TSG-6基因在3T3-L1脂肪细胞诱导分化中表达水平的变化及TNF-α对其调节作用的研究  

Expression of TSG-6 gene during 3T3-L1 preadipocyte differentiation and regulative role of tumor necrosis factor-alpha

作　　者：郭锡熔[1] 丁胜利[1] 潘晓勤[2] 龚海霞[1] 费莉[2] 倪毓辉[1] 陈荣华[2] 

机构地区：[1]南京医科大学第二附属医院儿童保健研究中心,210011 [2]南京医科大学儿科医学研究所

出　　处：《中华儿科杂志》2004年第5期344-347,共4页Chinese Journal of Pediatrics

基　　金：国家自然科学基金 ( 3 9870 3 62 );江苏省教委自然科学基金 ( 98KJB3 2 0 0 0 2 )

摘　　要：目的　观察 3T3 L1脂肪细胞诱导分化过程中TSG 6基因mRNA表达水平的变化 ,探讨TNF α对成熟脂肪细胞中TSG 6基因表达水平的调节作用。方法　体外培养 3T3 L1脂肪细胞 ,在诱导 3T3 L1脂肪细胞分化成熟的基础上 ,应用重组TNF α干预分化成熟的脂肪细胞 ,采用RT PCR技术检测诱导分化不同时段及TNF α干预后不同时间脂肪细胞中TSG 6基因的表达水平。结果　 (1)随脂肪细胞逐渐分化成熟 ,TSG 6基因mRNA表达水平逐渐升高。TSG 6基因表达水平除在细胞分化第0～ 2天、第 3～ 5天、第 4～ 6天和第 7～ 10天各时段内差异无显著意义 (P >0 0 5 )外 ,其余各时段之间表达水平 ,差异均有显著意义 (P <0 0 5 ) ;(2 )不同浓度重组TNF α(0 1～ 10 0ng/ml)均能显著抑制成熟脂肪细胞中TSG 6基因mRNA的表达 ,除外TNF α浓度 1 0ng/ml时 6～ 2 4h时段 ,其抑制作用呈现随TNF α浓度增加和刺激时间延长而明显增强的总体趋势。TNF α浓度为 0 1ng/ml时 ,TSG 6基因表达水平 6h内下降 33 73% ,12h内下降 97 39% ;TNF α浓度为 1 0ng/ml时 ,TSG 6基因表达水平 6h内下降 78 6 8% ,并持续至 2 4h ;TNF α浓度达 10 0ng/ml时 ,TSG 6基因表达水平 2h内即下降96 2 7% ,TSG 6基因的表达几乎被完全抑制。结论　 (1)TSG 6基因可能参?Objective Tumor necrosis factor alpha-stimulated gene-6 (TSG-6 gene) differentially expressed in adipose tissue of obese and normal human subjects or rats. To explore the relationship between the differential expression of TSG-6 and adipocyte differentiation, adipogenesis and obesity, the present study aimed to investigate the changes of TSG-6 gene expression during 3T3-L1 preadipocyte differentiation and to analyze the regulative role of TNF-alpha on TSG-6 gene expression in matured 3T3-L1 adipocytes. Methods 3T3-L1 preadipocytes were cultured in vitro and differentiated into the matured adipocytes. TNF-alpha in different concentrations (0.1 ng/ml, 1.0 ng/ml, 10.0 ng/ml) was added into the culture medium of fully differentiated adipocytes (day 10) for various times (0.5 h, 2 h, 6 h, 12 h, 24 h). Total RNA from these adipocytes was extracted and the levels of TSG-6 gene mRNA expression were evaluated by RT-PCR. Results (1) In preadipocytes, the level of TSG-6 gene mRNA expression remained low. In the presence of dexamethasone (Dex), MIX and insulin, with the 3T3-L1 preadipocytes being differentiated into the matured adipocytes, the level of TSG-6 gene mRNA expression was upregulated and reached the higher level in fully differentiated adipocytes. There is a significant difference between any two detected phases in the levels of TSG-6 gene mRNA expression (P<0.05), except that the levels of TSG-6 gene mRNA expression did not increase obviously on day 0 to day 2, day 3 to day 5, day 4 to day 6 and day 7 to day 10 (P>0.05). (2) Treatment of day 10 3T3-L1 adipocytes with TNF-alpha of different concentrations (0.1 ng/ml, 1.0 ng/ml, 10.0 ng/ml) resulted in a significant decrease in the level of TSG-6 gene mRNA expression. The inhibition effect of TNF-alpha on TSG-6 gene mRNA expression generally tended to be reinforced with the increasing concentrations of TNF-alpha and the elongation of time course, except for the period of 6-24 h after the stimulation of 10.0 ng/ml TNF-alpha. When 0.1 ng/ml TNF-alpha was applied, the

关 键 词：肥胖症 细胞黏附分子 脂肪细胞 细胞分化 肿瘤坏死因子 基因表达 TNF-Α TSG-6基因 诱导分化 发病机制 

分 类 号：R723[医药卫生—儿科]