浏览全部资源
扫码关注微信
NaNO2对HDM预处理的气道上皮细胞氧化损伤的影响及其机制
Effect of NaNO2 on oxidative damage in house dust mite pretreated airway epithelial cells and its underlying mechanism
- 陆军军医大学学报 2024年46卷第12期 页码：1395-1402
- 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：
[彭东红，E-mail：pdhdxy@163.comPENG Donghong, E-mail: pdhdxy@163.com
- 基金信息：
DOI：10.16016/j.2097-0927.202401018
中图分类号： R363.21;R562.25;X511收稿：2024-01-04，
修回：2024-02-16，
纸质出版：2024-06-30
- 稿件说明：
移动端阅览
李熙炫, 彭东红. NaNO2对HDM预处理的气道上皮细胞氧化损伤的影响及其机制[J]. 陆军军医大学学报, 2024,46(12):1395-1402.
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.
李熙炫, 彭东红. NaNO2对HDM预处理的气道上皮细胞氧化损伤的影响及其机制[J]. 陆军军医大学学报, 2024,46(12):1395-1402. DOI： 10.16016/j.2097-0927.202401018.
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
2
探究NaNO
2
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
2
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
2
染毒处理
CCK-8检测细胞存活率
确定NaNO
2
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(
n
=3): 正常对照组、HDM组(HDM预处理)、NaNO
2
组(HDM预处理后进行NaNO
2
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
2
CCK-8实验确定NaNO
2
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(
P
<
0.01)、Notch1的蛋白(
P
<
0.05)、mRNA水平和ROS含量升高(
P
<
0.01)
MDA、TNF-α和IL-6含量均增高
GSH含量降低(
P
<
0.05);与HDM组比较
NaNO
2
组KLF2蛋白和mRNA水平明显减少
HIF-1α蛋白和mRNA水平(
P
<
0.01)、Notch1的蛋白(
P
<
0.05)和mRNA水平、ROS含量显著升高(
P
<
0.01)
MDA (
P
<
0.05)、TNF-α(
P
<
0.001)和IL-6含量显著增高
GSH含量明显减少(
P
<
0.05)。
结论
2
NaNO
2
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
2
To investigate the effects and underlying mechanisms of NaNO
2
on oxidative damage in human bronchial epithelial cells pretreated with house dust mite (HDM).
Methods
2
Human bronchial epithelial cell line 16HBE were pretreated with HDM and then exposed to varying concentrations of NaNO
2
.Cell viability was assessed using CCK-8 assay to determine the optimal concentration and treatment duration of NaNO
2
.Then 16HBE cells were randomly divided into normal control group
HDM group (pretreated with HDM)
and NaNO
2
group (pretreated with HDM followed by NaNO
2
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
2
CCK-8 assay determined that the optimal intervention of NaNO
2
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 (
P
<
0.01) expression of HIF-1α
and Notch1 at protein (
P
<
0.05) and mRNA levels
elevated production of ROS (
P
<
0.01)
and increased contents of MDA
TNF-α
and IL-6
with a decrease in GSH content (
P
<
0.05).Compared with the HDM group
the NaNO
2
group exhibited a significant reduction in KLF2 protein and mRNA levels
increases in HIF-1α protein and mRNA levels
Notch1 protein (
P
<
0.05) and mRNA levels
elevated ROS content (
P
<
0.01)
and significantly increased contents of MDA
TNF-α(
P
<
0.001)
and IL-6
with a notable decrease in GSH content (
P
<
0.05).
Conclusion
2
NaNO
2
may exacerbate HDM-induced oxidative stress injury in human bronchial epithelial cells by regulating the expression of KLF2.
关键词
Keywords
references
PAPI A, BRIGHTLING C, PEDERSEN S E, et al . Asthma[J ] . Lancet, 2018, 391(10122): 783-800. DOI: 10.1016/S0140-6736(17)33311-1.
YANG I V, LOZUPONE C A, SCHWARTZ D A. The environment, epigenome, and asthma[J]. J Allergy Clin Immunol, 2017, 140(1): 14-23. DOI: 10.1016/j.jaci.2017.05.011.
ACHAKULWISUT P, BRAUER M, HYSTAD P, et al . Global, national, and urban burdens of paediatric asthma incidence attributable to ambient NO 2 pollution: estimates from global datasets[J ] . Lancet Planet Health, 2019, 3(4): e166-e178. DOI: 10.1016/S2542-5196(19)30046-4..
KÜNZLI N, KAISER R, MEDINA S, et al . Public-health impact of outdoor and traffic-related air pollution: a European assessment[J ] . Lancet, 2000, 356(9232): 795-801. DOI: 10.1016/s0140-6736(00)02653-2.
POYNTER M E, PERSINGER R L, IRVIN C G, et al . Nitrogen dioxide enhances allergic airway inflammation and hyperresponsiveness in the mouse[J ] . Am J Physiol Lung Cell Mol Physiol, 2006, 290(1): L144-L152. DOI: 10.1152/ajplung.00131.2005.
PETIT P C, FINE D H, VASQUEZ G B, et al . The pathophysiology of nitrogen dioxide during inhaled nitric oxide therapy[J ] . ASAIO J, 2017, 63(1): 7-13. DOI: 10.1097/MAT.0000000000000425.
HODGKINS SR, ATHER JL, PAVEGLIO SA, et al . NO 2 inhalation induces maturation of pulmonary CD11c+ cells that promote antigenspecific CD4 + T cell polarization[J ] . Respir Res, 2010, 11(1): 102. DOI: 10.1186/1465-9921-11-102.
VAN VLIET J, CROFTS L A, QUINLAN K G R, et al . Human KLF17 is a new member of the Sp/KLF family of transcription factors[J ] . Genomics, 2006, 87(4): 474-482. DOI: 10.1016/j.ygeno.2005.12.011.
SAAVEDRA M T, PATTERSON A D, WEST J, et al . Abrogation of anti-inflammatory transcription factor LKLF in neutrophil-dominated airways[J ] . Am J Respir Cell Mol Biol, 2008, 38(6): 679-688. DOI: 10.1165/rcmb.2007-0282OC.
BOON R A, FLEDDERUS J O, VOLGER O L, et al . KLF2 suppresses TGF-beta signaling in endothelium through induction of Smad7 and inhibition of AP-1[J ] . Arterioscler Thromb Vasc Biol, 2007, 27(3): 532-539. DOI: 10.1161/01.ATV.0000256466.65450.ce.
ALI F, HAMDULAY S S, KINDERLERER A R, et al . Statin-mediated cytoprotection of human vascular endothelial cells: a role for Kruppel-like factor 2-dependent induction of heme oxygenase-1[J ] . J Thromb Haemost, 2007, 5(12): 2537-2546. DOI: 10.1111/j.1538-7836.2007.02787.x.
FLEDDERUS J O, BOON R A, VOLGER O L, et al . KLF2 primes the antioxidant transcription factor Nrf2 for activation in endothelial cells[J ] . Arterioscler Thromb Vasc Biol, 2008, 28(7): 1339-1346. DOI: 10.1161/ATVBAHA.108.165811.
GARN H, SIESE A, STUMPF S, et al . Shift toward an alternatively activated macrophage response in lungs of NO 2 - exposed rats[J ] . Am J Respir Cell Mol Biol, 2003, 28(3): 386-396. DOI: 10.1165/rcmb.4888..
INOUE H, AKIMOTO K, HOMMA T, et al . Airway epithelial dysfunction in asthma: relevant to epidermal growth factor receptors and airway epithelial cells[J ] . J Clin Med, 2020, 9(11): 3698. DOI: 10.3390/jcm9113698.
POSTLETHWAIT E M, MUSTAFA M G. Fate of inhaled nitrogen dioxide in isolated perfused rat lung[J]. J Toxicol Environ Health, 1981, 7(6): 861-872. DOI: 10.1080/15287398109530029.
杨丽丽, 王亚苹, 薛坤娇, 等. KIF3A通过结合Wnt/β-catenin通路中的β-arrestin参与哮喘气道上皮的炎症调控[J]. 第三军医大学学报, 2021, 43(6): 496-502. DOI: 10.16016/j.1000-5404.202009180.
YANG L L, WANG Y P, XUE K J, et al . Kinesin family member 3A is involved in inflammatory regulation of asthmatic airway epithelial by binding to β-arrestin in the Wnt/β-catenin pathway[J ] . J Third Mil Med Univ, 2021, 43(6): 496-502. DOI: 10.16016/j.1000-5404.202009180.
ATHARI S S. Targeting cell signaling in allergic asthma[J]. Sig Transduct Target Ther, 2019, 4: 45. DOI: 10.1038/s41392-019-0079-0.
MICHAELOUDES C, ABUBAKAR-WAZIRI H, LAKHDAR R, et al . Molecular mechanisms of oxidative stress in asthma[J ] . Mol Aspects Med, 2022, 85: 101026. DOI: 10.1016/j.mam.2021.101026.
GREGORY L G, LLOYD C M. Orchestrating house dust mite-associated allergy in the lung[J]. Trends Immunol, 2011, 32(9): 402-411. DOI: 10.1016/j.it.2011.06.006.
罗南, 王玲, 袁小平, 等. 重庆地区变应性鼻炎和哮喘患儿的吸入性变应原的异同分析[J]. 陆军军医大学学报, 2023, 45(24): 2547-2555. DOI: 10.16016/j.2097-0927.202306016.
LUO N, WANG L, YUAN X P, et al . Characteristics of inhaled all ergen spectrum between allergic rhinitis and asthma of children in Chongqing[J ] . J Army Med Univ, 2023, 45(24): 2547-2555. DOI: 10.16016/j.2097-0927.202306016.
SUN X Y, HOU T H, CHEUNG E, et al . Anti-inflammatory mechanisms of the novel cytokine interleukin-38 in allergic asthma[J ] . Cell Mol Immunol, 2020, 17(6): 631-646. DOI: 10.1038/s41423-019-0300-7.
LU C, WANG F M, LIU Q, et al . Effect of NO 2 exposure on airway inflammation and oxidative stress in asthmatic mice[J ] . J Hazard Mater, 2023, 457: 131787. DOI: 10.1016/j.jhazmat.2023.131787..
KAWANAMI D, MAHABELESHWAR G H, LIN Z Y, et al . Kruppel-like factor 2 inhibits hypoxia-inducible factor 1alpha expression and function in the endothelium[J ] . J Biol Chem, 2009, 284(31): 20522-20530. DOI: 10.1074/jbc.M109.025346.[PubMed ]
林千愉, 张慧, 梁栋, 等. 缺氧诱导因子-1α/Notch1通路在缺氧致病机制中的研究进展[J]. 上海医学, 2022, 45(7): 509-514. DOI: 10.19842/j.cnki.issn.0253-9934.2022.07.016.
LIN Q Y, ZHANG H, LIANG D, et al . Research progress of hypoxia inducible factor-1α/Notch1 pathway in the pathogenesis of hypoxia[J ] . Shanghai Med J, 2022, 45(7): 509-514. DOI: 10.19842/j.cnki.issn.0253-9934.2022.07.016.
0
浏览量
76
下载量
0
CSCD
- 网络PDF下载
- Meta-XML下载
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 技术支持由北京北大方正电子有限公司提供 京ICP备09064830号-19
京公网安备11010802024621 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies
