Li Nan, Fang DeYu, Ge Feng, Zhang Lin, Liu Ying, Gao Yan, Jin HongXu
Department of Emergency Medicine, General Hospital of Northern Theater Command, Shenyang City, Liaoning Province, China.
Department of Chemistry, School of Information Engineering, Liaoning University of Traditional Chinese Medicine, Shenyang City, Liaoning Province, China.
J Biochem Mol Toxicol. 2025 May;39(5):e70241. doi: 10.1002/jbt.70241.
Exposure of the lungs to high O2 levels, can lead to a noninfectious lung damage known as hyperoxia-induced lung injury (HILI). Melatonin stimulation can enhance the efficacy of stem cells in some diseases. This study aims to investigate the mechanism of exosomes secreted by mesenchymal stem cells (MSCs) stimulated by melatonin in HILI. The MSCs-derived exosomes were isolated and identified after stimulation with melatonin, and the neonatal rat model of HILI was constructed. After injection of exosomes and related lentiviruses, the ratio of wet lung to dry lung was calculated to evaluate pulmonary edema. Inflammatory factors in medium or serum were measured by ELISA. HE staining was used to evaluate the pathological status of lung tissue. Masson staining was used to evaluate collagen deposition in lung tissue. Lung cell apoptosis was detected by Tunel staining. In vitro model of HILI was established, CCK-8 and EDU staining were used to detect cell viability and proliferation, and flow cytometry was used to detect cell apoptosis. The binding relationship between LINC00052, miR-152-3p, and KLF4 was verified through bioinformatics websites, dual luciferase reporter experiments, RIP experiments, and RNA pull down experiments. Melatonin-stimulated MSCs-derived exosomes could alleviate HILI. Exosomes had a therapeutic effect on HILI neonatal rats by carrying LINC00052. Inhibition of LINC00052 reversed the therapeutic effect of exosomes on HILI, while low expression of miR-152-3p or inducing KLF4 negated the effect of sh-LINC00052. LINC00052 bound to miR-152-3p. miR-152-3p targeted KLF4. In vitro, melatonin-stimulated MSC-derived exosomes alleviated the cytotoxicity and cell viability inhibition of AEC-II cells induced by hyperoxia. KLF4 overexpression activated NRF2 signaling in AEC-II cells. LINC00052 in MSCs-derived exosomes stimulated by melatonin activates the Nrf2 pathway through the miR-152-3p/KLF4 axis to alleviate HILI, which may be a potential therapeutic approach for HILI.
肺部暴露于高氧水平会导致一种称为高氧诱导性肺损伤(HILI)的非感染性肺损伤。褪黑素刺激可增强干细胞在某些疾病中的疗效。本研究旨在探讨褪黑素刺激的间充质干细胞(MSC)分泌的外泌体在HILI中的作用机制。用褪黑素刺激后分离并鉴定MSC来源的外泌体,并构建HILI新生大鼠模型。注射外泌体和相关慢病毒后,计算湿肺与干肺的比值以评估肺水肿。通过ELISA检测培养基或血清中的炎症因子。用HE染色评估肺组织的病理状态。用Masson染色评估肺组织中的胶原沉积。用Tunel染色检测肺细胞凋亡。建立HILI体外模型,用CCK-8和EDU染色检测细胞活力和增殖,用流式细胞术检测细胞凋亡。通过生物信息学网站、双荧光素酶报告实验、RIP实验和RNA下拉实验验证LINC00052、miR-152-3 p和KLF4之间的结合关系。褪黑素刺激的MSC来源的外泌体可减轻HILI。外泌体通过携带LINC00052对HILI新生大鼠具有治疗作用。抑制LINC00052可逆转外泌体对HILI的治疗作用,而miR-152-3 p的低表达或诱导KLF4可消除sh-LINC00052的作用。LINC00052与miR-152-3 p结合。miR-152-3 p靶向KLF4。在体外,褪黑素刺激的MSC来源的外泌体减轻了高氧诱导的AEC-II细胞的细胞毒性和细胞活力抑制。KLF4过表达激活了AEC-II细胞中的NRF2信号通路。褪黑素刺激的MSC来源的外泌体中的LINC00052通过miR-152-3 p/KLF4轴激活Nrf2途径以减轻HILI,这可能是HILI的一种潜在治疗方法。