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福莫特罗和ICI118551对大鼠成熟破骨细胞骨吸收功能的影响
The Effects of Formoterol and ICI118551 on Rat Ostoeclast in Vitro
【作者】 杨向东；
【作者基本信息】 南京医科大学 ， 骨科学， 2010， 硕士
【摘要】 [目的]:骨质疏松尤其是绝经后骨质疏松是常见老年性疾病,易造成骨折等相关并发症,对个人和社会造成的危害巨大。近年来相关基础研究也有很多新的进展,发现交感神经系统对骨代谢的调节作用是近年来在骨代谢研究方面取得的成就之一。瘦素-交感神经系统对骨代谢的影响是近年来研究的热点,瘦素可通过激活中枢下丘脑神经元,继而激活外周交感神经系统,再通过激活骨组织细胞表面的β2肾上腺素能受体而抑制骨形成。本实验通过研究选择性β2肾上腺素能受体激动剂福莫特罗（Formoterol）和选择性β2肾上腺素能受体阻滞剂ICI118551对体外大鼠成熟破骨细胞（osteoclast,OC）功能的影响,探讨β2肾上腺素能受体信号对骨代谢的影响。[方法]:取清洁级出生24h内的SD乳大鼠,长骨干骨髓腔内壁机械分离成熟OC后分别加入不同浓度(1×10-5mol/L,1×10-6mol/L,1×10-7mol/L,1×10-8mol/L,1×10-9mol/L)的Formoterol和ICI118551,以抗酒石酸酸性磷酸酶（TRAP）染色法观察破骨细胞形态,甲苯胺蓝染色法在100和250倍倒置相差显微镜下观察计数并拍照骨片上的骨吸收陷窝,Image-Pro Plus 6.0图像软件分析骨片上骨吸收陷窝面积。[结果]:破骨细胞与骨片共培养6天,Formoterol在1×10-5mol/L～1×10-9mol/L浓度时与空白对照组比明显增加骨片上OC的骨吸收陷窝数目和面积; ICI118551在1×10-5mol/L～1×10-9mol/L浓度时与空白对照组相比明显降低骨片上骨吸收陷窝的数目和面积,且随着ICI118551浓度的提高骨吸收陷窝的数目和面积逐渐减小。[结论]:β2肾上腺素能受体激动剂可促进体外培养OC的骨吸收功能,阻滞剂对OC的骨吸收功能有抑制作用。
【Abstract】 【Objectives】: To evaluate the effects of Formoterol （β2-adrenergic receptor-specific agonist） and ICI118551 （β2-adrenergic receptor-specific antagonist） on rat matured ostoeclast cells in vitro.【Methods】: Mature OC were isolated directly from neonatal 24-hour-SD rat long bones. The OC was treated with different concentrations of Formoterol and ICI118551 from 10-5mol/L～10-9mol/L respectively. Then the OC was stained with tartrate-resistant acid phosphatase （TRAP） to observe cellular morphology. The bone slices were stained with toluidine blue to count the number of the bone resorption pits and analyse the bone resorption pits by Image-Pro Plus 6.0.【Results】: The number and area of the bone resorption pits increased in different concentrations of Formoterol treatment compared that in control treatment after osteoclasts and bone slices co-cultured for 6 days. Furthermore, the number and area of the bone resorption pits decreased with the dose of ICI118551increased gradually.【Conclusions】: Theβ2-adrenergic receptor-specific agonist could promote the bone absorbion of OC in vitro. Theβ2-adrenergic receptor-specific antagonist inhibits the bone resorption of OC in a dose-dependent manner.