TIMP-3基因转染的血管平滑肌细胞移植对心肌梗死后心脏形态和功能的影响
《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]
- 期数:
- 2009年第6期
- 页码:
- 765-769
- 栏目:
- 基础研究
- 出版日期:
- 2009-11-05
文章信息/Info
- Title:
- Effects of transplantation of tissue inhibitor-3 of matrix metalloproteinase gene-transfected vascular smooth muscle cells on heart structure and function after myocardial infarction
- 作者:
- 贾智博1; 田海1; 苗宏志2; 刘开宇1; 孙露1; 蒋树林1; 李仁科3
- 1.哈尔滨医科大学附属第二医院心外科，黑龙江 哈尔滨 150086；2.齐齐哈尔市第一医院胸心外科， 黑龙江 齐齐哈尔，161005；3.加拿大多伦多大学多伦多总医院心血管外科研究所，加拿大 多伦多
- Author(s):
- JIA Zhi-bo1; TIAN Hai1; MIAO Hong-zhi2; LIU Kai-yu1; SUN Lu1; JIANG Shu-lin1; LI Ren-ke3
- 1.Department of Cardiovascular Surgery, Second Affiliated Hospital, Harbin Medical University, Harbin 150086, Heilongjiang, China; 2.Department of Cardiothoracic Surgery, First Hospital of Qiqihaer, Qiqihaer 161005, Heilongjiang, China; 3.Department of Cardiovascular Surgery, Toronto General Hospital, University Health Network, Toronto, Ontario, Canada
- Keywords:
- tissue inhibitor-3 of matrix metalloproteinases gene; myocardial infarction; heart structure; heart function; cell transplantation; rat
- 分类号:
- R542.22；Q789
- DOI:
- -
- 文献标识码:
- A
- 摘要:
- 目的: 观察基质金属蛋白酶组织抑制因子3 (tissue inhibitor-3 of matrix metalloproteinases，TIMP-3)基因转染的血管平滑肌细胞(vascular smooth muscle cells，VSMCs)移植，对大鼠心肌梗死后心脏形态和功能的影响。 方法: 取Wistar大鼠胸主动脉采用组织块贴壁法培养VSMCs。应用脂质体转染技术，将TIMP-3-pcDNA3.0重组质粒瞬时转染至VSMCs。另取雌性Wistar大鼠121只（体质量200～250 g），其中93只建立急性心肌梗死模型。将存活的84只随机分成A、B、C 3组（每组28只），即于心肌梗死后3 d，再次开胸并在梗死的心肌中分别注射含有3×106个TIMP-3基因转染的VSMCs（A组）、3×106个VSMCs（B组）或改良Eagle培养基（Dulbeccos modification of Eagles medium Dulbecco，DMEM培养基）（C组）各0.3 ml，121只中剩余的28只大鼠作为正常对照组（D组），不做任何处置。基因转移4周后，进行超声心动图检测心脏的功能，然后获取心脏标本，计算心脏左室容积(left ventricular volume，LVV)和左室容积指数(left ventricular volume index，LVVI)，最后进行心脏组织学观察。 结果: 成功分离、培养了VSMCs，纯度达98%，TIMP-3基因成功转入VSMCs中。基因转移4周，超声心动图检查发现，心脏功能的各项指标A组优于B组和C组，B组优于C组；但A、B、C 3组均差于D组（P均<0.05）。A、B、C和D组心脏LVV分别为(894.4±158.3)mm3、(1 126.5±284.7)mm3、(1 372.8±181.9)mm3和(420.2±39.6)mm3，LVVI分别为(2.957±0.362)mm3/g、(3.604±0.710)mm3/g、(4.538±0.259)mm3/g和(1.009±0.134)mm3/g，A、B、C 3组均明显大于D组，A、B组较C组明显减小，A组较B组明显减小（P均<0.05）。组织学观察显示，C组呈心肌梗死表现，B组梗死心肌内可见成簇的肌细胞，室壁较C组增厚；A组较B、C组梗死区缩小、室壁增厚，D组无异常。 结论: 大鼠心肌梗死后3 d，将TIMP-3基因转染的VSMCs移植入梗死心肌内，可明显改善心脏的形态及功能。
- Abstract:
- AIM: To investigate the effects of transplantation of tissue inhibitor-3 of matrix metalloproteinase (TIMP-3) gene-transfected vascular smooth muscle cells (VSMCs) on heart structure and function after myocardial infarction (MI) in rats. METHODS: VSMCs were obtained from rat thoracic aorta by tissue-piece inoculation. TIMP-3 gene was transfected into VSMCs by lipidosome-mediated method. One hundred and twenty-one female Wistar rats, aged 12 weeks and weighing between 200 and 250 g, were prepared. Acute MI models were established in 93 rats by ligating the descending left coronary artery; 84 survived and were divided randomly into three groups (n=28). Three days after MI, rat chests were opened again and 0.3 ml Dulbecco’s modified Eagle’s medium (DMEM) containing 3×106 TIMP-3 gene-transfected VSMCs (group A), 3×106 VSMCs (group B) or 0.3 ml DMEM (group C) was immediately injected into the infarcted myocardium. The control group (group D) consisted of 28 rats without any treatment. Heart function was detected by ultrasonic echocardiogram. Heart samples were harvested and heart left ventricular volume and volume index were measured. Heart structure was also observed by tissue morphologic examination. RESULTS: VSMCs were cultivated and had a high purity (98%). TIMP-3 gene was transfected into VSMCs successfully. Four weeks after gene transfer, heart function in group A was improved over group B and group C (P<0.05), group B was improved over group C (P<0.05), but they were all poorer than group D (P<0.05. Left ventricular volume was (94.4±158.3) mm3, (1 126.5±284.7) mm3, (1 372.8±181.9) mm3 and (420.2±39.6) mm3 in the four groups, and volume index was (2.957±0.362) mm3/g, (3.604±0.710) mm3/g, (4.538±0.259) mm3/g and (1.009±0.134) mm3/g. The results of histological observation showed MI in group C. Clustering muscle cells were seen in infarction field in group B and heart ventricle wall was thicker than in group C. Infarction field in group A was smaller than in groups B and C. Heart ventricle wall in group A was thicker than in groups B and C. CONCLUSION: Implanted TIMP-3 gene transfected VSMCs in infarcted myocardium at 3 days after acute MI noticeably improves heart structure and function in rats.
参考文献/References
［1］ 贾智博，田海，刘开宇，等. 基质金属蛋白酶组织抑制因子3基因转染血管平滑肌细胞移植对急性心肌梗死后早期心肌重塑的影响［J］. 中国修复重建外科杂志, 2007, 21(5):512-516.
［2］ Nakamura A, Isoyama S, Watanabe T, et al. Heterogeneous smooth muscle cell population derived from small and larger arteries［J］. Microvasc Res, 1998, 55(1):14-28.
［3］ 尹倪，陈胜喜，罗万俊，等. 冠状动脉前降支结扎法制作大鼠急性心肌梗死模型［J］. 中国医师杂志， 2006, 8(9):1193-1195.
［4］ 周欣，李玉明. 心肌梗死后创伤修复与心室重塑［J］. 中华心血管病杂志, 2004, 32(4):375-377.
［5］ Gelfand EV, Cannon CP. Myocardial infarction: contemporary management strategies［J］. J Intern Med, 2007, 262(1):59-77.
［6］ Murry CE, Field LJ, Menasche P. Cell-based cardiac repair: reflections at the 10-year point［J］. Circulation, 2005, 112(20):3174-3183.
［7］ Yasuda T, Weisel RD, Kiani C, et al. Quantitative analysis of survival of transplanted smooth muscle cells with real-time polymerase chain reaction［J］. J Thorac Cardiovasc Surg, 2005, 129(4):904-911.
［8］ Li RK, Jia ZQ, Weisel RD, et al. Smoothmuscle cell transplantation into myocardial scar tissue improves heart function［J］. J Mol Cell Cardiol, 1999, 31(3):513-522.
［9］ Cohn JN, Ferrari R, Sharpe N, et al. Cardiac remodeling concepts and clinical implications: a consensus paper from an international forum on cardiac remodeling［J］. J Am Coll Cardiol, 2000, 35(3):569-582.
［10］朱勇，李玉明. 基质金属蛋白酶及其抑制剂与心肌梗死后心室重塑［J］. 心血管病学进展, 2004, 25(Z1)：105-108.
［11］郝玉明，朱文玲. 心肌梗死后的心室重构［J］. 中华心血管病杂志, 2005, 33(2):196-197.
［12］Sternlicht MD, Werb Z. How matrix metalloproteinases regulate cell behavior［J］. Annu Rev Cell Dev Biol, 2001, 17(4):463-516.
［13］Fedak PW, Altamentova SM, Weisel RD, et al. Matrix remodeling in experimental and human heart failure: a possible regulatory role for TIMP-3［J］. Am J Physiol Heart Circ Physiol, 2003, 284(2):H626-H634.
［14］Fedak PW, Smookler DS, Kassiri Z, et al. TIMP-3 deficiency leads to dilated cardiomyopathy［J］. Circulation, 2004, 110(16):2401-2409.
备注/Memo
- 备注/Memo:
- 收稿日期：2009-5-27.基金项目：国家自然科学基金项目资助（30640052）；黑龙江省科技计划项目资助（GB06C40304）；黑龙江省普通高等学校青年学术骨干支持计划项目资助（1153G018）；哈尔滨市科技创新人才研究专项资金项目资助（2006RFLXS024）；哈医大二院博士科研基金项目资助（BS2008-18） 通讯作者：田海，副教授，主要从事细胞移植、基因治疗研究Email:doctor_tianhai@yahoo.com.cn 作者简介：贾智博，主治医师，博士Email:doctor_jiazhibo@yahoo.com.cn
常用功能
统计/Statistics
- 摘要浏览/Viewed1605
- 评论/Comments
