CAJ | 학술논문

目的应用MRI评价经冠状动脉途径移植猪自体骨髓间质干细胞(BM-MSCs)治疗急性心肌梗死(AMI)的治疗效果.方法 8月龄中华小型猪14只[(27±3)kg],平均分成移植组与对照组.胶圈套扎左前降支第一对角支分叉以远90 min后松开,建成AMI模型.心肌梗死后1周进行细胞移植,将干细胞悬液(移植组:1×106/ml × 10 ml)或无血清培养液(DMEM)(对照组10 ml)经微导管注入前降支.AMI术后1周(基线)和MSCs移植后6周(终点)各行1次MR扫描,评价移植前后心脏形态、功能、心肌灌注及延迟增强.第2次MR扫描后立即处死动物,分别行冰冻切片、石蜡切片和电镜观察.结果与对照组比较,移植组在移植6周后左心室整体功能较前明显改善,实验组与对照组比较,左心室平均射血分数(EF)值从(42.7±7.5)%升至(50.1±10.1)%(P＜0.01),左心室节段运动异常数平均减少4个(P＜0.01)、梗死面积减少3.2 cm2(P＜0.01),心脏重量指数增加4.1 g/m2(P＜0.05).病理证实移植组梗死区和梗死周边的病变情况显著轻于对照组,有大量存活心肌,纤维化程度显著减轻;并且可见核大、边集,胞质丰富的幼稚细胞;在梗死区和梗死周边区组织的冰冻切片上可见4-6-二脒基二苯基吲哚(DAPI)阳性的移植细胞存活.免疫荧光检测进一步表明大部分DAPI阳性细胞表达心肌特异性肌钙蛋白T(troponin T),并且表达间隙连接蛋白43(connexin 43).部分DAPI阳性细胞表达平滑肌肌动蛋白(smooth muscle actin)和血管性血友病因子(Von Willebrand),移植组梗死周边区毛细血管密度显著高于对照组[分别为(8.7±2.0)、(4.9±1.3)个/高倍镜](P＜0.01).结论 MRI可作为猪BM-MSCs在体移植前后评价其治疗效果的可靠影像检查方法 .
목적 응용MRI평개경관상동맥도경이식저자체골수간질간세포(BM-MSCs)치료급성심기경사(AMI)적치료효과.방법 8월령중화소형저14지[(27±3)kg],평균분성이식조여대조조.효권투찰좌전강지제일대각지분차이원90 min후송개,건성AMI모형.심기경사후1주진행세포이식,장간세포현액(이식조:1×106/ml × 10 ml)혹무혈청배양액(DMEM)(대조조10 ml)경미도관주입전강지.AMI술후1주(기선)화MSCs이식후6주(종점)각행1차MR소묘,평개이식전후심장형태、공능、심기관주급연지증강.제2차MR소묘후립즉처사동물,분별행빙동절편、석사절편화전경관찰.결과 여대조조비교,이식조재이식6주후좌심실정체공능교전명현개선,실험조여대조조비교,좌심실평균사혈분수(EF)치종(42.7±7.5)%승지(50.1±10.1)%(P＜0.01),좌심실절단운동이상수평균감소4개(P＜0.01)、경사면적감소3.2 cm2(P＜0.01),심장중량지수증가4.1 g/m2(P＜0.05).병리증실이식조경사구화경사주변적병변정황현저경우대조조,유대량존활심기,섬유화정도현저감경;병차가견핵대、변집,포질봉부적유치세포;재경사구화경사주변구조직적빙동절편상가견4-6-이미기이분기신타(DAPI)양성적이식세포존활.면역형광검측진일보표명대부분DAPI양성세포표체심기특이성기개단백T(troponin T),병차표체간극련접단백43(connexin 43).부분DAPI양성세포표체평활기기동단백(smooth muscle actin)화혈관성혈우병인자(Von Willebrand),이식조경사주변구모세혈관밀도현저고우대조조[분별위(8.7±2.0)、(4.9±1.3)개/고배경](P＜0.01).결론 MRI가작위저BM-MSCs재체이식전후평개기치료효과적가고영상검사방법 .
Objective To investigate the effects of autologous bone marrow-derived mesenchymal stem cells (MSCs)transplantation on acute myocardial infarction in swine models using MRI. Methods Fourteen Chinese mini-pigs(27±3 kg)were divided into control group(n=7)and transplantation group(n=7).Acute myocardial infarction(AMI)model was made by occlusion of the left anterior descending coronary artery for 90 minutes,and then 10 ml autologous MSCs(3 × 106 cell/ml)were injected into LAD by over-wire-balloon catheter after one week. MRl was performed to assess the cardiac function and myocardial perfusion 1 week after AMI and 6 weeks after transplantation.The implanted cells in vitro were analyzed by immunofluorescence.Resuits The left ventricular ejection fraction(LVEF)in transplantation group was increased from(42.7 ±7.5)%to(50.1±10.1)%,which was significantly different from that in control group(P＜0.01).In addition,the dyskinetic segments in infarcted region and the infareted area were decreased by 4 and 3.2 cm2 respectively(P＜0.01),and the left ventricular weight index was increased by 4.1 g/m2 in transplantation group(P＜0.05)compared with control group.The DAPI-labeled cells in infarcted and peri-infarcted region indicated the survived MSCs.Immunofluoreseence also confirmed that those cells expressed cardiomyocyte-specific troponin T,connexin 43 and vessel-specific smooth muscle actin.Capillary density in both infarcted and peri-infarcted region were higher in transplantation group than the control group(P＜0.01).Conclusion MRI is a reliable imaging method for assessing the effects of stem cell transplantation in acute myocardial infartion of swine models.