骨桥蛋白13肽抑制球囊内皮剥脱术后血管狭窄的实验研究

目的：利用含有骨桥蛋白（OPN）多种功能位点的13肽（Gly^158-Lys^170）,从细胞和整体水平观察其对VSMC和单核巨噬细胞黏附、浸润以及内膜增生的影响,并初步探讨其作用机制。方法：用不同浓度OPN13肽（0,100,200,300mg／L）检测其对体外培养的平滑肌细胞（VSMc）与OPN黏附的抑制作用;以不合黏附序列的6肽分子为对照组,用以确定13肽抑制黏附特异性。用球囊内皮剥脱法建立大鼠内膜增生模型。实验动物分为4组：治疗组大鼠自术前1h及术后静脉滴注13肽,连续给药7d;对照组大鼠给予非特异性对照6肽分子;模型组大鼠给予相同剂量的生理盐水;正常对照组大鼠施假手术。并利用免疫组织化学染色和Western印迹分析方法,检测血管壁中OPN、FAK、ILK的表达变化。结果：OPN13肽能特异性的及浓度依赖性的抑制VSMC与OPN的相互作用,血管内膜剥脱后给予13肽治疗组血管壁单核／巨噬细胞浸润减少,OPN及其下游信号分子ILK,FAK表达下调,内膜增生被明显抑制。结论：含有OPN多功能位点的13肽可通过阻断OPN与膜受体的相互作用而抑制血管炎症的进展和内膜增生。

Experimental study on inhibition of restenosis by osteopontin oligopeptide antagonist after de-endothelium

Aim:Osteopontin 13-peptide(Gly158-Lys170),containing multi-fuction domains was used to inhibit the VSMC adhesion,migration.The mechanism of 13-peptide inhibiting neointima formation was investigated.Methods:The effect of 13-peptide on VSMC adhesion was tested by adhesion assay.The restenosis model was prepared balloon injury after administration of 13-peptide for 1 h,and then the 13-peptide was given by an intravenous drip for 7 days.The expression changes of OPN,FAK,ILK in vessel wall were detected by immuohistochemistry and Western blot.Results:The 13-peptide dose-dependently reduced adhesion of VSMC in OPN matrix,and the infiltration of macrophage in vessel wall also was reduced in the treatment group after balloon injury.The expression of OPN,FAK,ILK was down-regulated following with the inhibition of neointima thickening.Conclusion:The OPN 13-peptide can inhibit inflammation and neointima formation by blocking the binding of OPN to it's receptors.