染料木黄酮对乳鼠颅盖骨成骨细胞增殖分化的影响及其与白细胞介素6的关系  

Role of interleukin-6 in the effect of genistein on the proliferation and differentiation of neonatal rat calvaria osteoblasts

作　　者：张月红[1] 金宏[2] 许志勤[2] 南文考[2] 王先远[2] 薛长勇 高兰兴[2] 

机构地区：[1]解放军总医院营养科,北京市100853 [2]卫生学环境医学研究所,天津市300050

出　　处：《中国组织工程研究与临床康复》2007年第36期7142-7145,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

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。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 cellagenase, 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 A570nm： 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 mot/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 pr

关 键 词：染料木黄酮 成骨细胞 白细胞介素6 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]