Effect of NaNO<sub>2</sub> on oxidative damage in house dust mite pretreated airway epithelial cells and its underlying mechanism

Xixuan LI; Donghong PENG

doi:10.16016/j.2097-0927.202401018

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Effect of NaNO₂ on oxidative damage in house dust mite pretreated airway epithelial cells and its underlying mechanism

Basic Medicine | 更新时间：2026-01-09

- Effect of NaNO₂ on oxidative damage in house dust mite pretreated airway epithelial cells and its underlying mechanism
- Journal of Army Medical University Vol. 46, Issue 12, Pages: 1395-1402(2024)
- 作者机构：
  
  1.400014 重庆，重庆医科大学附属儿童医院呼吸内科，国家儿童健康与疾病临床医学研究中心，儿童发育疾病研究教育部重点实验室，儿童发育重大疾病国家国际科技合作基地，儿童代谢与炎症性疾病重庆市重点实验室
  2.330108 南昌，重庆医科大学附属儿童医院江西医院
- 作者简介：
- 基金信息：
- DOI：10.16016/j.2097-0927.202401018
  CLC： R363.21;R562.25;X511
- Received：04 January 2024，
  
  Revised：2024-02-16，
  
  Published：30 June 2024
- 稿件说明：
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Xixuan LI, Donghong PENG. Effect of NaNO₂ on oxidative damage in house dust mite pretreated airway epithelial cells and its underlying mechanism[J]. Journal of Army Medical University, 2024, 46(12): 1395-1402.
DOI：

Xixuan LI, Donghong PENG. Effect of NaNO₂ on oxidative damage in house dust mite pretreated airway epithelial cells and its underlying mechanism[J]. Journal of Army Medical University, 2024, 46(12): 1395-1402. DOI： 10.16016/j.2097-0927.202401018.

摘要

目的

探究NaNO

对经屋尘螨(house dust mite

HDM)预处理的人支气管上皮细胞氧化损伤的影响及其机制。

方法

体外培养人支气管上皮细胞16HBE

经HDM预处理后用不同浓度NaNO

染毒处理

CCK-8检测细胞存活率

确定NaNO

最适浓度及处理时间; 将细胞接种于6孔板中随机分为3组(

=3): 正常对照组、HDM组(HDM预处理)、NaNO

组(HDM预处理后进行NaNO

染毒处理)。Western blot和RT-PCR检测各组细胞Krüppel样锌指转录因子2(Krüppel-like transcription factors 2

KLF2)、低氧诱导因子-1α(hypoxia inducible factor-1α

HIF-1α)和Notch1的表达水平; ELISA法检测细胞炎症因子白介素-6(interleukin-6

IL-6)和肿瘤坏死因子-α(tumor necrosis factor

TNF-α)的表达水平; 荧光探针法检测细胞活性氧(reactive oxygen species

ROS)水平; 微量法测定丙二醛(malondialdehyde

MDA)与谷胱甘肽(glutathione

GSH)含量。

结果

CCK-8实验确定NaNO

最适干预方式为500 μmol/L处理24 h。与正常对照组比较

HDM组KLF2蛋白和mRNA水平减少

HIF-1α蛋白和mRNA水平(

0.01)、Notch1的蛋白(

0.05)、mRNA水平和ROS含量升高(

0.01)

MDA、TNF-α和IL-6含量均增高

GSH含量降低(

0.05);与HDM组比较

NaNO

组KLF2蛋白和mRNA水平明显减少

HIF-1α蛋白和mRNA水平(

0.01)、Notch1的蛋白(

0.05)和mRNA水平、ROS含量显著升高(

0.01)

MDA (

0.05)、TNF-α(

0.001)和IL-6含量显著增高

GSH含量明显减少(

0.05)。

结论

NaNO

可能通过调控KLF2的表达加重HDM诱导的人支气管上皮细胞氧化应激损伤。

Abstract

Objective

To investigate the effects and underlying mechanisms of NaNO

on oxidative damage in human bronchial epithelial cells pretreated with house dust mite (HDM).

Methods

Human bronchial epithelial cell line 16HBE were pretreated with HDM and then exposed to varying concentrations of NaNO

.Cell viability was assessed using CCK-8 assay to determine the optimal concentration and treatment duration of NaNO

.Then 16HBE cells were randomly divided into normal control group

HDM group (pretreated with HDM)

and NaNO

group (pretreated with HDM followed by NaNO

exposure).The expression of Krüppel-like transcription factors 2(KLF2)

hypoxia-inducible factor-1α(HIF-1α)

and Notch1 at protein and mRNA levels in each group were measured by Western blotting and RT-PCR

respectively.The contents of the inflammatory cytokines IL-6 and TNF-α were assessed using ELISA.The production of reactive oxygen species (ROS) were measured using a fluorescence probe detection.The contents of malondialdehyde (MDA) and glutathione (GSH) were quantified by micro-methods.

Results

CCK-8 assay determined that the optimal intervention of NaNO

was 500 μmol/L for 24 h.Compared with the normal control group

the HDM group showed reduced protein and mRNA expression of KLF2

but increased protein and mRNA (

0.01) expression of HIF-1α

and Notch1 at protein (

0.05) and mRNA levels

elevated production of ROS (

0.01)

and increased contents of MDA

TNF-α

and IL-6

with a decrease in GSH content (

0.05).Compared with the HDM group

the NaNO

group exhibited a significant reduction in KLF2 protein and mRNA levels

increases in HIF-1α protein and mRNA levels

Notch1 protein (

0.05) and mRNA levels

elevated ROS content (

0.01)

and significantly increased contents of MDA

TNF-α(

0.001)

and IL-6

with a notable decrease in GSH content (

0.05).

Conclusion

NaNO

may exacerbate HDM-induced oxidative stress injury in human bronchial epithelial cells by regulating the expression of KLF2.

关键词

Keywords

references

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⁰

Effect of NaNO2 on oxidative damage in house dust mite pretreated airway epithelial cells and its underlying mechanism

DOI：10.16016/j.2097-0927.202401018

摘要

Abstract

关键词

Keywords

references

Effect of NaNO₂ on oxidative damage in house dust mite pretreated airway epithelial cells and its underlying mechanism