Huang Yanhui, Hong Haifa, Li Minghui, Liu Jinfen, Jiang Chuan, Zhang Haibo, Ye Lincai, Zheng Jinghao
Department of anesthesiology, Shanghai Children's Medical Center, Shanghai Jiaotong University School of Medicine, Shanghai, China.
Department of Thoracic and Cardiovascular Surgery, Shanghai Children's Medical Center, Shanghai Jiaotong University School of Medicine, Shanghai, China.
PLoS One. 2017 Jan 18;12(1):e0170351. doi: 10.1371/journal.pone.0170351. eCollection 2017.
Postnatal human cardiomyocyte proliferation declines rapidly with age, which has been suggested to be correlated with increases in oxidative DNA damage in mice and plays an important role in regulating cardiomyocyte proliferation. However, the relationship between oxidative DNA damage and age in humans is unclear.
Sixty right ventricular outflow myocardial tissue specimens were obtained from ventricular septal defect infant patients during routine congenital cardiac surgery. These specimens were divided into three groups based on age: group A (age 0-6 months), group B (age, 7-12 months), and group C (>12 months). Each tissue specimen was subjected to DNA extraction, RNA extraction, and immunofluorescence.
Immunofluorescence and qRT-PCR analysis revealed that DNA damage markers-mitochondrial DNA copy number, oxoguanine 8, and phosphorylated ataxia telangiectasia mutated-were highest in Group B. However immunofluorescence and qRT-PCR demonstrated that two cell proliferation markers, Ki67 and cyclin D2, were decreased with age. In addition, wheat germ agglutinin-staining indicated that the average size of cardiomyocytes increased with age.
Oxidative DNA damage of cardiomyocytes was not correlated positively with age in human beings. Oxidative DNA damage is unable to fully explain the reduced proliferation of human cardiomyocytes.
出生后人类心肌细胞增殖随年龄迅速下降,这被认为与小鼠氧化DNA损伤增加相关,且在调节心肌细胞增殖中起重要作用。然而,人类氧化DNA损伤与年龄之间的关系尚不清楚。
在常规先天性心脏手术期间,从室间隔缺损婴儿患者获取60份右心室流出道心肌组织标本。这些标本根据年龄分为三组:A组(0 - 6个月)、B组(7 - 12个月)和C组(>12个月)。对每个组织标本进行DNA提取、RNA提取和免疫荧光检测。
免疫荧光和qRT-PCR分析显示,DNA损伤标志物——线粒体DNA拷贝数、8-氧鸟嘌呤和磷酸化共济失调毛细血管扩张突变蛋白在B组中最高。然而,免疫荧光和qRT-PCR表明,两种细胞增殖标志物Ki67和细胞周期蛋白D2随年龄下降。此外,小麦胚凝集素染色表明心肌细胞的平均大小随年龄增加。
人类心肌细胞的氧化DNA损伤与年龄无正相关。氧化DNA损伤不能完全解释人类心肌细胞增殖减少的现象。