姜黄素对人脐静脉血来源内皮祖细胞HO-1表达的影响及机制探讨  

Effect of curcumin on HO-1 expression in endothelial progenitor cells from human umbilical vein blood

作　　者：丁红[1,2] 黄维义[1] 沈桂冬[2] 

机构地区：[1]泸州医学院附属医院,四川泸州646000 [2]安康市中心医院

出　　处：《山东医药》2015年第7期17-19,共3页Shandong Medical Journal

基　　金：泸州市人民政府-泸州医学院科技战略合作科技项目(2013LZLY-J15)

摘　　要：目的观察姜黄素（CMN）对人脐静脉血来源内皮祖细胞（EPCs）血红素氧合酶1（HO-1）蛋白表达的影响,并探讨其机制。方法采用密度梯度离心法体外分离、培养人脐静脉血单个核细胞,以免疫荧光法观察细胞吸收Dil-ac-LDL和FITC-UAE-1情况并同时进行CD133免疫荧光鉴定EPCs。将EPCs分为0、5、10、15μmol/L CMN组,分别以终浓度为0、5、10、15μmol/L CMN的培养基孵育24 h,检测各组HO-1蛋白表达、细胞增殖能力及培养液中乳酸脱氢酶（LDH）活力。将EPCs分为单纯组、CMN组、CMN＋ATRA组,分别在单纯培养基、加15μmol/L CMN的培养基、加15μmol/L CMN＋1μmol/L ATRA的培养基中孵育24 h,检测各组HO-1蛋白表达。结果从脐静脉血分离出来的单核细胞培养3~4 d开始贴壁,7 d后有一定方向性,呈簇状生长,免疫荧光检测显示CD133表面标记阳性率及Di I-ac LDL和FITC-UEA-1双染色阳性率均〉90%。随CMN浓度升高,EPCs的HO-1蛋白表达逐渐升高（P均〈0.05）,细胞增殖能力逐渐增强（P均〈0.05）,细胞培养液中LDH活力则无明显改变（P均〉0.05）。与单纯组比较,CMN组、CMN＋ATRA组HO-1蛋白表达量增加（P均〈0.05）,且后两组比较,P〈0.05。结论 CMN可诱导人脐静脉血来源EPCs高表达HO-1蛋白,这可能与Nrf2/ARE信号途径有关。Objective To observe the effects of curcumin （ CMN） on the HO-1 protein expression in endothelial pro-genitor cells （ EPCs ） from human umbilical vein blood and to investigate the mechanisms .Methods The mononuclear cells were isolated and cultured by density gradient centrifugation from human umbilical vein blood .The morphology of EPCs was observed with inverted microscope ,EPCs were also identified as adherent cells which were double positive for Dil -ac-LDL/FITC-UAE-1 by immumofluorescence method , and their expression of CD 133 was detected .EPCs were divided into four groups and incubated with 0, 5, 10 and 15μmol/L CMN respectively for 24 h.Then the HO-1 protein expression, the cell proliferation activity and the Lactate dehydrogenase （ LDH） activity in the supernatant were detected .EPCs were divid-ed into three groups and incubated with 0 μmol/L CMN （simple group）, 15 μmol/L CMN （CMN group） and 15μmol/L CMN＋1 μmol/L all-trans retinoic acid （ATRA） （CMN＋ATRA group） respectively for 24 h.The HO-1 protein expres-sion was detected .Results Attached cells were observed after 3-4 days and cluster growth was noticed after 7 days.The immunofluorescence showed that the positive rate of CD 133 , the double staining positive rates of DiI-acLDL and FITC-UEA-1 were all more than 90%.With the increase of CMN concentration , the expression of HO-1 protein in the EPCs increased gradually （all P〈0.05）, and the cell proliferation increased significantly （all P〈0.05）, but the LDH activity in the su-pernatant did not change obviously （all P〉0.05）.Compared with simple medium group , the expression of HO-1 protein in the CMN and CMN＋ATRA group increased （all P〈0.05）, and significant difference was found between the CMN and CMN＋ATRA groups （ P〈0.05）.Conclusion CMN may effectively induce the high protein expression of HO-1 in EPCs, which is related with Nrf2/ARE signal pathway.

关 键 词：姜黄素 血红素氧合酶1 内皮祖细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]