Department of Neonatology, The Affiliated Wuxi Maternity and Child Health Care Hospital of Nanjing Medical University, Wuxi, China.
Translational Medicine Laboratory, Research Institute for Reproductive Health and Genetic Diseases, The Affiliated Wuxi Maternity and Child Health Care Hospital of Nanjing Medical University, Wuxi, China.
Biomed Res Int. 2021 Jan 5;2021:4051504. doi: 10.1155/2021/4051504. eCollection 2021.
Neonatal respiratory distress syndrome (NRDS) is a common disease that occurs in premature infants. However, the mechanisms underlying the disease remain unclear. microRNAs (miRNAs) have been indicated to play a crucial role in the development of NRDS. In this study, we aimed to explore the regulatory mechanisms of miR-296-5p in NRDS. The expression levels of miR-296-5p in preterm infants with NRDS were determined using quantitative reverse-transcription polymerase chain reaction (RT-qPCR). A549 cells were transfected with lentiviral vectors encoding miR-296-5p, and the transfection efficiency was determined using RT-qPCR. Flow cytometry and CCK8 assay were performed to measure apoptosis and proliferation of A549 cells, respectively. The protein levels of pulmonary surfactant SP-A (SFTPA1), SP-B, Wnt7b, and -catenin were measured using western blotting. We demonstrated an upregulation of miR-296-5p in NRDS. The miR-296-5p was successfully overexpressed in A549 cells via lentivirus transfection, and the upregulation of miR-296-5p inhibited cell proliferation and secretion of SP-A and SP-B and also induced downregulation of the Wnt7b/-catenin . Therefore, miR-296-5p inhibits cell proliferation and secretion of pulmonary surfactants in A549 cells via downregulation of Wnt7b/-catenin signaling.
新生儿呼吸窘迫综合征（NRDS）是一种常见于早产儿的疾病。然而，其发病机制尚不清楚。微小 RNA（miRNA）在 NRDS 的发生发展中起着至关重要的作用。本研究旨在探讨 miR-296-5p 在 NRDS 中的调控机制。采用实时定量逆转录聚合酶链反应（RT-qPCR）检测 NRDS 早产儿 miR-296-5p 的表达水平。通过慢病毒转染 A549 细胞，采用 RT-qPCR 检测转染效率。流式细胞术和 CCK8 法分别检测 A549 细胞凋亡和增殖。采用 Western blot 法检测肺表面活性蛋白 A（SFTPA1）、SP-B、Wnt7b 和 β-catenin 的蛋白水平。结果表明，NRDS 中 miR-296-5p 表达上调。通过慢病毒转染成功上调 A549 细胞中的 miR-296-5p，上调 miR-296-5p 抑制细胞增殖和 SP-A、SP-B 的分泌，并诱导 Wnt7b/β-catenin 下调。因此，miR-296-5p 通过下调 Wnt7b/β-catenin 信号通路抑制 A549 细胞的增殖和肺表面活性物质的分泌。
