摘要
目的:通过培养3T3-L1前脂肪细胞,并诱导其分化至成熟,研究游离脂肪酸对脂肪细胞糖代谢的影响。方法:培养诱导3T3-L1脂肪细胞,用油红O染色鉴定并比较其形态结构的变化。LPS、EPA、SA、PA干预成熟脂肪细胞,收集不同时间的培养基,葡萄糖氧化酶法算出各组脂肪细胞的葡萄糖消耗量。用Western blot检测不同时间各组干预后细胞AMPK、GLUT4蛋白含量。结果:油红O染色鉴定成熟脂肪细胞胞浆中的脂滴染成红色,并出现"戒环"样结构;诱导分化第8天,90%以上细胞均分化成熟。含LPS、EPA、SA、PA的培养基作用于成熟脂肪细胞,随着时间的延长,显著抑制脂肪细胞对葡萄糖的吸收(P<0.05),同时,脂肪细胞AMPK、GLUT4蛋白含量在减少(P<0.05)。结论:游离脂肪酸可以诱导胰岛素抵抗的分子机制可能是通过胰岛素信号通路激活蛋白激酶(AMPK),进而影响GLUT4的蛋白表达,使脂肪细胞的葡萄糖吸收率减低,影响脂肪细胞的糖代谢。
Objective：3T3-L1 cells were induced into adipocytes,to study the effects of free fatty acid on glucose metabolism in 3T3-L1 adipocytes. Methods：3T3-L1 cells were induced into adipocytes identified by Oil Red O staining,observe the morphologic transformation of 3T3-L1 adipocytes. 3T3-L1 adipocytes were treated for LPS,EPA,SA,PA,collect adipocyte culture medium at different time point,Glucose Oxidase method was employed to measure the glucose consumption in the medium. Western blot was used to determine the protein expression of AMPK,GLUT4 after the treatment at different time point. Results：3T3-L1 cells were induced into adipocytes identified by Oil Red O staining,the lipid droplets were red,the cytoplasm had a stack fat and the cells looked seem as finger ring. On the 8th day,more than 90% cells were induced into adipocytes. After the treatment with LPS,EPA,SA,PA,the glucose consumption of 3T3-L1 adipose cells were decreased in time-dependent manner（P0.05） . Meantime,both the protein expression of AMPK,GLUT4 were decreased（P0.05） . Conclusions：FFA can induce insulin resistance,the molecular mechanism of which might be correlated with an AMPK activation by insulin signaling pathway,effects the protein expression of GLUT4,decreased the glucose consumption of 3T3-L1 adipose cells,and effects of free fatty acid on glucose metabolism in 3T3-L1 adipocytes.
出处
《现代生物医学进展》
CAS
2010年第2期212-215,F0002,共5页
Progress in Modern Biomedicine
基金
973计划前期研究专项课题(2009CB526505)
