槟榔碱对3T3-L1脂肪细胞脂代谢的影响

The effects of arecoline on the lipid metabolism of 3T3-L1 adipocytes

目的:观察槟榔碱对3T3-L1脂肪细胞脂代谢的影响并探讨其可能机制。方法:采用经典的"鸡尾酒"法诱导3T3-L1前脂肪细胞分化成熟,随后用不同浓度的槟榔碱(0、25、50、100μmol/L)处理成熟脂肪细胞72 h。72 h后,四甲基偶氮唑盐(MTT)法检测细胞的活性;油红O染色观察胞浆内脂滴情况;Western blot检测脂肪酸合成酶(FAS)、甘油三酯脂肪酶(ATGL)、激素敏感性脂肪酶(HSL)蛋白表达。结果:诱导分化成熟的脂肪细胞胞浆内可见大量脂滴;MTT显示:0~100μmol/L槟榔碱对脂肪细胞活力无显著影响;油红O染色后脂质含量测定结果表明槟榔碱能减少成熟脂肪细胞中脂质含量;Western blot结果显示:与0μmol/L组(对照组)相比,槟榔碱可显著降低脂肪细胞内FAS的蛋白表达,增加ATGL和HSL的蛋白表达;其中以50μmol/L组最为显著。结论:槟榔碱使脂肪细胞脂解增强,可能与降低脂质合成关键酶FAS的表达,增加脂质分解代谢关键酶ATGL和HSL的表达有关。

Objective： To observe the effects of arecoline on lipid metabolism in 3T3-L1 adipocytes and explore its possible mechanisms. Methods： 3T3-L1 pre-adipocytes were induced into adipocytes with the classic＂cocktail＂method,subsequently,adipocytes were treated with arecoline at the concentrations of 0,25,50 and 100 μmol / L for 72 hours. After 72 hours,cell vability was measured with MTT method,lipid droplet accumulation in the cytoplasm was observed with oil red O staining,the protein expression of fatty acid synthase（ FAS）,adipose triglyceride lipase（ ATGL） and hormone-sensitive lipase（ HSL） were detected by Western blot assay. Results： There were a large number of lipid droplets in the cytoplasm in the differentiated 3T3-L1 adipocytes. MTT results showed that 0 - 100 μmol / L arecoline had no significant effect on cell vability; oil red O staining found arecoline reduced lipid amount in 3T3-L1 adipocytes; Western blot results showed that compared with 0 μmol / L arecoline group（ the control group）,arecoline significantly reduced the protein level of FAS and increased the protein levels of ATGL and HSL,and 50 μmol / L arecoline group was the most significant. Conclusion： Arecoline significantly increased lipolysis of 3T3-L1 adipocyte,which might be associated with decreased the FAS expression of key enzyme of lipid synthesis and increased the ATGL and HSL expression of key enzyme of adipolysis.