The effect of endothelial cell overexpression of plasminogen activator inhibitor-1 on smooth muscle cell migration.
The effect of endothelial cell overexpression of plasminogen activator inhibitor-1 on smooth muscle cell migration.
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内皮细胞过表达纤溶酶原激活剂抑制剂-1对平滑肌细胞迁移的影响。
DOI：
10.1067/mva.2002.123687
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发表时间：
2002
影响因子：
4.3
通讯作者：
Powell,RichardJ
中科院分区：
文献类型：
--
作者：
Proia,RichardR;Nelson,PeterR;Mulligan-Kehoe,MaryJo;Wagner,RobertJ;Kehas,ArthurJ;Powell,RichardJ
纤溶酶原激活剂抑制剂-1 （PAI-1）是一种已知的纤溶酶原激活剂抑制剂，可能通过改变基质金属蛋白酶（MMP）活性来调节平滑肌细胞迁移（SMC）。方法为了研究内皮细胞PAI-1过表达对SMC迁移的影响，采用RT-PCR方法克隆PAI-1全长基因，并将其连接到pCMV/myc/ER表达载体上。采用电穿孔法，用PAI-1构建物或空载体转染牛主动脉内皮细胞作为对照。特异性PAI-1活性测定和酶联免疫吸附测定显示EC PAI-1过表达。采用改进的Boyden-chamber法检测EC PAI-1过表达对SMC迁移的影响。用酶谱法测定SMC的MMP表达。结果选择的克隆（EC9, EC21）与未转染的EC和空载体EC （ECC）相比，PAI-1活性增加了3 - 5倍。同样，酶联免疫吸附试验结果显示，与ECC相比，EC9和EC21的PAI-1水平增加了3.5至5.5倍。未转染的EC和ECC对SMC迁移模式有相似的影响。与ECC相比，暴露于PAI-1过表达EC的SMC迁移受到了35%至57%的抑制。这种抑制作用在添加外源性尿激酶型纤溶酶原激活剂（uPA）后被逆转。酶谱分析显示，暴露于PAI-1过表达EC的SMCs中，MMP-2和MMP-9表达下调。结论转染EC后pai -1过表达可抑制SMC迁移。这种作用可能是通过降低SMC MMP活性介导的。（中华外科杂志2002；36:164-71）
IntroductionPlasminogen activator inhibitor-1 (PAI-1), a known inhibitor of plasminogen activators, may regulate smooth muscle cell migration (SMC) through alteration in matrix metalloproteinase (MMP) activity.MethodsTo study the effect of endothelial cell (EC) PAI-1 overexpression on SMC migration, RT-PCR was used to clone the full length PAI-1 gene, which was ligated into the pCMV/myc/ER expression vector. With electroporation, bovine aortic ECs were transfected with either the PAI-1 construct or the empty vector as control. EC PAI-1 overexpression was shown with a specific PAI-1 activity assay and enzyme-linked immunosorbent assay. The effect of EC PAI-1 overexpression on SMC migration was measured with a modified Boyden-chamber assay. SMC MMP expression was measured with zymography.ResultsSelected clones (EC9, EC21) had a three-fold to five-fold increase in PAI-1 activity compared with untransfected EC and empty vector EC (ECC). Similarly, enzyme-linked immunosorbent assay results showed a 3.5-fold to 5.5-fold increase in PAI-1 levels in EC9 and EC21 versus ECC. Untransfected EC and ECC had similar effects on SMC migratory patterns. Migration of SMC exposed to PAI-1 overexpressing EC was inhibited by 35% to 57% compared with ECC. This inhibitory effect was reversed with addition of exogenous urokinase-type plasminogen activator (uPA). Zymography showed downregulation of MMP-2 and MMP-9 in SMCs exposed to PAI-1 overexpressing EC.ConclusionPAI-1 overexpression with transfected EC inhibits SMC migration. This effect may be mediated through decreased SMC MMP activity. (J Vasc Surg 2002;36:164-71.)