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

作　　者：刘毅[1] 娄少颖[1] 何燕铭[1] 陈伟华[1] 应健[1] 王文健[1] 

机构地区：[1]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,上海200040

出　　处：《中国中西医结合杂志》2008年第11期1005-1009,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：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)

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。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 （PPART） , 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 〈 O. 05 or P 〈 0.01 ）. The pre-adipocyte treated with 10μmol/L BER showed that the lipid droplets in the cytoplasm significantly lessened, so did the expression of differentiation related factor PPAR γ mR- NA 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 PPAR-γ, 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.

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 

分 类 号：R285.5[医药卫生—中药学]