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

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.

• Authors: Jiao Li, Xue Liang, Yuanyuan Wang, Zhao Xu, Guangping Li
• Journal: Molecular medicine reports (Mol Med Rep) Vol. 16 Issue 2 Pg. 1817-1825 (Aug 2017) ISSN: 1791-3004 [Electronic] Greece
• PMID: 28656218 (Publication Type: Journal Article)

Chemical References

• Lipoproteins, LDL
• RNA, Small Interfering
• bcl-2-Associated X Protein
• oxidized low density lipoprotein
• Proprotein Convertase 9
• Caspase 3

Topics

• Animals
• Apoptosis (drug effects)
• Caspase 3 (metabolism)
• Disease Models, Animal
• Down-Regulation (drug effects)
• Human Umbilical Vein Endothelial Cells (drug effects, enzymology, pathology)
• Humans
• Lipoproteins, LDL (pharmacology)
• MAP Kinase Signaling System (drug effects)
• Mice
• Plaque, Atherosclerotic (enzymology, pathology)
• Proprotein Convertase 9 (metabolism)
• RNA, Small Interfering (metabolism)
• Transfection
• Up-Regulation (drug effects)
• bcl-2-Associated X Protein (metabolism)