脂肪膜细胞相关蛋白对3T3-L1脂肪细胞糖代谢及细胞分化的影响

Effects of adipocyte plasma membrane-associated protein on the glycometabolism and cell differentiation in 3T3-L1 adipocytes

目的观察APMAP对脂肪细胞糖代谢和细胞分化的影响,并分析其对于糖代谢的作用机制。方法通过慢病毒干预3T3-L1脂肪细胞APMAP表达,观察对3H-葡萄糖的摄取并进行油红O染色定量分析脂肪细胞分化程度,检测培养上清中甘油和游离脂肪酸(NEFA)浓度以及细胞内三酰甘油(TG)的含量。脂肪细胞分化相关基因,过氧化物体增殖剂活化受体γ(PPARγ)和CAAT/增强子结合蛋白A(C/EBPα)mRNA的表达通过实时定量PCR检测。结果APMAP过表达组和沉默组葡萄糖摄取率分别为正常对照的176.8%和67.4%,同时过表达组油红O染色OD值明显高于沉默组和对照组(P<0.01)。APMAP过表达组明显促进脂肪细胞分化及PPARγ和C/EBPαmRNA表达。结论APMAP表达量直接影响葡萄糖摄入量,过表达的APMAP能促进细胞内脂质的积聚,促进细胞分化,对于脂肪细胞糖代谢有着重要调控作用。

Objective To investigate the effects of adipocyte plasma membrane-associated protein(APMAP)on the glycometabolism and cell differentiation in 3T3-L1 adipocytes.Methods The expression and APMAP in 3T3-L1 adipocyte was interfered by entivirus to observe the 3H-glucose uptake rate,and the degree of adipocyte differentiation was observed by means of oil red O staining quantitative analysis.The levels of glycerol,non-esterified fatty acid(NEFA)in culture supernatant and intracellular triglyceride(TG)levels were detected.Moreover the expression levels of PPARγand C/EBPαwere detected by real-time fluorescence quantitative polymerase chain reaction(RT-PCR).Results The 3H-glucose uptake rate in APMAP overexpression group and silencing group was 176.8%and 67.4%,respectively,moreover,the OD value of oil red O staining in APMAP overexpression group was significantly higher than that in silencing group and that in control group(P<0.01).In addition the APMAP overexpression could significantly promote the differentiation of adipocytes and the expressions of PPARγand C/EBPα(P<0.05).Conclusion The expression of APMAP can affect directly the glucose up-take,and the APMAP overexpression can promote intracellular lipidic accumulation and cell differentiation,which plays an important regulating role in glycometabolism of adipocyte.