上海交通大学学报(医学版)

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氯化钴诱导低氧对3T3-L1脂肪细胞脂连蛋白表达的影响

林 宁1,张珠华2,李晓永1,张洪梅1,苏 青1   

  1. 1.上海交通大学 医学院附属新华医院内分泌科, 上海 200092; 2.普陀区中心医院呼吸科, 上海 200062
  • 出版日期:2014-04-28 发布日期:2014-05-13
  • 通讯作者: 苏 青, 电子信箱: suqinxinhua@163.com。
  • 作者简介:林 宁(1984—), 男, 住院医师, 博士; 电子信箱: linning407@163.com。
  • 基金资助:

    国家自然科学基金(81000332)

Effects of hypoxia induced by CoCl2 on expression of adiponectin in 3T3-L1 adipocytes

LIN Ning1, ZHANG Zhu-hua2, LI Xiao-yong1, ZHANG Hong-mei1, SU Qing1   

  1. 1.Department of Endocrinology, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China; 2.Department Pneumology, Putuo District Central Hospital, Shanghai 200062, China
  • Online:2014-04-28 Published:2014-05-13
  • Supported by:

    National Natural Science Foundation of China, 81000332

摘要:

目的 研究体外模拟低氧对小鼠3T3-L1脂肪细胞脂连蛋白(ADPN)表达的影响。方法 体外培养小鼠3T3-L1前脂肪细胞,将其诱导分化成为成熟的脂肪细胞,油红O染色鉴定脂肪细胞分化程度和脂质积聚情况。氯化钴(CoCl2)化学模拟脂肪细胞低氧环境,采用实时PCR (Real-Time PCR) 检测低氧诱导因子-1α (HIF-1α)和ADPN mRNA的表达,蛋白质印迹法(Western blotting)检测低氧诱导因子-1α (HIF-1α)的蛋白表达;酶联免疫吸附测定(ELISA)分析细胞培养上清液中ADPN蛋白的分泌水平。结果 在CoCl2诱导3T3-L1脂肪细胞低氧的条件下,细胞内HIF-1α mRNA和蛋白的表达水平均显著上调;细胞内ADPN mRNA表达和细胞培养上清液中ADPN蛋白的分泌水平均显著下降;HIF-1α mRNA与ADPN mRNA的表达水平呈显著负相关(r=-0.854,P<0.001)。结论 CoCl2诱导低氧能够下调3T3-L1脂肪细胞ADPN的表达,该作用可能与脂肪细胞代谢功能障碍和胰岛素抵抗有关。

关键词: 3T3-L1脂肪细胞, 脂连蛋白, 氯化钴, 低氧诱导因子-1α

Abstract:

Objective To investigate the effects of hypoxia induced by CoCl2 on the expression of adiponectin (ADPN) in mouse 3T3-L1 adipocytes. Methods 3T3-L1 preadipocytes were cultured and differentiated to matured adipocytes in vitro. Cell differentiation and lipid accumulation were determined by oil red O staining. CoCl2 was used as a chemical hypoxia-inducible reagent to simulate hypoxic microenvironment. The expressions of hypoxia-inducible factor-1α (HIF-1α) and ADPN mRNA were detected by Real-Time PCR. The protein expressions of HIF-1α were determined by Western blotting and the secretion levels of ADPN protein in cell culture supernatant were detected by ELISA. Results Under the hypoxia condition induced by CoCl2, the expression levels of HIF-1α mRNA and protein in 3T3-L1 adipocytes were significantly increased, while the expressions of ADPN mRNA in 3T3-L1 adipocytes and the secretion levels of ADPN protein in cell culture supernatant were significantly decreased. The expression levels of HIF-1α mRNA and ADPN mRNA had significantly negative correlation (r=-0.854, P<0.001). Conclusion The hypoxia induced by CoCl2 can decrease the expression of ADPN in 3T3-L1 adipocytes. This effect may be relevant to the metabolic function impairment of adipose cells and insulin resistance.

Key words: 3T3-L1 adipocytes, adiponectin, CoCl2, hypoxia-inducible factor-1α