Tong Xin, Li Danni, Liu Na, Huang Wanjie, Zhao Xinyi, Zhang Dan, Xue Xindong, Fu Jianhua
Department of Pediatrics, Shengjing Hospital of China Medical University, Shenyang, China.
Mol Med. 2023 May 24;29(1):70. doi: 10.1186/s10020-023-00660-3.
Bronchopulmonary dysplasia (BPD) is the most common and serious chronic lung disease in preterm infants with pathological characteristics of arrested lung development. DNA double-strand breaks (DSBs) are a serious manifestation of oxidative stress damage, but little is known about the role of DSBs in BPD. The current study set out to detect DSB accumulation and cell cycle arrest in BPD and study the expression of genes related to DNA damage and repair in BPD through DNA damage signaling pathway-based PCR array to determine a suitable target to improve arrested lung development associated with BPD.
DSB accumulation and cell cycle arrest were detected in a BPD animal model and primary cells, then a DNA damage signaling pathway-based PCR array was used to identify the target of DSB repair in BPD.
DSB accumulation and cell cycle arrest were shown in BPD animal model, primary type II alveolar epithelial cells (AECII) and cultured cells after exposure to hyperoxia. Of the 84 genes in the DNA damage-signaling pathway PCR array, eight genes were overexpressed and 11 genes were repressed. Rad1, an important protein for DSB repair, was repressed in the model group. Real-time PCR and western blots were used to verify the microarray results. Next, we confirmed that silencing Rad1 expression aggravated the accumulation of DSBs and cell cycle arrest in AECII cells, whereas its overexpression alleviated DSB accumulation and cell cycle arrest.
The accumulation of DSBs in AECII might be an important cause of alveolar growth arrest associated with BPD. Rad1 could be an effective target for intervention to improve this arrest in lung development associated with BPD.
支气管肺发育不良（BPD）是早产儿中最常见且最严重的慢性肺部疾病，具有肺发育停滞的病理特征。DNA双链断裂（DSB）是氧化应激损伤的一种严重表现，但关于DSB在BPD中的作用知之甚少。本研究旨在检测BPD中DSB的积累和细胞周期停滞，并通过基于DNA损伤信号通路的PCR芯片研究BPD中与DNA损伤和修复相关基因的表达，以确定改善与BPD相关的肺发育停滞的合适靶点。
在BPD动物模型和原代细胞中检测DSB积累和细胞周期停滞，然后使用基于DNA损伤信号通路的PCR芯片鉴定BPD中DSB修复的靶点。
在BPD动物模型、原代II型肺泡上皮细胞（AECII）和暴露于高氧后的培养细胞中均显示出DSB积累和细胞周期停滞。在DNA损伤信号通路PCR芯片的84个基因中，8个基因过度表达，11个基因受到抑制。Rad1是DSB修复的一种重要蛋白，在模型组中受到抑制。使用实时PCR和蛋白质免疫印迹法验证芯片结果。接下来，我们证实沉默Rad1表达会加重AECII细胞中DSB的积累和细胞周期停滞，而其过表达则会减轻DSB积累和细胞周期停滞。
AECII中DSB的积累可能是与BPD相关的肺泡生长停滞的一个重要原因。Rad1可能是改善与BPD相关的肺发育停滞的有效干预靶点。
