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在机械应力和矿化刺激下培养的成骨细胞中血小板反应蛋白-1的表达及动态变化

Expression and Dynamics of Podoplanin in Cultured Osteoblasts with Mechanostress and Mineralization Stimulus.

作者信息

Takenawa Tomohiro, Kanai Takenori, Kitamura Tetsuya, Yoshimura Yoshitaka, Sawa Yoshihiko, Iida Junichiro

机构信息

Department of Orthodontics, Faculty of Dental Medicine and Graduate School of Dental Medicine, Hokkaido University, Kita 13 Nishi 7, Kita-ku, Sapporo 060-8586, Japan.

Department of Oral Pathology and Biology, Faculty of Dental Medicine and Graduate School of Dental Medicine, Hokkaido University, Kita 13 Nishi 7, Kita-ku, Sapporo 060-8586, Japan.

出版信息

Acta Histochem Cytochem. 2018 Feb 27;51(1):41-52. doi: 10.1267/ahc.17031. Epub 2018 Jan 30.

Abstract

This study investigates the significance of the expression and dynamics of podoplanin in mechanostress and mineralization in cultured murine osteoblasts. Podoplanin increased in osteoblasts subjected to straining in non-mineralization medium, suggesting that the mechanostress alone is a podoplanin induction factor. In osteoblasts subjected to vertical elongation straining in the mineralization medium, the mRNA amounts of podoplanin, osteopontin, and osteocalcin were significantly larger than those in cells not subjected to straining, suggesting that mechanostress is the cause of a synergistic effect in the expression of these proteins. In osteoblasts in the mineralization medium, significant increases in osteocalcin mRNA occurred earlier in cells subjected to straining than in the cells not subjected to straining, suggesting that the mechanostress is a critical factor to enhance the expression of osteocalcin. Western blot and ELISA analysis showed increased podoplanin production in osteoblasts with longer durations of straining. There was significantly less mineralization product in osteoblasts with antibodies for podoplanin, osteopontin, and osteocalcin. There was also less osteopontin and osteocalcin produced in osteoblasts with anti-podoplanin. These findings suggest that mechanostress induces the production of podoplanin in osteoblasts and that podoplanin may play a role in mineralization in cooperation with bone-associated proteins.

摘要

本研究调查了血小板反应蛋白-1在培养的小鼠成骨细胞机械应力和矿化过程中的表达及动态变化的意义。在非矿化培养基中施加应变的成骨细胞中血小板反应蛋白-1增加,这表明单纯的机械应力是血小板反应蛋白-1的诱导因子。在矿化培养基中经受垂直伸长应变的成骨细胞中,血小板反应蛋白-1、骨桥蛋白和骨钙素的mRNA量显著高于未施加应变的细胞,这表明机械应力是这些蛋白质表达协同效应的原因。在矿化培养基中的成骨细胞中,施加应变的细胞中骨钙素mRNA的显著增加比未施加应变的细胞更早出现,这表明机械应力是增强骨钙素表达的关键因素。蛋白质免疫印迹和酶联免疫吸附测定分析显示,应变持续时间较长的成骨细胞中血小板反应蛋白-1的产生增加。用针对血小板反应蛋白-1、骨桥蛋白和骨钙素的抗体处理的成骨细胞中矿化产物明显减少。用抗血小板反应蛋白-1处理的成骨细胞中产生的骨桥蛋白和骨钙素也较少。这些发现表明,机械应力诱导成骨细胞中血小板反应蛋白-1的产生,并且血小板反应蛋白-1可能与骨相关蛋白协同作用在矿化过程中发挥作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/797e/5880802/d4340588eeaa/AHC17031f01.jpg

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