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-q PCR和Western blot检测DNA修复酶OGG1和APE1表达的变化。用ELISA检测DNA内8羟基脱氧鸟苷(8-OHd G)的含量变化。结果糖尿病大鼠心肌mt DNA出现明显损伤(P<0.05),n DNA无明显损伤,DNA内8-OHd G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mt DNA损伤,虽然OGG1和APE1的表达是增加的,但可能并不足以修复mt DNA的损伤,导致mt DNA的损伤累积,引起心脏功能的损伤。
Objective To identify if there was increased DNA damage in cardiomyocytes of rats with diabetic cardio- myopathy and also to evaluate the change 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 fails to repair the damage of the mtDNA. The increased mtDNA damage may contributes to myocardium damage.
出处
《基础医学与临床》
CSCD
2017年第9期1281-1285,共5页
Basic and Clinical Medicine
基金
黑龙江省青年科学基金(QC2015106)