Jin Hong1, Rongle Liu1, Lewen Chen1, Bangwei Wu1, Jia Yu1, Wen Gao1, Junjie Pan2, Xinping Luo3, Haiming Shi1. 1. Department of Cardiology, Huashan Hospital, Fudan University, Wulumuqi Zhong Road 12, Shanghai 200040, China. 2. Department of Cardiology, Huashan Hospital, Fudan University, Wulumuqi Zhong Road 12, Shanghai 200040, China. Electronic address: junjie112@163.com. 3. Department of Cardiology, Huashan Hospital, Fudan University, Wulumuqi Zhong Road 12, Shanghai 200040, China. Electronic address: luoxp2007@aliyun.com.
Abstract
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-α-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-α and PPAR-γ 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-α and PPAR-γ may contribute to this phenotype.
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-α-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-α and PPAR-γ 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-α and PPAR-γ may contribute to this phenotype.
Authors: Robert Seifert; Michael T Kuhlmann; Sarah Eligehausen; Friedemann Kiefer; Sven Hermann; Michael Schäfers Journal: PLoS One Date: 2018-10-10 Impact factor: 3.240