硫化氢对内皮祖细胞数量和功能的影响

Effects of hydrogen sulfide on number and function of endothelial progenitor cells

目的:观察不同浓度的硫化氢(H2S)对体外培养内皮祖细胞(EPCs)数量和功能的影响。方法:以密度梯度离心法获取脐血单个核细胞,培养6 d后收集贴壁细胞。用流式细胞仪检测细胞表达CD34、CD133、血管内皮生长因子受体(VEGFR)-2、VE-Cadherin,并用荧光显微镜观察细胞吞噬Dil标记的乙酰化低密度脂蛋白(Dil-ac-LDL)和异硫氰酸荧光素-荆豆凝集素-1(FITC-UEA-1)来鉴定EPCs。以不同浓度的H2S(0、10μM、50μM、100μM或500μM)孵育EPCs(每天1 h),分别培养4 d观察细胞集落及6dEPCs数量。结果:与正常对照组相比,H2S在一定浓度下(<100μM)呈浓度依赖性促进EPCs增殖及细胞集落形成(P<0.05)。结论:一定浓度的硫化氢可能通过抗氧化发挥对内皮祖细胞的影响。

Objective： To study effects of different concentrations of hydrogen sulfide （H2 S） on number and function of endothelial progenitor cells （EPCs） cultured in vitro. Methods： Mononuclear cells were isolated from the umbilical cord using density gradient eentrifugation, and adherent cells were collected after six-day culture. Expression of CD34, CD133, vascular endothelial growth factor receptor （VEGFR） -2 and vascular endothelial （VE） -Cadherin were measured by flow cytometer. EPCs were identified as cells that swallowed Dil-acetyl-low density lipoprotein （Dil-ac-LDL） and/or fluorescein isothioeyanate Ulex Europaeus agglutinin-1 （FITC-UEA-1） under fluorescent microscope. EPCs were exposed to different concentrations of H2S （0, 10μM, 50μM, 100μM or 500μM, 1h/d）, cell colony was observed after four-day culture, and number of EPCs was observed after six days. Results：Compared with normal control group, certain concentration （〈 100μM） of H2 S with concentration-dependence promoted propagation of EPCs and formation of cell colonies （P〈0.05）. Conclusion： Certain concentration of H2S may have effects on EPCs through anti-oxidation.