Substance P stimulates late-stage rat osteoblastic bone formation through neurokinin-1 receptors
Substance P stimulates late-stage rat osteoblastic bone formation through neurokinin-1 receptors
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DOI：
10.1016/j.npep.2006.11.002
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发表时间：
2007-02-01
期刊：
影响因子：
2.9
通讯作者：
Tanaka, T.
中科院分区：
文献类型：
--
作者：
Goto, T.;Nakao, K.;Tanaka, T.
P物质（SP）是一种广泛分布的神经肽，作为神经递质和神经调质发挥作用。最近，SP受体，特别是对SP具有高亲和力的神经激肽-1受体（NK 1-Rs），不仅在神经元和免疫细胞中，而且在包括骨细胞在内的其它外周细胞中也被观察到。为了明确SP在骨形成中的作用，我们研究了NK 1-Rs在成骨细胞中的表达以及SP对大鼠颅骨成骨细胞骨形成的影响。分离大鼠颅骨成骨细胞，并在含有10%血清、抗坏血酸、地塞米松和β-甘油磷酸盐的α-MEM中培养3周。然后我们研究了NK 1-R表达、SP对成骨细胞骨形成的影响以及成骨细胞中骨钙素mRNA的表达。RT-PCR和免疫细胞化学结果显示，培养14天的颅骨成骨细胞中有NK 1-R mRNA表达，而培养7天的颅骨成骨细胞中无NK 1-R表达。培养的成骨细胞在加入10（-8）-10（-6）M SP后，骨形成显著增加。在培养的3周期间，在第一周加入SP没有显著增加骨形成，而在第一周和第二周或全部3周加入SP显著增加颅骨成骨细胞骨形成。此外，半定量RT-PCR表明，SP刺激骨钙素mRNA的表达在成骨细胞在第14天或21天，而SP没有刺激RunX 2或I型胶原mRNA的表达在第7天，但刺激他们在第14天。这些结果表明，在骨形成后期，SP通过NK 1-Rs刺激成骨细胞的骨形成。这些作用依赖于成骨细胞中NK 1-R的表达。我们的研究结果表明，从感觉神经元分泌的SP可能调节骨形成后，SP受体的表达。(c)2006爱思唯尔有限公司版权所有。
Substance P (SP) is a widely distributed neuropeptide that works as a neurotransmitter and neuromodulator. Recently, SP receptors, particularly neurokinin-1 receptors (NK1-Rs) that have a high affinity for SP, have been observed not only in neuron and immune cells, but also in other peripheral cells, including bone cells. To identify the role of SP in bone formation, we investigated the expression of NK1-Rs in osteoblastic cells and the effects of SP on bone formation by rat calvarial osteoblastic cells. Rat calvarial osteoblastic cells were isolated and cultured for 3 weeks in alpha-MEM containing 10% serum, ascorbic acid, dexamethasone, and beta-glycerophosphate. We then investigated NK1-R expression, SP effects on ostcoblastic bone formation, and osteocalcin mRNA expression in ostcoblastic cells. RT-PCR and immunocytochemistry showed that NK1-R mRNA was expressed and NK1-R was present in 14-day, but not 7-day, cultured calvarial osteoblasts. Bone formation by cultured osteoblastic cells significantly increased after the addition of 10(-8)-10(-6) M SP. During 3 weeks of culture, the addition of SP in the first week did not significantly increase bone formation, whereas adding SP during the first and second week or all 3 weeks significantly increased calvarial osteoblastic bone formation. Furthermore, semi-quantitative RT-PCR indicated that SP stimulated osteocalcin mRNA expression in the osteoblasts at day 14 or day 21, whereas SP did not stimulated the runX2 or type I collagen mRNA expression at day 7 but stimulated them at day 14. These results indicate that SP stimulates bone formation by osteoblastic cells via NK1-Rs at late-stage bone formation. These effects were dependent on the expression of NK1-R in osteoblastic cells. Our findings suggest that SP secreted from sensory neurons may modulate bone formation after the expression of SP receptors. (c) 2006 Elsevier Ltd. All rights reserved.