Hiramoto M, Aizawa S, Iwase O, Nakano M, Toyama K, Hoque M, Nabeshima R, Kaidow A, Imai T, Hoshi H, Handa H
Department of Anatomy, Nihon University School of Medicine, Itabashi-ku, Tokyo 173, Japan.
Int J Mol Med. 1998 Feb;1(2):347-54. doi: 10.3892/ijmm.1.2.347.
Substance P (SP) is a neuropeptide widely distributed in the nervous system. Extensive study has shown SP stimulates production of various cytokines by bone marrow stromal cells, although, the role of SP in hematopoietic phenomena is still unclear. Recently, we established a human cloned stromal cell line, HAS303, which can support hematopoietic stem cell proliferation and differentiation in vitro. We used this culture system to examine the effects of SP. Expression of the mRNAs of neurokinin (NK)-1R, NK-2R and NK-3R, specific SP receptors, on HAS303 cells was demonstrated by the RT-PCR. CD34+ cells isolated from bone marrow were co-cultivated with HAS303 cells in the presence and absence of SP and the total hematopoietic cells and progenitors were counted every 5 days. Introducing SP (10(-8) M) to the co-cultures significantly increased the number of total cells and progenitors compared with control cultures. SP showed no enhancing activity on CD34+ cells cultured alone. SP also stimulated IL-3-dependent colony formation of whole bone marrow MNCs in a soft agar culture system, but showed no such activity on isolated CD34+ cells in this system. These observations suggest that SP stimulated HAS303 cells, activated HAS303 cells, and stimulated the proliferation and differentiation of CD34+ cells. Treating HAS303 cells with SP increased the intracellular Ca2+ concentration and stimulated production of G-CSF, GM-CSF, SCF and IL-6, but not IL-1alpha, IL-1beta and TNF-alpha, but did not enhance proliferation. All these findings suggest that SP mediates hematopoietic cell proliferation and differentiation in vitro by activating stromal cell function.
P物质（SP）是一种广泛分布于神经系统的神经肽。大量研究表明，SP可刺激骨髓基质细胞产生多种细胞因子，尽管SP在造血现象中的作用仍不清楚。最近，我们建立了一种人克隆基质细胞系HAS303，它能够在体外支持造血干细胞的增殖和分化。我们利用这个培养系统来研究SP的作用。通过逆转录聚合酶链反应（RT-PCR）证实了神经激肽（NK）-1R、NK-2R和NK-3R（特异性SP受体）的mRNA在HAS303细胞上的表达。从骨髓中分离出的CD34+细胞在有或无SP的情况下与HAS303细胞共培养，每5天对总造血细胞和祖细胞进行计数。与对照培养相比，向共培养物中加入SP（10^(-8) M）可显著增加总细胞数和祖细胞数。SP对单独培养的CD34+细胞没有增强活性。在软琼脂培养系统中，SP还刺激了全骨髓单个核细胞（MNCs）依赖白细胞介素-3（IL-3）的集落形成，但在该系统中对分离的CD34+细胞没有这种活性。这些观察结果表明，SP刺激了HAS303细胞，激活了HAS303细胞，并刺激了CD34+细胞的增殖和分化。用SP处理HAS303细胞可增加细胞内钙离子浓度，并刺激粒细胞集落刺激因子（G-CSF）、粒细胞-巨噬细胞集落刺激因子（GM-CSF）、干细胞因子（SCF）和白细胞介素-6的产生，但不包括白细胞介素-1α、白细胞介素-1β和肿瘤坏死因子-α，且不增强增殖。所有这些发现表明，SP通过激活基质细胞功能在体外介导造血细胞的增殖和分化。
