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Pharmacological Study on Neuronal Regulation of Osteoclast Formation

破骨细胞形成的神经调节药理学研究

基本信息

批准号：
14571782
负责人：
TOGARI Akifumi
金额：
$ 1.92万
依托单位：
Aichi Gakuin University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

项目摘要

In the present study, to prove the physiological role of sympathetic nerves in bone metabolism in vivo, we examined by RT-PCR analysis the effects of a restraint stress (30 min) and intracerebroventricular (i.c.v.) injection of IPS (50 ng/mouse) on COX-2 and IL-6 mRNAs expression in mouse calvaria. LPS (i.c.v.) is well known to increase the outflow of the peripheral sympathetic nervous system. The expression of COX-2 and IL-6 mRNAs were increased by the treatment with the neurotoxin 6-hydroxydopamine (6-OHDA, 100 mg/kg/day, i.p., for 3 days) or β-blocker, propranolol (25 mg/kg, i.p.). Similarly, a restraint stress induced the expression of IL-6 mRNA in mouse calvaria. The induction was not influenced by 6-OHDA, but inhibited by propranolol. In addition, the treatment of calvaria with isoprenaline (Isp ; 100 μM) increased PGE2 and IL-6 synthesis in the organ culture system. These findings show that gene expressions increased by a restraint stress and i.c.v. injection of LPS was mediated … More by the activation of sympathetic nerve fibers and β-adrenoceptor in mouse calvaria and suggests that in vivo activation of the sympathetic nervous system modulates bone metabolism. Secondly, to elucidate the mode of action of Isp on osteoclast formation and to characterize the effect of the calcitonin gene-related peptide (CGRP) on osteoclast formation induced by Isp in a mouse bone marrow culture system. Treatment of mouse bone marrow cells with Isp generated tartrate-resistant acid phosphatase-positive multinuclear cells capable of excavating resorptive pits on dentine slices, and caused an increase in receptor activator of NF-κB ligand (RANKL) and a decrease in osteoprotegerin (OPG) production by the marrow cells. The osteoclast formation was significantly inhibited by OPG, suggesting the involvement of the RANKL-RANK system. CGRP inhibited the osteoclast formation caused by Isp or soluble RANKL but had no influence on RANKL or OPG production by the bone marrow cells treated with Isp, suggesting that CGRP inhibited the osteoclast formation by interfering with the action of RANKL produced by the Isp-treated bone marrow cells without affecting RANKL or OPG production. Since the mRNA encoding the calcitonin receptor-like receptor has been reported to express in mature osteoclast precursors, pre-osteoclasts, and mature osteoclast. This in vitro data suggest the physiological interaction of sympathetic and sensory nerves in osteoclastogenesis in vivo. Less

在本研究中，为了证明交感神经在体内骨代谢中的生理作用，我们通过RT-PCR分析了约束应激（30 min）和脑室内注射IPS （50 ng/小鼠）对小鼠颅骨COX-2和IL-6 mrna表达的影响。众所周知，LPS （i.c.v.）能增加外周交感神经系统的流出。用神经毒素6-羟多巴胺（6-OHDA， 100 mg/kg/天，i.p，连用3天）或β受体阻滞剂普萘洛尔（25 mg/kg, i.p）治疗后，COX-2和IL-6 mrna的表达增加。同样，抑制应激诱导小鼠颅骨中IL-6 mRNA的表达。6-OHDA不影响其诱导作用，而心得安有抑制作用。此外，异丙肾上腺素（Isp; 100 μM）处理颅骨可增加器官培养系统中PGE2和IL-6的合成。这些结果表明，在小鼠颅骨中，抑制应激和体外注射LPS介导了基因表达的增加，更多的是通过激活交感神经纤维和β-肾上腺素能受体，提示在体内交感神经系统的激活调节了骨代谢。其次，阐明Isp对破骨细胞形成的作用模式，表征降钙素基因相关肽（CGRP）对小鼠骨髓培养系统中Isp诱导的破骨细胞形成的影响。用Isp处理小鼠骨髓细胞可产生抗酒石酸酸性磷酸酶阳性的多核细胞，这些多核细胞能够在牙本质片上挖掘吸收凹坑，并引起骨髓细胞NF-κB配体受体激活剂（RANKL）的增加和骨保护素（OPG）的产生减少。OPG明显抑制破骨细胞的形成，提示RANKL-RANK系统的参与。CGRP抑制Isp或可溶性RANKL引起的破骨细胞形成，但对Isp处理的骨髓细胞产生RANKL或OPG没有影响，提示CGRP通过干扰Isp处理的骨髓细胞产生RANKL的作用来抑制破骨细胞的形成，而不影响RANKL或OPG的产生。由于编码降钙素受体样受体的mRNA已被报道在成熟的破骨细胞前体、破骨前细胞和成熟的破骨细胞中表达。这一体外数据表明，在体内，交感神经和感觉神经在破骨细胞形成过程中的生理相互作用。少