GRK2可使成骨细胞的血流诱导反应脱敏。

Yanghui Xing, Y. Gu, X. Shan, L. Wang, J. You
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引用次数: 1

摘要

机械加载后的骨脱敏是骨适应机械环境的必要条件。然而,脱敏机制尚不清楚。以往的研究表明,G蛋白偶联受体(gpcr),包括P2Y和甲状旁腺激素受体,在成骨细胞的机械生物学中发挥重要作用。因此,在本研究中,我们研究了G蛋白偶联受体激酶2 (GRK2)在机械刺激后成骨细胞脱敏中的作用。我们首先通过细胞质Ca2+和磷酸化的ERK1/2活性,分别使用荧光Ca2+敏感染料和western blotting检测机械刺激后成骨细胞脱敏的存在。然后,我们证明了GRK2在MC3T3-E1细胞中的过表达抑制了血流诱导的ERK1/2磷酸化,而GRK2的siRNA敲低增强了ERK1/2磷酸化。此外,我们发现GRK2在MC3T3-E1细胞中过表达短期抑制环氧化酶-2 mRNA的表达,长期抑制碱性磷酸酶的活性。更重要的是,我们发现GRK2在血流刺激后不久就转移到细胞膜上,这是GPCR脱敏的必要步骤。先前,我们已经证明P2Y2嘌呤能受体,一种gpcr,参与各种血流诱导的成骨细胞反应。在本研究中,我们还发现GRK2过表达不影响ATP的释放。因此,GRK2可能通过使P2Y2受体脱敏来抑制血流诱导的成骨细胞反应。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
GRK2 desensitizes flow-induced responses in osteoblasts.
Bone desensitization after mechanical loading is essential for bone to adapt to its mechanical environment. However, the desensitization mechanism is unknown. Previous studies suggest that G protein-coupled receptors (GPCRs), including P2Y and parathyroid hormone receptors, play important roles in osteoblast mechanobiology. Thus, for the present research, we examined the role of G protein-coupled receptor kinase 2 (GRK2) in osteoblast desensitization after exposure to mechanical stimulation. We first showed the existence of osteoblast desensitization after mechanical stimulation based on cytosol Ca2+ and phosphorylated ERK1/2 activities, detected using a fluorescent Ca2+-sensitive dye and western blotting, respectively. We then demonstrated that GRK2 overexpression in MC3T3-E1 cells inhibits flow-induced ERK1/2 phosphorylation, while siRNA knockdown of GRK2 enhances ERK1/2 phosphorylation. Additionally, we found that GRK2 overexpression in MC3T3-E1 cells inhibits cyclooxygenase-2 mRNA expression in the short term and alkaline phosphatase activity in the long term. More importantly, we discovered that GRK2 translocated to the cell membrane shortly after flow stimulation - a step necessary for GPCR desensitization. Previously, we have demonstrated that P2Y2 purinergic receptors, one type of GPCRs, are involved in various flow-induced osteoblastic responses. In this research, we also showed that GRK2 overexpression does not affect ATP release. Accordingly, GRK2 is able to inhibit flow-induced osteoblast responses possibly through desensitizing P2Y2 receptors.
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