TNFα对人脂肪细胞STEAP4基因表达的调控

Regulative role of TNFα on STEAP4 gene in matured human adipocytes

摘要:

目的：人STEAP4基因是一个参与胰岛素敏感性调节的肥胖相关差异表达基因，本研究旨在探讨胰岛素抵抗相关脂源性细胞因子TNFα对人成熟脂肪细胞中STEAP4基因的调节作用。方法：体外培养人前体脂肪细胞，在诱导人前体脂肪细胞分化成熟的基础上，应用不同浓度（0、5、10、25、50 ng/mL）人重组TNFα干预成熟脂肪细胞24 h，采用实时荧光定量RT-PCR技术、Western blot技术检测干预后人成熟脂肪细胞STEAP4基因在核酸和蛋白表达水平的变化。结果：不同浓度TNFα（5、10、25、50 ng/mL）干预24 h能显著上调人成熟脂肪细胞中STEAP4基因的mRNA表达,与未干预对照组差异有显著性（P<0.05）；50 ng/mL TNFα干预时STEAP4基因在人成熟脂肪细胞中的mRNA表达水平最高。与基因表达结果相一致，不同浓度TNFα（5、10、25、50 ng/mL）干预24 h能显著上调人成熟脂肪细胞中STEAP4蛋白的表达,与未干预对照组差异有显著性（P<0.05）；干预浓度提高到25 ng/mL 时干预效果最为明显。结论：TNFα能显著上调人成熟脂肪细胞中STEAP4基因核酸和蛋白的表达。［中国当代儿科杂志，2009，11（12）：1008-1011］

Abstract:

OBJECTIVE: Human STEAP4, a novel obesity-related gene, is associated with insulin sensitivity regulation in human adipocytes. This study aimed to explore the regulative role of TNFα on STEAP4 gene in matured human adipocytes. METHODS: Human preadipocytes were cultured and differentiated into matured adipocytes in vitro. Fully differentiated adipocytes (Day 17) were treated with different concentrations of TNFα (0, 5, 10, 25 and 50 ng/mL) for 24 hrs. Total RNA and protein were extracted from the adipocytes. Levels of STEAP4 mRNA and protein expression were determined by real-time quantitative RT-PCR and Western blot respectively. RESULTS: Different concentrations (5, 10, 25 and 50 ng/mL) of TNFα treatment for 24 hrs resulted in a significant increase in the STEAP4 mRNA expression of human matured adipocytes.The maximal effect was seen in the 50 ng/mL of TNFα treatment group. In parallel, STEAP4 protein synthesis in matured adipocytes increased in response to TNFα treatment of different concentrations (5, 10, 25 and 50 ng/mL) for 24 hrs. The maximal up-regulated effect was seen in the 25 ng/mL of TNFα treatment group. CONCLUSIONS: TNFα can up-regulate STEAP4 mRNA expression in human matured adipocytes.［Chin J Contemp Pediatr, 2009, 11 (12):1008-1011］