Organ-specific endothelial cell uptake of cationic liposome-DNA complexes in mice
Organ-specific endothelial cell uptake of cationic liposome-DNA complexes in mice
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DOI：
10.1152/ajpheart.1997.273.1.h387
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发表时间：
1997-07-01
影响因子：
4.8
通讯作者：
McDonald, DM
中科院分区：
文献类型：
--
作者：
McLean, JW;Fox, EA;McDonald, DM
这项研究表明。阳离子脂质体- dna复合物静脉注射到CD-1小鼠体内用于基因传递的器官和细胞分布。用荧光染料CM-DiI标记dotim -胆固醇脂质体，并与编码氯霉素乙酰转移酶报告基因的质粒DNA配合。注射后5 min ~ 24 h，用荧光显微镜、共聚焦显微镜和电子显微镜观察29个器官和组织中复合物的分布。这些复合物在血液中形成簇状，在20分钟内被清除。荧光显微镜下可见的复合物被内皮细胞、白细胞和巨噬细胞占据，除了脾脏外，没有离开血管系统。在5分钟时，复合物在内皮表面形成斑块状涂层，但在4小时时，它们以器官和血管特异性的模式内化到内体和溶酶体中。毛细血管内皮细胞在肺、卵巢和垂体前叶的摄取最多，在肌肉和心脏的摄取较少，在脑和胰岛几乎没有。在淋巴结和肠Peyer's斑块中，毛细血管中摄取较少，但在高内皮小静脉中摄取较多。在肝脏和脾脏中，摄取最多的是Kupffer细胞和巨噬细胞。氯霉素乙酰转移酶报告基因表达的测量结果与内皮细胞的摄取模式基本一致。摄取和基因表达伴随着循环白细胞和血小板的减少。总的来说，我们的研究结果表明，这些复合物以器官和血管特异性的模式被内皮细胞内化，这与之前发现的微血管特性不匹配。内皮细胞摄取的不寻常分布可能是由于膜受体的不均匀分布，其复合物是配体。
This study identified. the organ and cellular distribution of cationic liposome-DNA complexes injected intravenously into CD-1 mice for gene delivery. DOTIM-cholesterol liposomes were labeled with the fluorescent dye CM-DiI and complexed with plasmid DNA encoding the chloramphenicol acetyltransferase reporter gene. The distribution of the complexes was examined in 29 organs and tissues by fluorescence, confocal, and electron microscopy from 5 min to 24 h after injection. The complexes formed clusters in blood, which were cleared within 20 min. Complexes visible by fluorescence microscopy were taken up by endothelial cells, leukocytes, and macrophages and did not leave the vasculature except in the spleen. At 5 min, the complexes formed a patchy coating on the endothelial surface, but by 4 h, they were internalized into endosomes and lysosomes in organ- and vessel-specific patterns. Uptake by capillary endothelial cells was greatest in the lung, ovary, and anterior pituitary, less in muscle and the heart, and nearly absent in the brain and pancreatic islets. In lymph nodes and intestinal Peyer's patches, the uptake was sparse in capillaries but abundant in high endothelial venules. In the liver and spleen, most of the uptake was in Kupffer cells and macrophages. Measurements of chloramphenicol acetyltransferase reporter gene expression were generally consistent with the pattern of uptake by endothelial cells. The uptake and gene expression were accompanied by a decrease in circulating leukocytes and platelets. Overall, our results showed that the complexes were internalized by endothelial cells in organ-and vessel-specific patterns that did not match any previously identified properties of the microvasculature. The unusual distribution of endothelial cell uptake may be explained by a heterogeneously distributed membrane receptor for which the complexes are ligands.