C反应蛋白对外周血内皮祖细胞数量及功能的影响

目的研究C反应蛋白对人外周血内皮祖细胞数量及功能的影响。方法从外周血中分离出单个核细胞,体外培养7天,在贴壁细胞中加入不同浓度的C反应蛋白(1 mg/L、2.5 mg/L和5.0 mg/L)作用不同时间(24 h、48 h和72 h),用四唑盐比色试验(MTT)和细胞集落形成单位计数的方法评价C反应蛋白对内皮祖细胞增殖的影响;采用趋化试验评价不同浓度的C反应蛋白对血管内皮生长因子诱导的内皮祖细胞趋化能力的影响;检测细胞上清中一氧化氮的浓度变化;逆转录—聚合酶链式反应检测细胞内皮源性一氧化氮合酶表达强度变化。结果C反应蛋白减少内皮祖细胞的集落形成单位数量及抑制内皮祖细胞的增殖能力;随着C反应蛋白浓度的增加,内皮祖细胞的趋化能力受到抑制;同样细胞分泌的一氧化氮减少,内皮源性一氧化氮合酶表达减弱。结论C反应蛋白可能通过抑制内皮祖细胞的增殖和趋化能力促进内皮功能不全的发展。

Effects of C-Reactive Protein on Quantity and Activity of Endothelial Progenitor Cells From Peripheral Blood

Aim To investigate the effects of C-reactive protein(CRP) on the proliferation and chemotaxis of endothelial progenitor cells(EPC) of human peripheral blood. Methods Mononuclear cells were isolated from human peripheral blood and cultured for 7days.Attached cells were incubated with different concentration of CRP(1.0 mg/L,2.5 mg/L and 5.0 mg/L) for different times(24,48 and 72 h).MTT assay and quantified colony forming units(CFU) were used to assess the proliferation of EPC after treated with CRP.Meanwhile,chemotaxis assay was used to quntified EPC induced by VEGF.NO in the supernatant was measured by nitrate reductase assay.RT-PCR was employed to detect mRNA expression of endothelial nitric oxide synthase(eNOS). Results Incubation of EPC with CRP decreased the number and the function of EPC. Conclusions It is suggested that CRP can promote endothelial dysfunction by means of depressant EPC proliferation and migratory.