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首页> 外文期刊>World Journal of Gastroenterology >Hydrodynamics based transfection in normal and fibrotic rats.
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Hydrodynamics based transfection in normal and fibrotic rats.

机译：在正常和纤维化大鼠中基于流体力学的转染。

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AIM: Hydrodynamics based transfection (HBT), the injection of a large volume of naked plasmid DNA in a short time is a relatively simple, efficient and safe method for in vivo transfection of liver cells. Though used for quite some time, the mechanism of gene transfection has not yet been elucidated. METHODS: A luciferase encoding plasmid was injected using the hydrodynamics based procedure into normal and thioacetamide-induced fibrotic Sprague Dawley rats. Scanning and transmission electron microscopy images were taken. The consequence of a dual injection of Ringer solution and luciferase pDNA was followed. Halofuginone, an anti collagen type I inhibitor was used to reduce ECM load in fibrotic rats prior to the hydrodynamic injection. RESULTS: Large endothelial gaps formed as soon as 10' following hydrodynamic injection; these gradually returned to normal 10 d post injection. Hydrodynamic administration of Ringer 10 or 30 m prior to moderate injection of plasmid did not result in efficient transfection suggesting that endothelial gaps by themselves are not sufficient for gene expression. Gene transfection following hydrodynamic injection in thioacetamide induced fibrotic rats was diminished coinciding with the level of fibrosis. Halofuginone, a specific collagen type I inhibitor, alleviated this effect. CONCLUSION: The hydrodynamic pressure formed following HBT results in the formation of large endothelial gaps. These gaps, though important in the transfer of DNA molecules from the blood to the space of Disse are not enough to provide the appropriate conditions for hepatocyte transfection. Hydrodynamics based injection is applicable in fibrotic rats provided that ECM load is reduced.
机译：目的：基于流体动力学的转染（HBT），在短时间内注射大量裸质粒DNA是一种相对简单，有效和安全的肝细胞体内转染方法。尽管已经使用了一段时间，但基因转染的机制尚未阐明。方法：采用基于流体动力学的方法，将荧光素酶编码质粒注射到正常和硫代乙酰胺诱导的纤维化Sprague Dawley大鼠中。拍摄扫描和透射电子显微镜图像。跟踪了林格溶液和荧光素酶pDNA双重注射的结果。在进行流体动力注射之前，使用抗I型胶原蛋白氟丁酮来减轻纤维化大鼠的ECM负荷。结果：流体动力注射后10'形成了较大的内皮间隙。这些在注射后10天逐渐恢复正常。在适度注射质粒之前，对Ringer进行10或30 m的流体动力学给药不会导致有效的转染，这表明内皮间隙本身不足以进行基因表达。在硫代乙酰胺诱导的纤维化大鼠中，水动力注射后的基因转染与纤维化水平一致。 Halofuginone（一种特定的I型胶原抑制剂）减轻了这种作用。结论：HBT后形成的流体动力压力导致形成大的内皮间隙。这些缺口尽管对将DNA分子从血液转移到Disse的空间中很重要，但不足以为肝细胞转染提供适当的条件。如果减少ECM负荷，则基于水动力的注射适用于纤维化大鼠。

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