染料木黄酮对乳鼠颅骨成骨细胞增殖分化的影响及其与c-jun的关系

Role of c-jun in the effect of genistein on neonatal rat calvaria osteoblasts proliferation and differentiation

目的研究染料木黄酮对成骨细胞活性的影响及其相关机制。方法胰蛋白酶和Ⅱ型胶原酶分步消化法获得乳鼠盖骨成骨细胞,Ⅱ代细胞用于实验。MTT和3H-TdR测定成骨细胞增殖和DNA合成,原位杂交的方法测定c-jun表达。结果成骨细胞培养基中加入(10-5、10-6和10-7mol/L)染料木黄酮或10-9mol/L和10-10mol/L雌激素培养48h和72h后,MTT的吸光度值与对照组相比均明显升高,48h和72h后对照组、染料木黄酮组和雌激素组MTT值分别为0.19、0.15;0.39、0.45、0.46;0.29、0.32、0.37和0.35、0.38;0.50、0.49。3H-TdR掺入量均显著增加,对照组、染料木黄酮组和雌激素组3H-TdR掺入量分别为68.47;101、844、512和1108.8、1204.2。c-jun表达各组差异无显著性。结论染料木黄酮不是通过促进c-jun表达来促进成骨细胞增殖与分化。

Objective To study the role of the c-jun in the effects of genistein on osteoblasts in neonatal rat calvaria cultures in vitro. Methods Osteoblasts were isolated from neonatal rat calvaria through trypsin and collagenase digestion, and cultured in the medium at the doses of 10^-5 , 10^-6 and 10^-7 mol/L genistein and at the dose of 10^-9 mol/L and 10^-10 mol/L estrogen for 48h and 72h. The proliferation and DNA of osteoblasts were assayed by MTT method and 3 H-TdR incorporation, and c-jun was measured by in situ hybridization assay. Results Genistein significantly promoted osteoblast proliferation and DNA synthesis. The values of MTr in control and genistein and estrogen groups at 48h and 72h,were 0.19, 0.15, and 0.39, 0.45, 0.46, and 0.29, 0.32, 0.37 and 0.35,0.38, and 0.50, 0.49. The content of ^3H-TdR incorporation in control and genistein and estrogen groups were 68.47, and 101,844,512 and 1108.8,1204.2. There was no difference of expression of c-jun among control and genistein and estrogen groups. Conelusion Genistein could promoted osteoblast proliferation and differentiation not associated with c-jun.