sEHi对颈动脉狭窄患者晚期内皮祖细胞功能的影响

Effect of soluble epoxide hydrolase inhibitor on the function of late endothelial progenitor cell in patients with carotid stenosis

目的研究不同浓度可溶性环氧化物水解酶抑制剂(soluble epoxide hydrolase inhibitor,s EHi)AUDA对颈动脉狭窄(carotid stenosis,CS)患者外周血来源的晚期内皮祖细胞(late endothelial progenitor cell,late EPC)的影响。方法入选研究对象60例,分成颈动脉狭窄组(n=35)和对照组(n=25)。密度梯度离心法,从外周血获取单个核细胞培养至21 d后鉴定内皮祖细胞;以不同浓度AUDA(0,0.1,1,10μmol/L)和晚期EPC共培养24 h,分别采用MTT法,黏附能力测定实验和Transwell小室来观察其增殖,黏附,迁移能力。同时采用Western blot法观察AUDA处理后其VEGF的表达。结果体外培养21 d时,细胞呈典型长梭形,21 d时,呈铺路石样,并可摄取FITCUEA-I和Dil-ac LDL。与对照组相比,CS患者late EPC的增殖,黏附和迁移能力均显著下降(P<0.05);与处理前(0umol/L)相比,AUDA呈剂量依赖性地增强CS患者late EPC增殖,黏附和迁移能力并促进CS患者late EPC表达VEGF。结论 s EHi具有促进late EPC增殖,黏附和迁移等功能的作用,其有望成为一类治疗CS的新型药物。

Objective To investigate the effects of soluble epoxide hydrolase inhibitor（s EHi） AUDA on the function of late endothelia1 progenitor cell（late EPC） in patients with carotid stenosis.Methods Sixty cases were divided into CS group（n = 35） and control group（n = 25）.Mononuclear cells were isolated from the peripheral blood with density-gradient centrifugation and cultured.After 21 days,late EPC were identified,then incubated with AUDA in a series of concentrations（0,0.1,1 or 10 μmol/L） for 24 h.Late EPC proliferation,adhesion and migration were performed in MTT assay,adhesion assay and transwell chamber respectively.The expression of VEGF in late EPC was measured by Western blot.Results The early EPC became long spindle on day7,and then became cornerstone shape on day21.The proliferation,adhesion and migration activities of late EPC from CS patients were obviously damaged compared with those from healthy controls（P〈0.05）.The AUDA could dose-dependently increase the proliferation,adhesion and migration activities and increase the expression of VEGF in late EPC compared with those from CS patients without treatment.Conclusion s EHi can positively modulate the function of late EPC from CS patients,suggesting the potential predictive significance of s EHi in the therapy of CS.