4,5,6-三羟基异黄酮对原代培养大鼠成骨细胞增殖、分化、钙含量及矿化功能的影响(英文)

Effects of genistein on proliferation,differentiation,content of matrix calcium and mineralization in primary cultured rat osteoblasts

背景:骨质疏松症是人类老年时生活质量下降和寿命缩短的主要原因之一,而其发生与成骨细胞和破骨细胞的功能失偶联有关,因而在细胞水平上观察药物对成骨细胞功能的调整作用是评价骨质疏松症防治药物药效的方法之一。目的:了解4,5,6-三羟基异黄酮(Genistein)对体外培养成骨细胞的作用。设计:以大鼠成骨细胞为研究对象的随机对照单盲研究。单位:一所市级医院的检验科。材料:本实验于2001-02/2001-12在北京中日友好医院核医学科同位素室完成。选择10只出生24h的SD大鼠,(购自北京维通利华实验动物技术有限公司,许可证编号为SCXK11-00-0008),实验室级别为SPF级。方法:取小鼠头盖骨成骨细胞进行体外培养,应用四氮唑盐(MTT)法、对硝基苯磷酸盐法、原子吸收分光光度法及茜素红染色方法观察Genistein对体外培养成骨细胞的增殖、碱性磷酸酶表达、基质钙含量及矿化结节形成的影响。主要观察指标:形态观察、增值率测定、碱性磷酸酶染色、细胞基质钙含量测定、茜素红染色显示矿化结节数。结果:Genistein刺激成骨细胞的增殖,提高了碱性磷酸酶活性,细胞基质钙含量及矿化结节形成的数量增大。结论:Genistein具有刺激体外培养成骨细胞增殖、分化成熟及促进矿化的作用。

BACKGROUND:Osteoporosis has been one of the main reasons for old people's in decline living standard and shortening of lifespan.The onset of osteoporosis is related to the loss of functional coupling in osteoblast and osteoclast.So observation of the regulative effect of medicines on osteoblast is a good method to evaluate their preventive and therapeutical efficacy in osteoporosis. OBJECTIVE:To investigate the effects of genistein on osteoblast culture in vitro. DESIGN:A randomined,controlled and single-blind study based on rat osteoblasts. SETTING:Department of laboratory in amunicipal hospital. MATERIALS:The experiment was completed in the Room of Isotope,Department of Nuclear Medicine,China-Japan Friendship Hospital of Beijing from February to December 2001.Ten SD rats of 24 hours old were provided from Beijing Weitong Lihua Experimental Animal Ltd.Co.(license: SCXK11-00-0008).The laboratory was SPF grade. METHODS:Skull osteoblasts of rats were selected to culture in vitro. Microculture tetrozolium,p-nitrophenyl phosphate,atomic absorptiometry and alizarin red were used to observe the effect of genistein on proliferation, activity of alkaline phosphatase(ALP), contents of matrix calcium and the number of mineral nodes of osteoblasts cultured in vitro. MAIN OUTCOME MEASURES:Morphological observation, determination of proliferation ratio,ALP stain,determination of matrix calcium accumulation and number of mineralized nodes with alizarin red stain. RESULTS:It was found that genistein stimulated the proliferation of osteoblasts,improved the ALP activity,and increased the contents of matrix calcium and the number of mineral nodes in cultured osteoblasts. CONCLUSION:Genistein has the effects on stimulating the proliferation, differentiation, maturation and mineralization of osteoblasts cultured in vitro.