Sueblinvong Viranuj, Fan Xian, Guo Justin, Tao Hui, Guidot David M
Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine, Department of Medicine, Emory University School of Medicine, Atlanta, Georgia, USA.
Alcohol Clin Exp Res (Hoboken). 2025 Jul;49(7):1424-1434. doi: 10.1111/acer.70088. Epub 2025 May 29.
Alcohol use disorders (AUD) increase susceptibility to lung diseases. Ethanol suppresses nuclear factor erythroid 2-related factor 2 (Nrf2), impairing pulmonary antioxidant and immune defenses. We showed that HIV-mediated Nrf2 suppression in the lung is driven by miR-144. We hypothesized that ethanol similarly suppresses Nrf2 by inducing miR-144 in the lung.
miR-144 expression was quantified in lungs from rats chronically fed an ethanol-containing diet and in rat alveolar epithelial cells (AEC), alveolar macrophages (AMs), and lung fibroblasts (PLF) treated with ethanol. The levels of the AP-1 subunits c-Fos and c-Jun, both total and phosphorylated, were quantified by western immunoblotting in rat PLF. The link between ethanol-induced AP-1 activation and miR-144 expression on Nrf2 and Nrf2-regulated antioxidants was then assessed using c-Fos silencing RNA and AP-1 inhibitors. The impact of manipulating miR-144 expression and/or activity on the expression of Nrf2 and two key Nrf2-dependent antioxidants in ethanol-treated PLF was evaluated.
miR-144 expression was increased in the lungs of chronic ethanol-fed rats, and ethanol exposure increased miR-144 expression in AEC and PLF, with a trend toward increased expression in AM. Ethanol induced both total and phosphorylated c-Fos protein and total c-Jun protein in PLF. Inhibiting AP-1 with c-Fos silencing RNA or AP-1 inhibitors blocked ethanol-induced miR-144 expression in PLF. Furthermore, RNA silencing of c-Fos or inhibiting miR-144 restored the expression of Nrf2 and the Nrf2-dependent antioxidants GCLC and NQO-1 in ethanol-treated PLF. In contrast, direct overexpression of miR-144 suppressed Nrf2, GCLC, and NQO-1, thereby reproducing the pathophysiological effects of ethanol.
Ethanol induces miR-144 expression in the lung, mediated by AP-1 activation. These steps can be implicated in the ethanol-mediated inhibition of Nrf2 and the downstream suppression of Nrf2-dependent antioxidant and immune defenses. These results suggest that miR-144 could be a novel therapeutic target to mitigate susceptibility to acute inflammatory lung diseases in individuals with AUD.
酒精使用障碍（AUD）会增加患肺部疾病的易感性。乙醇会抑制核因子红细胞2相关因子2（Nrf2），损害肺部的抗氧化和免疫防御功能。我们发现，HIV介导的肺部Nrf2抑制是由miR-144驱动的。我们推测，乙醇同样通过在肺部诱导miR-144来抑制Nrf2。
对长期喂食含乙醇饮食的大鼠的肺组织以及用乙醇处理的大鼠肺泡上皮细胞（AEC）、肺泡巨噬细胞（AM）和肺成纤维细胞（PLF）中的miR-144表达进行定量分析。通过蛋白质免疫印迹法对大鼠PLF中AP-1亚基c-Fos和c-Jun的总水平及磷酸化水平进行定量分析。然后使用c-Fos沉默RNA和AP-1抑制剂评估乙醇诱导的AP-1激活与miR-144表达对Nrf2及Nrf2调节的抗氧化剂的影响。评估了操纵miR-144表达和/或活性对乙醇处理的PLF中Nrf2及两种关键的Nrf2依赖性抗氧化剂表达的影响。
长期喂食乙醇的大鼠肺组织中miR-144表达增加，乙醇暴露使AEC和PLF中的miR-144表达增加，AM中的表达有增加趋势。乙醇诱导PLF中c-Fos蛋白的总水平和磷酸化水平以及c-Jun蛋白的总水平升高。用c-Fos沉默RNA或AP-1抑制剂抑制AP-1可阻断乙醇诱导的PLF中miR-144表达。此外，c-Fos的RNA沉默或miR-144的抑制可恢复乙醇处理的PLF中Nrf2以及Nrf2依赖性抗氧化剂GCLC和NQO-1的表达。相反，miR-144的直接过表达会抑制Nrf2、GCLC和NQO-1，从而重现乙醇的病理生理效应。
乙醇通过AP-1激活介导在肺部诱导miR-144表达。这些步骤可能与乙醇介导的Nrf2抑制以及Nrf2依赖性抗氧化和免疫防御的下游抑制有关。这些结果表明，miR-144可能是减轻AUD个体对急性炎症性肺部疾病易感性的新治疗靶点。
