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Of Mice and Men: Neuropeptide Y and Its Receptors Are Associated with Atherosclerotic Lesion Burden and Vulnerability

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Abstract

Neuropeptide Y (NPY), a sympathetic and platelet-derived vascular mitogen and angiogenic factor, has been implicated in atherosclerosis in animal and human genetic studies. Here we evaluate its association with human and murine atherosclerosis, and assess the role of platelet-derived NPY in lesion vulnerability. NPY immunoreactivity (NPY-ir) was measured in the platelet-poor and platelet-rich (PRP) plasmas, and NPY receptors (mitogenic Y1R and angiogenic Y2 and Y5Rs), CD26/DPPIV (a protease forming Y2/Y5-selective agonist), CD31-positive vascularity, and lesion morphology assessed by histo- and immunocyto-chemistry—in patients with peripheral artery disease (PAD) and healthy volunteers, and in lard-fed ApoE−/− mice. NPY and NPY-R immunostaining was greater in lesions from PAD patients compared to normal vessels of healthy volunteers (p < 0.001), and localized to smooth muscle cells, macrophages, and adventitial/neovascular endothelial cells. CD26/DPPIV staining co-localized with CD31-positive endothelial cells only in atherosclerotic lesions. NPY-ir in PRP (but not plasma) and vascular immunostaining was higher (p < 0.05 and 0.001, respectively) in men (not women) with PAD compared to healthy subjects. A similar gender specificity was observed in mice. PRP NPY-ir levels correlated with lesion area (p = 0.03), necrotic core area, and the necrotic core-to-lesion area ratio (p < 0.01) in male, but not female, mice. Also males with neovascularized lesions had higher PRP NPY-ir levels than those lacking lesion microvessels (p < 0.05). NPY and its Rs are up-regulated in human and murine atherosclerotic lesions suggesting pathogenic role. DPPIV expression by microvascular endothelium in atherosclerotic tissue may shift NPY’s affinity toward angiogenic Y2/Y5Rs, and thus enhance angiogenesis and lesion vulnerability. Remarkably, plaque neovascularization was associated with increased NPY-ir in PRP in males but not females, suggesting that platelet NPY may be a novel mediator/marker of lesion vulnerability particularly in males, for reasons that remain to be determined. Both animal and human data suggest that NPY is an important contributor to, and platelet NPY-ir a marker of, atherosclerotic lesion burden and vulnerability but only in males, perhaps due to androgen-dependent up-regulation of NPY, previously shown in rats.

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Acknowledgments

The authors wish to thank research assistants, Anna Yu and Ennika Holley for their excellent technical assistance. This work was supported by NIH grants: the MERIT award HL055310 and HL067357 awarded to Zofia Zukowska.

Author Contributions

Conceived and designed the experiments for human and murine study—ZZ, with assistance from SEE and MSB, the latter playing a major role in designing the murine study. Performed experiments—LL, AHN, JBK, and MSB. Contributed materials—RN and JL. Wrote the paper—LL, AHN, and ZZ. Extensive reviewing and editing of the paper—MSB and ZZ.

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None to be disclosed.

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Authors and Affiliations

  1. Department of Physiology and Biophysics, Georgetown University Medical Center, 3900 Reservoir Rd NW, Washington, DC, 20057, USA

    Lijun Li & Joanna B. Kitlinska

  2. Department of Surgery, Division of Vascular Surgery, Georgetown University Medical Center, Washington, DC, USA

    Richard Neville & James Laredo

  3. Cardiovascular Research Institute, MedStar Health Research Institute, Washington, DC, USA

    Amir H. Najafi, Stephen E. Epstein & Mary Susan Burnett

  4. Department of Integrative Biology & Physiology, University of Minnesota, 321 Church Str. SE, Minneapolis, MN, 55455, USA

    Zofia Zukowska

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  1. Lijun Li
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Correspondence to Zofia Zukowska.

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Supplemental Figure 1

Negative controls for immunostaining for NPY, Y2, and Y5 receptors and cd38 using the same dilutions of secondary antibodies as before but no primary antibodies. Note: negative control for Y1 immunostaining shown in Figure 3k (PPT 317 kb)

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Li, L., Najafi, A.H., Kitlinska, J.B. et al. Of Mice and Men: Neuropeptide Y and Its Receptors Are Associated with Atherosclerotic Lesion Burden and Vulnerability. J. of Cardiovasc. Trans. Res. 4, 351–362 (2011). https://doi.org/10.1007/s12265-011-9271-5

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