为探讨小檗碱对前体脂肪细胞增殖和分化的影响,本文通过MTT法检测3T3-L1前体脂肪细胞增殖和油红O染色与酶标仪检测细胞内甘油三酯含量等分析小檗碱对其细胞形态及功能的影响;通过实时荧光定量PCR方法检测前体脂肪细胞成脂分化相关基因的表达水平,分析小檗碱影响其成脂分化可能的作用途径。结果表明,小檗碱浓度高于20μmol·L^-1时抑制3T3-L1细胞增殖,甚至出现部分细胞死亡,而5μmol·L^-1小檗碱对其抑制作用较弱;小檗碱浓度越大对3T3-L1细胞内脂滴堆积的抑制作用越明显;实时荧光定量PCR检测表明,不同浓度的小檗碱明显抑制C/EBPβ和SREBP-1c的基因表达,且其能够通过影响mi RNA表达而发挥更广泛的作用。
To investigate the effects of berberine on the proliferation and differentiation of preadipocytes, the multiplication of 3T3-L1 preadipocytes was detected by MTT method, and lipid droplets accumulated in preadipocytes were observed by oil O staining and quantified by colorimetry, and the expression of genes related to preadipocytes differetiation was analyzed by realtime RT-PCR. The results showed that the concentration of berberine over 20 μmol · L^-1 could lead to the inhibitation of 3T3-L1 preadipocytes proliferation, cells death, and the inhibitory effect of 5 μmol · L-1berberine was weak on its proliferation; Meanwhile, the accumulation of intracellular lipid droplets was more suppressed when 3T3-L1 preadipocytes treated with higher concentration of berberine; The expression of differentiation related factor C/EBPβ and SREBP-1c was downregulated and the effect on miR-143 gene expression displayed a dose-dependent manner by real-time PCR analysis. BBR weakens adipogenesis by impairing miR-143 expression maybe indirectly and provokes comprehensive changes in function.