQ干预组Nrf2、HO-1蛋白表达量分别为0.40±0.06、0.72±0.05,较高糖组增加,差异均有统计学意义(t=7.847、7.947,均P<0.001);tBHQ干预组HIF-1α、VEGF蛋白表达量分别为0.18±0.04、0.26±0.07,较高糖组降低,但较正常对照组增加,差异均有统计学意义(t=13.215、8.444,均P=0.000)。实时荧光定量PCR测定结果显示,正常对照组、高糖组和tBHQ干预组Müller细胞中Nrf2、HO-1、HIF-1α和VEGF mRNA相对表达量总体比较,差异均有统计学意义(F=340.317、1582.911、488.852、185.699,均P<0.001);其中高糖组Nrf2、HO-1、HIF-1α和VEGF mRNA相对表达量分别是1.53±0.06、1.50±0.04、2.56±0.09和3.04±0.19,较正常对照组的1.07±0.07、0.95±0.05、0.99±0.02和1.09±0.08明显增加,差异均有统计学意义(t=7.292、15.014、30.550、18.573,均P<0.001);tBHQ干预组Nrf2、HO-1mRNA相对表达量分别为2.68±0.09、2.94±0.05,较高糖组增Objective To expore the impact of teniary butyl hydroquinone(tBHQ)on Müller cells in SD rats retina under high glucose condition,and discuss the mechanism of tBHQ.Methods The Müller cells of SD rats were cultured in vitro and the experiment was divided into normal control group,high glucose group and tBHQ intervention group.Western blot and quantitative real time PCR were used to determine the expression of nuclear factor erythroid 2-related factor 2(Nrf2),hemeoxygenase-1(HO-1),hypoxia inducible factor-1α(HIF-1α)and vascular endothelial growth factor(VEGF)protein and mRNA in each group.Results Müller cells cultured in vitro were flat with irregularly sharp.The nucleus was oval,while the cytoplasm was abundant.Adjacent cells were interwoven to a network.Western blot assay showed the overall expression of Nrf2,HO-1,HIF-1α,and VEGF in Müller cells of normal control group,high glucose group,and tBHQ intervention group were significantly different(F=73.831,148.618,152.269,91.217,all at P<0.001);Among them,the relative expressions of Nrf2,HO-1,HIF-1αand VEGF proteins in the high glucose group were 0.17±0.02,0.47±0.02,0.67±0.07,and 0.6±0.05,which were increased in comparion with 0.06±0.01,0.19±0.03,0.06±0.00 and 0.07±0.02 in the normal control group,with statistically significant differences(t=4.114,9.275,16.479,13.353,all at P<0.001);the relative expressions of Nrf2 and HO-1 proteins in the tBHQ intervention group were 0.40±0.06 and 0.72±0.05,which were increased by comparion with those in the higher glucose group,with statistically significant differences(t=7.847,7.947,both at P<0.001);the relative expressions of HIF-1αand VEGF proteins in the tBHQ intervention group were 0.18±0.04,0.26±0.07,which were decreased in comparion with those in the higher glucose group,but were increased in comparion with those in normal control group,with statistically significant differences(t=13.215,8.444,both at P<0.001).Quantitative real time PCR showed that the relative mRNA expressions of Nrf2,HO-1,HIF-1α,and VE

关 键 词：糖尿病视网膜病变 MÜLLER细胞 叔丁基对苯二酚 氧化应激 保护机制 

分 类 号：R774.1[医药卫生—眼科]