Cui Yanhui, Wang Xueqin, Lin Fengyu, Li Wen, Zhao Yuhao, Zhu Fei, Yang Hang, Rao Mingjun, Li Yi, Liang Huaying, Dai Minhui, Liu Ben, Chen Lingli, Han Duoduo, Lu Rongli, Peng Wenzhong, Zhang Yan, Song Chao, Luo Yanwei, Pan Pinhua
1Respiratory and critical care medicine, Xiangya Hospital, Central South University, Hunan 410000, China.
2Center for neuroscience and behavior, Changsha medical university, Hunan 410219, China.
Aging Dis. 2022 Jun 1;13(3):899-909. doi: 10.14336/AD.2021.1023. eCollection 2022 Jun.
Alveolar epithelial cell damage is an important determinant of the severity of acute lung injury/acute respiratory distress syndrome (ALI/ARDS). However, the molecular mechanisms of alveolar epithelial death during the development of ALI/ARDS remain unclear. In this study, we explore the role of miR-29a-3p in ALI/ARDS and its molecular mechanism. Plasma samples were collected from healthy controls and ARDS patients. Mice were intratracheally instilled with lipopolysaccharide (LPS) to establish acute lung injury. N6-adenosine (m6A) quantification, RNA-binding protein immunoprecipitation, cell viability assay, quantitative real-time polymerase chain reaction, and western blotting were performed. We found that miR-29a-3p was down-regulated in plasma of ARDS patients and lung tissue of ALI model mice, and miR-29a-3p agomir injection down-regulated the levels of the inflammatory factors, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) in the lungs, reducing alveolar epithelial cell PANoptosis as evaluated by the downregulation of Z-DNA binding protein 1 (ZBP1), gasdermin D (GSDMD), caspase-3, caspase-8, and mixed lineage kinase domain-like protein (MLKL), ultimately improving lung injury in the ALI model mice. Mechanism studies demonstrated that the knockout of methyltransferase 3 (N6-adenosine-methyltransferase complex catalytic subunit) removed the m6A modification of miR-29a-3p and reduced miR-29a-3p expression. Our findings suggest that miR-29a-3p is a potential target that can be manipulated for ALI/ARDS.
肺泡上皮细胞损伤是急性肺损伤/急性呼吸窘迫综合征(ALI/ARDS)严重程度的重要决定因素。然而,ALI/ARDS发生过程中肺泡上皮细胞死亡的分子机制仍不清楚。在本研究中,我们探讨了miR-29a-3p在ALI/ARDS中的作用及其分子机制。收集健康对照者和ARDS患者的血浆样本。通过气管内注入脂多糖(LPS)建立小鼠急性肺损伤模型。进行N6-腺苷(m6A)定量、RNA结合蛋白免疫沉淀、细胞活力测定、定量实时聚合酶链反应和蛋白质免疫印迹分析。我们发现,ARDS患者血浆和ALI模型小鼠肺组织中miR-29a-3p表达下调,注射miR-29a-3p激动剂可降低肺组织中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)等炎症因子水平,通过下调Z-DNA结合蛋白1(ZBP1)、gasdermin D(GSDMD)、半胱天冬酶-3(caspase-3)、半胱天冬酶-8(caspase-8)和混合谱系激酶结构域样蛋白(MLKL)来减少肺泡上皮细胞全程序性死亡,最终改善ALI模型小鼠的肺损伤。机制研究表明,敲除甲基转移酶3(N6-腺苷甲基转移酶复合体催化亚基)可消除miR-29a-3p的m6A修饰并降低其表达。我们的研究结果表明,miR-29a-3p是一个可用于治疗ALI/ARDS的潜在靶点。