摘要
目的观察体外培养的不同月龄雌性大鼠成骨细胞内雌激素受体水平的差异及地塞米松(1×10-8mol/L)对不同月龄雌性大鼠成骨细胞雌激素受体表达的影响。方法采用酶消化法分别提取、纯化和培养新生(24h以内)、2、4、8、12月龄雌性Wistar大鼠成骨细胞,并且用Western-blot、ECL法检测体外培养成骨细胞雌激素受体的水平。结果提取的大鼠成骨细胞生长情况良好,碱性磷酸酶染色阳性,具有成骨细胞特性。Western-blot结果显示:雌性大鼠成骨细胞内雌激素受体水平在新生鼠和8月龄鼠较高,在2、4、12月龄鼠较低;地塞米松组成骨细胞雌激素受体表达水平在新生和2月龄低于对照组,在4、8、12月龄高于对照组。结论不同月龄雌性大鼠体外培养的成骨细胞雌激素受体的表达水平不同;1×10-8mol/L地塞米松对体外培养的雌性大鼠成骨细胞雌激素受体水平有影响且与大鼠的月龄有关。
Objective To study the effects of dexamethasone on the expression of estrogen receptor in osteoblast of different months old female rats. Methods A large amount of osteoblasts were isolated and cultured from the trabecular bone of different months old female Wister rats with trypsin and collagenase Ⅱ digestion method. Osteoblast was confirmed by the morphology alkaline phosphatase staining and estrogen receptor was detected by immunohistochemisty and western-blot. Results The osteoblasts isolated by the method above mentioned showed positive by the methods of morphology alkaline phosphatase staining and immunohistochemisty. The result of western-blot showed that the ER in osteoblast of female rat expression was higher on the newborn and 8-month-old rats than on the 2-,4- and 12-month-old ones. The level of ER expression in the osteoblasts which was affected by the dexamethasone was higher on the 4-,8- and 12-month-old female rats and lower on the newborn and 2-monthold ones, compared with the ER level of the osteoblasts without the effect of dexamethasone. Conclusions The level of ER expression in the osteoblast of female rat is different in rats of different age. ER expression＇s upregulation or down-regulation in osteoblast of rat by dexamethasone at the concentration of 1×10^-1 mol/L is correlated with the age of rats.
出处
《中国骨质疏松杂志》
CAS
CSCD
2007年第6期385-389,共5页
Chinese Journal of Osteoporosis
基金
黑龙江省"十五"攻关项目(G00C190702)
