hTERT基因对移植的内皮祖细胞在体内的增殖和血管新生的影响  

Effect of hTERT Gene on Proliferation,Migration and Angiogenesis of Transplanted Endothelial Progenitor Cells in Vivo

作　　者：柯伟良 李日健 闫海 冯锦山 邵芳芳 李兴岳 李上海 KE Weiliang;LI Rijian;YAN Hai;FENG Jinshan;SHAO Fangfang;LI Xingyue;LI Shanghai(Department of Cardiology,The Affiliated Hospital of Guangdong Medical University,Guangzhou 524001,Guangdong,China)

机构地区：[1]广东医科大学附属医院心内科

出　　处：《心血管病学进展》2019年第5期822-827,共6页Advances in Cardiovascular Diseases

基　　金：广东省医学科学技术研究基金项目(A2017494);广东省医学科学技术研究基金项目(A2016267)

摘　　要：目的 研究h TERT基因对移植的内皮祖细胞(EPC)在体内的增殖和血管新生影响的机制。方法 将90只健康雄性Sprague-Dawley大鼠(200~300 g)随机分为3组:对照组(n=30)、EPC转染组(n=30)和h TERT-EPC转染组(n=30)。选择三个梗死病灶区域通过心肌内注射移植EPC和h TERT-EPCs,对照组的大鼠注射磷酸缓冲盐溶液;通过苏木精-曙红染色的心肌的石蜡切片,观察心肌梗死区域的毛细血管再生;通过RT-qPCR和蛋白质免疫印迹检测血管内皮细胞标记物(CD31、CD34、CD105和CD146) mRNA和蛋白的表达量;通过检测对照组和模型组小鼠体外骨髓EPCs的增殖评估h TERT对EPC功能的影响;使用酶标仪检测NO浓度和乳酸脱氢酶(LDH)浓度,通过商业测定试剂盒检测诱导型一氧化氮合酶(i NOS)活性水平;通过酶联免疫吸附试验试剂盒检测血管内皮生长因子、血管内皮生长因子受体和丝裂原活化蛋白激酶含量。结果 与EPC转染组相比,h TERT-EPC转染组毛细血管在心肌梗死区增生较多;与EPC转染组相比,h TERT-EPC转染组CD31、CD34、CD105、CD146 mRNA和蛋白表达水平增加,差异有统计学意义(P<0. 05);与对照组(173. 52±20. 17) EPC相比,模型组(32. 14±12. 33)小鼠的EPC显示受损的增殖,h TERTEPC转染组(122. 14±13. 44)增加了受损EPC的增殖,差异有统计学意义(P<0. 05);h TERT-EPC转染组中NO(18. 23±3. 15)、LDH(4. 03±0. 44)和i NOS(4. 52±1. 13)的分泌均高于EPC转染组[(14. 62±2. 18)、(3. 24±0. 67)、(3. 83±0. 82)],差异有统计学意义(P<0. 05);h TERT-EPC转染组血管内皮生长因子、血管内皮生长因子受体和丝裂原活化蛋白激酶与EPC转染组相比升高,差异有统计学意义(P<0. 05)。结论 h TERT基因促进移植的内皮祖细胞在体内的增殖和血管新生。To study the mechanism of hTERT gene on the proliferation and angiogenesis of transplanted endothelial progenitor cell(EPC)in vivo.Ninety healthy male Sprague-Dawley rats(200~300 g)were randomly divided into three groups:control group(n=30),EPC transfection group(n=30)and hTERT-EPC transfection group(n=30).Three infarct lesion areas were selected.Transplanted EPC and hTERT-EPC were injected into myocardium,and PBS was injected into rats in the control group.The capillary regeneration of myocardial infarction area was observed by paraffin section of myocardium stained with hematoxylin-eosin.The mRNA and protein expression levels of vascular endothelial cell markers(CD31,CD34,CD105 and CD146)were detected by RT-qPCR and western blotting.The effect of hTERT on EPC function was assessed by detecting the proliferation of bone marrow EPCs in vitro in mice of control group and model group.NO concentration and LDH concentration were detected by the enzyme labelling instrument,and the iNOS activity level was detected by commercial assay kits.VEGF,VEGFR and MAPK protein contents were detected by ELISA Kit.Compared with EPC transfection group,capillaries in hTERT-EPC transfection group proliferated was more in myocardial infarction group;compared with EPC transfection group,the expression levels of CD31,CD34,CD105 and CD146 mRNA and protein in hTERT-EPC transfection group increased,the differences were statistically significant(P<0.05);compared with EPC of the control group(173.52±20.17),the model group(32.14±12.33)showed impaired proliferation and migration,hTERT-EPC group(122.14±13.44)increased the proliferation and migration of damaged EPC,the difference was statistically significant(P<0.05);in the hTERT-EPC transfection group,the secretion of NO(18.23±3.15),LDH(4.03±0.44)and iNOS(4.52±1.13)was higher than those of EPC transfection group[(14.62±2.18),(3.24±0.67),(3.83±0.82)],the differences were statistically significant(P<0.05);the levels of VEGF,VEGFR and MAPK hTERT-EPC transfection group were significantly hi

关 键 词：HTERT 内皮祖细胞 增殖 血管新生 

分 类 号：R73[医药卫生—肿瘤]