Zhang J C, Woo Y J, Chen J A, Swain J L, Sweeney H L
Department of Medicine, Division of Cardiovascular Medicine, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-6100, USA.
J Mol Cell Cardiol. 1999 Apr;31(4):721-32. doi: 10.1006/jmcc.1998.0905.
An efficient cardiac gene transfer technique in murine models would greatly facilitate the elucidation of the pathophysiology of cardiomyopathies and the development of genetic therapies. Direct myocardial injection or catheter-based intracoronary infusion is not easily achievable in mice and resultant transgene expression is often limited in distribution. A replication-defective, recombinant adenovirus encoding luciferase (5x10(9)pfu) or lacZ (4-5x10(10)particles/animal) was injected percutaneously into the pericardial cavity of 4-5 day old mice. Chemiluminescence assay for luciferase activity at 3 days post-injection revealed the highest activity in the heart (heart=288+/-110, lungs=19+/-5, liver=11+/-5 ng/gm tissue, n=11). X-gal staining of cryostat sections demonstrated widespread transmural lacZ expression in the left ventricle, interventricular septum, right ventricle, and atrial appendages, and the average fractional area of X-gal staining in a left ventricle was 66+/-16% (range 40-92%, n=21 sections). However, the long-term survival of these mice was compromised. Reduction in the injectate volume by 50% significantly improved survival but concurrently reduced lacZ expression. Significant lacZ expression was observed in the right ventricle and interventricular septum but left ventricular expression was predominantly epicardial, with variable myocardial penetration. At 2 months post-injection, lacZ expression persisted only in atrial tissues, pulmonary veins, and great vessels. Despite lack of persistent transgene expression in ventricular tissues, the high degree of transgene expression achieved may be sufficient to allow evaluation of short-term effects of specific genetic manipulations in the heart.
在小鼠模型中，一种高效的心脏基因转移技术将极大地促进心肌病病理生理学的阐明以及基因治疗的发展。在小鼠中，直接心肌注射或基于导管的冠状动脉内输注不易实现，并且由此产生的转基因表达在分布上通常受到限制。将编码荧光素酶（5×10⁹ pfu）或β-半乳糖苷酶（4 - 5×10¹⁰颗粒/动物）的复制缺陷型重组腺病毒经皮注射到4 - 5日龄小鼠的心包腔内。注射后3天对荧光素酶活性进行化学发光测定，结果显示心脏中的活性最高（心脏 = 288±110，肺 = 19±5，肝脏 = 11±5 ng/g组织，n = 11）。冰冻切片的X - gal染色显示左心室、室间隔、右心室和心耳中广泛存在透壁的β-半乳糖苷酶表达，左心室中X - gal染色的平均分数面积为66±16%（范围40 - 92%，n = 21个切片）。然而，这些小鼠的长期存活受到影响。将注射体积减少50%可显著提高存活率，但同时降低了β-半乳糖苷酶的表达。在右心室和室间隔中观察到显著的β-半乳糖苷酶表达，但左心室表达主要位于心外膜，心肌穿透程度不一。注射后2个月，β-半乳糖苷酶表达仅在心房组织、肺静脉和大血管中持续存在。尽管心室组织中缺乏持续的转基因表达，但所实现的高度转基因表达可能足以允许评估心脏中特定基因操作的短期效果。
