衰老对缺氧刺激下心脏干细胞生物学功能的影响  

Effect of ageing on biological function of cardiac stem cells under hypoxic stress

作　　者：潘宇[1] 肖静[1] 李晓红[1] 蔡飒[2] 余细勇[1] 

机构地区：[1]广东省人民医院广东省医学科学院医学研究中心,广州510080 [2] 深圳大学医学院组织学与胚胎学教研室

出　　处：《中国医药》2015年第7期933-937,共5页China Medicine

基　　金：国家自然科学基金（81000835、81120108003、813300445）

摘　　要：目的 探讨衰老对缺氧损伤刺激下心脏干细胞（CSC）生物学功能的影响.方法 原代分离小鼠干细胞抗原1 （Sca1）＋ CSC,培养6代后采用衰老β半乳糖苷酶（SA-β-Gal）染色鉴定细胞衰老情况.比较正常培养和缺氧培养第1代和第6代Sca1＋ CSC增殖速度,血管内皮生长因子（VEGF）、血小板衍生生长因子（PDGF）-A、PDGF-B mRNA水平,及VEGF和PDGF-BB分泌水平.将12只青年小鼠和12只老年小鼠各均分为心肌缺血组和假手术组,心肌缺血组采用冠状动脉前降支结扎法建立心肌缺血模型,术后3d取心脏分离细胞,采用流式细胞仪检测Sca1＋ CSC水平.结果 第6代Sca1＋ CSC与第1代Sca1＋ CSC比较体积明显增大,SA-β-Gal蓝染,第1代Sca1＋ CSC未见SA-3-Gal蓝染细胞.第6代Sca1＋ CSC增殖率明显低于第1代Sca1＋ CSC[（18±5）％比（103±27）％]（P＜0.01）.缺氧培养后,第1代Sca1＋ CSC增殖率[（58±11）％]明显降低（P＜0.05）,第6代Sca1＋ CSC增殖率[（26±8）％]无明显改变（P＞0.05）.缺氧培养与正常培养比较,第1代CSC中VEGF、PDGF-A和PDGF-B mRNA水平明显增加,第6代Sca1＋ CSC中VEGF、PDGF-A和PDGF-B mRNA水平无明显变化,第1代Sca1＋ CSC中VEGF和PDGF-BB分泌水平明显增加[（150±37） μg/L比（18±6）μg/L、（288±37） μg/L比（35±6）μg/L],差异有统计学意义（均P＜0.01）,第6代Sca1＋ CSC中VEGF和PDGF-BB分泌水平无明显变化,差异无统计学意义（均P＞0.05）.心肌缺血组老年和青年小鼠Sca1＋ CSC水平均明显高于假手术组[（13.2±4.2）％比（1.5±0.4）％、（31.7±6.5）％比（5.0±0.6）％]（P＜0.05）,2组老年小鼠Sca1＋ CSC水平均明显低于青年小鼠（P＜0.05）.结论 衰老可致缺氧损伤刺激下Sca1＋ CSC增殖能力、分泌能力以及损伤后的动员能力下降.Objective To investigate the effect of ageing on biological function of cardiac stem cells （CSC） under stress of hypoxia.Methods Primitive mouse Sca1 ＋ CSC were isolated and SA-β-Gal staining was used to confirm the ageing features of the 6th passage （P6） Sca1 ＋ CSC.Before and after hypoxic treatment,the proliferation abilities,the mRNA levels of vascular endothelial growth factor （VEGF） and platelet-derived growth factor （PDGF）-A/B,the secretive levels of VEGF and PDGF-BB in P1 and P6 Sca1 ＋ CSC were detected and compared.Ischemic models were established by ligation of the coronary artery in 6 young mice and 6 old mice （ischemic group）;in addition,6 young mice and 6 old mice were set as sham group.Three days after establishment of ischemic model,the Sca1 ＋ CSC pool in heart was detected through flow cytometry.Results P6 Sca1 ＋ CSC were positively stained by SA-β-Gal with larger volume compared with P1 Sca1 ＋ CSC,while no positive staining was observed in P6 Sca1 ＋ CSC.The proliferation rate of P6 Sca1 ＋ CSC was significantly lower than that of P1 Sca1 ＋ CSC [（18 ± 5）% vs （103 ± 27）%,P 〈 0.01）];after hypoxic treatment,the proliferation rate of P1 Sca1 ＋ CSC [（58 ± 11） %] was significantly reduced （P 〈 0.05）;no obvious change was found in P6 Sca1 ＋ CSC [（26 ± 8） %] （P 〉 0.05）.The mRNA levels of VEGF,PDGF-A and PDGF-B were significantly increased in P1 Sca1 ＋ CSC after hypoxic treatment,while no significant changes were found in P6 Sca1 ＋ CSC.After hypoxic treatment,the secretive VEGF and PDGF-BB were significantly increased in P1 Sca1 ＋ CSC [（150 ± 37） μg/L vs （18 ± 6） μg/L,（288 ± 37） μg/L vs （35 ± 6） μg/L,P 〈 0.01],while no significant changes were found in P6 Sca1 ＋ CSC （P 〉 0.05）.In experiment in vivo,the population of Sca1 ＋ CSC in ischemic group was significantly larger than that in sham group [old mice：（13.2 ± 4.2） % vs （1.5 ± 0.4%） %,young mice：（31.7 ± 6.5） % vs （5.0

关 键 词：衰老 心脏干细胞 缺氧 

分 类 号：R255.1[医药卫生—中医内科学]