课题背景：国内外文献均在临床上和动物实验上证实了交感神经系统在骨量调控中发挥关键的作用。交感神经系统在骨上的效应器是成骨细胞上的β2肾上腺素能受体，该受体的激活可增加细胞核因子кB受体活化因子配基(RANKL)的表达，从而刺激破骨细胞生成进而导致骨量的下降。相反，β受体拮抗剂则能削弱破骨细胞的分化从而增加骨量。

应用要点：如果在体外交感神经分泌的神经递质也能够促进破骨细胞形成，这无疑为破骨细胞诱导培养增加了新方法。实验证实此方法获得的破骨细胞在形态上及特异酶分泌及其骨片嗜骨的程度上跟正常细胞所差无几，并且数量上高于传统的诱导方法。解决了目前各种培养方法获得的破骨细胞数量少这一难题，为建立破骨细胞系奠定基础。

重要的概念：破骨细胞分化因子(ODF/OPGL)：是肿瘤坏死因子家族的一员，属于Ⅱ型跨膜蛋白，有较短的N-端的细胞内区域和较长的C-端细胞外区域。C-端细胞外区从赖氨酸(lysine 158)至C-末端的活性配基区域与肿瘤坏死因子相关蛋白具有高度同源性。其氨基酸49~60之间的序列编码了疏水性的跨膜区。

摘要
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24 h Wistar大鼠，体质量5.0~6.0 g，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，以异丙肾上腺素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。
关键词：破骨细胞；培养；异丙肾上腺素；鉴定；组织构建

颜南，王正东，金韵，刘自力，叶丽杰. 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[J].中国组织工程研究与临床康复，2008，12(20):3818-3821 [www.zglckf.com/zglckf/ejournal/upfiles/08-20/20k-3818(ps).pdf]

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

Abstract

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.

Yan N, Wang ZD, Jin Y, Liu ZL, Ye LJ. Effect of isoprel on the transformation of rat marrow mononuclear cells into osteoclasts.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2008;12(20):3818-3821(China)
[www.zglckf.com/zglckf/ejournal/upfiles/08-20/20k-3818(ps).pdf]