ANGPTL4基因在3T3-L1前脂肪细胞诱导分化过程中表达水平的变化  

Angiopoietin-like protein 4(ANGPTL4) gene expression during the period of 3T3-L1 preadipocyte differentiation

作　　者：黄颖[1] 曹岩[1] 刘阳[1] 郑永晨[1] 

机构地区：[1]吉林大学第二医院中心实验室,吉林长春130041

出　　处：《中国实验诊断学》2009年第10期1332-1335,共4页Chinese Journal of Laboratory Diagnosis

摘　　要：目的观察血管生成素样蛋白-4(Angiopoietin-like protein 4,ANGPTL4)基因在3T3-L1前脂肪细胞诱导分化过程中表达水平的变化,探讨ANGPTL4基因与脂肪细胞分化的关系。方法体外培养3T3-L1前脂肪细胞,在3T3-L1前脂肪细胞分化成熟过程中(0-10 d),采用油红O染色,观察脂肪细胞分化及脂质积聚情况;采用Real-time PCR、逆转录PCR技术检测脂肪细胞分化标志基因(LEPTIN,ADIPONECTIN)和ANGPTL4 mRNA表达;采用Western blot检测ANGPTL4蛋白表达。结果随着脂肪细胞诱导分化逐步深入,油红O染色显示,细胞中脂滴形成逐渐增多,达到占全视野95%以上;同时瘦素(LEPTIN)和脂联素(ADIPONECTIN)基因在诱导后的第2天开始表达并逐步增多,均提示脂肪细胞分化逐步成熟。ANGPTL4 mRNA和蛋白在前脂肪细胞中低表达,分化第2-6天ANGPTL4基因表达显著上调,分化第8-10天ANGPTL4基因表达保持在较高水平并趋于稳定。结论在3T3-L1前脂肪细胞分化过程中ANGPTL4基因表达上调,ANGPTL4基因可能参与了脂肪细胞分化。Objective To investigate ANGPTL4 gene expression during the period of 3T3-L1 preadipocyte differentiation and to explore the relationship between the ANGPTL4 gene and adipocytes differentiation.Methods 3T3-L1 preadipocytes were cultured in vitro and differentiated into the matured adipocytes.Cell differentiation and lipid accumulation was determined by Oil Red O staining.Differentiation marker genes（ADIPONECTIN,LEPTIN） and ANGPTL4 gene mRNA expressions were evaluated by reverse transcription PCR and Real-time PCR.ANGPTL4 protein was detected by Western Blot.Results With the progression of 3T3-L1 cell differentiation,oil red O staining showed that the lipid droplets increased gradually to 95% of the visible field,meanwhile,the expression of ADIPONECTIN and LEPTIN mRNA detected from day 2 post-induction,suggesting the differentiation and maturation of the preadipocytes.ANGPTL4 mRNA and protein was detected before the induction of differentiation.From the second day to the sixth day of differentiation,the level of ANGPTL4 gene mRNA expression was obviously up-regulated.From the eighth day to the tenth day,the mRNA expression of ANGPTL4 gene was kept in higher level and remained unchanged.Conclusion The level of ANGPTL4 gene expression was up-regulated during the differentiation of preadipocyte,which means ANGPTL4 gene may participate in the adipocyte differentiation.

关 键 词：ANGPTL4基因 3T3-L1前脂肪细胞 细胞分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]