姜黄素对慢性阻塞性肺疾病患者外周血内皮祖细胞数量及功能的影响

Effects of curcumin on number and function of endothelial progenitor cells derived from peripheral blood of chronic obstructive pulmonary disease patients

目的观察不同浓度姜黄素对慢性阻塞性肺疾病(COPD)患者外周血内皮祖细胞(EPCs)数量、增殖功能、一氧化氮(NO)分泌功能及内皮型一氧化氮合酶(e NOS)表达的影响,探讨姜黄素修复和保护COPD患者受损的血管内皮功能作用的可能机制。方法选择30例住院的COPD患者,密度梯度离心法获取外周血单个核细胞,加入不同浓度姜黄素(0、5、10、15μmol·L-1)刺激并诱导其分化为EPCs,荧光双染色鉴定EPCs并进行细胞计数。收集贴壁细胞,再加入各浓度姜黄素,相同条件下培养一定时间(6、12、24、48 h)。采用CCK-8法检测EPCs的增殖能力,硝酸还原酶法(Griess法)检测NO分泌量,Western Blot法检测e NOS蛋白的表达。结果与未加入姜黄素的对照组相比,姜黄素可显著增加COPD患者EPCs的数量,并呈浓度依赖性;姜黄素显著改善了EPCs的增殖功能及NO分泌功能,呈浓度时间依赖性;姜黄素也可显著上调EPCs的e NOS表达,与姜黄素浓度之间具有相关性。结论姜黄素增加了EPCs的数量,改善了EPCs增殖功能及NO分泌功能,并上调了e NOS的表达。提示姜黄素可能通过增加EPCs的数量及改善EPCs的功能来修复COPD患者的血管内皮功能损伤。

AIM To investigate curcumin influence of curcumin on endothelial progenitor cells( EPCs) of chronic obstructive pulmonary diseases( COPD) patients and discuss the possible mechanism of the protective effects curcumin on vascular endothelial function inpatients with COPD. METHODS The peripheral blood mononuclear cells isolated from peripheral blood of 30 COPD in patients were induced into EPCs and stimulated with curcumin( to make a series of concentrations: 0,5,10,15 μmol·L-1). EPCs were characterized as adherent cells double positive by direct fluorescent staining and number of EPCs were counted. Attached EPCs were collected and cultured with various concentrations of curcumin for the respective time points( 6,12,24 and 48 h). Proliferation of EPCs was tested with CCK-8 assay. Nitric oxide secretion was assayed with Griess Reagent System. Western Blot was used to measure the expression of endothelial nitric oxide synthase. RESULTS Compared with control group,curcumin increased the number of EPCs with concentration dependently. And curcumin significantly improved the proliferation nitric oxide( NO) secretion of endothelial progenitor cells within concentration and time dependence.The expression of endothelial nitric oxide synthase( e NOS) was also be significantly up-regulated. CONCLUSION Curcumin increases the number of EPCs and enhances the proliferation,NO secretion and e NOS expression as well,suggesting that curcumin could repair the injured vascular endothelial function of COPD patients by promote the number and function of EPCs.