Multiple components in serum contribute to hepatic transgene expression by lipoplex in mice
Multiple components in serum contribute to hepatic transgene expression by lipoplex in mice
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血清中的多种成分有助于脂质复合物在小鼠体内表达肝脏转基因
DOI：
10.1002/jgm.1618
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发表时间：
2011
期刊：
影响因子：
3.5
通讯作者：
Naoki Yoshikawa
中科院分区：
文献类型：
--
作者：
藁谷美香;東顕二郎;森部久仁一;山本恵司;Naoki Yoshikawa
背景阳离子脂质体/质粒DNA复合物（lipoplex）与血清的相互作用不是体内转染的限制因素。门脉内注射lipoplex后，小鼠肝脏转基因表达通过与血清相互作用而增强。在本研究中，我们分析了增强的肝脏转基因表达的lipoplex的机制，通过与血清components.MethodsLipoplexes的相互作用，在37 °C下孵育5分钟，几个血清成分前管理。lipoplexes的转染效率测定后6小时intraportal注射lipoplex在mice.ResultsDepletion从血清中的二价阳离子降低肝转基因表达。向二价阳离子耗尽的血清中添加钙离子恢复了转基因表达。热灭活血清和牛血清白蛋白减弱了血清对肝转基因表达的增强作用。另一方面，使用阴离子交换柱从血清中去除阴离子蛋白对于血清对转基因表达的增强作用至关重要。在测试的血清成分中，纤连蛋白和补体成分C3增强肝转基因expression. ConclusionsLiver转基因表达lipoplex增强与血清中的多个组件的相互作用。脂质复合物与血清的相互作用可能是体内基因转移成功的重要因素。因此，在本研究中获得的信息是有价值的有效的基因载体的未来发展。版权所有© 2011约翰威利父子有限公司.
BackgroundInteraction of cationic liposome/plasmid DNA complex (lipoplex) with serum was not a limiting factor forin vivotransfection. After intraportal injection of lipoplex, hepatic transgene expression was enhanced by interaction with serum in mice. In the present study, we analyzed the mechanism of enhanced hepatic transgene expression of lipoplex by interaction with serum components.MethodsLipoplexes were incubated with several serum components for 5 min at 37 °C before administration. Transfection efficiency of lipoplexes was measured 6 h after intraportal injection of lipoplex in mice.ResultsDepletion of divalent cation from serum decreased hepatic transgene expression. The addition of calcium ion to divalent cation‐depleted serum restored transgene expression. Heat‐inactivated serum and bovine serum albumin diminished the enhancing effect of serum on hepatic transgene expression. On the other hand, removal of anionic proteins from serum using an anion‐exchanging column was critical for the enhancing effect of serum on transgene expression. Among the serum components tested, fibronectin and complement component C3 enhanced hepatic transgene expression.ConclusionsHepatic transgene expression by lipoplex was enhanced by interaction with multiple components in serum. Interaction of lipoplex with serum could be an important factor for successfulin vivogene transfer. Hence, the information obtained in the present study is valuable for the future development of effective gene carriers. Copyright © 2011 John Wiley & Sons, Ltd.
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