糖尿病模型大鼠视网膜PGC-1α表达和表观遗传修饰的变化  

The alterations of PGC-1α expression and epigenetic modifications in the retina of streptozotocin-induced diabetic rats

作　　者：耿爽[1] 陈有信[1] 姚翔 徐海燕[1] 张古沐阳 夏松[1] 刘子扬[1] Geng Shuang;Chen Youxin;Yao Xiang;Xu Haiyan;Zhang Gumuyang;Xia Song;Liu Ziyang(Department of Ophthalmology,Peking Union Medical College Hospital,Peking Union Medical College,Chinese Academy of Medical Sciences,Beijing 100730,China)

机构地区：[1]中国医学科学院北京协和医院眼科,北京100730

出　　处：《中华实验眼科杂志》2018年第6期410-416,共7页Chinese Journal Of Experimental Ophthalmology

基　　金：国家自然科学基金项目（81371026）

摘　　要：目的 通过检测糖尿病模型大鼠视网膜过氧化物酶体增生物激活受体γ辅助激活因子1α（PGC-1α）mRNA和蛋白表达以及PPARGC1A启动子区DNA甲基化的改变,探讨糖尿病发生和发展过程中PGC-1α的变化趋势、表观遗传学修饰改变及其在糖尿病视网膜病变（DR）代谢记忆中的作用.方法 取清洁级6～7周龄雄性SD大鼠80只,其中60只采用腹腔内注射链脲佐菌素（STZ）的方法建立糖尿病大鼠模型.将60只糖尿病模型大鼠应用随机数字表法随机分为3个组：血糖控制不佳组大鼠造模后4个月内血糖水平控制不佳;血糖半控制组大鼠造模后2个月内血糖水平控制不佳,2个月后维持正常血糖水平;血糖控制组大鼠造模后4个月内均保持正常血糖水平,每组各20只.对照组为周龄匹配的正常饲养大鼠20只.分离各实验组大鼠视网膜组织,应用实时荧光定量PCR法检测PGC-1α及超氧化物歧化酶2（SOD2） mRNA的表达,应用Western blot法检测PGC-1α及锰超氧化物歧化酶（MnSOD）蛋白的表达,应用亚硫酸氢钠测序法（BSP）检测PPARGC1A启动子区DNA甲基化状态的变化.结果 造模后4个月,对照组大鼠体质量明显高于血糖控制不佳组、血糖半控制组和血糖控制组,血糖控制不佳组血糖水平显著高于对照组,差异均有统计学意义（均P=0.000）.血糖控制组、血糖半控制组和血糖控制不佳组大鼠视网膜组织中PGC-1 αmRNA相对表达量依次下降,均低于对照组,差异均有统计学意义（均P=0.000）;血糖控制组PGC-1αmRNA相对表达量明显高于血糖控制不佳组,差异有统计学意义（P=0.002）.血糖控制组、血糖半控制组和血糖控制不佳组大鼠视网膜组织中SOD2 mRNA表达依次增加,均高于对照组,差异均有统计学意义（P=0.006、0.000、0.000）;血糖控制组与血糖控制不佳组SOD2 mRNA相对表达量比较,差异有统计学意义（P=0.001）.血糖控制不佳组、血糖半控制组�Objective To investigate the role of epigenetic regulations of peroxisome proliferator-activated receptor γ coactivator 1α （PGC-1α） in the development of diabetic retinopathy and the metabolic memory phenomenon after hyperglycemia was terminated.Methods Diabetic rat model was established by intraperitoneal injection of streptozotocin （STZ）.Sixty diabetic rats were randomly divided into 3 groups,poor glycemic control group rats were maintained in poor glycemic control for 4 months;semi glycemic control group rats were maintained in poor glycemic control for 2 months,followed by good glycemic control for 2 additional months;good glycemic control group rats were maintained in good glycemic control for 4 months.Twenty normal rats served as control group.The mRNA expression of PGC-1α and superoxide dismutase 2 （SOD2） of retina were measured by real-time PCR;the expression of PGC-1α and manganese superoxide dismutase （MnSOD） protein were measured by Western blot;the situation of DNA methylation in the promotor region of PPARGC1A was measured by bisulfite sequencing.Results The body-weight in the control group was significantly higher than that in the poor glycemic control group,semi glycemic control group and good glycemic control group （all at P =0.000）.The blood glucose value in the poor glycemic control group was significantly higher than that in the control group （P =0.000）.The expression levels of PGC-1 α mRNA were significantly lower and the expression levels of SOD2 mRNA were significantly higher in the good glycemic control group,semi glycemic control group and poor glycemic control group than those in the control group （all at P＜0.05）.The expression levels of PGC-1α and SOD2 mRNA were significantly different between the good glycemic control group and poor glycemic control group （both at P＜0.05）.Compared with the control group,the expression levels of PGC-1α and MnSOD protein were decreased in the diabetic model groups,with significant differences between them （all

关 键 词：糖尿病视网膜病变 过氧化物酶体增生物激活受体γ辅助激活因子1α 代谢记忆 表观遗 传修饰 锰超氧化物歧化酶 

分 类 号：R-332[医药卫生] R587.2R774.1