Chen Xiao-Hui, Zhao Ya-Ping, Zhu Chun, Ji Chen-Bo, Zhang Chun-Mei, Zhu Jin-Gai, Gao Chun-Lin, Guo Xi-Rong
Nanjing Maternal and Child Health Hospital Affiliated to Nanjing Medical University, Nanjing 210004, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2009 Dec;11(12):1008-11.
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 TNFalpha on STEAP4 gene in matured human adipocytes.
Human preadipocytes were cultured and differentiated into matured adipocytes in vitro. Fully differentiated adipocytes (Day 17) were treated with different concentrations of TNFalpha (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.
Different concentrations (5, 10, 25 and 50 ng/mL) of TNFalpha 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 TNFalpha treatment group. In parallel, STEAP4 protein synthesis in matured adipocytes increased in response to TNFalpha 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 TNFalpha treatment group.
TNFalpha can up-regulate STEAP4 mRNA expression in human matured adipocytes.
人STEAP4是一种新型肥胖相关基因，与人类脂肪细胞中的胰岛素敏感性调节有关。本研究旨在探讨肿瘤坏死因子α（TNFα）对成熟人类脂肪细胞中STEAP4基因的调节作用。
体外培养人前脂肪细胞并将其分化为成熟脂肪细胞。用不同浓度的TNFα（0、5、10、25和50 ng/mL）处理完全分化的脂肪细胞（第17天）24小时。从脂肪细胞中提取总RNA和蛋白质。分别通过实时定量逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法测定STEAP4 mRNA和蛋白质表达水平。
用不同浓度（5、10、25和50 ng/mL）的TNFα处理24小时导致人类成熟脂肪细胞中STEAP4 mRNA表达显著增加。在50 ng/mL TNFα处理组中观察到最大效应。同时，用不同浓度（5、10、25和50 ng/mL）的TNFα处理24小时后，成熟脂肪细胞中STEAP4蛋白合成增加。在25 ng/mL TNFα处理组中观察到最大上调效应。
TNFα可上调人类成熟脂肪细胞中STEAP4 mRNA的表达。
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