天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (9): 1124-1127.doi: 10.11958/20160041

• 临床研究 • 上一篇    下一篇

不同程度间歇低氧对 3T3-L1 脂肪细胞 NF-κB、IL-10 和内脂素的影响

韩苗苗1,周芹2,冯靖3,牛文彦4,5,何庆6   

  1. 1 天津市西青医院内分泌科（邮编 300380）；2天津医科大学总医院内分泌科， 3 呼吸科；4天津医科大学
  • 收稿日期:2016-01-28 修回日期:2016-06-23 出版日期:2016-09-15 发布日期:2016-09-28
  • 通讯作者: 何庆 E-mail:icy__baby@163.com
  • 作者简介:韩苗苗（1987）， 女， 硕士研究生， 住院医师， 主要从事糖尿病研究

Effects of different degrees of intermittent hypoxia on NF-κB, IL-10 and visfatin in 3T3-L1 adipocytes

HAN Miaomiao1, ZHOU Qin2, FENG Jing3, NIU Wenyan4, HE Qing2△   

  1. 1 Department of Endocrinology, Tianjin Xiqing Hospital, Tianjin 300380, China; 2 Department of Endocrinology, 3 Department of Respiratory Medicine, General Hospital of Tianjin Medical University; 4 Tianjin Medical University
  • Received:2016-01-28 Revised:2016-06-23 Published:2016-09-15 Online:2016-09-28
  • Contact: HE Qing E-mail:icy__baby@163.com

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摘要： 目的 测定不同程度间歇低氧（IH）处理的脂肪细胞中核因子（NF）-κB、白细胞介素（IL）-10 及内脂素水平的变化， 探讨 IH 导致胰岛素抵抗的机制。 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， 将分化成熟的脂肪细胞 3T3-L1 随机分为 10 组， 包括 4 个不同程度 IH 组（IH1、IH2、IH3、IH4， 先充入 1.5%O245 s， 各组分别充入 21%O22 min 15 s、4 min 15 s、5 min 45 s、8 min 45 s， 每组 60 个循环）及各自的正常氧对照组（SC1、SC2、SC3、SC4， 将各 IH 组中 1.5%O2改为 21%O2， 其余同 IH 组处理）、持续低氧组（CH， 10%O26 h）及持续正常氧对照组（CC， 21%O2 6 h）。 采用双抗夹心酶联免疫吸附试验（ELISA）法测定 3T3-L1 上清液中 IL-10 和内脂素水平； 采用 Western Blotting 法测定脂肪细胞内脂素及细胞核 NF-κB p65 的蛋白水平； 采用 real-time PCR 方法测定脂肪细胞 IL-10、内脂素的 mRNA 水平。 结果 IH 组和 CH 组 IL-10 蛋白及 mRNA 的表达水平明显低于各自对照组（P < 0.01）。 IH 和 CH 组 NF-κB p65 蛋白水平高于各自对照组。 IH1、IH2组和 CH 组内脂素蛋白及 mRNA 的表达水平高于各自对照组（P < 0.01）。 结论 IH 作为 OSA 的主要病生理特征， 其可能通过导致脂肪细胞 NF-κB、IL-10 和内脂素的分泌异常， 参与 OSA 患者胰岛素抵抗的发生。

关键词: 睡眠呼吸暂停, 阻塞性, 间歇低氧, 胰岛素抵抗, NF-κB, 烟酰胺磷酸核糖基转移酶, 白细胞介素 10, 内脂素

Abstract: Objective To determine levels of nuclear factor (NF)-κB, interleukin (IL)-10, and visfatin in adipocytes treated by different degrees of intermittent hypoxia (IH), and to investigate the mechanism of IH leading to insulin resistance (IR). Methods The cell model of intermittent hypoxia/re-oxygenation (IH/ROX) in obstructive sleep apnea (OSA) was established. Differentiation mature 3T3- L1 adipocytes, were randomly divided into 10 groups including four differentfrequency intermittent hypoxia groups（IH1-4, fixed intermittent hypoxia scheme for 1.5% O2 45 s and then re-oxygen 21% O2 for 2 min 15 s, 4 min 15 s, 5 min 45 s and 8 min 45 s, 60 times circulation), and their normal oxygen control groups (SC 1-4, instead each IH group 1.5% O2 to 21% O2, the rest groups were treated as same as IH group), continuous hypoxia group (CH, 10% O2 for 6 h) and normal oxygen control group (CC, 21%O2 for 6 h). ELISA method was used to determine the levels of IL- 10 and visfatin in the supematant of adipocytes. Western blot method was used to determine the protein levels of NF-κB p65 and visfatin. Real-time PCR method was used to determine the mRNA levels of IL-10 and visfatin. Results The protein and mRNA expressions of IL-10 were significantly lower in IH group and CH group than those of control groups (P＜ 0.01). The levels of NF-κB p65 protein were significantly increased in IH group and CH group than those of control group. The protein and mRNA expressions of visfatin were significantly higher in IH1, IH2 and CH groups than those of control group (P＜ 0.01). Conclusion As a prominent feature of OSA pathophysiology, IH may take part in insulin resistance of OSA patients by abnormally secreting NF-κB, IL-10 and visfatin in adipocytes.

Key words: sleep apnea, obstructive, intermittent hypoxia, insulin resistance, NF-kappa B, nicotinamide phosphoribo syltransferase, interleukin-10, visfatin