Molecular and Cellular Biochemistry

, Volume 431, Issue 1–2, pp 67–74 | Cite as

HMGB1 induces endothelial progenitor cells apoptosis via RAGE-dependent PERK/eIF2α pathway

  • Qun Huang
  • Zhen Yang
  • Ji-Peng ZhouEmail author
  • Ying LuoEmail author


Studies have demonstrated that the high-mobility group 1B protein (HMGB1) could regulate endothelial progenitor cell (EPC) homing, but the effect of HMGB1 on EPC apoptosis and associated mechanisms are still unclear. The aim of this study was to investigate the effects of HMGB1 on EPC apoptosis and the possible involvement of the endoplasmic reticulum (ER) stress pathway. EPC apoptosis was determined by flow cytometry. The expressions of PERK, eIF2α, and CHOP were detected by western blotting. Additionally, the effects of PERK shRNA on the biological behaviors of EPCs were assessed. Our results showed that incubation of EPCs with HMGB1 (0.1–1 μg/ml) for 12–48 h induced apoptosis as well as activated ER stress transducers, as assessed by up-regulating PERK protein expression and eIF2α phosphorylation in a dose or time-dependent manner. Moreover, HMGB1-mediated EPC apoptosis and CHOP expression were dramatically suppressed by PERK shRNA or a specific eIF2α inhibitor (salubrinal). Importantly, a blocking antibody specifically targeted against RAGE (anti-RAGE antibody) markedly inhibited HMGB1-induced EPC apoptosis and ER stress marker protein (PERK, eIF2α, and CHOP) expression levels. Our novel findings suggest that HMGB1 triggered EPC apoptosis in a manner of RAGE-mediated activation of the PERK/eIF2α pathway.


Endothelial progenitor cell High-mobility group 1B Endoplasmic reticulum stress Apoptosis 



This study was supported by the National Natural Science Foundation of China (No. 31270992 and No. 81670453) and China Postdoctoral Science Foundation (2015M582350).

Compliance with ethical standards

Conflict of interest

The authors declare no conflicts of interest.


  1. 1.
    Ruan C, Shen Y, Chen R, Wang Z, Li J, Jiang Y (2013) Endothelial progenitor cells and atherosclerosis. Front Biosci (Landmark 18:1194–1201CrossRefGoogle Scholar
  2. 2.
    Lin CP, Lin FY, Huang PH, Chen YL, Chen WC, Chen HY, Huang YC, Liao WL, Huang HC, Liu PL, Chen YH (2013) Endothelial progenitor cell dysfunction in cardiovascular diseases: role of reactive oxygen species and inflammation. Biomed Res Int 2013:845037Google Scholar
  3. 3.
    Du F, Zhou J, Gong R, Huang X, Pansuria M, Virtue A, Li X, Wang H, Yang XF (2012) Endothelial progenitor cells in atherosclerosis. Front Biosci (Landmark 17:2327–2349CrossRefGoogle Scholar
  4. 4.
    Werner N, Kosiol S, Schiegl T, Ahlers P, Walenta K, Link A, Böhm M, Nickenig G (2005) Circulating endothelial progenitor cells and cardiovascular outcomes. N Engl J Med 353(10):999–1007CrossRefPubMedGoogle Scholar
  5. 5.
    Peplow PV (2014) Influence of growth factors and cytokines on angiogenic function of endothelial progenitor cells: a review of in vitro human studies. Growth Factors 32(3–4):83–116CrossRefPubMedGoogle Scholar
  6. 6.
    Chiang CH, Huang PH, Chiu CC, Hsu CY, Leu HB, Huang CC, Chen JW, Lin SJ (2014) Reduction of circulating endothelial progenitor cell level is associated with contrast-induced nephropathy in patients undergoing percutaneous coronary and peripheral interventions. PLoS One 9(3):e89942CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Du G, Song Y, Zhang T, Ma L, Bian N, Chen X, Feng J, Chang Q, Li Z (2014) Simvastatin attenuates TNF–α–induced apoptosis in endothelial progenitor cells via the upregulation of SIRT1. Int J Mol Med 34(1):177–182PubMedGoogle Scholar
  8. 8.
    Asavarut P, Zhao H, Gu J, Ma D (2013) The role of HMGB1 in inflammation-mediated organ injury. Acta Anaesthesiol Taiwan 51(1):28–33CrossRefPubMedGoogle Scholar
  9. 9.
    Mitola S, Belleri M, Urbinati C, Coltrini D, Sparatore B, Pedrazzi M, Melloni E, Presta M (2006) Cutting edge: extracellular high mobility group box-1 protein is a proangiogenic cytokine. J Immunol 176(1):12–15CrossRefPubMedGoogle Scholar
  10. 10.
    Hayakawa K, Pham LD, Katusic ZS, Arai K, Lo EH (2012) Astrocytic high-mobility group box 1 promotes endothelial progenitor cell-mediated neurovascular remodeling during stroke recovery. Proc Natl Acad Sci U S A 109(19):7505–7510CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Meng E, Guo Z, Wang H, Jin J, Wang J, Wang H, Wu C, Wang L (2008) High mobility group box 1 protein inhibits the proliferation of human mesenchymal stem cells and promotes their migration and differentiation along osteoblastic pathway. Stem Cells Dev 17(4):805–813CrossRefPubMedGoogle Scholar
  12. 12.
    Kaufman RJ (2002) Orchestrating the unfolded protein response in health and disease. J Clin Invest 110(10):1389–1398CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Kim I, Xu W, Reed JC (2008) Cell death and endoplasmic reticulum stress: disease relevance and therapeutic opportunities. Nat Rev Drug Discov 7(12):1013–1030CrossRefPubMedGoogle Scholar
  14. 14.
    Rutkowski DT, Kaufman RJ (2004) A trip to the ER: coping with stress. Trends Cell Biol 14(1):20–28CrossRefPubMedGoogle Scholar
  15. 15.
    Minamino T, Kitakaze M (2010) ER stress in cardiovascular disease. J Mol Cell Cardiol 48(6):1105–1110CrossRefPubMedGoogle Scholar
  16. 16.
    Iurlaro R, Muñoz-Pinedo C (2016) Cell death induced by endoplasmic reticulum stress. FEBS J 283(14):2640–2652CrossRefPubMedGoogle Scholar
  17. 17.
    Marciniak SJ, Yun CY, Oyadomari S, Novoa I, Zhang Y, Jungreis R, Nagata K, Harding HP, Ron D (2004) CHOP induces death by promoting protein synthesis and oxidation in the stressed endoplasmic reticulum. Genes Dev 18(24):3066–3077CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Zheng H, Fu G, Dai T, Huang H (2007) Migration of endothelial progenitor cells mediated by stromal cell-derived factor-1alpha/CXCR4 via PI3K/Akt/eNOS signal transduction pathway. J Cardiovasc Pharmacol 50(3):274–280CrossRefPubMedGoogle Scholar
  19. 19.
    Chen J, Song M, Yu S, Gao P, Yu Y, Wang H, Huang L (2010) Advanced glycation endproducts alter functions and promote apoptosis in endothelial progenitor cells through receptor for advanced glycation endproducts mediate overpression of cell oxidant stress. Mol Cell Biochem 335(1–2):137–146CrossRefPubMedGoogle Scholar
  20. 20.
    Rouschop KM, van den Beucken T, Dubois L, Niessen H, Bussink J, Savelkouls K, Keulers T, Mujcic H, Landuyt W, Voncken JW, Lambin P, van der Kogel AJ, Koritzinsky M, Wouters BG (2010) The unfolded protein response protects human tumor cells during hypoxia through regulation of the autophagy genes MAP1LC3B and ATG5. J Clin Invest 120(1):127–141CrossRefPubMedGoogle Scholar
  21. 21.
    Zhang M, Malik AB, Rehman J (2014) Endothelial progenitor cells and vascular repair. Curr Opin Hematol 21(3):224–228CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Dzau VJ, Gnecchi M, Pachori AS, Morello F, Melo LG (2005) Therapeutic potential of endothelial progenitor cells in cardiovascular diseases. Hypertension 46(1):7–18CrossRefPubMedGoogle Scholar
  23. 23.
    Edelberg JM, Tang L, Hattori K, Lyden D, Rafii S (2002) Young adult bone marrow-derived endothelial precursor cells restore aging-impaired cardiac angiogenic function. Circ Res 90(10):E89–E93CrossRefPubMedGoogle Scholar
  24. 24.
    Aragona CO, Imbalzano E, Mamone F, Cairo V, Lo Gullo A, D’Ascola A, Sardo MA, Scuruchi M, Basile G, Saitta A, Mandraffino G (2016) Endothelial progenitor cells for diagnosis and prognosis in cardiovascular disease. Stem Cells Int 2016:8043792CrossRefPubMedGoogle Scholar
  25. 25.
    Malhotra JD, Kaufman RJ (2011) ER stress and its functional link to mitochondria: role in cell survival and death. Cold Spring Harb Perspect Biol 3(9):a004424CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Rouhiainen A, Kuja-Panula J, Tumova S, Rauvala H (2013) RAGE-mediated cell signaling. Methods Mol Biol 963:239–263CrossRefPubMedGoogle Scholar
  27. 27.
    Xie J, Méndez JD, Méndez-Valenzuela V, Aguilar-Hernández MM (2013) Cellular signalling of the receptor for advanced glycation end products (RAGE). Cell Signal 25(11):2185–2197CrossRefPubMedGoogle Scholar
  28. 28.
    Chavakis E, Hain A, Vinci M, Carmona G, Bianchi ME, Vajkoczy P, Zeiher AM, Chavakis T, Dimmeler S (2007) High-mobility group box 1 activates integrin-dependent homing of endothelial progenitor cells. Circ Res 100(2):204–212CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Department of Child Health CareHunan Provincial Maternal and Child Health Care HospitalChangshaChina
  2. 2.Department of Hypertension and Vascular Disease, The First Affiliated HospitalSun Yat-Sen UniversityGuangzhouChina
  3. 3.Department of Cardiovascular Medicine, Xiangya HospitalCentral South UniversityChangshaChina
  4. 4.Department of Geriatric Medicine, Xiangya HospitalCentral South UniversityChangshaChina

Personalised recommendations