中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2015年
4期
640-646
,共7页
高青%李树仁%荀丽颖%苑可心%谢悦陶%张倩辉%郝清卿%党懿%齐晓勇
高青%李樹仁%荀麗穎%苑可心%謝悅陶%張倩輝%郝清卿%黨懿%齊曉勇
고청%리수인%순려영%원가심%사열도%장천휘%학청경%당의%제효용
骨髓间充质干细胞%基因治疗%心肌梗死%bcl-2基因
骨髓間充質榦細胞%基因治療%心肌梗死%bcl-2基因
골수간충질간세포%기인치료%심기경사%bcl-2기인
Bone marrow mesenchymal stem cells%Gene therapy%Myocardial infarction%bcl-2 gene
目的:探讨经bcl-2基因修饰的骨髓间充质干细胞( BMSCs)移植对急性心肌梗死家兔心肌细胞凋亡、血管再生及心功能的影响。方法:体外分离、培养、纯化兔BMSCs,分别转染腺病毒及重组腺病毒-Bcl-2。结扎兔冠状动脉前降支制作心肌梗死(MI)模型,2周后于心梗边缘区分别注射等量的腺病毒-Bcl-2-BMSCs(MI+Bcl-2-BM-SCs组)、腺病毒-BMSCs( MI+BMSCs组)及DMEM液( MI组)。细胞移植4周后经超声测定心功能;荧光显微镜观察BMSCs的存活及分布;TUNEL法检测心肌细胞凋亡;real-time PCR检测VEGF mRNA表达;免疫组化染色法检测CD31表达,计算新生毛细血管密度。以上数据分别与心功能进行相关性分析。结果:与MI组相比,MI+Bcl-2-BM-SCs组和MI+BMSCs组的心功能改善、细胞凋亡率降低、VEGF mRNA表达增多、毛细血管密度增加,其中MI+Bcl-2-BMSCs组的变化更为显著( P<0.05)。相关性分析显示左室射血分数与心肌细胞凋亡率呈负相关;与VEGF mR-NA的表达量及毛细血管密度呈正相关(P<0.01)。结论:经bcl-2基因修饰的BMSCs移植可显著减少缺血性心功能不全兔心肌细胞凋亡、促进血管再生、改善心功能。
目的:探討經bcl-2基因脩飾的骨髓間充質榦細胞( BMSCs)移植對急性心肌梗死傢兔心肌細胞凋亡、血管再生及心功能的影響。方法:體外分離、培養、純化兔BMSCs,分彆轉染腺病毒及重組腺病毒-Bcl-2。結扎兔冠狀動脈前降支製作心肌梗死(MI)模型,2週後于心梗邊緣區分彆註射等量的腺病毒-Bcl-2-BMSCs(MI+Bcl-2-BM-SCs組)、腺病毒-BMSCs( MI+BMSCs組)及DMEM液( MI組)。細胞移植4週後經超聲測定心功能;熒光顯微鏡觀察BMSCs的存活及分佈;TUNEL法檢測心肌細胞凋亡;real-time PCR檢測VEGF mRNA錶達;免疫組化染色法檢測CD31錶達,計算新生毛細血管密度。以上數據分彆與心功能進行相關性分析。結果:與MI組相比,MI+Bcl-2-BM-SCs組和MI+BMSCs組的心功能改善、細胞凋亡率降低、VEGF mRNA錶達增多、毛細血管密度增加,其中MI+Bcl-2-BMSCs組的變化更為顯著( P<0.05)。相關性分析顯示左室射血分數與心肌細胞凋亡率呈負相關;與VEGF mR-NA的錶達量及毛細血管密度呈正相關(P<0.01)。結論:經bcl-2基因脩飾的BMSCs移植可顯著減少缺血性心功能不全兔心肌細胞凋亡、促進血管再生、改善心功能。
목적:탐토경bcl-2기인수식적골수간충질간세포( BMSCs)이식대급성심기경사가토심기세포조망、혈관재생급심공능적영향。방법:체외분리、배양、순화토BMSCs,분별전염선병독급중조선병독-Bcl-2。결찰토관상동맥전강지제작심기경사(MI)모형,2주후우심경변연구분별주사등량적선병독-Bcl-2-BMSCs(MI+Bcl-2-BM-SCs조)、선병독-BMSCs( MI+BMSCs조)급DMEM액( MI조)。세포이식4주후경초성측정심공능;형광현미경관찰BMSCs적존활급분포;TUNEL법검측심기세포조망;real-time PCR검측VEGF mRNA표체;면역조화염색법검측CD31표체,계산신생모세혈관밀도。이상수거분별여심공능진행상관성분석。결과:여MI조상비,MI+Bcl-2-BM-SCs조화MI+BMSCs조적심공능개선、세포조망솔강저、VEGF mRNA표체증다、모세혈관밀도증가,기중MI+Bcl-2-BMSCs조적변화경위현저( P<0.05)。상관성분석현시좌실사혈분수여심기세포조망솔정부상관;여VEGF mR-NA적표체량급모세혈관밀도정정상관(P<0.01)。결론:경bcl-2기인수식적BMSCs이식가현저감소결혈성심공능불전토심기세포조망、촉진혈관재생、개선심공능。
[ ABSTRACT] AIM:To investigate the effects of transplantation of bone marrow mesenchymal stem cells ( BMSCs) modified by bcl-2 gene on myocardial cell apoptosis, angiogenesis and cardiac function in the rabbit after acute myocardial in-farction ( MI) .METHODS:The rabbit BMSCs were isolated, cultured and purified in vitro.The BMSCs were transfected with adenovirus or adenovirus-Bcl-2.The rabbit model of MI was established by ligation of left anterior descending branch. The rabbits were injected with Ad-Bcl-2-BMSCs ( MI+Bcl-2-BMSCs group) , Ad-BMSCs ( MI+BMSCs group) and DMEM ( MI group) in infarction marginal zone 2 weeks after ligation.The cardiac function was evaluated by echocardiography.The apoptosis of myocardial cells was measured by TUNEL.The mRNA expression of VEGF was detected by real-time PCR.The expression of CD31 was examined by immunohistochemical staining, and new blood capillaries were counted at 4 weeks after BMSCs transplantation.The correlation of the above values with cardiac function was analyzed.RESULTS: The cardiac function was better, the apoptotic rate was lower, the mRNA expression of VEGF and the capillary density were higher in both MI+Bcl-2-BMSCs group and the MI+BMSCs group than those in MI group, and those in MI+Bcl-2-BMSCs group in-creased more obviously .The left ventricular ejection fraction ( LVEF) had a negative correlation with the myocardial cell ap-optosis rate.A positive correlation with the mRNA expression level of VEGF and the capillary density was also observed. CONCLUSION:The transplantation of BMSCs modified by bcl-2 gene significantly reduces the myocardial cell apoptosis, promotes angiogenesis, improves heart function of the rabbits with MI.