异丙肾上腺素对大鼠骨髓单核细胞向破骨细胞转化的影响

Effect of isoprel on the transformation of rat marrow mononuclear cells into osteoclasts

背景：交感神经系统在骨量的调控中发挥关键的作用，假设体外交感神经分泌的神经递质能够促进破骨细胞形成，无疑为破骨细胞诱导培养增加了新方法。目的：实验拟观察异丙肾上腺素对大鼠骨髓单核细胞向破骨细胞转化的影响。设计、时间及地点：横断面设计实验，于2007—03／06在沈阳医学院实验中心完成。材料：新生24h Wistar大鼠，体质量5．0～6．0g，雌雄不拘，用于分离培养大鼠成熟破骨细胞。异丙肾上腺素由武汉福鑫化工有限公司提供。方法：分离培养大鼠成熟破骨细胞为原代组，以异丙肾上腺索作用于骨髓单核细胞为肾上腺素组，无异丙肾上腺素为对照组，用苏木精-伊红染色、抗酒石酸酸性磷酸酶染色加以鉴定。主要观察指标：倒置显微镜观察第3，6，9天3组细胞的计数，扫描电镜观察破骨细胞在骨片上的嗜骨能力。结果：肾上腺素组破骨细胞数目为原代组数目的3倍，其形态与原代组消化获得的细胞形态相同，抗酒石酸酸性磷酸酶染色均阳性。扫描电镜显示肾上腺素组破骨样细胞在骨片上培养时产生骨陷凹。结论：在体外异丙肾上腺素对破骨样细胞形成有促进作用，形成的破骨样细胞在形态、特异酶及噬骨能力方面与经典分离培养的破骨细胞无差异。

BACKGROUND： Sympathetic nervous system plays an important role in the regulation of bone mass. Provided the neurotransmitters secreted by sympathetic nerve promote the formation of osteoclast in vitro, thus a new method to induce and culture the osteoclasts could be found. OBJECTIVE： To study the effect of isoprel on the transformation of osteoclasts from the mononuclear cell in rat bone marrow. DESIGN, TIME AND SETTING： A cross section design experiment was carried out between March to June in 2007 in the center laboratory of Shenyang Medical College. MATERIALS： Wistar neonatal rats within 24 hours, weighing 5.0-6.0 g, either gender, were used to culture osteoclasts. Isoprel was offered by Wuhan Fuxin Chemical Industry Co.,Ltd. METHODS： The mature isolated osteoclasts after isolation and culture were taken as primary group, the rat marrow mononuclear cells with the presence of isoprel as isoprel group, and rat marrow mononuclear cells without isoprel as control group. Cells were identified by hematoxylin-eosin staining and tartaric-resistant acid phosphatase staining. MAIN OUTCOME MEASURES： All cells were counted by phase-contrast microscope on the third, sixth and ninth days. Scanning electron microscope was applied to observe the resorption capacity of osteoclasts in bone slice. RESULTS： The cell amount of isoprel group was 3 times to that of primary group, and the cell morphology of isoprel group was similar with that of primary group. Cells in isoprel group and primary group were both positive for tartaric-resistant acid phosphatase staining. Under scanning electron microscope, the resorption lacuna was observed in the osteoclast-like cells of isoprel group. CONCLUSION： Isoprel can promote the formation of osteoclast-like cells in vitro, the osteoclast-like cells have no difference from the classically isolated and cultured osteoclasts in morphology, special enzyme and bone resorption.