芪红合剂含药血清对体外培养人外周血内皮祖细胞NO及NOS的影响

目的观察芪红合剂含药血清对人外周血内皮祖细胞（EPCs）一氧化氮（NO）、一氧化氮合酶（NOS）的影响,探讨其对人外周血内皮祖细胞的保护机制。方法12只雄性Wistar大鼠随机分为芪红合剂组（低剂量组、中剂量组、高剂量组）和对照组,制备含药血清和空白血清。采集健康成人外周血,分离单核细胞,体外培养7d后,收集贴壁细胞,激光共聚焦显微镜及流式细胞仪鉴定正在分化的EPCs,加入含不同浓度芪红合剂药物血清及空白血清的M199培养基孵育24h,将其在倒置荧光显微镜下计数,硝酸还原酶法测定NO,化学比色法测定NOS。结果芪红合剂药物组的血清NO水平和NOS活性明显高于空白血清对照组（P〈0.05）,并且呈剂量依赖性。结论芪红合剂可能通过增加EPCsNOS活性,促进NO分泌,防止血管内皮损伤及动员血管内皮祖细胞修复损伤血管来发挥血管内皮保护作用。

Influence of Qihong Decoction-containing Serum on Nitric Oxide and Nitric Oxide Synthase of Blood-derived Endothelial Progenitor Cells

Objective To investigate the effects of Qihong decoction-containing serum on nitric oxide（NO） and nitric oxide synthase（NOS） of endothelial progenitor cells（EPCs）derived from peripheral blood.Methods Twelve Wistar male rats were randomly divided into four groups：Qihong decoction low dosage group,medium dosage group,high dosage group and control group.The mononuclear cells were isolated from human peripheral blood and cultured with Medium 199,that were identified to be EPCs on day 7 by the laser scanning confocal microscope and the flow cytometry.The cultured EPCs were incubated with different concentration of Qihong decoction-containing serum for 24 h.The activity of EPCs was observed under reverse microscope.The activity of NOS was measured with the NOS detecting kit and the concentrations of NO in the cell supernatant were measured with the nitrate reductase method.Results The contents of nitric oxide in Qihong decoction-containing serum group（low,medium and high dose group） were significantly higher than that in control group.The activity of NOS in the Qihong decoction-containing serum group（low,medium and high dose group）was significantly higher than that in control group（P 0.05）.Qihong decoction-containing serum promoted the release of NO from EPCs and enhanced the activity of NOS in a dose-dependent manner.Conclusion Qihong decoction could promote the release of NO and enhance the activity of NOS of the endothelial progenitor cells derived from peripheral blood.