sEHi对颈动脉狭窄患者外周血早期内皮祖细胞血管内皮生长因子表达的影响

Effect of Soluble Epoxide Hydrolase Inhibitor on Vascular Endothelial Growth Factor Expression in Early Endothelial Progenitor Cells from Patients with Carotid Stenosis

目的探讨可溶性环氧化物水解酶抑制剂(s EHi)AUDA对颈动脉狭窄(CS)患者外周血来源的早期内皮祖细胞血管内皮生长因子(VEGF)表达的影响。方法采用密度梯度离心法,从CS患者外周血获取单个核细胞,培养至7天,收集贴壁细胞,采用激光共聚焦显微镜和流式细胞仪进行早期内皮祖细胞鉴定。分别用不同浓度(0、0.1、1、10μmol/L)AUDA干预24 h,采用免疫印迹法观察AUDA处理后其VEGF的表达。取年龄性别匹配的健康体检者的早期内皮祖细胞作为对照组。结果与对照组相比,CS患者早期内皮祖细胞VEGF表达显著下降;与处理前(0μmol/L AUDA)相比,AUDA呈剂量依赖性地增强CS患者早期内皮祖细胞VEGF表达。结论 s EHi通过环氧二十碳三烯酸介导上调早期内皮祖细胞VEGF的表达,并呈浓度依赖性,其有望成为一类治疗CS的新型药物。

Aim To investigate the effect of soluble epoxide hydrolase inhibitor（ s EHi） 12-（ 3-adamantan-1-y1-ureido）-dodecanoic acid（ AUDA） on vascular endothelial growth factor（ VEGF） expression of early endothelia1 progenitor cells in patients with carotid stenosis（ CS）. Methods Mononuclear cells from the peripheral blood of CS patients were isolated by ficoll density gradient centrifugation and cultured. After 7 days of culture in vitro,attached cells were collected. Early endothelia1 progenitor cells were identified by double staining and flow cytometry. Early endothelia1 progenitor cells were then stimulated by 0,0. 1,1,10 μmol / L of AUDA for 24 h. The expression of VEGF in early endothelia1 progenitor cells was measured by Western blot. Early endothelial progenitor cells from age and gender matched healthy subjects were also cultured as controls. Results The expression of VEGF in early endothelia1 progenitor cells from CS patients was obviously damaged compared with those from healthy controls. The AUDA could dose-dependently increase the expression of VEGF in early endothelia1 progenitor cells compared with those from CS patients without treatment. Conclusion It is suggested that s EHi can positively induce VEGF protein secretion via epoxyeicosatrienoic acid（ EET）,and s EHi may become a new drug for the therapy of CS.