黄连素对3T3-L1前脂肪细胞增殖、过氧化物酶体增殖物激活受体γ、CAAT/增强子结合蛋白mRNA及蛋白表达的影响

Liu Yi, Lou Shao-Ying, He Yan-Ming

Institute of Integrative Medicine, Fudan University, Shanghai.

Zhongguo Zhong Xi Yi Jie He Za Zhi. 2008 Nov;28(11):1005-9.

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 gamma (PPARgamma), CAAT/enhancer binding protein alpha (C/EBPalpha) mRNA and protein were detected by Real-time PCR and Western blotting respectively.

RESULTS

Intervention with BER in concentration below 10 micromol/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 micromol/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 micromol/L BER showed that the lipid droplets in the cytoplasm significantly lessened, so did the expression of differentiation related factor PPAR gamma mRNA as well as the expressions of C/EBPalpha 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 PPARgamma, C/EBPalpha mRNA and protein. The study provides a basis for applying BER on the prevention and treatment of such metabolic related diseases as obesity.

目的

研究黄连素（BER）对脂肪细胞增殖和分化的影响及其机制。

方法

采用XTT法检测3T3-L1前脂肪细胞的增殖情况。用油红O染色观察脂肪细胞分化过程中细胞质内脂滴的积聚情况，并通过比色法进行定量分析。分别采用实时荧光定量PCR和蛋白质免疫印迹法检测过氧化物酶体增殖物激活受体γ（PPARγ）、CCAAT/增强子结合蛋白α（C/EBPα）mRNA及蛋白的表达。

结果

与对照组相比，浓度低于10 μmol/L的BER干预24 h对脂肪细胞增殖无明显影响（P>0.05）；而浓度为20、40和80 μmol/L时则显示出显著的抑制作用；不同浓度作用48 h和72 h均可影响增殖，且呈剂量依赖性，即BER浓度越高，抑制作用越明显，与对照组相比差异有统计学意义（P<0.05或P<0.01）。用10 μmol/L BER处理的前脂肪细胞，其细胞质内脂滴明显减少，分化相关因子PPARγ mRNA的表达以及C/EBPα mRNA和蛋白的表达也减少，与空白对照组和罗格列酮干预组相比，差异有统计学意义（P<0.05或P<0.01）。