CaN-NFAT信号传导通路参与EPCs凋亡作用的研究  

Study for CaN-NFAT signal transduction pathway participating in EPCs apoptosis

作　　者：杨军珂[1] 杨新春[2] 杨龙[3] 胡桃红[1] 谢晓春[1] 赵贵锋[1] 赵明星[4] 

机构地区：[1]中国人民解放军第二炮兵总医院心血管内科,北京100088 [2]首都医科大学附属朝阳医院心脏中心 [3]贵州省人民医院心血管内科 [4]中国人民解放军第二炮兵礼士路门诊部保健科

出　　处：《心血管康复医学杂志》2013年第3期208-212,F0002,共6页Chinese Journal of Cardiovascular Rehabilitation Medicine

基　　金：全军"十一五"医药卫生科研基金(06Q008)

摘　　要：目的:研究钙调神经磷酸酶-活化T细胞核因子(CaN-NFAT)信号传导通路参与内皮祖细胞(EPCs)的凋亡过程。方法:采用密度梯度离心法分离外周血单个核细胞,贴壁培养获得EPCs。实验分为4组:空白对照组、苯肾上腺素(PE)组、环孢素A(CsA)组、CsA+PE组。采用TUNEL染色测定EPCs凋亡状况。逆转录聚合酶链反应法(RT-PCR)检测Bax、Bcl-2、Caspase-3及NFAT4的mRNA的转录水平。免疫荧光染色检测NFAT4亚细胞水平定位。结果:(1)EPCs凋亡检测:与空白对照组及PE组比较,CsA组和CsA+PE组EPCs凋亡数明显增加[(3.41±0.89)比(4.39±1.92)比(23.68±1.14)比(25.92±2.62),P<0.05];(2)与空白对照组和PE组比较,CsA组和CsA+PE组的Bcl-2mRNA水平、Bcl-2/Bax mRNA比值明显下降(P均<0.05),Caspase-3mRNA水平明显升高(P<0.05)。NFAT4mRNA水平:与空白对照组比较,PE组的明显升高(P<0.05),CsA组的明显下降(P<0.05);与PE组比较,CsA组和CsA+PE组的NFAT4mRNA水平均明显下降(P<0.05)。四组之间Bax mRNA水平均无显著差异;(3)与空白对照组比较,PE组NFAT4核转移率明显增加[(25.33±2.08)%比(95±2)%,P<0.05];与空白对照组及PE组比较,CsA组和CsA+PE组NFAT4核转移率[(8.00±2.65)%、(7.00±1.73)%]明显下降(P均<0.05)。结论:CaN-NFAT信号传导通路参与EPCs凋亡的调节作用。Objective：To investigate action of calcineurin enzyme-nuclear factor activated T（CaN-NFAT）signal transduction pathway participating in apoptosis of endothelial progenitor cells（EPCs）.Methods：Peripheral blood mononuclear cells were isolated by density gradient centrifugation and EPCs were obtained by adherent culture.There were four groups：blank control group,phenylephrine（PE）group,cyclosporine A（CsA）group and CsA＋ PE group.Apoptotic status of EPCs was determined using TUNEL staining.Reverse transcription-polymerase chain reaction（RT-PCR）was used to measure mRNA transcription levels of Bax,Bcl-2,Caspase-3and NFAT4.Immunofluorescence staining method was used to measure subcell level location of NFAT4.Results：（1） Detection of EPCs apoptosis：compared with blank control group and PE group,there was significant increase in number of apoptotic EPCs[（3.41±0.89）vs.（4.39±1.92）vs.（23.68±1.14）vs.（25.92±2.62）,P〈0.05]in CsA group and CsA＋PE group（P〈0.05）;（2）Compared with blank control group and PE group,there were significant decrease in Bcl-2mRNA level and ratio of Bcl-2/Bax mRNA（P〈0.05both）,and significant increase in Caspase-3 mRNA level in CsA group and CsA＋PE group（P〈0.05）.Compared with blank control group,NFAT4mRNA level significantly increased（P〈0.05）in PE group and significantly decreased in CsA group（P 0.05）;compared with PE group,NFAT4mRNA level significantly decreased in CsA group and CsA＋PE group（P 0.05both）.There was no significant difference in Bax mRNA level among the four groups;（3）Compared with blank control group,nuclear transfer rate of NFAT4significant increased[（25.33±2.08）% vs.（95±2）%,P〈0.05 ] in PE group（P〈0.05）;compared with blank control group and PE group,nuclear transfer rate of NFAT4 [（8.00±2.65） %,（7.00±1.73） % ] significantly decreased in CsA group and CsA＋PE group（P〈0.05both）.Conclusion：CaN-NFAT signal transduction pathway participates in regulation of EPCs

关 键 词：内皮细胞 信号传导 细胞凋亡 

分 类 号：Q813.110.9[生物学—生物工程]