A remarkable permeability of canalicular tight junctions might facilitate retrograde, non-viral gene delivery to the liver via the bile duct
A remarkable permeability of canalicular tight junctions might facilitate retrograde, non-viral gene delivery to the liver via the bile duct
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DOI：
10.1136/gut.2005.070904
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发表时间：
2005-10-01
期刊：
影响因子：
24.5
通讯作者：
Fabre, JW
中科院分区：
文献类型：
--
作者：
Hu, J;Zhang, X;Fabre, JW
目的：在超微结构水平上确定逆行胆管输注的程度，作为评估通过胆管分支向大鼠肝脏传递基因的功效之前的第一步。方法：在将 10 nm 金颗粒输注到右侧叶后，通过光学和电子显微镜确定逆行输注到胆管树的程度。通过输注 67 kDa 蛋白质进一步评估小管通透性。对于基因传递，裸露 DNA 和合成肽载体系统均进行了评估。由于受损肝脏中小管紧密连接通透性会受到损害，因此对正常大鼠和从肝毒素 D-半乳糖胺中恢复的大鼠进行了研究。结果：金颗粒穿透了肝小叶外周三分之一，令人惊讶的是，到达了正常大鼠的 Disse 间隙。同样令人惊讶的是，胆管输注和门静脉注射后 67 kDa 蛋白质的血液水平相同。在用 D-半乳糖胺治疗的大鼠中，用肽/DNA 复合物进行基因传递要有效得多。然而，使用裸露 DNA 进行基因传递对于正常肝脏和受损肝脏同样有效。基因表达的定位显示阳性肝细胞仅限于右侧叶的分散。结论：逆行输注到胆管中可以很好地进入肝小叶，并显示正常大鼠中小管或胆管紧密连接的显着通透性。这是一种将基因传递至少量（大约 1%）肝细胞的有效方法。
Aims: To establish the extent of retrograde bile duct infusion at an ultrastructural level, as a preliminary step before evaluating the efficacy of gene delivery to the rat liver via a branch of the bile duct.Methods: The extent of retrograde infusion into the biliary tree was established by light and electron microscopy, following infusion of 10 nm gold particles into the right lateral lobe. Canalicular permeability was further assessed by the infusion of a 67 kDa protein. For gene delivery, both naked DNA and a synthetic peptide vector system were evaluated. Because canalicular tight junction permeability can be compromised in damaged livers, both normal rats and rats recovering from the hepatotoxin D-galactosamine were studied.Results: The gold particles penetrated the peripheral one third of the hepatic lobules and, surprisingly, reached the space of Disse in normal rats. Equally surprisingly, blood levels of a 67 kDa protein were identical after bile duct infusion and portal vein injection. Gene delivery with peptide/DNA complexes was much more effective in rats treated with D-galactosamine. However, gene delivery with naked DNA was equally effective in normal and damaged livers. Localisation of gene expression showed a scattering of positive hepatocytes restricted to the right lateral lobe.Conclusions: Retrograde infusion into the bile duct advances well into the hepatic lobule and reveals a remarkable permeability of the canalicular or cholangiole tight junctions in normal rats. It is an effective approach for delivering genes to a small population (similar to 1%) of hepatocytes.