Department of Neonatology, The First Affiliated Hospital of Shaoyang University, No. 39, Tongheng Street, Shuangqing District, Shaoyang City, 422000, Hunan Province, China.
Mol Biotechnol. 2024 Aug;66(8):2078-2086. doi: 10.1007/s12033-023-00849-1. Epub 2023 Aug 16.
miR-146b-5p has been studied to be highly expressed in bronchopulmonary dysplasia (BPD), but whether it is involved in regulating the process of BPD in premature infants remains unclear. This study was to explore miR-146b-5p in premature BPD and reveal its molecular mechanism. BPD mouse model and high-oxygen MLE-12 cell model were established. HE staining, TUNEL staining, and IF staining were conducted to evaluate the pathological injury and protein expression in mouse lung tissue. LDH assay, MMT assay, and flow cytometry were achieved to evaluate cytotoxicity, cell viability, and apoptosis. ELISA and immunoblotting were performed to evaluate inflammatory cytokines and Wnt pathway proteins in lung tissues and cells. Dual-luciferase reporter assay and RIP assay were needed to examine the targeting relationship between miR-146b-5p and KDM6B. miR-146b-5p was abundantly expressed in BPD and KDM6B was lowly expressed. miR-146b-5p knockdown improved hyperoxia-induced lung epithelial cell inflammation and apoptosis in both models. miR-146b-6p upregulation or KDM6B downregulation aggravated hyperoxia-induced inflammation and apoptosis of lung epithelial cells. This effect of overexpressing miR-146b-5p was rescued by forcing KDM6B. MiR-146b-5p activated Wnt signaling by regulating KDM6B. miR-146b-5p activates the Wnt pathway through targeted regulation of KDM6B, thereby aggravating hyperoxia-induced inflammation and apoptosis of lung epithelial cells.
miR-146b-5p 在支气管肺发育不良（BPD）中被研究为高度表达，但它是否参与调节早产儿的 BPD 过程尚不清楚。本研究旨在探讨 miR-146b-5p 在早产儿 BPD 中的作用，并揭示其分子机制。建立了 BPD 小鼠模型和高氧 MLE-12 细胞模型。进行 HE 染色、TUNEL 染色和 IF 染色评估小鼠肺组织的病理损伤和蛋白表达。进行 LDH 测定、MMT 测定和流式细胞术评估细胞毒性、细胞活力和细胞凋亡。进行 ELISA 和免疫印迹法评估肺组织和细胞中的炎症细胞因子和 Wnt 通路蛋白。需要进行双荧光素酶报告基因检测和 RIP 实验来检测 miR-146b-5p 和 KDM6B 之间的靶向关系。miR-146b-5p 在 BPD 中大量表达，而 KDM6B 表达水平较低。miR-146b-5p 敲低改善了两种模型中高氧诱导的肺上皮细胞炎症和凋亡。miR-146b-6p 的上调或 KDM6B 的下调加重了高氧诱导的肺上皮细胞炎症和凋亡。过表达 miR-146b-5p 的这种作用通过强制表达 KDM6B 得到挽救。miR-146b-5p 通过调节 KDM6B 激活 Wnt 信号通路。miR-146b-5p 通过靶向调节 KDM6B 激活 Wnt 通路，从而加重高氧诱导的肺上皮细胞炎症和凋亡。
