持续性压力通过影响ERK1/2和GIT1的结合促进大鼠成骨细胞的迁移
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国家自然科学基金(30670876);江苏省自然科学基金(BK2006249);江苏省青蓝工程(JX10210535);江苏省六大人才(07-B-043)


The role of GIT1 and ERK1/2 of osteoblast migration induced by durative pressure
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    摘要:

    目的:探讨持续压力对体外培养的大鼠成骨细胞内ERK1/2及GIT1蛋白相互关系及对大鼠成骨细胞迁移能力的影响-方法:取1~2天龄SD大鼠颅盖顶骨进行成骨细胞原代培养,采用压缩空气在密闭容器内对第2代细胞施以300 kPa的静压力,分别于未加压,加压后0.5-1.0-2.0 h提取细胞总蛋白,Western blot 检测各组成骨细胞在Src抑制剂PP2不干预和干预下ERK1/2 和GIT1 的表达及其磷酸化,免疫共沉淀分析GIT1同活化的ERK1/2(phospho-ERK1/2,pERK1/2)相互作用的变化-Image-Pro Plus 5.0软件检测成骨细胞在Src抑制剂PP2不干预和干预下加压前后面积-结果:Western blot结果显示pERK1/2和pGIT1的表达在持续加压组较对照组明显增加;PP2干预下持续加压组的成骨细胞内的pERK1/2和pGIT1的表达较对照组无统计学差异;免疫共沉淀结果显示GIT1-pERK1/2结合力在持续加压组较对照组明显增加;持续加压后成骨细胞面积明显增加;PP2干预下持续加压后成骨细胞面积较对照组无统计学差异-结论:300 kPa左右的持续性压力能通过激活Src的活性,从而诱导ERK1/2,GIT1的磷酸化,同时增加pERK1/2和GIT1的相互结合,促进大鼠成骨细胞的迁移,Src-GIT1/ERK1/2途径可能是成骨细胞力学信号转导过程中的重要通路之一-

    Abstract:

    Objective:To investigate the role and mechanism of G-protein-coupled receptor kinase interacting protein 1(GIT1)and extracellular signal-regulated kinases 1 and 2(ERK1/2)in osteoblasts induced by durative pressure. Methods:The osteoblasts were mechanically stimulated on coverslips. The phosphorylation of ERK1/2 and GIT1 of osteobasts treated with or without the Src kinase inhibitor PP2 was measured by Western blot. The interaction between GIT1 and pERK1/2 was detected by coimmunoprecipitation. Cell area of groups treated with or without the Src kinase inhibitor PP2 was caculated by image-pro plus 5.0. Results:The phosphorylation of ERK1/2 and GIT1 and the interaction between GIT1 and pERK1/2 of osteoblasts treated with durative pressure significantly increased. The cell size increased significantly after durative pressure. Conclusion:These data suggests that 300 kPa durative pressure stimuates Src,consequently pathway induces the phosphorylation of ERK1/2 and GIT1 in osteoblasts,further increases the combination of pERK1/2 and GIT1 to migrate osteoblasts. The Src-GIT1/ERK1/2 pathway may play a important role in osteoblast migration induced by durative pressure.

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史亮亮,张柏江,林俊安,张 宁,任永信,殷国勇.持续性压力通过影响ERK1/2和GIT1的结合促进大鼠成骨细胞的迁移[J].南京医科大学学报(自然科学版),2009,29(7):934-938

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  • 收稿日期:2009-02-24
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