[1]杨娟,李晓栩,陈建,等.植物雌激素通过缺氧诱导因子-1α与核因子-κB下调缺氧肺泡上皮细胞膜联蛋白A1的表达[J].第三军医大学学报,2013,35(19):2019-2023.
 Yang Juan,Li Xiaoxu,Chen Jian,et al.Phytoestrogen attenuates hypoxia-induced expression of annexin A1 in alveolar epithelial cells through down-regulating NF-κB and HIF-1α[J].J Third Mil Med Univ,2013,35(19):2019-2023.
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植物雌激素通过缺氧诱导因子-1α与核因子-κB下调缺氧肺泡上皮细胞膜联蛋白A1的表达(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
35卷
期数:
2013年第19期
页码:
2019-2023
栏目:
论著
出版日期:
2013-10-15

文章信息/Info

Title:
Phytoestrogen attenuates hypoxia-induced expression of annexin A1 in alveolar epithelial cells through down-regulating NF-κB and HIF-1α
作者:
杨娟李晓栩陈建官立彬黄瑊白志川
西南大学园艺园林学院;第三军医大学高原军事医学系高原生理学教研室,高原医学教育部重点实验室,全军高原医学重点实验室
Author(s):
Yang Juan Li Xiaoxu Chen Jian Guan Libing Huang Jian Bai Zhichuan
College of Horticulture and Landscape Agriculture, Southwest University, Chongqing, 400716; Department of High Altitude Physiology, Key Laboratory of High Altitude Medicine of Ministry of Education, Key Laboratory of High Altitude Medicine of PLA, College of High Altitude Military Medicine, Third Military Medical University, Chongqing, 400038, China
关键词:
缺氧肺泡上皮细胞植物雌激素膜联蛋白A1
Keywords:
hypoxia human pulmonary alveolar epithelial cells phytoestrogen genistein daidzein Annexin A1
分类号:
R722.35; R394.3; R977.12
文献标志码:
A
摘要:
目的      从人肺泡上皮细胞甲酰肽受体激动剂——膜联蛋白A1(Annexin A1,AnxA1)的表达变化,探讨植物雌激素影响缺氧肺部炎症浸润的机制。      方法       人肺泡Ⅱ型上皮细胞(A549)接种于60 mm培养皿中常规培养,待培养皿中细胞长满约70%,根据是否用药分为:对照组(溶剂DMSO对照)、染料木黄酮(genistein,Gen)组、大豆苷元(daidzein,DD)组,每组细胞再根据是否缺氧再分为常氧组(N)、缺氧组(H)。Gen组、DD组于缺氧处理前分别加入Gen及DD,终浓度均为15 μmol/L。缺氧组细胞置于缺氧培养箱(1%O2)24 h。取细胞培养上清及细胞裂解离心上清,结合甲酰肽受体特异性抑制剂tBoc2,通过趋化实验,检测其甲酰肽受体激动活性。用Western blot分析AnxA1、缺氧诱导因子-1α(HIF-1α)、核因子-κB(NF-κB)在蛋白水平的变化,用RT2-PCR法分析其mRNA水平的变化。      结果       缺氧后的细胞冻融上清及培养上清趋化活性显著高于常氧组(P<0.05),而植物雌激素处理后,两种上清的趋化活性均显著高于对照组(P<0.05)。Western blot与RT2-PCR检测结果显示,缺氧组AnxA1在蛋白水平和mRNA水平均高于常氧组(P<0.05),两种植物雌激素不同程度地削弱了缺氧对AnxA1的上调效果,同时对HIF-1α、NF-κB的蛋白水平也产生同样的影响。      结论       植物雌激素削弱了缺氧对AnxA1基因表达的上调效应,其机制与HIF-1α、NF-κB信号途径有关。
Abstract:
Objective      To determine the effects of phytoestrogen on the expression of formyl peptide receptor (FPR) and Annexin A1 (AnxA1) in human alveolar epithelial cells under hypoxia exposure.       Methods       Human alveolar type Ⅱ epithelial cells (A549) were cultured and divided into normoxia (5%CO2 and 95%O2) and hypoxia (1%O2 5%CO2, and 94%N2 for 24 h) groups. Each group was further divided into the control, genistein treatment group (15 μmol/L before hypoxia) and daidzein treatment group (15 μmol/L before hypoxia). Chemotaxis assay was used to detect the activity of FPR agonists in the supernatants of culture media and lysates of the treated cells in presence or absence of FPR specific inhibitor tBoc2. The protein levels of AnxA1, hypoxia inducible factor 1α (HIF-1α) and nuclear factor κB (NF-κB) were detected by Western blotting. The mRNA level of AnxA1 was assayed by RT2-PCR.       Results      Hypoxia induced both cultural supernatants and lysate supernatants had significantly higher chemotactic activity than normoxia (P<0.05). Phytoestrogen treatment also induced higher chemotactic activity than the control groups (P<0.05). Western blotting and RT2-PCR indicated that hypoxia up-regulated the expression of AnxA1 significantly at the levels of protein and mRNA (P<0.05). The elevation was inhibited in the presence of phytoestrogen so as well NF-κB and HIF-1α.       Conclusion       Phytoestrogen attenuates hypoxia-induced up-regulated expression of AnxA1 through NF-κB and HIF-1α signal pathways.

参考文献/References:

杨娟,李晓栩,陈建,等. 植物雌激素通过缺氧诱导因子-1α与核因子-κB下调缺氧肺泡上皮细胞膜联蛋白A1的表达[J].第三军医大学学报,2013,35(19):2019-2023.

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更新日期/Last Update: 2013-09-30