目的 观察Beclin 1及其相互作用蛋白在脂肪细胞分化过程中的表达,同时比较在内脏脂肪和皮下脂肪组织的表达差异.方法 3T3-L1脂肪细胞分化过程中,于第0、2、4、6、8天分别收取细胞,获取细胞蛋白或mRNA后通过Western印迹和实时定量PCR技术分析相关基因表达;正常喂养C57小鼠处死后取皮下和内脏脂肪组织,观察相关基因及蛋白的表达.结果 随着3T3-L1脂肪细胞的分化成熟,细胞自噬水平增强,LC3、Beclin 1和TNFAIP3的基因表达逐渐增高,BIRC5呈现先升后降的变化趋势,FEZ1基因表达从分化一开始即显著降低;基础条件下,内脏脂肪自噬水平高于皮下脂肪,LC3、Beclin 1、YWHAQ、FEZ1、BAD、WAC、TNFAIP3、GOPC在内脏脂肪的表达水平显著高于皮下脂肪,SLAMF1皮下脂肪的表达水平明显高于内脏脂肪表达.结论 自噬及相关基因在脂肪细胞分化中起着重要的作用,且在不同脂肪组织存在差异.
Objective To compare the expressions of Beclin 1 and its 11 interacting proteins in subcutaneous fat and visceral fat,and to observe the expressions of these genes in 3T3-L1 adipocytes differentiation process.Methods In 3T3-L1 adipocytes,cells were harvested at day 0,2,4,6,8 in differentiation process,and then proteins or mRNAs were obtained and followed with Western blot and realtime PCR.Mice tissue proteins or mRNAs were detected with Western blot and realtime PCR.Results In 3T3-L1 adipocytes differentiation process,expressions of LC3,Beclin 1,and TNFAIP3 continued to rise,expression of BIRC5 was high to low,FEZ1 continued at a low level.In mice,expressions of LC3,Beclin 1,YWHAQ,FEZI,BAD,WAC,TNFAIP3,and GOPC in visceral fat were significantly higher than those in subcutaneous fat,while expression of SLAMF1was higher in subcutaneous fat.Conclusion Autophagy plays an important role in 3T3-L1 differentiation process and may be tissue-specific in visceral fat and subcutaneous fat.