[1]薛坤娇,阮玲瑛,胡婕,等.糖皮质激素调控Keap1-Nrf2通路减轻哮喘小鼠肺氧化应激反应的作用研究[J].第三军医大学学报,2020,42(08):807-814.
 XUE Kunjiao,RUAN Lingying,HU Jie,et al.Glucocorticoid regulates Keap1-Nrf2 pathway to relieve pulmonary oxidative stress in asthmatic mice[J].J Third Mil Med Univ,2020,42(08):807-814.
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糖皮质激素调控Keap1-Nrf2通路减轻哮喘小鼠肺氧化应激反应的作用研究(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
42卷
期数:
2020年第08期
页码:
807-814
栏目:
基础医学
出版日期:
2020-04-30

文章信息/Info

Title:
Glucocorticoid regulates Keap1-Nrf2 pathway to relieve pulmonary oxidative stress in asthmatic mice
作者:
薛坤娇阮玲瑛胡婕应林燕邹文静符州
400014 重庆市, 重庆医科大学附属儿童医院儿科研究所,儿童发育疾病研究教育部重点实验室1,国家儿童健康与疾病临床医学中心(重庆),儿科发育重大疾病国家国际科技合作基地1;儿科学重庆市重点实验室2
Author(s):
XUE Kunjiao12 RUAN Lingying12 HU Jie12 YING Linyan1 ZOU Wenjing12 FU Zhou12

1Ministry of Education of Pediatric Research, Key Laboratory of Child Development and Disorders of Institute; National Clinical Research Center for Child Health and Disorders (Chongqing), China International Science and Technology Cooperation Base of Child development and Critical Disorders, Children’s Hospital of Chongqing Medical University, Chongqing, 400014; 2Chongqing Key Laboratory of Pediatrics, Chongqing, 400014, China 

关键词:
哮喘地塞米松氧化应激Nrf2-Keap1信号通路
Keywords:
asthma dexamethasone oxidative stress Nrf2-Keap1 pathway
分类号:
R562.25;R963;R977.1
文献标志码:
A
摘要:

目的探讨哮喘小鼠肺氧化应激水平以及地塞米松对其影响及可能的机制,为哮喘治疗提供新的方向。方法将30只6-8周龄C57BL/6雌鼠随机分为对照组、哮喘组及地塞米松组,利用屋尘螨(House dust mites, HDM)建立哮喘小鼠模型,检测哮喘激发后或地塞米松干预后,各组小鼠脂质过氧化物代谢产物丙二醛(Malondialdehyde,MDA)、氧化型谷胱甘肽(Oxidized glutathione,GSSG)的表达水平。利用qPCR检测小鼠肺部氧化应激转录因子NF-E2相关因子2(Nuclear factor erythroid 2-related factor2,Nrf2)和血红素加氧酶1(Heme oxygenase-1,HO-1)表达情况。免疫组化和免疫荧光检测小鼠肺部Nrf2蛋白表达水平。Western blot检测Nrf2-Keap1信号通路改变情况。结果哮喘组小鼠肺组织中MDA含量较对照组明显升高(P<0.01),地塞米松组MDA较哮喘组明显降低(P<0.05);哮喘小鼠的GSSG含量较对照组升高(P<0.01),地塞米松组小鼠GSSG含量较哮喘组降低(P<0.05)。qPCR结果显示哮喘组小鼠Nrf2和HO-1mRNA表达均较对照组明显增加(P<0.05), 地塞米松组小鼠Nrf2和HO-1 mRNA表达较哮喘组降低(P<0.05)。免疫组化和免疫荧光检测结果显示哮喘组小鼠肺组织中Nrf2蛋白表达较对照组明显增加(P<0.01), 地塞米松组小鼠肺组织Nrf2蛋白表达较哮喘明显减少(P<0.01)。Western blot结果显示地塞米松组肺组织Nrf2蛋白表达较哮喘组明显降低(P<0.05),Keap1蛋白表达较哮喘组明显增加(P<0.05)。结论哮喘小鼠中氧化应激水平增加,地塞米松可能通过调节 Keap1-Nrf2通路,影响下游抗氧化反应元件的表达,从而抑制哮喘小鼠肺部氧化应激水平。

Abstract:

ObjectiveTo investigate the level of oxidative stress in asthmatic mice and the mechanism of dexamethasone forrelieving oxidative stress. MethodsThirty female C57BL/6 mice were randomly divided into control group, asthma group and dexamethasone group. Following challenges with house dust mites in the latter 2 groups and prior dexamethasone treatment in dexamethasone group, the mice were examined for expression levels of lipid peroxide metabolites malondialdehyde (MDA) and oxidized glutathione (GSSG). Quantitative real-time PCR was used to detect the expression of nuclear factor erythroid 2-related factor 2 (NF-E2) and heme oxygenase 1 (HO-1) in the lung tissues. Immunohistochemistry was used to detect the expression of Nrf2 protein, and Western blotting was performed to detect the changes of Nrf2-Keap1 signaling pathway in the lungs of the mice. ResultsMDA and GSSG levels were significantly higher in the asthmatic mice than in control group (P<0.01), and were significantly lowered by dexamethasone treatment in the asthmatic mice (P<0.05). Compared with the control group, the asthmatic mice showed significantly higher pulmonary expression levels of Nrf2 and HO-1 mRNA (P<0.05), which were obviously lowered by dexamethasone treatment (P<0.05). Immunohistochemistry and immunofluorescence assay revealed a significantly higher expression of Nrf2 protein in the lung tissue of asthmatic mice (P<0.01), which was significantly lowered by dexamethasone treatment (P<0.01). The asthmatic mice had a significantly lower expression of Keap1 protein in the lungs (P<0.05), which was obviously increased by dexamethasone treatment (P<0.05). ConclusionAsthmatic mice have an increased level of oxidative stress. Dexamethasone inhibits the level of pulmonary oxidative stress in asthmatic mice possibly by inhibiting the expression of the downstream antioxidant elements via regulating the Keap1-Nrf2 pathway.

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更新日期/Last Update: 2020-04-24