Molecular Medicine

, Volume 19, Issue 1, pp 195–202 | Cite as

Suppression of Coronary Atherosclerosis by Helix B Surface Peptide, a Nonerythropoietic, Tissue-Protective Compound Derived from Erythropoietin

  • Hiroto Ueba
  • Masashi Shiomi
  • Michael Brines
  • Michael Yamin
  • Tsutomu Kobayashi
  • Junya Ako
  • Shin-ichi Momomura
  • Anthony Cerami
  • Masanobu Kawakami
Research Article


Erythropoietin (EPO), a type I cytokine originally identified for its critical role in hematopoiesis, has been shown to have non-hematopoietic, tissue-protective effects, including suppression of atherosclerosis. However, prothrombotic effects of EPO hinder its potential clinical use in nonanemic patients. In the present study, we investigated the antiatherosclerotic effects of helix B surface peptide (HBSP), a nonerythropoietic, tissue-protective compound derived from EPO, by using human umbilical vein endothelial cells (HUVECs) and human monocytic THP-1 cells in vitro and Watanabe heritable hyperlipidemic spontaneous myocardial infarction (WHHLMI) rabbits in vivo. In HUVECs, HBSP inhibited apoptosis (≈70%) induced by C-reactive protein (CRP), a direct mediator of atherosclerosis. By using a small interfering RNA approach, Akt was shown to be a key molecule in HBSP-mediated prevention of apoptosis. HBSP also attenuated CRP-induced production of tumor necrosis factor (TNF)-α and matrix metalloproteinase-9 in THP-1 cells. In the WHHLMI rabbit, HBSP significantly suppressed progression of coronary atherosclerotic lesions as assessed by mean cross-sectional stenosis (HBSP 21.3 ± 2.2% versus control peptide 38.0 ± 2.7%) and inhibited coronary artery endothelial cell apoptosis with increased activation of Akt. Furthermore, TNF-α expression and the number of M1 macrophages and M1/M2 macrophage ratio in coronary atherosclerotic lesions were markedly reduced in HBSP-treated animals. In conclusion, these data demonstrate that HBSP suppresses coronary atherosclerosis, in part by inhibiting endothelial cell apoptosis through activation of Akt and in association with decreased TNF-α production and modified macrophage polarization in coronary atherosclerotic lesions. Because HBSP does not have the prothrombotic effects of EPO, our study may provide a novel therapeutic strategy that prevents progression of coronary artery disease.



We thank Takashi Ito, Satoshi Yamada, Nobue Hirayama, Harue Fukaya, Kimiko Aoki, Chie Ishikawa and Kazuko Futaka for expert technical assistance.

This work was funded in part by Grant-in-Aid for Scientific Research 20590887 from the Ministry of Education, Culture, Sports, Science and Technology of Japan (to H Ueba and M Kawakami) and by a Research Grant for Health Science from the Ministry of Health, Labor and Welfare of Japan (to M Kawakami).

Supplementary material

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  1. 1.
    Jelkmann W. (2007) Erythropoietin after a century of research: younger than ever. Eur. J. Haematol. 78:183–205.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Brines M, Cerami A. (2008) Erythropoietin-mediated tissue protection: reducing collateral damage from the primary injury response. J. Intern. Med. 264:405–32.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Buemi M, et al. (1998) Does erythropoietin administration affect progression of atherosclerosis in Watanabe heritable hyperlipaemic rabbits? Nephrol. Dial. Trans-plant. 13:2706–8.CrossRefGoogle Scholar
  4. 4.
    Pawlak K, Pawlak D, Mysliwiec M. (2006) Long-term erythropoietin therapy decreases CC-chemokine levels and intima-media thickness in hemodialyzed patients. Am. J. Nephrol. 26:497–502.CrossRefPubMedGoogle Scholar
  5. 5.
    Siamopoulos KC, et al. (2006) Long-term treatment with EPO increases serum levels of high-density lipoprotein in patients with CKD. Am. J. Kidney Dis. 48:242–9.CrossRefPubMedGoogle Scholar
  6. 6.
    Lu KY, et al. (2010) Erythropoietin suppresses the formation of macrophage foam cells: role of liver X receptor alpha. Circulation. 121:1828–37.CrossRefPubMedGoogle Scholar
  7. 7.
    Brines M, et al. (2004) Erythropoietin mediates tissue protection through an erythropoietin and common beta-subunit heteroreceptor. Proc. Natl. Acad. Sci. U. S. A. 101:14907–12.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Khorana AA, Francis CW, Culakova E, Lyman GH. (2005) Risk factors for chemotherapy-associated venous thromboembolism in a prospective observational study. Cancer. 104:2822–9.CrossRefPubMedGoogle Scholar
  9. 9.
    Corwin HL, et al. (2007) Efficacy and safety of epoetin alfa in critically ill patients. N. Engl. J. Med. 357:965–76.CrossRefPubMedGoogle Scholar
  10. 10.
    Aapro M, Scherhag A, Burger HU. (2008) Effect of treatment with epoetin-beta on survival, tumour progression and thromboembolic events in patients with cancer: an updated meta-analysis of 12 randomised controlled studies including 2301 patients. Br. J. Cancer. 99:14–22.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Brines M, et al. (2008) Nonerythropoietic, tissue-protective peptides derived from the tertiary structure of erythropoietin. Proc. Natl. Acad. Sci. U. S. A. 105:10925–30.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Erbayraktar Z, Erbayraktar S, Yilmaz O, Cerami A, Coleman T, Brines M. (2009) Nonerythropoietic tissue protective compounds are highly effective facilitators of wound healing. Mol. Med. 15:235–41.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Ueba H, et al. (2010) Cardioprotection by a non-erythropoietic, tissue-protective peptide mimicking the 3D structure of erythropoietin. Proc. Natl. Acad. Sci. U. S. A. 107:14357–62.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Ahmet I, et al. (2011) A small nonerythropoietic helix B surface peptide based upon erythropoietin structure is cardioprotective against ischemic myocardial damage. Mol. Med. 17:194–200.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Shiomi M, Ito T, Yamada S, Kawashima S, Fan J. (2003) Development of an animal model for spontaneous myocardial infarction (WHHLMI rabbit). Arterioscler. Thromb. Vasc. Biol. 23:1239–44.CrossRefPubMedGoogle Scholar
  16. 16.
    Nabata A, et al. (2008) C-reactive protein induces endothelial cell apoptosis and matrix metalloproteinase-9 production in human mononuclear cells: implications for the destabilization of atherosclerotic plaque. Atherosclerosis. 196:129–35.CrossRefPubMedGoogle Scholar
  17. 17.
    Ueba H, et al. (2005) Glimepiride induces nitric oxide production in human coronary artery endothelial cells via a PI3-kinase-Akt dependent pathway. Atherosclerosis. 183:35–9.CrossRefPubMedGoogle Scholar
  18. 18.
    Shiomi M, Ito T, Hirouchi Y, Enomoto M. (2001) Fibromuscular cap composition is important for the stability of established atherosclerotic plaques in mature WHHL rabbits treated with statins. Atherosclerosis 157:75–84.CrossRefPubMedGoogle Scholar
  19. 19.
    Khreiss T, Jozsef L, Potempa LA, Filep JG. (2004) Conformational rearrangement in C-reactive protein is required for proinflammatory actions on human endothelial cells. Circulation. 109:2016–22.CrossRefPubMedGoogle Scholar
  20. 20.
    Verma S, Szmitko PE, Ridker PM. (2005) C-reac-tive protein comes of age. Nat. Clin. Pract. Cardiovasc. Med. 2:29–36.CrossRefPubMedGoogle Scholar
  21. 21.
    Pepys MB, et al. (2006) Targeting C-reactive protein for the treatment of cardiovascular disease. Nature. 440:1217–21.CrossRefPubMedGoogle Scholar
  22. 22.
    Calvillo L, et al. (2003) Recombinant human erythropoietin protects the myocardium from ischemia-reperfusion injury and promotes beneficial remodeling. Proc. Natl. Acad. Sci. U. S. A. 100:4802–6.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Fiordaliso F, et al. (2005) A nonerythropoietic derivative of erythropoietin protects the myocardium from ischemia-reperfusion injury. Proc. Natl. Acad. Sci. U. S. A. 102:2046–51.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Salahudeen AK, et al. (2008) Antiapoptotic properties of erythropoiesis-stimulating proteins in models of cisplatin-induced acute kidney injury. Am. J. Physiol. Renal Physiol. 294:F1354–65.CrossRefPubMedGoogle Scholar
  25. 25.
    Agnello D, et al. (2002) Erythropoietin exerts an anti-inflammatory effect on the CNS in a model of experimental autoimmune encephalomyelitis. Brain Res. 952:128–34.CrossRefPubMedGoogle Scholar
  26. 26.
    Villa P, et al. (2003) Erythropoietin selectively attenuates cytokine production and inflammation in cerebral ischemia by targeting neuronal apoptosis. J. Exp. Med. 198:971–5.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Valgimigli M, et al. (2003) Endothelial dysfunction in acute and chronic coronary syndromes: evidence for a pathogenetic role of oxidative stress. Arch. Biochem. Biophys. 420:255–61.CrossRefPubMedGoogle Scholar
  28. 28.
    McKellar GE, McCarey DW, Sattar N, McInnes IB. (2009) Role for TNF in atherosclerosis? Lessons from autoimmune disease. Nat. Rev. Cardiol. 6:410–7.CrossRefPubMedGoogle Scholar
  29. 29.
    de Nooijer R, et al. (2006) Lesional overexpression of matrix metalloproteinase-9 promotes intraplaque hemorrhage in advanced lesions but not at earlier stages of atherogenesis. Arterioscler. Thromb. Vasc. Biol. 26:340–6.CrossRefPubMedGoogle Scholar
  30. 30.
    Siren AL, et al. (2001) Erythropoietin prevents neuronal apoptosis after cerebral ischemia and metabolic stress. Proc. Natl. Acad. Sci. U. S. A. 98:4044–9.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Bittorf T, Buchse T, Sasse T, Jaster R, Brock J. (2001) Activation of the transcription factor NF-kappaB by the erythropoietin receptor: structural requirements and biological significance. Cell Signal. 13:673–81.CrossRefPubMedGoogle Scholar
  32. 32.
    Parsa CJ, et al. (2003) A novel protective effect of erythropoietin in the infarcted heart. J. Clin. Invest. 112:999–1007.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Watanabe Y, et al. (1988) Preventive effect of pravastatin sodium, a potent inhibitor of 3-hy-droxy-3-methylglutaryl coenzyme A reductase, on coronary atherosclerosis and xanthoma in WHHL rabbits. Biochim. Biophys. Acta. 960:294–302.CrossRefPubMedGoogle Scholar
  34. 34.
    Chinetti-Gbaguidi G, Staels B. (2011) Macrophage polarization in metabolic disorders: functions and regulation. Curr. Opin. Lipidol 22:365–72.CrossRefPubMedPubMedCentralGoogle Scholar

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Authors and Affiliations

  • Hiroto Ueba
    • 1
  • Masashi Shiomi
    • 2
  • Michael Brines
    • 3
  • Michael Yamin
    • 3
  • Tsutomu Kobayashi
    • 2
  • Junya Ako
    • 1
  • Shin-ichi Momomura
    • 1
  • Anthony Cerami
    • 3
  • Masanobu Kawakami
    • 1
  1. 1.Department of Integrated Medicine 1, Saitama Medical CenterJichi Medical UniversitySaitama CityJapan
  2. 2.Institute for Experimental AnimalsKobe University Graduate School of MedicineKobeJapan
  3. 3.Araim PharmaceuticalsOssiningUSA

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