【摘要】 目的:观察染料木黄酮对成骨细胞活性的影响及其相关机制。方法:实验于2001-05/2003-05在卫生学环境医学研究所实验室完成。①成骨细胞培养:无菌条件下分离出生3d的乳鼠颅盖骨,剪碎,加入胰蛋白酶和Ⅱ型胶原酶,用含体积分数为0.1的胎牛血清的F-12培养基重悬,调整细胞浓度后接种于25mL细胞培养瓶,Ⅱ代细胞用于实验。②实验方法:实验分为染料木黄酮组、雌激素组和对照组(吐温20),染料木黄酮剂量分别为10-5,10-6、10-7mol/L,雌激素剂量分别为10-9,10-10mol/L,培养48,72h后,应用MTT和3H-TdR掺入实验观察不同剂量的染料木黄酮和雌激素对成骨细胞活性影响,免疫组化的方法测定白细胞介素6表达。结果:①A570nm值:培养48h,染料木黄酮10-5,10-6,10-7mol/L组分别比对照组增加105%,136%和143%;10-9,10-10mol/L雌激素组分别比对照组增加84%和100%;培养72h,染料木黄酮3个剂量组分别比对照组增加93%,113%和146%,各剂量组与对照组相比均有显著性差异(P<0.05)。②3H-TdR掺入量:10-5,10-6,10-7mol/L染料木黄酮组分别比对照组增加0.47,11.29,6.45倍;10-9和10-10mol/L雌激素组增加15.4倍和16.5倍(P<0.05)。③白细胞介素6表达:各组成骨细胞周围均有较强的呈棕色的阳性颗粒,但染料木黄酮组组与对照组相比无明显差别。结论:①10-5～10-7mol/L染料木黄酮和雌激素一样可促进成骨细胞增殖和DNA合成。②染料木黄酮不是通过促进白细胞介素6表达来促进成骨细胞增殖与分化。更多还原

【Abstract】 AIM: To investigate the effects of genistein on osteoblasts in neonatal rat calvaria,and explore the related mechanism. METHODS: The study was finished at the laboratory in Institute of Health and Environment Medicine from May 2001 to May 2003.①Culture of osteoblasts: Calvarias from newborn 3 days rats were aseptically isolated and cut into pieces,then digested through trypsin and collagenase,precipitation was suspended with F-12 culture medium containing 10% fetal bovine serum and then cultured in 25 mL culture bottle after adjusting cell density. The second generation cells were used in the experiment.②The experiment was divided into three groups: control (tween 20),genistein and estrogen. Doses of genistein were 10-5,10-6 and 10-7 mol/L,the concentration of estrogen was 10-9 and 10-10 mol/L,respectively. The proliferation of osteoblasts was assayed by MTT method and 3H-TdR incorporation. Interleukin (IL)-6 was measured by immunohistochemical assay after culturing 48 and 72 hours. RESULTS: ①The value of A570 nm: Compared with control group,the value in the genistein of 10-5,10-6,10-7 mol/L groups was significantly increased 105%,136%,143% and 93%,113%,146%,respectively after culturing 48,72 hours; the estrogen of 10-9,10-10 mol/L groups increased 84% and 100% after culturing 48 hours; the difference was significant between treatment groups and control group (P < 0.05).②The quantity of 3H-TdR incorporation: The genistein of 10-5,10-6,10-7 mol/L groups significantly increased 0.47,11.29,6.45 folds respectively,while the estrogen of 10-9 and 10-10 mol/L groups increased 15.4 and 16.5 folds (P < 0.05) compared with control group.③The expression of IL-6: There were a lot of positive brown particles around osteoblasts of all groups,but no significant difference was found between genistein group and control group. CONCLUSION: ①Genistein and estrogen can promote osteoblast proliferation and DNA synthesis.②Genistein promotes the proliferation and differentiation of osteoblasts not by the expression of IL-6.更多还原