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目的 探讨小檗碱对脂肪细胞分化的影响和其机制。方法 培养3T3-L1前脂肪细胞。以四甲基偶氮唑盐(MTT)方法检测3T3-L1前脂肪细胞的增殖,以油红O染色检测3T3-L1脂肪细胞分化过程中胞浆脂肪的堆积,同时采用逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹技术检测小檗碱对过氧化脂质体增殖激活受体γ2(PPARγ2)mRNA和蛋白质表达的影响。结果 10μmol/L小檗碱使3T3-L1前脂肪细胞的MTT值增加36%(P<0.001),小檗碱处理的脂肪细胞胞浆中,脂肪的堆积明显少于对照组,仅10％～20％的细胞出现较大脂滴。RT-PCR结果显示,在3T3-L1脂肪细胞分化过程中,小檗碱组脂肪细胞的PPARγ2 mRNR较对照组低48％(P<0.01),蛋白质免疫印迹结果也显示小檗碱抑制PPARγ2蛋白质的表达。结论 小檗碱促进前脂肪细胞的增殖,小檗碱减少脂肪细胞分化过程中脂质的堆积、抑制脂肪细胞的分化,可能与其降低PPARγ2mRNA和蛋白质的表达有关,这提示小檗碱在临床上可能适合于治疗肥胖的2型糖尿病患者。
Objective To investigate the effect of berberine on adipocyte differentiation and its mechanism. Methods 3T3-L1 preadipocytes were cultured. The proliferation of 3T3-L1 preadipocytes was detected by MTT assay. The accumulation of cytoplasmic fat during the differentiation of 3T3-L1 adipocytes was detected by oil red O staining, and the reverse transcriptase-polymerase chain (RT-PCR) and Western blotting were used to detect the effect of berberine on the expression of PPARγ2 mRNA and protein in peroxisome proliferator-activated receptors. Results Berberine 10 μmol / L increased the MTT of 3T3-L1 preadipocytes by 36% (P <0.001). The accumulation of fat in cytoplasm of berberine-treated adipocytes was significantly less than that of the control group (10% 20% of the cells appear larger lipid droplets. RT-PCR results showed that the PPARγ2 mRNR of berberine adipocytes was 48% lower than that of the control group (P <0.01) during the differentiation of 3T3-L1 adipocytes, and western blot also showed that berberine inhibited PPARγ2 protein expression. Conclusion Berberine can promote the proliferation of preadipocytes. Berberine can reduce the accumulation of lipids and inhibit the differentiation of adipocytes during adipocyte differentiation, which may be related to the decrease of PPARγ2 mRNA and protein expression, suggesting that berberine may be clinically possible Type 2 diabetes suitable for the treatment of obesity.