小檗碱激活AMPK抑制3T3-L1脂肪细胞分化  

Berberine Activates AMPK and Inhibits 3T3-L1 Adipocyte Differentiation

作　　者：王宁[1,2,3] 张娟[1,2,3] 建方方[1,2,3] 邓儒元[1,2,3] 唐红菊[1,2,3] 刘赟[1,2,3] 李凤英[1,2,3] 王晓[1,2,3] 周丽斌[1,2,3] 

机构地区：[1]上海市内分泌代谢病研究所 [2]上海内分泌代谢病临床医学中心 [3]上海交通大学医学院附属瑞金医院,200025

出　　处：《放射免疫学杂志》2012年第3期273-275,共3页Journal of Radioimmanology

基　　金：国家自然科学基金项目(81070652;81070617;81170720)

摘　　要：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 the effect of berberine （BBR） on 3T3-L1 adipocyte differentiation is related to AMP activated protein （AMPK） activation. Methods The accumulation of lipid in the cytoplasm of differentiated 3T3-L1 adipoeytes was observed by oil red O staining. Realtime-PCR was used to detect the mRNA expressions of PPAR~/2, CEBPot, and AMPK. The phosphorylation levels of AMPK and acetyl CoA carboxylase （ACC） were detected by Western blot. Result Berberine inhibited 3T3- L1 adipocyte differentiation in a dose-dependent manner. At the concentration of 10taanol/L berberine, the accumulation of lipid in the cytoplasm of adipocytes was almost inhibited. CEBPot mRNA expression was inhibited by 51μmol/L berberine after 1,3,5, and 7day induction differentiation （ P 〈 0.05 or P 〈 0.01 ） and PPARγ2 mRNA expression was decreased by berberine after induction differentiation of 3,5, and 7 day （ P 〈 0.05 or P 〈 0.01 ）. There were no changes of AMPK mRNA level after 3＇1＂3-L1 cells were incubated with berberine during differentiation. However, the phosphorylation levels AMPK and its downstream target gene ACC were significantly in- creased by berberine. Conclusion Berberine seems to inhibit 3T3-LI adipocyte differentiation via stimulating AMPK activity.

关 键 词：小檗碱 腺苷酸活化蛋白激酶 脂肪细胞分化 过氧化物酶体增殖物激活受体Γ CCAAT增强子结合蛋白α 

分 类 号：R153.2[医药卫生—营养与食品卫生学]