Protection against vascular leak in neprilysin transgenic mice with complex overexpression pattern.
Protection against vascular leak in neprilysin transgenic mice with complex overexpression pattern.
复制标题
具有复杂过度表达模式的脑啡肽酶转基因小鼠免受血管渗漏的保护。
DOI：
10.1007/s11248-016-9969-x
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发表时间：
2016
影响因子：
3
通讯作者：
Dempsey,EdwardC
中科院分区：
文献类型：
--
作者：
Wick,MarileeJ;Loomis,ZoeL;Harral,JulieW;Le,Mysan;Wehling,CarolA;Miller,YorkE;Dempsey,EdwardC
NEP是一种存在于多种组织中的细胞表面金属肽酶。主要基于药理学操作，NEP被认为可以保护血管免受血浆外渗。我们认为NEP可能对肺血管损伤有保护作用。然而，这些先前的研究没有使用过表达NEP的小鼠。本研究的目的是建立和鉴定普遍和有条件地过表达NEP的双转基因（DT）小鼠，并研究过表达NEP对血管内血浆外渗的保护作用。十二指肠常用于评估血管通透性，NEP蛋白在我们的DT小鼠中过表达了两倍，我们选择了十二指肠作为我们的实验准备。我们发现，经强力霉素处理的DT小鼠十二指肠p物质诱导的血浆外渗显著减少（3.5倍），这为NEP对血浆外渗的保护作用提供了独立证据。由于转基因肺NEP蛋白在DT小鼠体内的表达不稳定，因此我们无法检测NEP过表达对肺的影响。虽然最初在该部位过表达近9倍，但肺NEP蛋白过表达最终消失。令人惊讶的是，在没有肺转基因NEP蛋白过表达的情况下，肺NEP mRNA的表达仍然增加了23倍，表明表达缺陷可能不是转录性的。这些研究有助于表征NEP过表达的复杂转基因模型，并进一步证明NEP对血浆外渗的保护作用。
Neprilysin (NEP) is a cell surface metallopeptidase found in many tissues. Based mostly on pharmacological manipulations, NEP has been thought to protect blood vessels from plasma extravasation. We have suggested that NEP may protect against pulmonary vascular injury. However, these prior studies did not utilize mice which overexpress NEP. The aims of the present investigation were to develop and characterize doubly transgenic (DT) mice that overexpress NEP universally and conditionally, and to investigate the protective effect that overexpressed NEP may have against plasma extravasation in the vasculature. The duodenum, which is often used to assess vascular permeability, and in which the NEP protein was overexpressed in our DT mice two-fold, was selected as our experimental preparation. We found that substance P-induced plasma extravasation was decreased substantially (3.5-fold) in the duodenums of our doxycycline-treated DT mice, giving independent evidence of NEP’s protective effects against plasma extravasation. Transgenic lung NEP protein was not stably expressed in the DT mice, so we were not able to test the effect of NEP overexpression in the lung. Although initially overexpressed nearly nine-fold at that site, pulmonary NEP protein overexpression eventually dissipated. Surprisingly, at a time when there was no lung transgenic NEP protein overexpression, lung NEP mRNA expression was still increased 23-fold, indicating that the expression defect probably is not transcriptional. These studies help to characterize our complex transgenic model of NEP overexpression and further demonstrate NEP’s protective effects against plasma extravasation.