You Position: Home > Paper

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

( views:8, downloads:0 )
Author:
No author available
Journal Title:
Acta Physiologica Sinica
Issue:
3
DOI:
10.13294/j.aps.2021.0084
Key Word:
核呼吸因子1;转录调控;NF-κB;急性肺损伤;肺上皮炎症

Abstract: 本文旨在研究核呼吸因子1(nuclear respiratory factor 1,NRF1)对脂多糖(lipopolysaccharide,LPS)诱导的肺上皮细胞炎症应答中关键分子核因子-κB(nuclear factor kappa B,NF-κB)的影响,以阐明NRF1对肺上皮细胞炎性应答的调控作用及机制.在体内水平上,雄性BALB/c小鼠经呼吸道转染NRF1小干扰RNA,LPS(4 mg/kg)或生理盐水经呼吸道雾化给药,48 h后取肺组织.采用免疫印迹(Western blot,WB)及real-time PCR法检测肺组织NRF1、NF-κB p65及其靶基因表达变化,免疫荧光染色法检测NRF1及NF-κB p65核转位情况.体外培养L132肺上皮细胞,转染NRF1小干扰RNA,或给予BAY 11-7082(5 μmol/L)处理24 h后,给予1 mg/LLPS刺激6h,用real-time PCR法检测NRF1、NF-κB p65及其靶基因表达变化.采用染色质免疫沉淀技术(chromatin immunoprecipitation assay,ChIP)验证NRF1的靶基因RELA(NF-κB p65编码基因).结果显示:LPS处理组小鼠肺组织或L132细胞NRF1表达水平明显上升,炎症因子p65及其磷酸化水平显著升高,靶基因白介素-1β(interleukin-1β,IL-1β)与白介素-6(IL-6)显著上升.肺组织NRF1与p65均发生明显的核转位现象.在体内水平干扰NRF1后,上述症状显著减轻.在细胞水平干扰NRF1的表达后,p65表达水平显著下调,且NF-κB抑制剂BAY 11-7082有效抑制了LPS诱导的炎症反应,但对NRF1表达无影响.ChIP实验证实,NRF1可有效结合p65编码基因RELA的启动子区.上述结果提示,NRF1有可能介导了LPS诱导的急性肺损伤,其机制为:NRF1转录调控NF-κBp65,并参与LPS诱导的肺上皮细胞炎症.

WanfangData CO.,Ltd All Rights Reserved
About WanfangData | Contact US
Healthcare Department, Fuxing Road NO.15, Haidian District Beijing, 100038 P.R.China
Tel:+86-010-58882616 Fax:+86-010-58882615 Email:yiyao@wanfangdata.com.cn