Investigation of highly expressed PCSK9 in atherosclerotic plaques and ox-LDL-induced endothelial cell apoptosis

Li,Jiao; Liang,Xue; Wang,Yuanyuan; Xu,Zhao; Li,Guangping

doi:10.3892/mmr.2017.6803

Investigation of highly expressed PCSK9 in atherosclerotic plaques and ox-LDL-induced endothelial cell apoptosis

Authors:
- Jiao Li
- Xue Liang
- Yuanyuan Wang
- Zhao Xu
- Guangping Li
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Affiliations: Tianjin Key Laboratory of Ionic‑Molecular Function of Cardiovascular Disease, Department of Cardiology, Tianjin Institute of Cardiology, The Second Hospital of Tianjin Medical University, Tianjin 300211, P.R. China
Published online on: June 19, 2017 https://doi.org/10.3892/mmr.2017.6803
Pages: 1817-1825
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The present study aimed to explore the direct toxicity of proprotein convertase subtilisin/kexin type 9 (PCSK9) to atherosclerosis (AS) and its association with apoptotic endothelial cells. Apolipoprotein E‑/‑ mice were randomly divided into two groups, control and experimental. The control group was administered a normal diet and the experimental group was administered a high‑fat diet. After 20 weeks, the aorta was isolated and dissected. Hematoxylin and eosin staining, and immunohistochemical analysis were performed. Human umbilical vein endothelial cells were incubated with varied concentrations of oxidized low‑density lipoprotein (ox‑LDL) for different times. The apoptotic rate was detected by flow cytometry. Western blotting and reverse transcription‑quantitative polymerase chain reaction analysis were conducted to detect the expression of PCSK9, B‑cell lymphoma 2 (Bcl‑2), bcl‑2‑like protein 4 (Bax) and caspase-3. Short hairpin (sh) RNA‑PCSK9 was transfected into endothelial cells using lentiviral transfection. The expression levels of PCSK9, Bax, Bcl‑2, caspase-3 and the mitogen‑activated protein kinase (MAPK) pathway proteins were detected. The high‑fat group was successfully established as an AS model and PCSK9 was highly expressed in the AS plaque. Treatment with ox‑LDL induced apoptosis and increased mRNA and protein levels of PCSK9. PCSK9 mRNA and proteins levels were downregulated by shRNA‑PCSK9. The deficiency of PCSK9 markedly inhibited the expression of pro‑apoptotic proteins and promoted anti‑apoptotic proteins. In addition, phosphorylation of p38 and c‑Jun N‑terminal kinases was altered by shRNA‑PCSK9. Targeting of PCSK9 by shRNA‑PCSK9 may repress endothelial cell apoptosis through MAPK signaling in AS, providing a novel direction for understanding the mechanism and treatment of AS.

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