小檗碱对3T3-L1前脂肪细胞增殖及分化相关基因PPARγ、C/EBPα mRNA和蛋白表达的影响

Effects of Berberine on Cell Proliferation,Peroxisome Proliferation Activated Receptor γ,CAAT/Enhancer Binding Protein mRNA and Protein Expression in 3T3-L1 Pre-adipocytes

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DOI：

中文关键词: 小檗碱脂肪细胞过氧化物体增殖剂活化受体γ CAAT/增强子结合蛋白α mRNA表达蛋白表达

英文关键词:berberine adipocyte peroxisome proliferator activated receptor CAAT/enhancer binding protein α mRNA expression protein expression

基金项目:国家自然科学基金资助项目(No.30672735);教育部985工程资助项目(No.2001-13);国家教委高校博士点基金资助项目(No.20050246041)

作者	单位
刘毅	复旦大学中西医结合研究所,复旦大学附属华山医院中西医结合科
娄少颖	复旦大学中西医结合研究所,复旦大学附属华山医院中西医结合科
何燕铭	复旦大学中西医结合研究所,复旦大学附属华山医院中西医结合科
陈伟华	复旦大学中西医结合研究所,复旦大学附属华山医院中西医结合科
应健	复旦大学中西医结合研究所,复旦大学附属华山医院中西医结合科
王文健	复旦大学中西医结合研究所,复旦大学附属华山医院中西医结合科

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中文摘要:

目的探讨小檗碱对脂肪细胞增殖、分化的影响及其机制。方法以XTT法检测3T3-Ll前脂肪细胞的增殖;油红O染色并通过比色定量分析检测3T3-L1前脂肪细胞分化过程中胞浆脂质的堆积;采用Real-time PCR和蛋白质免疫印迹(Western blotting)技术检测脂肪细胞分化相关基因过氧化物体增殖剂活化受体γ(peroxisome proliferator activated receptor,PPARγ)、CAAT/增强子结合蛋白α(CAAT/enhancer bindingprotein,C/EBPα)mRNA以及蛋白的表达。结果浓度低于10μmol/L小檗碱干预24h,对脂肪细胞增殖的影响不明显(与空白对照组比较,P>0.05),20、40、80μmol/L小檗碱在作用24h后即表现出明显的抑制效应;不同浓度的小檗碱作用48、72h后对脂肪细胞的增殖亦表现出抑制效应,且有一定的量效关系,即小檗碱浓度越高抑制作用越明显,与空白对照组比较差异有统计学意义(P<0.05,P<0.01);10μmol/L小檗碱处理的前脂肪细胞,分化后胞浆中脂滴明显减少,分化相关基因PPARγmRNA、C/EBPαmRNA和蛋白的表达亦减少,与空白对照组和罗格列酮干预组比较差异有统计学意义(P<0.05,P<0.01)。结论小檗碱能够抑制前脂肪细胞的增殖和分化,减少脂肪细胞分化过程中脂质的堆积,机制可能与其抑制脂肪细胞分化相关基因PPARγ、C/EBPαmRNA和蛋白表达有关,实验为小檗碱防治肥胖等代谢相关性疾病提供了依据。

英文摘要:

Objective To study the effect and mechanism of berberine(BER)on the proliferation and differentiation of adipocytes.Methods The proliferation of 3T3-L1 pre-adipocytes was detected by XTT method.Lipid droplets accumulated in the cytoplasm of adipocytes in the differentiating process were observed by oil red O staining and quantified by colorimetry.The expressions of peroxisome proliferation activated receptor γ(PPARγ),CAAT/enhancer binding protein α(C/EBPα)mRNA and protein were detected by Real-time PCR and Western blotting respectively.Results Intervention with BER in concentration below 10 μmol/L for 24 h showed insignificant effect on the proliferation of adipocytes,as compared with that in the control group(P>0.05);but that in concentrations 20,40 and 80 μmol/L revealed significant suppressive effect;that in different concentrations acting for 48 h and 72 h could affect the proliferation and the effect displayed a dose-dependent manner,i.e.the higher the concentration of BER,the more apparent the suppression,showing significant difference as compared with those in the control group(P<0.05 or P<0.01).The pre-adipocyte treated with 10 μmol/L BER showed that the lipid droplets in the cytoplasm significantly lessened,so did the expression of differentiation related factor PPAR γ mRNA as well as the expressions of C/EBPα mRNA and protein,as compared with those in the blank control group and the group intervened with rosiglitazone,the difference was significant(P<0.05 or P<0.01).Conclusions BER can suppress the proliferation and differentiation of 3T3-L1 pre-adipocytes,reduce the accumulation of lipid drops in the adipocyte differentiating process,which may be associated with its effects in decreasing the expressions of adipocyte differentiation related gene PPARγ,C/EBPα mRNA and protein.The study provides a basis for applying BER on the prevention and treatment of such metabolic related diseases as obesity.

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