全反式维甲酸对人脐静脉内皮细胞增殖和分泌NO的影响

Effect of All-trans Retinoic Acid on Endothelial Cell Proliferation and Nitric Oxide Production in Human Umbilical Vein

【目的】探讨观察全反式维甲酸（ATRA）对内皮细胞的增殖和分泌一氧化氮（NO）功能的影响。【方法】将体外培养人脐静脉内皮细胞株（ECV-304）随机分为5组：空白对照组（加入ATRAR溶剂二甲亚砜，DMSO）、另4组ATRA浓度分别为10^-9 mol/L、10^-8 mol/L、10^-7 mo1/L、10^-6 mol/L。在24 h、48 h、72 h收集细胞，通过流式细胞仪、四氮唑（MTT）法检测不同浓度的 ATRA对ECV-304增殖的影响，观察ATRA对内皮细胞分泌NO功能的影响，并测定细胞培养液内NO含量。【结果】① ATRA在较低浓度（10^-9～10^-7 mol/L），对血管内皮细胞ECV-304有增殖作用，并呈浓度依赖性增加，与空白对照组比有显著差异（ P ＜0．01），且随 ATRA作用时间延长而增加。在较高浓度时（10^-6 mol/L），对血管内皮细胞ECV-304却产生轻度抑制作用，但与空白对照组比较无显著差异（ P＞0．05）。②ATRA浓度在较低浓度（10^-9～10^-7 mol/L）可促进内皮细胞分泌 NO ，随ATRA浓度升高促进NO分泌的作用增强，但ATRA浓度在10-6 mol/L与对照组无明显差异（ P ＞0．05）。【结论】一定浓度的ATRA对内皮细胞有明显的增殖作用，并促进其合成NO。

Objective To explore the effect of all-trans retinoic acid（ATRA） on proliferation and nitric oxide （NO） production of endothelial cells .[Methods]Human umbilical vein endothelial cells （ECV-304） in vitro were randomly divided into 5 groups including blank control group（added by ATRA solvent dimethyl sulfoxide ,DMSO） and other 4 groups（added by ATRA 10^-9 ,10^-8 ,10^-7 and 10^-6 mol/L ,respectively） .The cells were collected at 24h ,48h and 72h .Flow cytometry and MTT method were used to analyze the effect of different concentration of ATRA on ECV-304 proliferation .The effect of ATRA on NO production of endothelial cells was observed .NO level in cell culture fluid was measured .[Results]Low concentration（10^-9 mol/L～10^-7 mol/L） of ATRA could promote the proliferation of ECV-304 in a concentration-dependent manner ,and there was significant difference between ATRA groups and control group（ P 〈0 .01） ,and the effect was increased with the prolongation of action time .High concentration（10^-6mol/L） of ATRA showed slight inhibition effect on ECV-304 ,but there was no significant difference（ P 〉0 .05） .Low concentration（10^-9 mol/L～10^-7mol/L） of ATRA could promote the NO production of endothelial cells ,and the effect was increased with the rising of ATRA concentrations ,but there was no significant difference between ATRA 10-6 mol/L group and control group（ P 〉 0 .05） .[Conclusion]ATRA with proper concentration can promote the proliferation and nitric oxide production of endothelial cells .