染料木黄酮对成骨细胞I型胶原表达及转化生长因子β_1的影响

EFFECTS OF GENISTEIN ON THE EXPRESSION OF COLLAGEN I AND TRANSFORMING GROWTH FACTOR-β_1 OF OSTEOBLAST

目的染料木黄酮(genistein,GEN)与新生SD大鼠颅骨成骨细胞(osteoblast,OB)共同孵育,研究GEN对OBI型胶原表达及转化生长因子β1(transforminggrowthfactor-β1,TGF-β1)的影响。方法采用第二继代OB进行实验,实验分为对照组,GEN各浓度组(10-8,10-7,10-6,10-5mol/L),雌激素组(17-βestradiol,E2,10-10mol/L)。测定指标包括四甲基偶氮唑盐(methythiazolyltetrazoliumbromide,MTT)、胞浆内蛋白、碱性磷酸酶活性(activityofalkalinephosphatase,ALP)、以及I型胶原与TGFβ1表达。结果体外培养48h及72h,GEN各剂量组和E2组的MTT(OD)值均有显著性增高。与培养48h相比,对照组、10-8、10-7、10-6mol/LGEN剂量组72h后细胞增殖有所增加,差异有显著性。10-6、10-5mol/LGEN剂量组和E2组可增高OB内蛋白含量,差别有显著性。GEN各组和E2组均可增高OB内ALP活性,差别有显著性。上述指标与GEN剂量呈显著相关。10-7、10-6、10-5mol/LGEN组和E2组OB内I型胶原的表达及TGFβ-1的合成高于对照组。I型胶原表达和GEN剂量呈显著相关。结论GEN可刺激OB增殖、分化,增加OB内I型胶原的表达及TGFβ-1的分泌,在10-8M～10-5M范围内与剂量相关;高浓度GEN与E2相比作用相似。

Objective To investigate the effects of genistein （GEN） on the expression of collagen Ⅰ and transforming growth factor-β1（TGF-β1 ） of osteoblast. Method The secondary generation of skull osteoblast of newborn SD rat was incubated with GEN. The cells were divided into six groups： control group, different dose of GEN （10^-8, 10^-7, 10^-6, 10^-5mol/L, respectively） groups and EE group （10^-10mol/L）. MTT （OD）, the contents of cell protein, the activity of alkaline phosphatase （ALP）, the expression of collagen I and the content of TGF-β1 were detected. Results After 48h and 72h, the MTT（OD） of all GEN group and EE group were significantly higher than those in control group. The MTT（OD） of control group and 10^-8,10^-7,10^-6mol/L GEN groups in 72h were significantly higher than those in 48 h. The protein of 10^-5,10^-6 mol/L GEN group and EE group were significantly higher than those in control group. The ALP activity of all GEN groups and EE group were significantly higher than those in control group.The level of above indices were correlated with the dose of GEN. The expression of collagen I and the content of TGF-β1 in 10^-7, 10^-6, 10^-5mol/L GEN group and EE groups were higher than those in control group. They were correlated with the dose of GEN and TGF-β1. Conclusion GEN could stimulate proliferation and differentiation of osteoblast, and enhance the expression of collagen Ⅰ and content of TGFβ-1. Compared with E2,, there were similar effects with the higher dosage of GEN.