雌激素和来曲唑对鸡胚额骨成骨细胞影响的差异比较  

Effects of estrogen versus letrozole on chicken embryo frontal bone osteoblast

作　　者：邓益锋[1] 陈秀霞[1] 胡云峰[1] 侯加法[1] 

机构地区：[1]南京农业大学动物医学院,江苏省南京市210095

出　　处：《中国组织工程研究与临床康复》2010年第7期1157-1161,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30671546);南京农业大学青年创新基金资助项目(KJ0716)~~

摘　　要：背景:雌激素对哺乳动物成骨细胞影响研究较多,但其对禽类成骨细胞的影响还未见报道;来曲唑对骨代谢的不良反应报道较多,但其对成骨细胞的影响还未见报道。目的:实验创新性为通过观察雌激素、来曲唑对体外培养鸡胚成骨细胞增殖、周期及雌激素受体mRNA表达和碱性磷酸酶活性的影响,以阐明蛋鸡髓质骨的代谢机制。方法:取15日龄SPF鸡胚,酶消化法获取鸡胚额骨成骨细胞,分别采用不同质量浓度雌激素(0,5,10,20,100,200,400,800,2000,20000ng/L)和来曲唑(0,5,10,25,50,100,250,500,1000,5000μg/L)处理。以四甲基偶氮唑盐法测定细胞增殖率,以pNPP法测定碱性磷酸酶活性,以流式细胞术测定细胞周期情况,以用实时荧光定量PCR法测定雌激素受体mRNA的表达。结果与结论:雌激素可促进鸡胚成骨细胞增殖,且具有浓度和时间依赖性,雌激素能诱导成骨细胞雌激素受体mRNA表达,推动细胞周期进程,提高鸡胚成骨细胞碱性磷酸酶活性。来曲唑可促进成骨细胞增殖,推动细胞周期进程,抑制成骨细胞雌激素受体mRNA表达,但来曲唑对碱性磷酸酶的合成与分泌没有明显影响。BACKGROUND： There are plenty of studies of estrogen effects on mammalian osteoblast, but the studies of estrogen effects on bird osteoblast cannot be found. There are many reports about the side effects of letrozole on bone metabolism, but there are no reports about the effect of letrozole on osteoblast. OBJECTIVE： The effects of estrogen and letrozole on the proliferation, cell cycle, estrogen receptor mRNA expression and alkaline phosphatase （ALP） activity of chicken osteoblast in vitro were studied in order to illustrate the mechanism of medullary bone osteogenesis. METHODS： The osteoblasts were harvested from the frontal bone of 15-day SPF chicken embryos by the enzyme digestion, and treated with various mass concentrations of estrogen （0, 5, 10, 20, 100, 200, 400, 800, 2 000, 20 000 ng/L） and letrozole （0, 5, 10, 25, 50, 100, 250, 500, 1 000, 5 000 μg /L）. The proliferation rates of the osteoblast treated with estrogen or letrozole were measured through the MTT method. The ALP activities of osteoblast were measured by the pNPP method. The cell cycle was measured by flow cytometry. The expression of estrogen receptor mRNA was detected using real-time fluorescent quantitative polymerase chain reaction （PCR）. RESULTS AND CONCLUSION： The estrogen could promote proliferation of osteoblast in concentration-and time-dependent fashion. Estrogen could increase the expression of estrogen receptor mRNA, impulse cell cycle, and elevate ALP activities. Letrozole could increase the cell population, impulse cell cycle, inhibit estrogen receptor mRNA expression, but letrozole has no effects on ALP synthesis and secretion.

关 键 词：鸡胚 成骨细胞 雌激素 来曲唑 雌激素受体 增殖 碱性磷酸酶 

分 类 号：R318[医药卫生—生物医学工程]