吲哚昔芬对大鼠成骨细胞增殖和Ⅰ型胶原合成的影响

Effect of idoxifene on the proliferation of osteoblasts and typeⅠcollagen synthesis in rats

目的研究吲哚昔芬对大鼠成骨细胞增殖和Ⅰ型胶原合成的影响。方法分离、培养大鼠头盖骨成骨细胞,分别采用四唑盐(MTT)比色实验和3H-Proline(3氢-脯氨酸)法测定成骨细胞增殖和Ⅰ型胶原合成的情况。结果吲哚昔芬作用大鼠成骨细胞48、72h,1×10-10 mol/L吲哚昔芬与对照组比较,MTTA490nm值显著升高(P<0.05),1×10-9、1×10-8 mol/L的吲哚昔芬使MTTA490nm值进一步升高(P<0.01)。经1×10-10 mol/L吲哚昔芬作用72、96h,3H-Proline掺入值显著升高(P<0.05),1×10-9、1×10-8 mol/L的吲哚昔芬使3H-Proline掺入值进一步升高(P<0.05)。分别用1×10-10 mol/L吲哚昔芬和雌二醇作用大鼠成骨细胞72h,二者的MTTA490nm值和3H-Proline掺入值均显著高于对照组,且吲哚昔芬MTTA490nm值和3H-Proline掺入值显著高于雌二醇(P<0.05)。结论吲哚昔芬发挥抗骨质疏松作用在短期主要促进成骨细胞增殖,中期既促进成骨细胞增殖又促进Ⅰ型胶原合成,而长期效应主要是通过促进胶原合成实现,并且其效应显著强于雌二醇,提示吲哚昔芬对骨质疏松的防治作用可能优于雌二醇。

Objective To investigate the effect of idoxifene on the proliferation of osteoblasts and typeⅠcollagen synthesis in culture from rat calvarium. Methods Calvarial osteoblasts were isolated and cultured from fetal rats. The growth of osteoblasts and typeⅠ collagen synthesis were measured by the means of MTT method and 3H-Proline incorporation respectively. Results In the proliferation of osteoblasts, MTT A490 nm value of osteoblasts was significantly increased by idoxifene (1×10-10 mol/L) for 48 h and 72 h (P< 0.05 vs control), and the values of osteoblasts were much higher in the presence of idoxifene (1×10-9 and 1×10-8 mol/L) and significantly difference( P< 0.01) comparing to that of control group. In the typeⅠcollagen synthesis typeⅠcollagen synthesis, 72-96 h after adding idoxifene (1×10-10 mol/L), 3H-Proline incorporation of osteoblasts was also enhanced with significant difference (P< 0.05) comparing to that of control group, and it was much higher in the presence of idoxifene (1×10-9,1×10-8 mol/L, P< 0.05 vs control). After treating with 1×10-10 mol/L idoxifene and estradiol 72 h respectively, the value of MTT A490 nm and 3H-Proline incorporation of osteoblasts were both significantly higher than those of control group, and the values of MTT A490 nm and 3H-Proline incorporation of osteoblasts treated with idoxifene were higher than that osteoblasts in the presence of estradiol (P< 0.05). Conclusion The short-term anti-osteoporosis effect of idoxifene is mainly achieved by enhancing the proliferation of osteoblasts, it's middle term effect is to stimulate both proliferation of osteoblasts and typeⅠcollagen synthesis, and the long term effect is the enhancement of the collagen synthesis, and its function is more potent than estradiol, the anti-osteoporosis effect of idoxifene is probablely better than estradiol.