Mi Liangyu, Zhou Yuankai, Ding Wenyan, Chen Xiangyu, Yang Yingying, Wang Qianlin, Wang Lu, Su Longxiang, Long Yun
State Key Laboratory of Complex Severe and Rare Disease, Department of Critical Care Medicine Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences Beijing China.
Pulm Circ. 2025 Aug 6;15(3):e70137. doi: 10.1002/pul2.70137. eCollection 2025 Jul.
Acute lung injury (ALI) involves inflammatory cytokines and chemokines, resulting in lung and multiple organ injuries. This study explored the mechanism of mitophagy and cGAS/STING pathway in oleic acid (OA)-induced ALI. Mice and pulmonary microvascular endothelial cells were divided into four groups: control group (Con), ALI group, control group (FCon), and ALI group (F-ALI). After 24 h of modeling, proceed with tissue collection. Lung tissues were stained using hematoxylin eosin. Autophagosomes were observed by electron microscope and mtDNA was detected by qPCR. Western blot was used to analyze protein expression of pathways cGAS, STING, pTBK1, pIRF3, and pNF-κB. Serum IFN-β expression was detected by ELISA. Cellular morphological changes were observed using microscopy. LDH level, cGAS, and STING in endothelial cells were observed. Compared with control group, pathological changes in ALI group were significantly aggravated. Expressions of serum IFN-β, cGAS, STING, pTBK1, pIRF3, and pNF-κB in lung tissues of ALI mice were significantly higher than control group. After OA, the morphology of lung microvascular endothelial cells changed and LDH expression increased. After FUNDC1 gene was knocked out to inhibit mitophagy, autophagosomes were significantly reduced and mtDNA increased. Expressions of pathway proteins in lung tissues and cells of ALI group were higher than those of wild-type ALI group. Serum IFN-β expression also increased. Silencing FUNDC1 inhibits mitophagy. Subsequently, accumulated mtDNA activates cGAS/STING pathway, aggravating ALI pathological damage and inflammation, suggesting that mitophagy may provide protection in OA-induced ALI through cGAS/STING pathway.
急性肺损伤（ALI）涉及炎症细胞因子和趋化因子，可导致肺和多器官损伤。本研究探讨了线粒体自噬和cGAS/STING通路在油酸（OA）诱导的ALI中的作用机制。将小鼠和肺微血管内皮细胞分为四组：对照组（Con）、ALI组、对照组（FCon）和ALI组（F-ALI）。建模24小时后，进行组织采集。肺组织用苏木精伊红染色。通过电子显微镜观察自噬体，并用qPCR检测线粒体DNA。采用蛋白质免疫印迹法分析cGAS、STING、pTBK1、pIRF3和pNF-κB通路的蛋白表达。用酶联免疫吸附测定法检测血清IFN-β表达。用显微镜观察细胞形态变化。观察内皮细胞中的乳酸脱氢酶（LDH）水平、cGAS和STING。与对照组相比，ALI组的病理变化明显加重。ALI小鼠肺组织中血清IFN-β、cGAS、STING.pTBK1、pIRF3和pNF-κB的表达明显高于对照组。OA作用后，肺微血管内皮细胞形态改变，LDH表达增加。敲除FUNDC1基因抑制线粒体自噬后，自噬体明显减少，线粒体DNA增加。ALI组肺组织和细胞中通路蛋白的表达高于野生型ALI组。血清IFN-β表达也增加。沉默FUNDC1可抑制线粒体自噬。随后，积累的线粒体DNA激活cGAS/STING通路，加重ALI的病理损伤和炎症，提示线粒体自噬可能通过cGAS/STING通路对OA诱导的ALI起到保护作用。
