基础医学与临床 ›› 2017, Vol. 37 ›› Issue (9): 1281-1285.

• 研究论文 • 上一篇    下一篇

糖尿病大鼠心肌DNA损伤及DNA修复酶表达的变化

张兰,李赫,刘娴,顾金霞,王立峰,王晓云   

  1. 哈尔滨医科大学附属第四医院
  • 收稿日期:2016-04-21 修回日期:2016-11-07 出版日期:2017-09-05 发布日期:2017-08-28
  • 通讯作者: 王晓云 E-mail:wxy@medmail.com.cn

Expression changes of DNA repair enzymes and DNA damage in diabetes rat hearts

  • Received:2016-04-21 Revised:2016-11-07 Online:2017-09-05 Published:2017-08-28

摘要: 目的:探讨大鼠糖尿病心肌病变的心肌细胞DNA损伤及DNA修复酶 OGG1和 APE1表达的变化。方法:提取糖尿病大鼠心肌组织中DNA、RNA及总蛋白。用Q-PCR检测心肌DNA损伤;用RT-qPCR和Western blot检测DNA修复酶OGG1和APE1表达的变化。用ELISA法检测DNA内8羟基脱氧鸟苷(8-OHdG)的含量变化。结果:糖尿病大鼠心肌mtDNA出现明显损伤(P<0.05),nDNA无明显损伤,DNA内8-OHdG的含量增加(P<0.05),OGG1和APE1的表达增加(P<0.01)。结论:糖尿病大鼠心肌组织内出现mtDNA损伤,虽然OGG1和APE1的表达是增加的,但可能并不足以修复mtDNA的损伤,导致mtDNA的损伤累积,引起心脏功能的损伤。

关键词: 脱氧核糖核酸, 脱氧核糖核酸修复酶, 心肌, 糖尿病, 氧化损伤

Abstract: Objective To identify if there was increased DNA damage in cardiomyocytes of rats with diabetic cardiomyopathy and also to evaluate the changes in gene expression of DNA repair enzymes 8-oxoguanine DNA glycosylase 1 (OGG1) and AP endonuclease 1 (APE1). Methods Total DNA,RNA and proteins of hearts were isolated in diabetes rat hearts. DNA damage was examined with quantitative polymerase chain reaction (Q-PCR) assay. mRNA and protein expressions of OGG1 and APE1 were detected with real time-polymerase chain reaction (RT–qPCR) and Western blot analysis. The levels of 8-hydroxy-2’-deoxy-guanosine (8-OHdG) in DNA were studied with ELISA. Results The damage of mtDNA was increased in rat hearts(P<0.05) and there was not significant damage in nDNA. The amount of 8-OHdG in DNA was significantly increased(P<0.05),mRNA and protein expressions of OGG1 and APE1 were increased in diabetic rat hearts(P<0.01). Conclusions The damage of mtDNA increased in diabetic rat hearts. Although the expression of OGG1 and APE1 increased in diabetic rat hearts,which can not repair the damage of the mtDNA. The increased mtDNA damage may contributes to myocardium damage.

Key words: DNA, DNA repair enzymes, Myocardium, Diabetic, Oxidative damage