P2X7R is involved in the progression of atherosclerosis by promoting NLRP3 inflammasome activation

Peng,Kuang; Liu,Lushan; Wei,Dangheng; Lv,Yuncheng; Wang,Gang; Xiong,Wenhao; Wang,Xiaoqing; Altaf,Afrasyab; Wang,Lili; He,Dan; Wang,Hongyan; Qu,Peng

doi:10.3892/ijmm.2015.2129

P2X7R is involved in the progression of atherosclerosis by promoting NLRP3 inflammasome activation

Authors:
- Kuang Peng
- Lushan Liu
- Dangheng Wei
- Yuncheng Lv
- Gang Wang
- Wenhao Xiong
- Xiaoqing Wang
- Afrasyab Altaf
- Lili Wang
- Dan He
- Hongyan Wang
- Peng Qu
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Affiliations: Department of Cardiology, The Second Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116023, P.R. China, Institute of Cardiovascular Disease, Key Laboratory for Arteriosclerology of Hunan Province, University of South China, Hengyang, Hunan 421001, P.R. China, Department of Cardiology, The First Affiliated Hospital of University of South China, Hengyang, Hunan 421001, P.R. China, Medical College, University of South China, Hengyang, Hunan 421001, P.R. China
Published online on: March 9, 2015 https://doi.org/10.3892/ijmm.2015.2129
Pages: 1179-1188
Copyright: © Peng et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Purinergic 2X7 receptor (P2X7R) and nucleotide‑binding oligomerization domain-like receptor protein 3 (NLRP3) are expressed in macrophages in atherosclerotic lesions. However, the mechanisms through which P2X7R participates in the inflammatory response in atherosclerosis remain largely unknown. The aim of the present study was to investigate the role of P2X7R in atherosclerosis and the mechanisms of action of the NLRP3 inflammasome following stimulation with oxidized low-density lipoprotein (oxLDL). We observed the expression and distribution of P2X7R in the atherosclerotic plaque in the coronary arteries from an autopsy specimen and in that of the aortic sinuses of apoE-/- mice by immunohistochemistry and immunofluorescence staining. The specificity of short interfering RNA (siRNA) was used to suppress P2X7R and NLRP3 mRNA expression. RT-qPCR and western blot analysis were used to analyze mRNA and protein expression, respectively. Co-immunoprecipitation was used to examine the interaction between protein kinase R (PKR) phosphorylation and NLRP3. P2X7R and NLRP3 were expressed at high levels in the atherosclerotic plaque in the coronary arteries. Stimulation with oxLDL upregulated P2X7R, NLRP3 and interleukin (IL)-1β expression. P2X7R knockdown by siRNA suppressed NLRP3 inflammasome activation by inhibiting the PKR phosphorylation mediated by oxLDL. In the atherosclerotic lesions in the aortic sinuses of apoE-/- mice, P2X7R expression was found at high levels. Moreover, P2X7R siRNA attenuated the development of atherosclerosis in the apoE-/- mice. In conclusion, our results demonstrate that P2X7R plays a significant role in the development of atherosclerosis and regulates NLRP3 inflammasome activation by promoting PKR phosphorylation.

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