骨髓间充质干细胞治疗小型猪急性心肌梗死的实验研究

中国临床药理学与治疗学 ›› 2007, Vol. 12 ›› Issue (7): 767-772.

骨髓间充质干细胞治疗小型猪急性心肌梗死的实验研究

方五旺¹, 林丙来¹, 张升霞², 刘玲玲², 陈绍良²

¹皖南医学院附属芜湖市第二人民医院, 芜湖 241000, 安徽;
²南京医科大学附属南京第一医院, 南京 210006, 江苏

收稿日期:2007-04-08 修回日期:2007-07-16 出版日期:2007-07-26 发布日期:2020-10-27
作者简介:方五旺,男,博士研究生,副主任医师,副教授,研究方向:缺血性心脏病血运重建。Tel:13705530450 E-mail:fangwuwang@sina.com
基金资助:
江苏省科技厅创新基金资助项目(H2003);江苏省南京市科研基金资助项目(200301090)

Autologous bone marrow mesenchymal stem cell transplantation for acute myocardial infarction in pigs

FANG Wu-wang¹, LIN bin-lai¹, ZHANG Sheng-xia², LIU Ling-ling², CHEN Shao-liang²

¹The Second People's Hospital of Wuhu, Wannan Medical Hospital, Wuhu 241000, Anhui, China;
²Department of Cardiology, Nanjing First Hospital Affiliated to Nanjing Medical University, Nanjing 210006, Jiangsu, China

Received:2007-04-08 Revised:2007-07-16 Online:2007-07-26 Published:2020-10-27

摘要/Abstract

摘要： 目的: 研究自体骨髓间充质干细胞(mesenchymalstem cell, MSC) 经冠脉内注射治疗急性心肌梗死的效果。方法: 先以球囊阻塞法制成小型猪急性心肌梗死模型。取骨髓体外培养并扩增MSC, 标记后经冠脉注入, 1 个月后核素心肌显像检测相对心肌梗死面积、心肌灌注评分和射血分数、经导管检测左室收缩压(LVSP)、左室舒张末期压(LVEDP)、压力升高最大速率(LV+dp/dt_max) 和压力下降最大速率(LV-dp/dt_min) 。并以NOGA 电机械标测细胞移植前和移植后心肌缺血面积, 免疫荧光法分析MSC 的植入和分化情况。结果: 和对照组相比, 细胞治疗组射血分数[(44.1±4.3)% vs (39.0±5.2)%, P<0.05]、心肌灌注评分[(30.0±5.3) vs (41.8±8.8),P<0.01]、相对心肌梗死面积[(28.6±4.7)% vs(33.5±3.4)%, P<0.05]、LV+dp/dt_max [(1818±117) mm Hg/s vs (1611±115) mm Hg/s, P<0.01,1 mm Hg=0.133 kPa]、LV-dp/dt_min [(1801±89)mm Hg/s vs (1618±66) mm Hg/s, P<0.01] 均显著增加, LVEDP[(9.4±1.7) mm Hg vs (12.6±2.8)mm Hg, P<0.05] 显著降低。病理检查发现细胞移植术后1 月, 原梗死心肌相关区域即可见带MSC 标记物的新生心肌和血管, 且该部分心肌表达阳性的肌凝蛋白重链、连接蛋白43、平滑肌肌动蛋白和Ⅷ因子相关抗原。结论: MSC 体外培养扩增后经冠脉植入可在心肌梗死小型猪心肌梗死相关区域分化为具有生理功能的新生心肌和血管组织, 可改善急性心肌梗死猪的心功能。

关键词: 骨髓间充质干细胞, 急性心肌梗死, 干细胞移植

Abstract: AIM: The preclinical study was designed to test the feasibility and safety of autologous bone mesenchymal stem cell (MSC) transplantation by intracoronary injection in pigs with acute myocardiac infarction. METHODS: Coronary obstruction with balloon was used to produce acute myocardial infarction (AMI).Labeled MSCs were implanted by intracoronary injection.Four weeks later, single photon emission computed tomography was used to evaluate the relative infarct size.The myocardial ventricular systolic pressure (LVSP), left end-diastolic pressure (LVEDP), peak of pressure rise (LV+dp/dt_max) and peak rate of pressure fall (LV-dp/dt_min) were assessed by catheterization.Immunofluorescence was performed to evaluate MSC engrafment and differentiation. RESULTS: Compared with control group, heart function parameters were all improved in pigs receiving MSCs transplantation, including EF [(44.1±4.3)% vs (39.0±5.2)%, P<0.05] perfusion score [(30.0±5.3) vs (41.8±8.8), P<0.01], relative infarct size [(28.6±4.7)% vs (33.5±3.4)%, P<0.05], LVSP (87±9 mm Hg vs 78±4 mm Hg, P<0.05), LV+dp/dt_max [(1818±117) mm Hg/s vs (1611±115) mm Hg/s, P<0.01, 1 mm Hg=0.133 kPa], LV-dp/dt_min [(1801±89) mm Hg/s vs (1618±66) mm Hg/s, P<0.01] and LVEDP [(9.4±1.7) mm Hg vs (12.6±2.8) mm Hg, P<0.05].MSCs implanted in myocardium were stained positively for myosin heavy chain, connexin 43, smooth muscle actin and von Willebrand factor. CONCLUSION: These results demonstrate that transplantation of MSCs can improve cardiac function in AMI pig, and that the improvement might result from myocardial regeneration and angiogenesis by MSC differentiation.

Key words: bone marrow mesenchymal stem cell, acute myocardial infarction, stem cell transplantation

中图分类号:

R965.2

方五旺, 林丙来, 张升霞, 刘玲玲, 陈绍良. 骨髓间充质干细胞治疗小型猪急性心肌梗死的实验研究[J]. 中国临床药理学与治疗学, 2007, 12(7): 767-772.

FANG Wu-wang, LIN bin-lai, ZHANG Sheng-xia, LIU Ling-ling, CHEN Shao-liang. Autologous bone marrow mesenchymal stem cell transplantation for acute myocardial infarction in pigs[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2007, 12(7): 767-772.

参考文献

1 Orlic D, Kajstura J, Chimenti S, et al.Bone marrow cells regenerate infarcted myocardium[J].Nature, 2001;410:701-705.
2 Jackson KA, Majka SM, Wang H, et al.Regeneration of ischemic cardiac muscle and vascular endothelium by adult stem cells[J].J Clin Invest, 2001;107:1395-1402.
3 Shintani S, Murohara T, Ikeda H, et al.Mobilization of endothelial progenitor cells in patients with acute myocardial infarction[J].Circulation, 2001;103:2776-2779.
4 Beltrami AP, Burbank K, Kajstura J, et al.Evidence that human cardiac myocytes divide after myocardial infarction[J].N Engl J Med, 2001;344:1750-1757.
5 Ye ZJ, Kluger Y, Lian Z, et al.Two types of precursor cells in multipotential hematopoietic cell line[J].Proc Natl Acad Sci USA, 2005;102:18461-18466.
6 Timmermans F, De Sutter J, Gillebert TC, et al.Stem cells for the heart, are we there yet[J]? Cardiology, 2003;100:176-185.
7 Perin EC, Geng YJ, Willerson JT.Adult stem cell therapy in perspective[J].Circulation, 2003;107:935-938.
8 Strauer BE, Kornowski R.Stem cell therapy in perspective[J]. Circulation, 2003;107:935-938.
9 Strauer BE, Brehm M, Zeus T, et al.Repair of infracted myocardium by autologous intracoronary mononuclear bone marrow cell transplantation in humans[J].Circulation, 2002;106:1913-1918.
10 呼莹, 马丽, 马冠杰, 等.成人和胎儿骨髓间充质干细胞的比较研究[J].中华血液学杂志, 2002;23645-23648.
11 Reyes M, Lund T, Lenvik T, et al.Purification and ex vivo expansion of postnatal human marrow mesodermal progenitor cells[J].Blood, 2001;98:2615-2625.
12 Kashiwakura Y, Katoh Y, Tamayose K, et al.Isolation of bone marrow stromal cell-derived smooth muscle cells by a human SM22αpromoter:in vitro differentiation of putative smooth muscle progenitor cells of bone marrow[J].Circulation, 2003; 107:2078-2081.
13 Makino S, Fukuda K, Miyoshi S, et al.Cardiomyocytes can be generated from marrow stromal cells in vitro[J].J Clin Invest, 1999;103:697-705.
14 Tomita S, Li PK, Weisel RD, et al.Autologous transplantation of bone marrow cells improves damaged heart function[J].Circulation, 1999;100:II247-256.

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