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不同程度间歇低氧对3T3-L1脂肪细胞炎性细胞因子和脂肪因子的影响

杨庆婵1,周芹2,王彦3,何庆2,冯靖3,陈宝元4   

  1. 1. 天津医科大学研究生院
    2. 天津医科大学总医院内分泌科
    3. 天津医科大学总医院呼吸科
    4. 天津医科大学总医院
  • 收稿日期:2013-08-01 修回日期:2013-08-27 出版日期:2013-12-15 发布日期:2013-12-15
  • 通讯作者: 冯靖

Effect of Different Degrees of Intermittent Hypoxia on inflammatory Cytokines and Adipokines in 3T3-L1 Adipocytes

YANG Qing chan1,ZHOU Qin 2,WANG Yan 3,HE Qing 4,FENG Jing 3,CHEN Bao yuan5   

  1. 1. Graduate School of Tianjin Medical University
    2. Endocrinology Department of Tianjin MedicalUniversity General Hospital
    3. Respiratory Department of Tianjin Medical University General Hospita
    4. Endocrinology Department of Tianjin Medical University General Hospital
    5. Respiratory Department of Tianjin Medical University General Hospital
  • Received:2013-08-01 Revised:2013-08-27 Published:2013-12-15 Online:2013-12-15
  • Contact: FENG Jing

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摘要：

【摘要】目的 测定不同程度间歇低氧（IH）处理的3T3-L1脂肪细胞中炎性细胞因子和脂肪因子水平的变化。  方法  建立阻塞性睡眠呼吸暂停（OSA）模式间歇低氧/再氧合（IH/ROX）细胞模型，将分化成熟的3T3-L1脂肪细胞分为5组，即3个不同程度IH组（5%O2，7.5%O2，10%O2，编号为IH-1，IH-2，IH-3）、持续低氧（10%O2，CH）组和正常氧对照（21%O2，IN）组。采用ELISA法测定脂肪细胞上清液中肿瘤坏死因子（TNF）-α、白细胞介素（IL）-6、瘦素和脂联素的水平，Western blot测定脂肪细胞中低氧诱导因子（HIF）-1α、葡萄糖转运蛋白（Glut）-1水平，Real-time-PCR测定脂肪细胞中HIF-1α,lut-1、TNF-α、IL-6、瘦素、脂联素的mRNA表达水平。  结果  IH组和CH组TNF-α、IL-6和瘦素蛋白及其mRNA水平均高于IN组，IH-1组TNF-α、IL-6和瘦素蛋白及瘦素mRNA水平高于IH-2、IH-3组（均
P＜0.05）；IH组和CH组脂联素蛋白及其mRNA水平均低于IN组，IH-1组低于IH-2、IH-3组（均P＜0.05）。  结论  OSA模式IH与脂肪细胞炎性细胞因子和脂肪因子的表达和释放有关，IH可能是脂肪细胞炎症反应的基础，参与OSA患者胰岛素抵抗的发生。

关键词: 间歇低氧, 胰岛素抵抗, 肿瘤坏死因子-α, 白细胞介素-6, 缺氧诱导因子1, α亚基, 葡萄糖转运体1型, 瘦素

Abstract:

[Abstract]   Objective   To study the effect of different degrees of intermittent hypoxia (IH) on inflammatory cytokines
and adipokines in3T3-L1adipocytes.  Methods   An intermittent hypoxia/reoxygenation (IH/ROX) from obstructive sleep apnea (OSA) adipocyte model was established.3T3-L1adipocytes were divided into five groups, three IH groups (5% O2,7.5% O2 and10% O2, referred to as IH-1, IH-2and IH-3), sustained hypoxia group (10% O2, CH) and the normal oxygen control group (21%O2, IN). ELISA method was used to detect values of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), leptin and adiponectin in cell supernatant. Western blot analysis was used to detect levels of hypoxia-inducible factor-1α(HIF-1α) and glucose transporter-1(Glut-1). RT-PCR assay was used to analyze HIF-1α, Glut-1, TNF-α, IL-6, leptin and adiponectin mRNA expression levels in adipocytes.   Results   The expression levels of TNF-α, IL-6and leptin  were significantly higher in IH and CH groups than those of IN group (P＜0.05). The expression levels of TNF-α, IL-6and leptin protein and leptin mRNA were significantly higher in IH-1group than those of IH-2and IH-3groups (P＜0.05). The adiponectin and its mRNA levels were significantly lower in IH and CH groups than those of IN group (P＜0.05). The adiponectin level was significantly lower in IH-1group than that of IH-2and IH-3groups (P＜0.05).  Conclusion   These results demonstrate that IH is related to the extensive changes in the expression and release of inflammation-related adipokines in cultured adipocytes. IH from OSA may underlie the development of the inflammatory response in adipocytes, which is involved in insulin resistance in patients with OSA.

Key words: intermittent hypoxia, Insulin Resistence, tumor necrosis factor-α, interleukin-6, hypoxia-induciblefactor1, alpha subunit, glucose transporter type 1, 瘦素