Wu Weijing, Zhang Jiamin, Su Xiaoshan, Lin Xiaoping, Zhu Li, Zhuang Zesen, Liu Chennan, Zhu Zhixing, Zeng Yiming
Department of Pulmonary and Critical Care Medicine, Respirology Medicine Center of Fujian Province, the Second Affiliated Hospital of Fujian Medical University, Quanzhou, China.
Department of Radiology, Fujian Medical University Affiliated First Quanzhou Hospital, Quanzhou, China.
Allergol Immunopathol (Madr). 2023 Jan 1;51(1):54-62. doi: 10.15586/aei.v51i1.732. eCollection 2023.
Acute lung injury causes severe inflammation and oxidative stress in lung tissues. In this study, we analyzed the potential regulatory role of nuclear factor erythroid-2-related factor 2 (Nrf2) on NADPH oxidase 1 (NOX1) in tumor necrosis factor-α (TNF-α)-induced inflammation and oxidative stress in human type II alveolar epithelial cells. In this study, A549 cells were transfected with Nrf2 siRNA and overexpression vectors for 6 h before being induced by TNF-α for 24 h. TNF-α upregulated the expression of NOX1 and Nrf2 in A549 cells. Furthermore, overexpression of Nrf2 could reduce TNF-α-induced NF-κB mRNA and protein expression after transfection with the Nrf2 siRNA vector, and the levels of IL-6, IL-8, ROS, and malondialdehyde (MDA) in TNF-α-induced A549 cells increased, while the level of total antioxidation capability (T-AOC) decreased. On the other hand, the overexpression of Nrf2 decreased the levels of IL-6, IL-8, ROS, and MDA, while increasing T-AOC. The mRNA and protein levels of NOX1 were dramatically increased by TNF-α, while those changes were notably suppressed by Nrf2 overexpression. Further studies demonstrated that Nrf2 suppressed NOX1 transcription by binding to the -1199 to -1189 bp (ATTACACAGCA) region of the NOX1 promoter in TNF-α-stimulated A549 cells. Our study suggests that Nrf2 may bind to and regulate NOX1 expression to antagonize TNF-α-induced inflammatory reaction and oxidative stress in A549 cells.
急性肺损伤会导致肺组织严重炎症和氧化应激。在本研究中，我们分析了核因子红细胞2相关因子2（Nrf2）对烟酰胺腺嘌呤二核苷酸磷酸氧化酶1（NOX1）在肿瘤坏死因子-α（TNF-α）诱导的人II型肺泡上皮细胞炎症和氧化应激中的潜在调控作用。在本研究中，A549细胞在被TNF-α诱导24小时前，用Nrf2小干扰RNA（siRNA）和过表达载体转染6小时。TNF-α上调了A549细胞中NOX1和Nrf2的表达。此外，在用Nrf2 siRNA载体转染后，Nrf2的过表达可降低TNF-α诱导的核因子κB（NF-κB）mRNA和蛋白表达，且TNF-α诱导的A549细胞中白细胞介素-6（IL-6）、白细胞介素-8（IL-8）、活性氧（ROS）和丙二醛（MDA）水平升高，而总抗氧化能力（T-AOC）水平降低。另一方面，Nrf2的过表达降低了IL-6、IL-8、ROS和MDA水平，同时提高了T-AOC。TNF-α显著增加了NOX1的mRNA和蛋白水平，而这些变化被Nrf2过表达显著抑制。进一步研究表明，在TNF-α刺激的A549细胞中，Nrf2通过与NOX1启动子的-1199至-1189碱基对（ATTACACAGCA）区域结合来抑制NOX1转录。我们的研究表明，Nrf2可能通过结合并调节NOX1表达来拮抗TNF-α诱导的A549细胞炎症反应和氧化应激。
