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目的探讨胰高糖素肽-1(GLP-1)对3T3-L1前脂肪细胞分化过程中游离脂肪酸结合蛋白4(FABP-4)、肉碱脂酰转移酶1A(CPT-1A)表达水平的影响。方法采用传统“鸡尾酒”法诱导3T3-L1前脂肪细胞分化,并在分化过程中加入1×10-8mol/L GLP-1进行干预。在诱导分化的第6、8天,用油红O染色检测细胞内脂质聚集,RT-PCR及Western blotting法检测FABP-4、CPT-1A的mRNA及蛋白水平。结果①在诱导分化第6天,大约40%的细胞出现脂滴聚集,第8天时大约80%的细胞已分化成熟;②油红O染色结果显示,与对照组相比,GLP-1组脂质总量在分化第6天略有增加,在分化第8天轻度下降(P>0.05);③RT-PCR结果显示,GLP-1组FABP-4的mRNA水平在分化第6、8天都明显增加;CPT1-A的mRNA水平在分化第8天增加明显(P<0.05);④Western blotting结果显示,GLP-1显著增加了FABP-4的蛋白表达(P<0.05)。对照组CPT-1A的蛋白水平在分化第6、8天无显著变化;而GLP-1组CPT-1A蛋白水平显著增加,尤其在分化第8天,与对照组相比增加了3.57倍(P<0.01)。结论在3T3-L1前脂肪细胞分化过程中,GLP-1显著促进了FABP-4和CPT-1A的mRNA及蛋白水平,提示GLP-1同时促进脂肪细胞对游离脂肪酸的吸收及分解,能有效改善游离脂肪酸介导的胰岛素抵抗。
Objective To investigate the expression of free fatty acid binding protein 4 (FABP-4) and carnitine acyltransferase 1A (CPT-1A) during the differentiation of 3T3-L1 preadipocytes induced by GLP-1 influences. Methods 3T3-L1 preadipocytes were induced by traditional “cocktail” method, and 1 × 10-8 mol / L GLP-1 was added during the differentiation process. On the 6th and 8th days after induced differentiation, the lipid aggregates were detected by oil red O staining. The mRNA and protein levels of FABP-4 and CPT-1A were detected by RT-PCR and Western blotting. Results ① At day 6, about 40% of the cells accumulated lipid droplets, and about 80% of the cells had differentiated and matured on day 8. ② Oil red O staining showed that compared with the control group, GLP-1 lipids The quantity of FABP-4 mRNA in GLP-1 group increased slightly on the 6th day of differentiation and slightly decreased on the 8th day of differentiation (P> 0.05) (P <0.05); ④Western blotting results showed that GLP-1 significantly increased the protein expression of FABP-4 (P <0.05). The protein level of CPT-1A in control group had no significant change on the 6th and 8th day of differentiation, while the level of CPT-1A protein in GLP-1 group increased significantly, especially on the 8th day of differentiation, increased 3.57 times (P <0.01). Conclusion GLP-1 can significantly promote the mRNA and protein levels of FABP-4 and CPT-1A during the differentiation of 3T3-L1 preadipocytes, suggesting that GLP-1 can improve the absorption and decomposition of free fatty acids Free fatty acid-mediated insulin resistance.