牛蒡子苷对分化的小鼠C2C12细胞的影响

EFFECT OF ARCTIIN ON CELL GROWTH IN MOUSE C2C12 CELLS

目的观察牛蒡子中木脂素成分牛蒡子苷对C2C12成肌细胞及分化后肌细胞生长的影响并探讨其作用机制。方法以0、0.01、0.1、1、10、100和1000 ng/ml共7种剂量大豆苷原和牛蒡子苷分别处理体外培养的C2C12成肌细胞和经2%马血清诱导分化的肌细胞。大豆苷元作为对照,采用四唑盐(MTT)比色法检测C2C12成肌细胞增殖能力,以诱导分化后的肌细胞内蛋白含量和磷酸肌酸激酶含量变化为指标观察大豆苷原和牛蒡子苷对分化后的肌细胞生长的影响,采用real-time PCR分析C2C12细胞中雌激素受体β(ERβ)mRNA表达量。结果牛蒡子苷和大豆苷原终浓度为10 ng/ml时均能极显著促进C2C12成肌细胞增殖(P<0.01),促进肌细胞总蛋白质的合成(P<0.01),促进肌细胞肌酸激酶的活性(P<0.01),促进肌细胞中ERβmRNA表达量极显著上升。结论牛蒡子苷可提高C2C12成肌细胞的增殖和分化后的肌细胞肥大,促进骨骼肌细胞肥大型生长。这种促生长作用是通过直接作用于肌细胞上的ERβ介导的,提示牛蒡子苷有望开发为一种新型的绿色有效的生理调节剂。[营养学报,2012,34(4):395-399]

Objective To determine whether arctiin from Fructus Arctii could affect the growth of C2C12 myoblasts and the differentiated myocytes and identify its mechanism.Method The growth of C2C12 myoblasts and the different-tiated myocytes induced by 2% horse serum was observed in vitro by treatment with different doses of of arctiin（0,0.01,0.1,1,10,100 and 1000 ng/ml）.Daidzein was used as control.C2C12 myoblasts＇ proliferation was assessed by MTT method,and the differentiated myoblasts＇ total protein,CK activities and the expression of estrogen receptor β（ER β） were detected by Coomassie brilliant blue method,colorimetric analysis and QRT-PCR,respectively.Results C2C12 myoblasts＇ proliferation was significantly upregulated（P0.01）.The differentiated myocytes＇ total protein（P0.01） and CK activities（P0.01） were markedly increased after treatment of actiin at the concentration of 10ng/ml.Meanwhile,the expression of estrogen receptor β（ER β） in differentiated myoblasts＇ was increased by actiin at the same concentration.Conclusion Arctiin can facilitate the proliferation of C2C12 myoblasts,and the hypertrophic growth of skeletal muscle cell after differentiation by binding to ERβ located within C2C12 myoblasts and differentiated myocytes.Consequently,arctiin might become potentially a new type of green and effective physiological modulator.