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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 Genistein (GEN) was co-incubated with neonatal SD rat osteoblast (OB) to investigate the effect of GEN on the expression of OBI-type collagen and transforming growth factor-β1 (TGF-β1). Methods The second passage OB was used for experiments. The experiment was divided into control group, GEN concentration groups (10-8, 10-7, 10-6, 10-5mol/L), estrogen group (17-βestradiol, E2, 10-10 mol/L). Measured indicators include methythiazolyltetrazoliumbromide (MTT), intracytoplasmic protein, activity of alkaline phosphatase (ALP), type I collagen and TGFβ1 expression. Results The MTT (OD) values of GEN in each dose group and E2 group were significantly increased at 48h and 72h in vitro. Compared with the culture for 48 hours, the proliferation of the cells in the control group, 10-8, 10-7, and 10-6mol/LGEN groups increased after 72 hours, and the difference was significant. 10-6, 10-5mol/LGEN dose group and E2 group can increase OB protein content, the difference was significant. Both GEN and E2 increased the ALP activity in OB, and the difference was significant. The above indicators were significantly associated with GEN dose. The expression of type I collagen and the synthesis of TGFβ-1 in OB in 10-7, 10-6, 10-5mol/LGEN and E2 groups were higher than those in the control group. There was a significant correlation between type I collagen expression and GEN dose. Conclusions GEN can stimulate the proliferation and differentiation of OB, increase the expression of type I collagen in OB and the secretion of TGFβ-1. It is related to dose in the range of 10-8M～10-5M. The effect of high concentration of GEN is similar to that of E2.