Inhibition of neuropeptide Y Y1 receptor induces osteoblast differentiation in MC3T3-E1 cells
Inhibition of neuropeptide Y Y1 receptor induces osteoblast differentiation in MC3T3-E1 cells
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DOI：
10.3892/mmr.2017.6866
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发表时间：
2017-09-01
影响因子：
3.4
通讯作者：
Tamura, Masato
中科院分区：
文献类型：
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作者：
Yahara, Motoki;Tei, Kanchu;Tamura, Masato
神经肽Y（NPY）是一种重要的神经信号分子。NPY由外周组织如成骨细胞产生，并与属于G蛋白偶联受体家族的相应Y1受体结合。成骨细胞特异性Y1受体敲除小鼠显示高骨量，表明NPY-Y1受体轴在骨稳态调节中的作用。在骨微环境中，周围神经纤维和成骨细胞产生NPY。然而，Y1受体对成骨细胞的影响仍未得到研究。在本研究中，RNA干扰的方法被用于靶向Y1受体，以确定它是否可以调节成骨细胞的生长，分化和活力。通过小干扰RNA（siRNA）敲低Y1受体导致小鼠MC 3 T3-E1成骨细胞中碱性磷酸酶（ALP）活性和矿化的诱导。此外，ALP、骨钙素、胶原（I）α 1和骨唾液蛋白的mRNA表达水平在Y1受体siRNA转染后显著增加。此外，Runx 2和osterix的mRNA表达水平显著增加;然而，与非靶向对照相比，在Y1受体siRNA转染的细胞中未观察到细胞增殖和caspase-3/7活性的显著改变。结果表明，Y1受体抑制可增加成骨细胞分化，这表明Y1受体在成骨细胞分化的调节中的作用。
Neuropeptide Y (NPY) is a major neural signaling molecule. NPY is produced by peripheral tissues, such as osteoblasts, and binds to the corresponding Y1 receptor that belongs to the G-protein-coupled receptor family. Osteoblast-specific Y1 receptor knockout mice exhibit high bone mass, indicating a role of the NPY-Y1 receptor axis in the regulation of bone homeostasis. In the bone microenvironment, peripheral nerve fibers and osteoblasts produce NPY. However, the effects of the Y1 receptor on osteoblasts remain unexplored. In the present study, an RNA interference approach was employed to target the Y1 receptor, in order to determine whether it may function to regulate the growth, differentiation and viability of osteoblasts. Knockdown of the Y1 receptor by small interfering RNA (siRNA) lead to induction of alkaline phosphatase (ALP) activity and mineralization in mouse MC3T3-E1 osteoblast cells. In addition, the mRNA expression levels of ALP, osteocalcin, collagen (I) alpha 1, and bone sialoprotein were significantly increased following transfection of a Y1 receptor siRNA. Furthermore, the mRNA expression levels of Runx2 and osterix were significantly increased; however, no significant alterations in cell proliferation and caspase-3/7 activity were observed in Y1 receptor siRNA-transfected cells when compared with non-targeting controls. The results demonstrate that Y1 receptor inhibition may increase osteoblastic differentiation, which indicates a role of the Y1 receptor in the regulation of osteoblastic differentiation.