Activation of Nrf2/HO-1 Pathway and Human Atherosclerotic Plaque Vulnerability:an In Vitro and In Vivo Study - PubMed (original) (raw)

doi: 10.3390/cells8040356.

Benedetta Porro 2, Nicola Cosentino 3, Alessandro Di Minno 4, Chiara Maria Manega 5, Franco Fabbiocchi 6, Giampaolo Niccoli 7, Francesco Fracassi 8, Simone Barbieri 9, Giancarlo Marenzi 10, Filippo Crea 11, Viviana Cavalca 12, Elena Tremoli 13, Sonia Eligini 14

Affiliations

Activation of Nrf2/HO-1 Pathway and Human Atherosclerotic Plaque Vulnerability:an In Vitro and In Vivo Study

Susanna Fiorelli et al. Cells. 2019.

Abstract

Reactive oxygen species (ROS) induce nuclear factor erythroid 2-related factor 2 (Nrf2) activation as an adaptive defense mechanism, determining the synthesis of antioxidant molecules, including heme-oxygenase-1 (HO-1). HO-1 protects cells against oxidative injury, degrading free heme and inhibiting ROS production. HO-1 is highly expressed in macrophages during plaque growth. Macrophages are morpho-functionally heterogeneous, and the prevalence of a specific phenotype may influence the plaque fate. This heterogeneity has also been observed in monocyte-derived macrophages (MDMs), a model of macrophages infiltrating tissue. The study aims to assess oxidative stress status and Nrf2/HO-1 axis in MDM morphotypes obtained from healthy subjects and coronary artery disease (CAD) patients, in relation to coronary plaque features evaluated in vivo by optical coherence tomography (OCT). We found that MDMs of healthy subjects exhibited a lower oxidative stress status, lower Nrf2 and HO-1 levels as compared to CAD patients. High HO-1 levels in MDMs were associated with the presence of a higher macrophage content, a thinner fibrous cap, and a ruptured plaque with thrombus formation, detected by OCT analysis. These findings suggest the presence of a relationship between in vivo plaque characteristics and in vitro MDM profile, and may help to identify patients with rupture-prone coronary plaque.

Keywords: heme-oxygenase-1; macrophages; nuclear factor erythroid 2–related factor 2; optical coherence tomography; oxidative stress; plaque vulnerability.

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Conflict of interest statement

Funding: This work was supported by the Italian Ministry of Health, Rome, Italy (Ricerca Corrente 2014: CC13; 2015: CC13; 2016: CC18). Co-funding provided by the contribution of the Italian “5 × 1000” tax (2010, 2011, 2012).

Figures

Figure 1

Figure 1

GSH/GSSG evaluation in monocyte-derived macrophages (MDMs). MDMs were obtained from CAD patients and healthy subjects. Data are expressed as mean ± SD and derive from independent cultures obtained from 10 healthy subjects and 30 CAD patients (SA n = 10; NSTEMI n = 10; STEMI n = 10. * p < 0.05 vs. healthy subjects.

Figure 2

Figure 2

HO-1 levels in CAD patients and healthy subjects. (a) The protein levels of HO-1 were detected by western blot analysis. β-actin was used as a control of protein loading. Densitometry is shown in the bar graph. Data are expressed as mean ± SD and derive from MDMs obtained from 10 healthy subjects and 17 CAD patients. (b) Representative images of HO-1 in round and spindle MDMs obtained from healthy subjects and CAD patients (400× original magnification), nuclei were visualized by Hoechst 33258. (c) Quantitative analysis of HO-1 in round and spindle MDMs. Data are expressed as mean ± SD of fluorescence intensity/µm2 (at least three fields, 400× magnification, were analyzed) and data derive from independent cultures obtained from 10 healthy subjects and 30 CAD patients (SA n = 10; NSTEMI n = 10; STEMI n = 10. # p < 0.05 vs. round; * p < 0.05, ** p < 0.01, *** p < 0.001 vs. healthy subjects.

Figure 3

Figure 3

Nrf2 levels in CAD patients and healthy subjects. (a,b) Nrf2 levels in (a) total cellular lysate, (b) nuclear and cytosolic compartments were detected by western blot analysis. β-actin was used as a control of protein loading. Densitometry is shown in the bar graph. Data are expressed as mean ± SD and derive (a) from MDMs obtained from 10 healthy subjects and 17 CAD patients; (b) from MDMs obtained from 5 healthy subjects and 5 CAD patients. (c) Representative images of Nrf2 in round and spindle MDMs obtained from healthy subjects and CAD patients (400× original magnification), nuclei were visualized by Hoechst 33258. (d) Quantitative analysis of Nrf2 in round and spindle MDMs. Data are expressed as mean ± SD of fluorescence intensity/µm2 (at least three fields, 400× magnification, were analyzed) and derive from independent cultures obtained from 10 healthy subjects and 30 CAD patients (SA n = 10; NSTEMI n = 10; STEMI n = 10). * p < 0.05, ** p < 0.01, vs. healthy subjects.

Figure 4

Figure 4

In vivo plaque features and HO-1 levels in MDM morphotypes. (a) Association between HO-1 levels and thin cap fibroatheroma (TCFA), (b) fibrous cap integrity, (c) presence of thrombi detected by means of optical coherence tomography (OCT). Data are expressed as median and IQR. * p < 0.05, ** p < 0.01.

Figure 5

Figure 5

Correlations between in vivo plaque features and HO-1 levels in MDM morphotypes. Correlation between HO-1 levels in round and spindle MDMs and (a) macrophage content (NSD) and (b) max lipid arc, detected by means of OCT in in vivo plaque.

References

    1. Schaar J.A., Muller J.E., Falk E., Virmani R., Fuster V., Serruys P.W., Colombo A., Stefanadis C., Ward Casscells S., Moreno P.R., et al. Terminology for high-risk and vulnerable coronary artery plaques. Report of a meeting on the vulnerable plaque, June 17 and 18, 2003, Santorini, Greece. Eur. Heart J. 2004;25:1077–1082. doi: 10.1016/j.ehj.2004.01.002. - DOI - PubMed
    1. Schwartz S.M., Galis Z.S., Rosenfeld M.E., Falk E. Plaque rupture in humans and mice. Arterioscler. Thromb. Vasc. Biol. 2007;27:705–713. doi: 10.1161/01.ATV.0000261709.34878.20. - DOI - PubMed
    1. Vancraeynest D., Pasquet A., Roelants V., Gerber B.L., Vanoverschelde J.L. Imaging the vulnerable plaque. J. Am. Coll. Cardiol. 2011;57:1961–1979. doi: 10.1016/j.jacc.2011.02.018. - DOI - PubMed
    1. Chinetti-Gbaguidi G., Colin S., Staels B. Macrophage subsets in atherosclerosis. Nat. Rev. Cardiol. 2015;12:10–17. doi: 10.1038/nrcardio.2014.173. - DOI - PubMed
    1. Yokoyama M. Oxidant stress and atherosclerosis. Curr. Opin. Pharmacol. 2004;4:110–115. doi: 10.1016/j.coph.2003.12.004. - DOI - PubMed

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