摘要
研究促酰化蛋白(acylation stimulating protein,ASP)在3T3-L1脂肪细胞分化中对脂滴相关蛋白TIP47(tail-interacting protein 47 kD)表达的影响,从而探讨ASP在成脂方面的重要意义.用免疫荧光染色法观察3T3-L1前脂肪细胞中TIP47的表达定位;采用经典激素鸡尾酒法诱导分化3T3-L1前脂肪细胞,用RT-PCR和Western印迹方法检测诱导分化的3T3-L1脂肪细胞中TIP47 mRNA和蛋白表达;在分化过程中不同时点,对诱导分化中的3T3-L1脂肪细胞分别给予胰岛素和ASP处理,并设立相应空白对照,用RT-PCR和Western印迹方法检测TIP47 mRNA和蛋白表达.结果显示,3T3-L1前脂肪细胞中TIP47主要在胞浆内表达;诱导分化过程中的3T3-L1脂肪细胞TIP47 mRNA和蛋白的表达水平呈时间依赖性降低;ASP对诱导分化的3T3-L1脂肪细胞中TIP47 mRNA和蛋白表达有显著的上调作用,但随着分化至48 h,其上调作用已不明显;胰岛素仅在分化的0 d对脂肪细胞中TIP47 mRNA和蛋白表达有上调作用,之后基本无影响.结果提示,ASP促成脂作用可能与其调节脂滴相关蛋白TIP47的表达密切相关,从而为认识及防治肥胖症开拓新的思路.
To study the effect of acylation stimulating protein （ASP） on the expression of lipid droplets associated protein TIP47 during preadipocyte differentiation and evaluate its significance in adipogenesis. The localization of TIP47 were analyzed by immunofluorescent staining in 3T3-L1 preadipocytes. 333-L1 preadipocytes were differentiated with the traditional hormone cocktail, then incubated with ASP and insulin respectively at indicated time point, the mRNA and protein levels of TIP47 were determined by semiquantitive-PT-PCR and Western blot analysis. The results showed that TIP47 located in the cytoplasm; and its expression decreased during the differentiation process in 333-L1 adipocytes. Exposure to ASP resulted in an increase of TIP47, but the effect of ASP was not significant after 48 hours of differentiation. Insulin could elevate the expression of TIP47 in 333-L1 adipocytes at 0 day of differentiation, but not lasting after then. Taken together, this study provided evidence that the promotion of adipogenesis by ASP might be associated with the up- regulation of lipid droplets associated protein TIP47 expression.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2009年第5期442-447,共6页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金(No.30600213)资助~~
关键词
促酰化蛋白
脂滴相关蛋白
TIP47
3T3-L1脂肪细胞
肥胖症
acylation stimulating protein
lipid droplets associated protein
tail-interacting protein47-TIPdT
3T3-L1 adipocytes
obesity
