呼吸机诱导性肺损伤中半胱天冬酶-1介导的肺泡巨噬细胞焦亡对干扰素调节因子-1的调控
The Modulation of Interferon Regulatory Factor-1 via Caspase-1-Mediated Alveolar Macrophage Pyroptosis in Ventilator-Induced Lung Injury.
作者信息
Dai Minhui, Li Qian, Pan Pinhua
机构信息
Respiratory Department, Xiangya Hospital, Central South University, China.
Central Hospital, Changsha, Hunan Province, China.
出版信息
Mediators Inflamm. 2022 Apr 15;2022:1002582. doi: 10.1155/2022/1002582. eCollection 2022.
BACKGROUND
To examine the role of interferon regulatory factor-1 (IRF-1) and to explore the potential molecular mechanism in ventilator-induced lung injury.
METHODS
Wild-type C57BL/6 mice and IRF-1 gene knockout mice/caspase-1 knockout mice were mechanically ventilated with a high tidal volume to establish a ventilator-related lung injury model. The supernatant of the alveolar lavage solution and the lung tissues of these mice were collected. The degree of lung injury was examined by hematoxylin and eosin staining. The protein and mRNA expression levels of IRF-1, caspase-1 (p10), and interleukin (IL)-1 (p17) in lung tissues were measured by western blot and quantitative real-time polymerase chain reaction, respectively. Pyroptosis of alveolar macrophages was detected by flow cytometry and western blotting for active caspase-1 and cleaved GSDMD. An enzyme-linked immunosorbent assay was used to measure the levels of IL-1, IL-18, IL-6, TNF-, and high mobility group box protein 1 (HMGB-1) in alveolar lavage fluid.
RESULTS
IRF-1 expression and caspase-1-dependent pyroptosis in lung tissues of wild-type mice were significantly upregulated after mechanical ventilation with a high tidal volume. The degree of ventilator-related lung injury in IRF-1 gene knockout mice and caspase-1 knockout mice was significantly improved compared to that in wild-type mice, and the levels of GSDMD, IL-1, IL-18, IL-6, and HMGB-1 in alveolar lavage solution were significantly reduced ( < 0.05). The expression levels of caspase-1 (p10), cleaved GSDMD, and IL-1 (p17) proteins in lung tissues of IRF-1 knockout mice with ventilator-related lung injury were significantly lower than those of wild-type mice, and the level of pyroptosis of macrophages in alveolar lavage solution was significantly reduced.
CONCLUSIONS
IRF-1 may aggravate ventilator-induced lung injury by regulating the activation of caspase-1 and the focal death of alveolar macrophages.
背景
探讨干扰素调节因子-1（IRF-1）的作用，并探索机械通气诱导的肺损伤潜在分子机制。
方法
将野生型C57BL/6小鼠及IRF-1基因敲除小鼠/半胱天冬酶-1敲除小鼠用高潮气量进行机械通气，建立机械通气相关性肺损伤模型。收集这些小鼠的肺泡灌洗液上清液及肺组织。通过苏木精-伊红染色检查肺损伤程度。分别采用蛋白质免疫印迹法和定量实时聚合酶链反应检测肺组织中IRF-1、半胱天冬酶-1（p10）和白细胞介素（IL）-1（p17）的蛋白质和mRNA表达水平。通过流式细胞术及蛋白质免疫印迹法检测活性半胱天冬酶-1和裂解的Gasdermin D（GSDMD），以检测肺泡巨噬细胞的焦亡。采用酶联免疫吸附测定法检测肺泡灌洗液中IL-1、IL-18、IL-6、肿瘤坏死因子-α（TNF-α）和高迁移率族蛋白B1（HMGB-1）的水平。
结果
野生型小鼠在高潮气量机械通气后，肺组织中IRF-1表达及半胱天冬酶-1依赖性焦亡显著上调。与野生型小鼠相比，IRF-1基因敲除小鼠和半胱天冬酶-1敲除小鼠的机械通气相关性肺损伤程度显著改善，肺泡灌洗液中GSDMD、IL-1、IL-18、IL-6和HMGB-1水平显著降低（P<0.05）。患有机械通气相关性肺损伤的IRF-1基因敲除小鼠肺组织中半胱天冬酶-1（p10）、裂解的GSDMD和IL-1（p17）蛋白表达水平显著低于野生型小鼠，肺泡灌洗液中巨噬细胞焦亡水平显著降低。
结论
IRF-1可能通过调节半胱天冬酶-1的激活及肺泡巨噬细胞的焦亡加重机械通气诱导的肺损伤。
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