| 关莹 续惠云瓮媛媛 商澎.二维回转培养对MLO-Y4骨样细胞PKD2表达定位
及胞内钙信号的影响[J].,2014,14(14):2601-2605 |
| 二维回转培养对MLO-Y4骨样细胞PKD2表达定位
及胞内钙信号的影响 |
| The Effects of 2D-Clinorotation on Expression and Location of PKD2Protein and the Intracellular Ca2+Concentration of MLO-Y4 |
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| DOI： |
| 中文关键词: 二维回转培养 模拟失重 PKD2 MLO-Y4 |
| 英文关键词: 2D-Clinorotation Weightlessness PKD2 MLO-Y4 |
| 基金项目:国家自然科学基金项目（30700152；31170812），西北工业大学基础研究基金（NPU-FFR-JC201160） |
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| 中文摘要: |
| 目的：PKD2(polycystin2，多囊肾病蛋白2)能够在细胞膜上形成无选择性的阳离子通道，在肾上皮细胞中PKD2 与初级纤毛
共定位，通过改变胞内的钙信号过程参与细胞对力学刺激的响应。本实验通过二维回转培养来模拟失重效应，旨在探讨二维回转
培养对MLO-Y4 骨样细胞PKD2 表达定位，及胞内钙信号的影响。初步了解PKD2 在小鼠骨样细胞MLO-Y4 响应力学刺激过程
中起的作用。方法：采用二维回转培养骨样细胞MLO-Y4，用RT-PCR和western blotting检测PKD2的表达，用荧光共聚焦显微镜
检测细胞中PKD2 与初级纤毛的定位及细胞内钙离子含量。结果：与对照组相比，在二维回转培养后，骨样细胞MLO-Y4 的PKD2
表达在mRNA和蛋白水平都有明显的下降，PKD2、PKD1(polycystin1，多囊肾病蛋白1)和乙酰化的α-tubulin 共定位，同时二维回
转培养降低了细胞内钙离子含量。结论：在二维回转培养下，PKD2可能通过调节自身表达来改变细胞膜上PKD 通道的数目和开
放情况来影响细胞内钙离子含量，参与骨细胞对细胞外应力的感受过程，其详细机制还有待进一步实验研究。这将对探讨骨细胞
响应力学刺激的具体机制提供重要的理论依据。 |
| 英文摘要: |
| Objective: PKD2 (polycystin2) could form nonspecific cation channel on the cytomembrane, which is responsible for
Ca2+ penetration when kidney epithelial cell transforms mechanical stimulation into intracellular chemical information. In this study,
2D-clinorotation was used to simulate weightlessness condition. The aim of this study was to investigate the effects of 2D-clinorotation
culture on expression and location of PKD2 protein and the intracellular Ca2+ concentration of mouse osteocyte-like MLO-Y4 cell. Also
we want to understand the role of PKD2 in the process of MLO-Y4 responsding to mechanical stimulation.Methods:Mouse
osteocyte-like MLO-Y4 cell were cultured under the condition of 2D-Clinorotation.The expression and location of PKD2 was detected by
RT-PCR, western-blotting and fluorescent staining, the intracellular Ca2+ concentration was detected by Ca2+ staining technique.Results:Under the condition of 2D-Clinorotation culture, the exspression of PKD2 and the intracellular Ca2+concentration were distinctly reduced.
At the same time, we found that PKD2 co-localized with PKD1 (polycystin1) and the specific ciliary axoneme marker-acetylated
α-tubulin. Conclusion: the results suggest under 2D-clinorotation culture, mouse osteocyte-like cell could respond to the mechanical
stimulation by regulating the expression of PKD2 and the intracellular Ca2+ concentration. It is helpful to investigate the cellular
mechanismof bone cells in responding to mechanical stimulation. |
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