Ling Hong-Yan, He Juan, Yang Si-Si, Zhang Kai-Fang, He Jian-Qin, Zhu Ze-Mei, Feng Shui-Dong
Department of Physiology, University of South China, Hengyang 421001, China.
Department of Physiology, Hunan Institute of Medicine, Hengyang 421001, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2017 Jan 8;33(1):22-25. doi: 10.12047/j.cjap.5448.2017.005.
To observe the effects of arecoline on lipid metabolism in 3T3-L1 adipocytes and explore its possible mechanisms.
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 mol/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.
There were a large number of lipid droplets in the cytoplasm in the differentiated 3T3-L1 adipocytes. MTT results showed that 0~100 mol/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 mol/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 mol/L arecoline group was the most significant.
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.
观察槟榔碱对3T3-L1脂肪细胞脂质代谢的影响,并探讨其可能机制。
采用经典的“鸡尾酒”法将3T3-L1前脂肪细胞诱导为脂肪细胞,随后分别用浓度为0、25、50和100 μmol/L的槟榔碱处理脂肪细胞72小时。72小时后,采用MTT法检测细胞活力,用油红O染色观察细胞质中脂滴的积累情况,通过蛋白质免疫印迹法检测脂肪酸合酶(FAS)、脂肪甘油三酯脂肪酶(ATGL)和激素敏感性脂肪酶(HSL)的蛋白表达。
分化后的3T3-L1脂肪细胞胞质内有大量脂滴。MTT结果显示,0~100 μmol/L槟榔碱对细胞活力无显著影响;油红O染色发现槟榔碱可减少3T3-L1脂肪细胞内脂质含量;蛋白质免疫印迹结果显示,与0 μmol/L槟榔碱组(对照组)相比,槟榔碱显著降低FAS蛋白水平,增加ATGL和HSL蛋白水平,其中50 μmol/L槟榔碱组作用最为显著。
槟榔碱可显著增加3T3-L1脂肪细胞的脂解作用,这可能与降低脂质合成关键酶FAS的表达以及增加脂解关键酶ATGL和HSL的表达有关。