Indirect intracoronary delivery of adenovirus encoding adenylyl cyclase increases left ventricular contractile function in mice
Indirect intracoronary delivery of adenovirus encoding adenylyl cyclase increases left ventricular contractile function in mice
复制标题
DOI：
10.1152/ajpheart.01009.2003
复制
发表时间：
2004-07-01
影响因子：
4.8
通讯作者：
Hammond, HK
中科院分区：
文献类型：
--
作者：
Roth, DM;Lai, NC;Hammond, HK
我们在小鼠中进行了腺病毒载体的间接冠状动脉内递送，并探索了包括低温和药理学手段在内的技术来增加心脏基因转移。将小鼠保持在正常体温状态或冷却至25 ℃。夹闭主动脉或肺动脉和主动脉两者，同时将针推进左心室腔以递送编码增强型绿色荧光蛋白（EGFP）或鼠腺苷酸环化酶VI型（AC（VI））的腺病毒载体和盐水、硝普钠、乙酰胆碱或5-羟色胺。腺病毒给药后，钳夹维持30秒（常温）或2分钟（25 ℃）。7天或21天后处死小鼠，并检查心脏的EGFP表达。与仅夹闭主动脉且不冷却相比，基因转移增加如下：1）低温延长停留时间增加1.3倍; 2）夹闭主动脉和肺动脉增加4.5倍; 3）硝普钠给药增加11.4倍; 4）5-羟色胺添加增加11.8倍，以及5）乙酰胆碱递送增加14.3倍。基因表达在21天时保持显著，并且没有观察到显著的炎症反应。通过进行编码AC（VI）的腺病毒的基因转移来测试该方法的功效。基因转移后14天，从接受编码AC（VI）的腺病毒的小鼠中分离的心脏显示出增强的收缩功能。在小鼠中间接冠状动脉内递送腺病毒载体与AC（VI）基因转移后有效的心脏基因转移和增加的左心室功能相关。
We performed indirect intracoronary delivery of adenovirus vectors in mice and explored techniques including hypothermia and pharmacological means to increase cardiac gene transfer. Mice were maintained in a normothermic state or cooled to 25degreesC. The aorta or both the pulmonary artery and aorta were clamped while a needle was advanced into the left ventricular cavity to deliver adenovirus vectors encoding enhanced green fluorescent protein (EGFP) or murine adenylyl cyclase type VI (AC(VI)) with saline, sodium nitroprusside, acetylcholine, or serotonin. Clamping was maintained for 30 s (normothermia) or 2 min (25degreesC) after adenovirus administration. Mice were killed 7 or 21 days later, and hearts were examined for EGFP expression. Compared with clamping the aorta alone and with no cooling, gene transfer was increased as follows: 1) 1.3-fold with hypothermia to extend dwell time; 2) 4.5-fold by clamping the aorta and the pulmonary artery; 3) 11.4-fold with nitroprusside administration; 4) 11.8-fold with serotonin addition, and 5) 14.3-fold with acetylcholine delivery. Gene expression remained substantial at 21 days, and no significant inflammatory response was seen. Efficacy of the method was tested by performing gene transfer of adenovirus encoding AC(VI). Fourteen days after gene transfer, hearts isolated from mice that received adenovirus encoding AC(VI) showed increased contractile function. Indirect intracoronary delivery of adenovirus vectors in mice is associated with efficient cardiac gene transfer and increased left ventricular function after AC(VI) gene transfer.