染料木黄酮对成骨细胞内游离钙、钙调素水平及ALP、ATP酶的影响

目的研究不同浓度的染料木黄酮（genistein，GEN）对体外培养大鼠成骨细胞游离钙、钙调素、碱性磷酸酶（alkaline phosphatase，ALP）、ATP酶的影响。方法采用新生大鼠颅骨分离培养成骨细胞，加入不同浓度的染料木黄酮（10^-8mol／L，10^-7mol／L，10^-6mol／L，10^-5mol／L），以妊马雌酮（商品名：倍美力）（10^-8mol／L，10^-7mol／L，10^-6mol／L，10^-5mol／L）作为阳性对照组，用PNPP（对硝基苯磷酸盐）法测定ALP活性、放射生物法测定钙调素水平、激光扫描共聚焦显微镜测定细胞游离钙、比色法测定ATP酶活性。结果染料木黄酮对成骨细胞钙调素、ALP、Ca^2＋-ATP、细胞内游离钙水平的影响呈正相关性，并与其剂量成正相关性，但这些作用均低于雌激素水平（P〈0．01）。结论染料木黄酮能增加成骨细胞ALP、Ca^2＋-ATP活性和钙调素水平、钙离子震荡波，并与其浓度有关，作用与雌激素相似。

Effect of genistein on the levels of intracellular calcium, calmodulin, ALP and ATP expressed in cultured rat osteoblasts

Objective To study the effect of different concentrations of genistein on the levels of calcium (Ca~(2+)), calmodulin, alkaline phosphatase(ALP) and ATP in cultured rat osteoblasts.Methods Newborn rat osteoblasts in calvarid were isolated and cultured with various concentrations of genistein (10 -8 mol/L, 10 -7 mol/L, 10 -6 mol/L and 10 -5 mol/L) and estrogen (10 -8 mol/L, 10 -7 mol/L, 10 -6 mol/L and 10 -5 mol/L). The ALP, calmodulin content and ATPases activities of the cell membranes were measured by biochemical assay, radiobiological analysis and colorimetric analysis, respectively. The changes of Ca~(2+) was determined with laser scanning confocal microscopy.Results The activity of ALP, calmodulin, Ca~(2+)-ATP and the level Ca~(2+) were all positively related to the concentration of genistein added to the medium. However, these effects were less significant than those to estrogen (P<0.01).Conclusions Genistein could promote the activity of ALP, calmodulin, Ca~(2+)-ATP and can enhanced the frequence of calcium oscillation dose-dependently which is similar to the effect of estrogen.