THE FATE OF PLASMID DNA AFTER INTRAVENOUS-INJECTION IN MICE - INVOLVEMENT OF SCAVENGER RECEPTORS IN ITS HEPATIC-UPTAKE
THE FATE OF PLASMID DNA AFTER INTRAVENOUS-INJECTION IN MICE - INVOLVEMENT OF SCAVENGER RECEPTORS IN ITS HEPATIC-UPTAKE
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DOI：
10.1023/a:1016248701505
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发表时间：
1995-06-01
影响因子：
3.7
通讯作者：
HASHIDA, M
中科院分区：
文献类型：
--
作者：
KAWABATA, K;TAKAKURA, Y;HASHIDA, M
目的。我们检测了裸质粒DNA pCAT作为模型基因在小鼠体内静脉注射后的稳定性和配置特性，以构建体内基因传递系统策略。方法。小鼠注射pCAT后，观察其稳定性、组织分布、肝细胞定位及部分多阴离子对肝脏摄取的影响。结果。体外研究表明，在100 μ g/ml浓度下，pCAT在小鼠全血中降解迅速，半衰期约为10 min。静脉注射后，pCAT的降解速度明显快于在全血中观察到的速度，这表明体内pCAT在其他隔室中也被降解。静脉注射[P-32] pCAT后，由于肝脏的广泛吸收，放射性迅速从血浆中消除。肝脏积聚优先发生在非实质细胞中。肝脏对来自[P-32] pCAT的放射性的摄取被预先给予多阴离子如多肌苷酸、硫酸葡聚糖、马来酰化和琥珀酰化牛血清白蛋白抑制，但不被多胞酸抑制。这些发现表明pCAT通过肝脏非实质细胞上的清道夫受体被肝脏吸收。药代动力学分析显示，肝摄取清除率与肝血浆流量相当接近。结论。这些发现为体内基因治疗递送系统的开发提供了有用的信息。
Purpose. We examined the stability and disposition characteristics of a naked plasmid DNA pCAT as a model gene after intravenous injection in mice to construct the strategy of in vivo gene delivery systems. Methods. After the injection of pCAT to the mice, stability, tissue distribution, hepatic cellular localization, and effect of some polyanions on the hepatic uptake were studied. Results. The in vitro study demonstrated that the pCAT was rapidly degraded in mouse whole blood with a half-life of approximately 10 min at a concentration of 100 mu g/ml. After intravenous injection, pCAT was degraded at a significantly faster rate than that observed in the whole blood, suggesting that pCAT in vivo was also degraded in other compartments. Following intravenous injection of [P-32] pCAT, radioactivity was rapidly eliminated from the plasma due to extensive uptake by the liver. Hepatic accumulation occurred preferentially in the non-parenchymal cells. The hepatic uptake of radioactivity derived from [P-32] pCAT was inhibited by preceding administration of polyanions such as polyinosinic acid, dextran sulfate, maleylated and succinylated bovine serum albumin but not by polycytidylic acid. These findings indicate that pCAT is taken up by the liver via scavenger receptors on the non-parenchymal cells. Pharmacokinetic analysis revealed that the apparent hepatic uptake clearance was fairly close to the liver plasma flow. Conclusions. These findings provide useful information for the development of delivery systems for in vivo gene therapy.