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目的:探讨小檗碱对3T3-L1脂肪分化的作用是否与激活腺苷酸活化蛋白激酶(AMPK)有关。方法:在3T3-L脂肪细胞分化全程加入小檗碱,以油红O染色检测3T3-L1脂肪细胞胞浆中脂肪的堆积,实时定量PCR检测过氧化物酶体增殖物激活受体γ2(PPARγ2)、CCAAT增强子结合蛋白α(CEBPα)和AMPK的mRNA表达,以Western印迹法检测AMPK和乙酰辅酶A羧化酶(ACC)的磷酸化水平。结果:小檗碱剂量依赖性地抑制3T3-L1脂肪细胞分化,10μmol/L小檗碱几乎完全抑制胞浆中脂肪的堆积。5μmol/L小檗碱在脂肪细胞诱导分化1、3、5、7d后均显著降低CEBPαmRNA表达(P<0.05或P<0.01),诱导分化3、5、7d时显著降低PPARγ2的mRNA表达(P<0.05或P<0.01)。AMPK的mRNA水平在分化过程中未受小檗碱的明显影响,而小檗碱明显增加其蛋白磷酸化水平,其下游靶基因ACC磷酸化水平也明显增加。结论:小檗碱抑制3T3-L1脂肪细胞的分化可能与其激活AMPK有关。
OBJECTIVE: To investigate whether Berberine’s role in 3T3-L1 adipogenic differentiation is related to the activation of AMPK. Methods: Berberine was added into the 3T3-L adipocytes throughout the course of differentiation. The accumulation of fat in cytoplasm of 3T3-L1 adipocytes was detected by oil red O staining. The levels of peroxisome proliferator-activated receptor γ2 (PPARγ2 ), CCAAT enhancer binding protein α (CEBPα) and AMPK mRNA expression were detected by Western blotting AMPK and acetyl-CoA carboxylase (ACC) phosphorylation level. Results: Berberine inhibited the differentiation of 3T3-L1 adipocytes in a dose-dependent manner. 10μmol / L berberine almost completely inhibited the accumulation of fat in the cytoplasm. 5μmol / L berberine significantly decreased the expression of CEBPα mRNA (P <0.05 or P <0.01) after induced by adipocytes for 1,3,5 and 7 days, and significantly decreased the mRNA expression of PPARγ2 (P <0.05 or P <0.01). The mRNA level of AMPK was not significantly affected by berberine during the differentiation process, while berberine significantly increased the phosphorylation level of its protein, and its downstream target gene ACC phosphorylation level was also significantly increased. Conclusion: Berberine inhibits the differentiation of 3T3-L1 adipocytes may be related to its activation of AMPK.