LRP16蛋白在3T3-L1前脂肪细胞诱导分化中表达量的变化

Expression of LRP16 protein during 3T3-L1 preadipocyte differentiation

目的 LRP16蛋白在3T3-L1前脂肪细胞诱导分化过程中表达量的变化及胰岛素对脂肪细胞LRP16蛋白表达的调控。方法经典脂肪细胞诱导方案诱导前脂肪细胞为成熟脂肪细胞,每天(0-8d)均留取蛋白标本。不同浓度(0、1、10、100nmol/L)胰岛素刺激脂肪细胞24h,分别留取蛋白标本。Western-Blot方法检测样本中LRP16蛋白的表达量。结果 LRP16蛋白在前脂肪细胞无表达或极低表达,从诱导的第3、4天开始表达,迅速升至高峰,此后维持在高表达水平。LRP16蛋白在第6-8天之间表达水平无统计学差异(P>0.05),其余各时段间表达水平均有显著差异(P<0.01)。不同浓度胰岛素刺激脂肪细胞24h,细胞中LRP16蛋白表达量与胰岛素浓度呈负相关(P<0.01)。结论 LRP16蛋白随着前脂肪细胞分化的开启,表达量从无到有逐渐增加至高峰,并一直维持在高表达水平;胰岛素(INS)负调控脂肪细胞LRP16蛋白的表达。

Objective To study the expression levels of LRP16 protein in 3T3-L1 preadipocyte differentiation and the effect of insulin on LRP16 protein expression in LRP16 cells.Methods 3T3-L1 preadipocytes were cultured in vitro and differentiated into matured adipocytes,then protein samples were collected everyday（0-8d）.Insulin was added into the culture medium of adipocytes for 24h at different concentrations（0,1,10,100nmol/L）,and then protein samples were collected.Expression levels of LRP16 protein in 3T3-L1 cells were measured by Western blot.Results LRP16 protein was not expressed or very weakly expressed in preadipocytes.The expression level of LRP16 protein increased rapidly from the day 3 or day 4 and reached its peak on day 7（P0.01） and then maintained there.No statistical difference was observed in expression of LRP16 protein on days 6-8（P0.05）.The insulin concentration was negatively correlated with the expression level of LRP16 protein in adipocytes after stimulated by insulin for 24h（P0.01）.Conclusion LRP16 protein expression level increases with the differentiation of 3T3-L1 adipocytes,gradually reaches its peak and then maintains there.Insulin down-regulates the expression of LRP16 protein in adipocytes.