Blockade of integrin β3-FAK signaling pathway activated by osteopontin inhibits neointimal formation after balloon injury
Blockade of integrin β3-FAK signaling pathway activated by osteopontin inhibits neointimal formation after balloon injury
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DOI：
10.1016/j.carpath.2007.04.002
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发表时间：
2007-09-01
影响因子：
3.7
通讯作者：
Chen, Yunhui
中科院分区：
文献类型：
--
作者：
Han, Mei;Wen, Jin-Kun;Chen, Yunhui
背景资料：骨桥蛋白（OPN）通过细胞表面受体整合素β 3促进血管平滑肌细胞（VSMCs）的迁移和粘附。为了阐明OPN参与新生内膜形成的信号通路，我们将重点放在整合素β 3-粘着斑激酶（FAK）对VSMC迁移的影响上。方法：采用免疫组化和Western blot方法检测VSMC和新生内膜中整合素3和FAK的表达。使用含有RGD基序的线性OPN 13肽和抗OPN抗体验证由OPN诱导的FAK磷酸化。通过检测FAK相关的非激酶区和整合素β 3胞浆区的过表达，检测整合素β 3-FAK通路在OPN诱导的VSMC粘附和迁移中的作用。结果：OPN可增加整合素β 3的表达，并诱导FAK快速、短暂的磷酸化。抑制FAK的磷酸化可显著抑制OPN诱导的VSMC迁移。同样，阻断整合素β 3与OPN的相互作用可以抑制OPN诱导的VSMC粘附。体内实验表明，OPN表达水平与新生内膜增厚一致。给予抗OPN抗体阻断OPN功能可抑制球囊损伤诱导的整合素β 3和FAK表达，并抑制新生内膜增厚。结论：这些数据表明，整合素β 3-FAK信号调节OPN诱导的VSMC迁移过程中的新生内膜形成。(C)2007年由Elsevier Inc.出版
Background: Osteopontin (OPN) promotes the migration and adhesion of vascular smooth muscle cells (VSMCs) through cell surface receptor, integrin beta 3. In order to elucidate the signaling pathway by which OPN is involved in neointimal formation, we focused on integrin beta 3-focal adhesion kinase (FAK) upon VSMC migration. Methods: The integrin 3 and FAK expression in VSMC and in neointima was detected by Western blot and immunohistochemistry staining. FAK phosphorylation induced by OPN was verified using a linear OPN 13 peptide containing RGD motif and anti-OPN antibody. The role of integrin beta 3-FAK pathway in VSMC adhesion and migration induced with OPN was tested by the overexpression of FAK-related nonkinase and integrin beta 3 cytoplasmic domain. Results: The results showed that OPN increased integrin beta 3 expression and induced rapid and transient FAK phosphorylation. Inhibition of the phosphorylation of FAK significantly suppressed VSMC migration induced by OPN. Similarly, blockade of the interaction of integrin beta 3 with OPN inhibited VSMC adhesion induced by OPN. The experiment, in vivo, demonstrated that OPN expression level was consistent with neointimal thickening. Administration of anti-OPN antibody for blocking OPN function suppressed integrin beta 3 and FAK expression induced by balloon injury, and neointimal thickening was inhibited. Conclusions: These data indicate that integrin beta 3-FAK signaling modulates OPN-induced VSMC migration during neointimal formation. (C) 2007 Published by Elsevier Inc.