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Nuclear respiratory factor 1 mediates LPS-induced acute lung injury through NF-κB

CHENG Kang, ZHU Li, WANG Xue-Ting^*

Institute of Special Environmental Medicine, Nantong University, Nantong 226019, China

Abstract

The purpose of this paper was to study the transcriptional regulation of nuclear respiratory factor 1 (NRF1) on nuclear factor kappa B (NF-κB), a key molecule in lipopolysaccharide (LPS)-induced lung epithelial inflammation, and to clarify the mechanism of NRF1-mediated inflammatory response in lung epithelial cells. In vivo, male BALB/c mice were treated with NRF1 siRNA, followed with LPS (4 mg/kg) or 0.9% saline through respiratory tract, and sacrificed 48 h later. Expression levels of NRF1, NF-κB p65 and its target genes were detected by Western blot and real-time PCR. Nuclear translocation of NRF1 or p65 was measured by immunofluorescent technique. In vitro, L132 cells were transfected with NRF1 siRNA or treated with BAY 11-7082 (5 μmol/L) for 24 h, followed with treatment of 1 mg/L LPS for 6 h. Cells were lysed for detections of NRF1, NF-κB p65 and its target genes as well as the binding sites of NRF1 on RELA (encoding NF-κB p65) promoter by chromatin immunoprecipitation assay (ChIP). Results showed that LPS stimulated NRF1 and NF-κB p65. Pro-inflammatory factors including interleukin-1β (IL-1β) and IL-6 were significantly increased both in vivo and in vitro. Obvious nuclear translocations of NRF1 and p65 were observed in LPS-stimulated lung tissue. Silencing NRF1 resulted in a decrease of p65 and its target genes both in vivo and in vitro. In addition, BAY 11-7082, an inhibitor of NF-κB, significantly repressed the inflammatory responses induced by LPS without affecting NRF1 expression. Furthermore, it was proved that NRF1 had three binding sites on RELA promoter region. In summary, NRF1 is involved in LPS-mediated acute lung injury through the transcriptional regulation on NF-κB p65.

Key words: nuclear respiratory factor 1; transcriptional regulation; NF-κB; acute lung injury; lung epithelial inflammation

Received: 　　Accepted:

Corresponding author: 王雪婷　　E-mail: wangxueting@ntu.edu.cn

DOI: 10.13294/j.aps.2021.0084

Citing This Article：

CHENG Kang, ZHU Li, WANG Xue-Ting. Nuclear respiratory factor 1 mediates LPS-induced acute lung injury through NF-κB. Acta Physiol Sin 2022; 74 (3): 401-410 (in Chinese with English abstract).