Substance P stimulates bone marrow stromal cell osteogenic activity, osteoclast differentiation, and resorption activity in vitro.
Substance P stimulates bone marrow stromal cell osteogenic activity, osteoclast differentiation, and resorption activity in vitro.
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DOI：
10.1016/j.bone.2009.04.203
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发表时间：
2009-08
期刊：
影响因子：
4.1
通讯作者：
Kingery, Wade S.
中科院分区：
文献类型：
--
作者：
Wang, Liping;Zhao, Rong;Shi, Xiaoyou;Wei, Tzuping;Halloran, Bernard P.;Clark, David J.;Jacobs, Christopher R.;Kingery, Wade S.
SP是一种神经肽，分布于支配骨的髓组织以及骨膜的感觉神经纤维中。以前，我们证明辣椒素治疗后神经肽信号的抑制导致骨量丢失，我们假设SP通过刺激骨生成促进骨完整性。培养来自C57 BL/6小鼠的成骨细胞前体（骨髓基质细胞，BMSC）和破骨细胞前体（骨髓巨噬细胞，BMSC）。免疫细胞化学染色和PCR检测SP受体（NK 1）的表达。采用酶免疫分析（EIA）和免疫印迹法（Western blot）检测SP对BMSCs增殖和分化的影响，并检测BMSCs中BrdU掺入、基因表达、碱性磷酸酶活性、骨钙素和Runx 2蛋白水平。采用BrdU法检测SP对BMP 2的影响，计数抗酒石酸酸性磷酸酶（TRAP+）染色阳性的多核细胞，测量窝蚀面积，并采用ELISA和western blot法评价RANKL蛋白产生和NF-κB活性。NK 1受体在BMSCs和BMSCs中均有表达。SP以浓度依赖性方式刺激BMSCs增殖。低浓度（10−12 M）的SP刺激碱性磷酸酶和骨钙素表达，增加碱性磷酸酶活性，上调Runx 2蛋白水平，高浓度的SP（10−8 M）增强分化的BMSCs的矿化。SP还刺激BMSCs产生RANKL，但在SP存在下与巨噬细胞共培养时，SP浓度太低而不能引起破骨细胞生成。SP还激活BMSCs中的NF-κB，并直接促进RANKL诱导的巨噬细胞破骨细胞生成和骨吸收活性。NK 1受体由成骨细胞和破骨细胞前体表达，SP在体外刺激成骨细胞和破骨细胞的分化和功能。感觉神经元释放的SP神经递质可能在体内调节局部骨转换。
SP is a neuropeptide distributed in the sensory nerve fibers that innervate the medullar tissues of bone, as well as the periosteum. Previously we demonstrated that inhibition of neuropeptide signaling after capsaicin treatment resulted in a loss of bone mass and we hypothesized that SP contributes to bone integrity by stimulating osteogenesis. Osteoblast precursors (bone marrow stromal cells, BMSCs) and osteoclast precursors (bone marrow macrophages, BMMs) derived from C57BL/6 mice were cultured. Expression of the SP receptor (NK1) was detected by using immunocytochemical staining and PCR. Effects of SP on proliferation and differentiation of BMSCs were studied by measuring BrdU incorporation, gene expression, alkaline phosphatase activity, and osteocalcin and Runx2 protein levels with EIA and western blot assays, respectively. Effects of SP on BMMs were determined using a BrdU assay, counting multinucleated cells staining positive for tartrate-resistant acid phosphatase (TRAP+), measuring pit erosion area, and evaluating RANKL protein production and NF-κB activity with ELISA and western blot. The NK1 receptor was expressed in both BMSCs and BMMs. SP stimulated the proliferation of BMSCs in a concentration-dependent manner. Low concentrations (10−12 M) of SP stimulated alkaline phosphatase and osteocalcin expression, increased alkaline phosphatase activity, and up-regulated Runx2 protein levels, and higher concentrations of SP (10−8 M) enhanced mineralization in differentiated BMSCs. SP also stimulated BMSCs to produce RANKL, but at concentrations too low to evoke osteoclastogenesis in co-culture with macrophages in the presence of SP. SP also activated NF-κB in BMMs and directly facilitate RANKL induced macrophage osteoclastogenesis and bone resorption activity. NK1 receptors are expressed by osteoblast and osteoclast precursors and SP stimulates osteoblast and osteoclast differentiation and function in vitro. SP neurotransmitter release from sensory neurons could potentially regulate local bone turnover in vivo.
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影响因子：
3.6
作者：
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通讯作者：
LEMBECK, F
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3.6
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