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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
  • View Affiliations / Copyright

    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
    Copyright: © Peng et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].
  • Pages: 1179-1188
    |
    Published online on: March 9, 2015
       https://doi.org/10.3892/ijmm.2015.2129
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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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1 

Piscopiello M, Sessa M, Anzalone N, et al: P2X7 receptor is expressed in human vessels and might play a role in atherosclerosis. Int J Cardiol. 168:2863–2866. 2013. View Article : Google Scholar : PubMed/NCBI

2 

Mehta N, Kaur M, Singh M, et al: Purinergic receptor P2X7: A novel target for anti-inflammatory therapy. Bioorg Med Chem. 22:54–88. 2014. View Article : Google Scholar

3 

Wang LL, Wang HY and Qu P: Role and relationships of pattern-recognition receptors in atherosclerosis (In Chinese). Chin J Arterioscler. 20:951–955. 2012.

4 

Jia Y, Zhou L and Li XH: Advance in the study of nod-like receptor protein-3 inflammasome and atherosclerosis (In Chinese). Chin J Arterioscler. 22:79–84. 2014.

5 

Yang CS, Shin DM and Jo EK: The role of NLR-related protein 3 inflammasome in host defense and inflammatory diseases. Int Neurourol J. 16:2–12. 2012. View Article : Google Scholar : PubMed/NCBI

6 

Grebe A and Latz E: Cholesterol crystals and inflammation. Curr Rheumatol Rep. 15:3132013. View Article : Google Scholar : PubMed/NCBI

7 

Duewell P, Kono H, Rayner KJ, et al: NLRP3 inflammasomes are required for atherogenesis and activated by cholesterol crystals. Nature. 464:1357–1361. 2010. View Article : Google Scholar : PubMed/NCBI

8 

Rajamäki K, Lappalainen J, Oörni K, et al: Cholesterol crystals activate the NLRP3 inflammasome in human macrophages: a novel link between cholesterol metabolism and inflammation. PLoS One. 5:e117652010. View Article : Google Scholar : PubMed/NCBI

9 

Riteau N, Baron L, Villeret B, et al: ATP release and purinergic signaling: a common pathway for particle-mediated inflammasome activation. Cell Death Dis. 3:e4032012. View Article : Google Scholar : PubMed/NCBI

10 

Niemi K, Teirilä L, Lappalainen J, et al: Serum amyloid A activates the NLRP3 inflammasome via P2X7 receptor and a cathepsin B-sensitive pathway. J Immunol. 186:6119–6128. 2011. View Article : Google Scholar : PubMed/NCBI

11 

Eklund KK, Niemi K and Kovanen PT: Immune functions of serum amyloid A. Crit Rev Immunol. 32:335–348. 2012. View Article : Google Scholar : PubMed/NCBI

12 

Gicquel T, Victoni T, Fautrel A, et al: Involvement of purinergic receptors and NOD-like receptor-family protein 3-inflammasome pathway in the adenosine triphosphate-induced cytokine release from macrophages. Clin Exp Pharmacol Physiol. 41:279–286. 2014. View Article : Google Scholar : PubMed/NCBI

13 

Gombault A, Baron L and Couillin I: ATP release and purinergic signaling in NLRP3 inflammasome activation. Front Immunol. 3:4142013. View Article : Google Scholar : PubMed/NCBI

14 

Kuroda E, Ishii KJ, Uematsu S, et al: Silica crystals and aluminum salts regulate the production of prostaglandin in macrophages via NALP3 inflammasome-independent mechanisms. Immunity. 34:514–526. 2011. View Article : Google Scholar : PubMed/NCBI

15 

Hornung V, Bauernfeind F, Halle A, et al: Silica crystals and aluminum salts activate the NALP3 inflammasome through phagosomal destabilization. Nat Immunol. 9:847–856. 2008. View Article : Google Scholar : PubMed/NCBI

16 

Tschopp J and Schroder K: NLRP3 inflammasome activation: The convergence of multiple signalling pathways on ROS production? Nat Rev Immunol. 10:210–215. 2010. View Article : Google Scholar : PubMed/NCBI

17 

Latz E, Xiao TS and Stutz A: Activation and regulation of the inflammasomes. Nat Rev Immunol. 13:397–411. 2013. View Article : Google Scholar : PubMed/NCBI

18 

Wen H, Miao EA and Ting JP: Mechanisms of NOD-like receptor-associated inflammasome activation. Immunity. 39:432–441. 2013. View Article : Google Scholar : PubMed/NCBI

19 

McGettrick AF and O’Neill LA: How metabolism generates signals during innate immunity and inflammation. J Biol Chem. 288:22893–22898. 2013. View Article : Google Scholar : PubMed/NCBI

20 

Masters SL, Dunne A, Subramanian SL, et al: Activation of the NLRP3 inflammasome by islet amyloid polypeptide provides a mechanism for enhanced IL-1β in type 2 diabetes. Nat Immunol. 11:897–904. 2010. View Article : Google Scholar : PubMed/NCBI

21 

Chávez-Sánchez L, Garza-Reyes MG, Espinosa-Luna JE, Chávez-Rueda K, Legorreta-Haquet MV and Blanco-Favela F: The role of TLR2, TLR4 and CD36 in macrophage activation and foam cell formation in response to oxLDL in humans. Hum Immunol. 75:322–329. 2014. View Article : Google Scholar : PubMed/NCBI

22 

Björkbacka H, Kunjathoor VV, Moore KJ, et al: Reduced atherosclerosis in MyD88-null mice links elevated serum cholesterol levels to activation of innate immunity signaling pathways. Nat Med. 10:416–421. 2004. View Article : Google Scholar : PubMed/NCBI

23 

West XZ, Malinin NL, Merkulova AA, et al: Oxidative stress induces angiogenesis by activating TLR2 with novel endogenous ligands. Nature. 467:972–976. 2010. View Article : Google Scholar : PubMed/NCBI

24 

Miller YI, Viriyakosol S, Binder CJ, Feramisco JR, Kirkland TN and Witztum JL: Minimally modified LDL binds to CD14, induces macrophage spreading via TLR4/MD-2, and inhibits phagocytosis of apoptotic cells. J Biol Chem. 278:1561–1568. 2003. View Article : Google Scholar

25 

Mullick AE, Tobias PS and Curtiss LK: Modulation of atherosclerosis in mice by Toll-like receptor 2. J Clin Invest. 115:3149–3156. 2005. View Article : Google Scholar : PubMed/NCBI

26 

Chávez-Sánchez L, Chávez-Rueda K, Legorreta-Haquet MV, et al: The activation of CD14, TLR4, and TLR2 by mmLDL induces IL-1β, IL-6, and IL-10 secretion in human monocytes and macrophages. Lipids Health Dis. 9:1172010. View Article : Google Scholar

27 

Wiley JS, Sluyter R, Gu BJ, Stokes L and Fuller SJ: The human P2X7 receptor and its role in innate immunity. Tissue Antigens. 78:321–332. 2011. View Article : Google Scholar : PubMed/NCBI

28 

Benetti E, Chiazza F, Patel NS and Collino M: The NLRP3 Inflammasome as a novel player of the intercellular crosstalk in metabolic disorders. Mediators Inflamm. 2013:6786272013. View Article : Google Scholar : PubMed/NCBI

29 

Solini A, Menini S, Rossi C, et al: The purinergic 2X7 receptor participates in renal inflammation and injury induced by high-fat diet: possible role of NLRP3 inflammasome activation. J Pathol. 231:342–353. 2013.PubMed/NCBI

30 

Hussen J, Düvel A, Koy M and Schuberth HJ: Inflammasome activation in bovine monocytes by extracellular ATP does not require the purinergic receptor P2X7. Dev Comp Immunol. 38:312–320. 2012. View Article : Google Scholar : PubMed/NCBI

31 

Dekali S, Divetain A, Kortulewski T, et al: Cell cooperation and role of the P2X7 receptor in pulmonary inflammation induced by nanoparticles. Nanotoxicology. 7:1302–1314. 2013. View Article : Google Scholar

32 

Usui F, Shirasuna K, Kimura H, et al: Critical role of caspase-1 in vascular inflammation and development of atherosclerosis in Western diet-fed apolipoprotein E-deficient mice. Biochem Biophys Res Commun. 425:162–168. 2012. View Article : Google Scholar : PubMed/NCBI

33 

Qiao Y, Wang P, Qi J, Zhang L and Gao C: TLR-induced NF-κB activation regulates NLRP3 expression in murine macrophages. FEBS Lett. 586:1022–1026. 2012. View Article : Google Scholar : PubMed/NCBI

34 

Matsuura E, Lopez LR, Shoenfeld Y and Ames PR: β2-glycoprotein I and oxidative inflammation in early atherogenesis: a progression from innate to adaptive immunity? Autoimmun Rev. 12:241–249. 2012. View Article : Google Scholar : PubMed/NCBI

35 

Lin J, Shou X, Mao X, et al: Oxidized low density lipoprotein induced caspase-1 mediated pyroptotic cell death in macrophages: implication in lesion instability? PLoS One. 8:e621482013. View Article : Google Scholar : PubMed/NCBI

36 

Ehara S, Ueda M, Naruko T, et al: Elevated levels of oxidized low density lipoprotein show a positive relationship with the severity of acute coronary syndromes. Circulation. 103:1955–1960. 2001. View Article : Google Scholar : PubMed/NCBI

37 

Lu B, Nakamura T, Inouye K, et al: Novel role of PKR in inflammasome activation and HMGB1 release. Nature. 488:670–674. 2012. View Article : Google Scholar : PubMed/NCBI

38 

Oury C: CD36: linking lipids to the NLRP3 inflammasome, atherogenesis and atherothrombosis. Cell Mol Immunol. 11:8–10. 2014. View Article : Google Scholar :

39 

Sheedy FJ, Grebe A, Rayner KJ, et al: CD36 coordinates NLRP3 inflammasome activation by facilitating intracellular nucleation of soluble ligands into particulate ligands in sterile inflammation. Nat Immunol. 14:812–820. 2013. View Article : Google Scholar : PubMed/NCBI

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Copy and paste a formatted citation
Spandidos Publications style
Peng K, Liu L, Wei D, Lv Y, Wang G, Xiong W, Wang X, Altaf A, Wang L, He D, He D, et al: P2X7R is involved in the progression of atherosclerosis by promoting NLRP3 inflammasome activation. Int J Mol Med 35: 1179-1188, 2015.
APA
Peng, K., Liu, L., Wei, D., Lv, Y., Wang, G., Xiong, W. ... Qu, P. (2015). P2X7R is involved in the progression of atherosclerosis by promoting NLRP3 inflammasome activation. International Journal of Molecular Medicine, 35, 1179-1188. https://doi.org/10.3892/ijmm.2015.2129
MLA
Peng, K., Liu, L., Wei, D., Lv, Y., Wang, G., Xiong, W., Wang, X., Altaf, A., Wang, L., He, D., Wang, H., Qu, P."P2X7R is involved in the progression of atherosclerosis by promoting NLRP3 inflammasome activation". International Journal of Molecular Medicine 35.5 (2015): 1179-1188.
Chicago
Peng, K., Liu, L., Wei, D., Lv, Y., Wang, G., Xiong, W., Wang, X., Altaf, A., Wang, L., He, D., Wang, H., Qu, P."P2X7R is involved in the progression of atherosclerosis by promoting NLRP3 inflammasome activation". International Journal of Molecular Medicine 35, no. 5 (2015): 1179-1188. https://doi.org/10.3892/ijmm.2015.2129
Copy and paste a formatted citation
x
Spandidos Publications style
Peng K, Liu L, Wei D, Lv Y, Wang G, Xiong W, Wang X, Altaf A, Wang L, He D, He D, et al: P2X7R is involved in the progression of atherosclerosis by promoting NLRP3 inflammasome activation. Int J Mol Med 35: 1179-1188, 2015.
APA
Peng, K., Liu, L., Wei, D., Lv, Y., Wang, G., Xiong, W. ... Qu, P. (2015). P2X7R is involved in the progression of atherosclerosis by promoting NLRP3 inflammasome activation. International Journal of Molecular Medicine, 35, 1179-1188. https://doi.org/10.3892/ijmm.2015.2129
MLA
Peng, K., Liu, L., Wei, D., Lv, Y., Wang, G., Xiong, W., Wang, X., Altaf, A., Wang, L., He, D., Wang, H., Qu, P."P2X7R is involved in the progression of atherosclerosis by promoting NLRP3 inflammasome activation". International Journal of Molecular Medicine 35.5 (2015): 1179-1188.
Chicago
Peng, K., Liu, L., Wei, D., Lv, Y., Wang, G., Xiong, W., Wang, X., Altaf, A., Wang, L., He, D., Wang, H., Qu, P."P2X7R is involved in the progression of atherosclerosis by promoting NLRP3 inflammasome activation". International Journal of Molecular Medicine 35, no. 5 (2015): 1179-1188. https://doi.org/10.3892/ijmm.2015.2129
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