Conditional knockout of tissue factor pathway inhibitor 2 in vascular endothelial cells accelerates atherosclerotic plaque development in mice
Conditional knockout of tissue factor pathway inhibitor 2 in vascular endothelial cells accelerates atherosclerotic plaque development in mice
复制标题
有条件地敲除血管内皮细胞中的组织因子途径抑制剂 2 可加速小鼠动脉粥样硬化斑块的发展。
DOI：
10.1016/j.thromres.2015.11.010
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发表时间：
2016-01-01
影响因子：
7.5
通讯作者：
Shi, Haiming
中科院分区：
文献类型：
--
作者：
Hong, Jin;Liu, Rongle;Shi, Haiming
背景：组织因子途径抑制剂-2 （TFPI-2）调节基质金属蛋白酶的活化和细胞外基质降解。TFPI-2的过表达增强了动脉粥样硬化斑块的稳定性。本研究的目的是探讨血管内皮细胞条件敲除（KO） TFPI-2对动脉粥样硬化斑块发生和发展的影响。方法：采用Cre/mloxP条件KO系统和Tek-Cre小鼠在内皮细胞中产生TFPI-2基因单等位基因缺失的后代。选取TFPI-2(fl/+)/Tek-Cre小鼠、TFPI-2（fl/+）小鼠和ApoE（-/-）小鼠（每组n = 6）。获得动脉。采用HE、EVG和抗- sma染色检测血管和斑块的形态。Western blot和免疫组织化学检测蛋白表达和磷酸化水平。结果：生成TFPI-2(fl/+)/Tek-Cre小鼠。TFPI-2(fl/+)/Tek-Cre组TFPI-2水平下降40.68%。TFPI-2(fl/+)/Tek-Cre在TFPI-2（fl/+）小鼠中未发现斑块时发生斑块。与ApoE（-/-）组相比，TFPI-2(fl/+)/Tek-Cre组斑块面积较小，脂质含量降低，埋藏纤维帽层数减少。TFPI-2(fl/+)/Tek-Cre组MMP-2、MMP-9水平高于TFPI-2（fl/+）组。TFPI-2(fl/+)/Tek-Cre组ppar - α和ppar - γ的磷酸化水平降低。结论：建立了一种新的小鼠模型，可用于研究TFPI-2在动脉粥样硬化过程中的作用。我们的研究结果表明，血管内皮细胞中TFPI-2基因的单等位基因缺失导致了TFPI-2的显著下调。TFPI-2缺乏可能加速小鼠动脉粥样硬化病变的发生。MMP-2和9的升高以及ppar - α和ppar - γ磷酸化的降低可能有助于这种表型。(C) 2015 Elsevier Ltd.版权所有。
Background: Tissue factor pathway inhibitor-2 (TFPI-2) regulates matrix metalloproteinases activation and extracellular matrix degradation. Over-expression of TFPI-2 enhances atherosclerotic plaque stability. The aim of this study is to investigate the effect of conditional knockout (KO) of TFPI-2 in vascular endothelial cells on the initiation and development of atherosclerotic plaque.Methods: A Cre/mloxP conditional KO system and Tek-Cre mice were used to generate offsprings with monoallelic deletion of the TFPI-2 gene in endothelial cells. TFPI-2(fl/+)/Tek-Cre mice, TFPI-2(fl/+) mice and ApoE(-/-) mice (n = 6 for each group) were included. Arteries were obtained. HE, EVG and anti-alpha-SMA staining were used to examine the morphology of vessel and plaque. Protein expression and phosphorylation were detected by Western blot or immunohistochemistry.Results: TFPI-2(fl/+)/Tek-Cre mice were generated. TFPI-2 level decreased to 40.68% in TFPI-2(fl/+)/Tek-Cre group. TFPI-2(fl/+)/Tek-Cre developed plaques when no plaque was found in TFPI-2(fl/+) mice. Compared with ApoE(-/-) group, TFPI-2(fl/+)/Tek-Cre group has smaller plaque area, decreased lipid content and less buried fibrous cap layers. MMP-2 and MMP-9 in TFPI-2(fl/+)/Tek-Cre group was higher than in TFPI-2(fl/+) group. The phosphorylation of PPAR-alpha and PPAR-gamma was decreased in TFPI-2(fl/+)/Tek-Cre group.Conclusions: A novel mouse model is presented and can be used to investigate the role of TFPI-2 in the process of atherosclerosis. Our findings suggest that monoallelic deletion of TFPI-2 gene in vascular endothelial cells leads to significant downregulation of TFPI-2. TFPI-2 deficiency may accelerate initiation of atherosclerotic lesion in mice. Elevated MMP-2 and 9 and decreased phosphorylation of PPAR-alpha and PPAR-gamma may contribute to this phenotype. (C) 2015 Elsevier Ltd. All rights reserved.