叔丁基对苯二酚对2型糖尿病大鼠视网膜细胞的保护作用及其机制  

Protecting effects and mechanism of tert-butyl hydroquinone on retinal cells in type 2 diabetic rats

作　　者：韩佩晏 张思远 李晶艳 黄棋 田敏 吕红彬 

机构地区：[1]西南医科大学附属第一医院眼科,泸州646000

出　　处：《中华实验眼科杂志》2016年第6期496-502,共7页Chinese Journal Of Experimental Ophthalmology

基　　金：四川省科技厅项目（14JC01723-LH36、2015SZ0086）

摘　　要：背景 细胞凋亡是糖尿病视网膜病变（DR）中的重要机制之一,氧化应激、高糖等可启动细胞凋亡通路,从而造成细胞损伤.叔丁基对苯二酚（tBHQ）是一种抗氧化应激药物,但其在DR的发生和发展过程中是否发挥视网膜细胞的保护作用尚不明确. 目的 观察tBHQ对2型糖尿病大鼠视网膜血管内皮生长因子（VEGF）及Bcl-2表达的影响,探讨tBHQ通过核因子-E2相关因子2/抗氧化反应元件（Nrf2/ARE）信号通路对2型糖尿病大鼠视网膜可能的保护机制.方法 选取50只清洁级健康雄性SD大鼠.应用随机数字表法随机选取其中10只大鼠为正常对照组,以正常饲料喂养;其余40只大鼠采用高脂高糖饲料喂养4周后空腹12h,然后大鼠腹腔内注射链脲佐菌素（STZ）诱导2型糖尿病模型.造模成功大鼠均衡分组,模型对照组大鼠继续给予高脂高糖饮食,tBHQ干预组大鼠于造模后1周在高脂高糖饲料中添加质量分数1％ tBHQ进行干预,各组大鼠分别于造模后4周和12周收集大鼠心脏全血,检测各组大鼠空腹血糖（FPG）,血浆总胆固醇（TC）、总三酰甘油（TG）、高密度脂蛋白胆固醇（HDL-C）和低密度脂蛋白胆固醇（LDL-C）含量;采用放射免疫分析法检测各组大鼠空腹血清胰岛素（FINs）含量.采用免疫组织化学法检测各组大鼠视网膜中VEGF、bcl-2蛋白的表达分布;采用实时荧光定量PCR（qRT-PCR）法定量检测各组大鼠视网膜中Bcl-2 mRNA和VEGF mRNA的相对表达量. 结果 35只大鼠成功建立2型糖尿病模型.各组大鼠造模后不同时间点FINs含量总体比较差异均有统计学意义（F分组=22.480,P=0.000;F时间=7.636,P=0.008）.各组间大鼠血浆FPG、TC、TG和LDL-C含量总体比较差异均有统计学意义（FPG：F分组=78.531,P=0.000;TC：F分组=28.049,P=0.000;TG：F分组=13.108,P=0.000;LDL-C：F分组=6.804,P＜0.05）.免疫组织化学检测显示,Bcl-2和VEGF蛋白主要表达于视网膜各层的血Background Apoptosis is a primary clinical pathological mechanism of diabetic retinopathy （DR）.Oxidative stress and high glucose can activate cell apoptosis pathway and thus leads to cellular damage.It is confirmed that tert-butyl hydroquinone （tBHQ） plays an antioxidation effect,however,whether it has a protective role on retinal cells in DR is still unelucidated.Objective This study was to investigate the effect of tBHQ on vascular endothelial growth factor （VEGF） and bcl-2 expressions in retina of type 2 diabetic rats and its possible mechanism via nuclear factor erythroid 2-related factor 2/antioxidant response element （Nrf2/ARE） signal pathway.Methods Fifty clean healthy male SD rats were included in this experimental study.Ten rats were fed with normal diet as the normal control group,and other rats were fed with high fatty and high sugar food for 4 weeks.After 12 hours of fasting,streptozotoin （STZ） （30 mg/kg） was intraperitoneally injected to induce the type 2 diabetic models.The model rats were randomly divided into the diabetic control group and tBHQ group and 1% tBHQ was added into the high fatty and sugar food 1 week after modeling in the tBHQ group.Fasting plasma glucose （FPG） level,blood total cholesterol （TC） level,blood triglyceride （TG） level,high density lipoprotein-cholesterol （HDL-C）,low density lipoproteincholesterol （LDL-C） and fasting serum insulin （FINs） were detected 4 and 12 weeks after modeling,respectively,and radio immunoassay was used to detect the FIN levels of the rats.The relative expression of VEGF and bcl-2 in retinas of the rats were assayed by immunohistochemistry and fluorescence real-time quantitative PCR （qRT-PCR）.The use of the animals complied with the Regulations for the Administration of Affairs Concerning Experimental Animals by State and Technology Commission.Results Type 2 diabetic models were successfully established in 35 rats with successful rate 92.1%.The FIN levels were significantly different among different g

关 键 词：糖尿病/并发症 视网膜病变 血管内皮生长因子 Bcl-2/代谢 动物模型 SD大鼠 叔丁基对苯二酚 

分 类 号：R587.2[医药卫生—内分泌] R774.1[医药卫生—内科学]