蛇床子素对骺板TGF-β1及其受体TGF-βRⅡ蛋白表达的影响  

Effects of osthole on expression of TGF-β1 and its receptor Ⅱ in epiphyseal plate

作　　者：张丽 李灵芝[1,2] 张永亮 龚海英[2] 

机构地区：[1]天津市职业与环境危害防制重点实验室,天津300309 [2]武警后勤学院药物化学教研室,天津300309

出　　处：《武警后勤学院学报（医学版）》2014年第12期985-988,992,F0003,共6页Journal of Logistics University of PAP（Medical Sciences）

基　　金：国家自然科学基金项目(81471823);天津市重点项目(12JCZDJC34700)

摘　　要：【目的】考察蛇床子素对小鼠长骨转化生长因子-β1（transforming growth factorβ,TGF-β1）及其受体TGF-βRⅡ蛋白表达的影响,初步探讨该化合物对骨的作用机制。【方法】取16 d胚胎小鼠前肢尺骨在BGJb培养基中培养48 h,培养基中分别含有1×10^-6mol/L雌酚酮、1×10^-7-1×10^-4mol/L四个不同梯度浓度的蛇床子素。采用免疫组织化学方法测定长骨骺板静止区、增殖区和肥大区TGF-β1和TGF-βRⅡ蛋白。【结果】（1）经不同浓度的蛇床子素和雌酚酮处理后,胎鼠长骨骨总长显著增加。与对照组相比,蛇床子素1×10^-7、1×10^-6、1×10^-5mol/L和雌酚酮1×10^-6mol/L组有显著性差异（P〈0.01）。蛇床子素1×10^-4mol/L组骨总长有增加的趋势,但无统计学意义（P〉0.05）。（2）与空白对照组,蛇床子素和雌酚酮组骺板各区TGF-β1和TGF-βRⅡ蛋白水平明显上调。当蛇床子素浓度由1×10^-7、1×10^-6、1×10^-5mol/L逐渐升高时,各区TGF-β1和TGF-βRⅡ蛋白表达升高,而1×10^-4mol/L组各区TGF-β1和TGF-βRⅡ蛋白表达降低。与雌酚酮组相比,蛇床子素组除1×10^-5mol/L组外,其他各组长骨骺板各区TGF-β1和TGF-βRⅡ表达与雌酚酮组相比有统计学意义（P〈0.05）。【结论】（1）蛇床子素能明显促进体外培养胚胎小鼠长骨生长。（2）培养基中加入蛇床子素可上调长骨TGF-β1及其受体TGF-βRⅡ表达,提示可能与其促进体外培养胚胎小鼠长骨生长有关。[ Objective ] To study the effect of osthole on expressions of TGF- β and its receptor Ⅱ （TGF- βR Ⅱ ）in the long bones of fetal mice in vitro for postulating the mechanism by which osthloe exerts its effect on bone. [ Methods ] The fetuses of pregnant mice were removed on day 16 of gestation, then the long bones of the forelimbs of female fetal mice were freed of muscle and soft tissues and cultured in a specifically manufactured device for 48 h in BGJb medium which contain 10^-6 mol/L estrone and 10^-7, 10^-6, 10^-5, 10^-4 mol/L osthole respectively. After cultured for 48 h, the long bones were harvested and immunohistochemical analysis was performed for determination of TGF-β1 and TGF-βⅡ proteins expression in epiphyseal plates. The number and their area of positive cells in the resting zone, the proliferative zone and the hypertrophic zone in epiphyseal plate were determined under image analytic system. [ Results] （1）After the bones were treated with different concentrations of osthole and estrone, the total length of the bones were longer significantly than that of blank. Compared with blank group, osthole at concentration of 1 ×10^-7, 1 ×10^-6, 1 ×10^-5 and 1 ×10^-6 mol/L estrone increased the length remarkably（P〈 0.05）. There is no difference between the osthole 1 ×10^4 mol/L （P 〉 0.05 ）. （2） Compared with the control group, the levels of TGF-β1 land TGF-βRⅡ proteins were significantly up-regulate .In the cartilaginous epiphyses, the expression of TGF-β1 and TGF-βR Ⅱ proteins were up-regulated by 1 ×10^-6 mol/L estrone and up-regulated dose-dependently by osthole in the range of 1 ×10^-7 mol/L to 1 ×10^5 mol/L, but down-regulated by osthole of 1 ×10^-4 mol/L. Compared with estrone group, the expression of TGF-β1 and TGF-βRⅡ in the groups treated with different concentration osthole excluding osthole 1×10^-5 mol/L decreased siginificantly（P 〈 0.05）. [ Conclusion ] Osthole could accelerate the bone formation in vitro. The expressions of TGF-β1 and

关 键 词：蛇床子素 雌酚酮 长骨 转化生长因子-β1 转化生长因子β受体Ⅱ 

分 类 号：R285.5[医药卫生—中药学]