Article | Published:

OxHDL controls LOX-1 expression and plasma membrane localization through a mechanism dependent on NOX/ROS/NF-κB pathway on endothelial cells


Systemic inflammatory diseases enhance circulating oxidative stress levels, which results in the oxidation of circulating high-density lipoprotein (oxHDL). Endothelial cell function can be negatively impacted by oxHDL, but the underlying mechanisms for this remain unclear. Some reports indicate that the lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is also a receptor for oxHDL. However, it is unknown if oxHDL induces increased LOX-1 expression at the plasma membrane, as an event that supports endothelial dysfunction. Therefore, the aims of this study were to determine if oxHDL induces plasma-membrane level changes in LOX-1 and, if so, to describe the underlying mechanisms in endothelial cells. Our results demonstrate that the incubation of arterial or vein endothelial cells with oxHDL (and not HDL) induces the increase of LOX-1 expression at the plasma membrane; effect prevented by LOX-1 inhibition. Importantly, same results were observed in endothelial cells from oxHDL-treated rats. Furthermore, the observed oxHDL-induced LOX-1 expression is abolished by the down-regulation of NOX-2 expression with siRNA (and no others NOX isoforms), by the pharmacological inhibition of NAD(P)H oxidase (with DPI or apocynin) or by the inhibition of NF-κB transcription factor. Coherently, LOX-1 expression is augmented by the incubation of endothelial cells with H2O2 or GSSG even in absence of oxHDL, indicating that the NOX-2/ROS/ NF-κB axis is involved. Interestingly, oxHDL incubation also increases TNF-α expression, cytokine that induces LOX-1 expression. Thus, our results suggest a positive feedback mechanism for LOX-1 receptor during inflammatory condition where an oxidative burst will generate oxHDL from native HDL, activating LOX-1 receptor which in turn will increase the expression of NOX-2, TNF-α and LOX-1 receptor at the plasma membrane. In conclusion, oxHDL-induced translocation of LOX-1 to the plasma membrane could constitute an induction mechanism of endothelial dysfunction in systemic inflammatory diseases.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1.

    Nestel PJ. Dietary cholesterol and plasma lipoproteins. Atherosclerosis. 1994;109:87.

  2. 2.

    Tall AR. Plasma high density lipoproteins: therapeutic targeting and links to atherogenic inflammation. Atherosclerosis. 2018;276:39–43.

  3. 3.

    Yan BP, Chiang F-T, Ambegaonkar B, et al. Low-density lipoprotein cholesterol target achievement in patients surviving an acute coronary syndrome in Hong Kong and Taiwan—findings from the Dyslipidemia International Study II. Int J Cardiol. 2018;265:1–5.

  4. 4.

    Miller GJ. High-density lipoprotein, low-density lipoprotein, and coronary heart disease. Thorax. 1978;33:137–9.

  5. 5.

    O’Connell BJ, Genest J. High-density lipoproteins and endothelial function. Circulation. 2001;104:1978–83.

  6. 6.

    Besler C, Heinrich K, Rohrer L, et al. Mechanisms underlying adverse effects of HDL on eNOS-activating pathways in patients with coronary artery disease. J Clin Invest. 2011;121:2693–708.

  7. 7.

    Ku IA, Imboden JB, Hsue PY, Ganz P. Rheumatoid arthritis: model of systemic inflammation driving atherosclerosis. Circ J. 2009;73:977–85.

  8. 8.

    Pinsky MR. Dysregulation of the immune response in severe sepsis. Am J Med Sci. 2004;328:220–9.

  9. 9.

    Pérez L, Muñoz-Durango N, Riedel CA, et al. Endothelial-to-mesenchymal transition: cytokine-mediated pathways that determine endothelial fibrosis under inflammatory conditions. Cytokine Growth Factor Rev. 2017;33:41–54.

  10. 10.

    Suboc TM, Dharmashankar K, Wang J, et al. Moderate obesity and endothelial dysfunction in humans: influence of gender and systemic inflammation. Physiol Rep. 2013;1:e00058.

  11. 11.

    Muller MM, Griesmacher A. Markers of endothelial dysfunction. Clin Chem Lab Med. 2000;38:77–85.

  12. 12.

    Zhang C. The role of inflammatory cytokines in endothelial dysfunction. Basic Res Cardiol. 2008;103:398–406.

  13. 13.

    Closa D, Folch-Puy E. Oxygen free radicals and the systemic inflammatory response. IUBMB Life. 2004;56:185–91.

  14. 14.

    Higashi Y, Noma K, Yoshizumi M, Kihara Y. Endothelial function and oxidative stress in cardiovascular diseases. Circ J. 2009;73:411–8.

  15. 15.

    Swindle EJ, Metcalfe DD. The role of reactive oxygen species and nitric oxide in mast cell-dependent inflammatory processes. Immunol Rev. 2007;217:186–205.

  16. 16.

    Pirillo A, Norata GD, Catapano AL. LOX-1, OxLDL, and atherosclerosis. Mediat Inflamm. 2013;2013:152786–12.

  17. 17.

    Tsimikas S, Miller YI. Oxidative modification of lipoproteins: mechanisms, role in inflammation and potential clinical applications in cardiovascular disease. Curr Pharm Des. 2011;17:27–37.

  18. 18.

    Shroff R, Speer T, Colin S, et al. HDL in children with CKD promotes endothelial dysfunction and an abnormal vascular phenotype. J Am Soc Nephrol. 2014;25:2658–68.

  19. 19.

    Speer T, Rohrer L, Blyszczuk P, et al. Abnormal high-density lipoprotein induces endothelial dysfunction via activation of toll-like receptor-2. Immunity. 2013;38:754–68.

  20. 20.

    Cominacini L, Pasini AF, Garbin U, et al. Oxidized low density lipoprotein (ox-LDL) binding to ox-LDL receptor-1 in endothelial cells induces the activation of NF-kappaB through an increased production of intracellular reactive oxygen species. J Biol Chem. 2000;275:12633–8.

  21. 21.

    Kataoka H, Kume N, Miyamoto S, et al. Expression of lectinlike oxidized low-density lipoprotein receptor-1 in human atherosclerotic lesions. Circulation. 1999;99:3110–7.

  22. 22.

    Honjo M, Nakamura K, Yamashiro K, et al. Lectin-like oxidized LDL receptor-1 is a cell-adhesion molecule involved in endotoxin-induced inflammation. Proc Natl Acad Sci USA. 2003;100:1274–9.

  23. 23.

    Mitra S, Goyal T, Mehta JL. Oxidized LDL, LOX-1 and atherosclerosis. Cardiovasc Drugs Ther. 2011;25:419–29.

  24. 24.

    Matsunaga T, Hokari S, Koyama I, et al. NF-kB activation in endothelial cells treated with oxidized high-density lipoprotein. Biochem Biophys Res Commun. 2003;303:313–9.

  25. 25.

    Pirillo A, Uboldi P, Ferri N, et al. Upregulation of lectin-like oxidized low density lipoprotein receptor 1 (LOX-1) expression in human endothelial cells by modified high density lipoproteins. Biochem Biophys Res Commun. 2012;428:230–3.

  26. 26.

    Kume N, Murase T, Moriwaki H, et al. Inducible expression of lectin-like oxidized LDL receptor-1 in vascular endothelial cells. Circ Res. 1998;83:322–7.

  27. 27.

    Edgell CJ, McDonald CC, Graham JB. Permanent cell line expressing human factor VIII-related antigen established by hybridization. Proc Natl Acad Sci USA. 1983;80:3734–7.

  28. 28.

    Scoccia AE, Molinuevo MS, McCarthy AD, Cortizo AM. A simple method to assess the oxidative susceptibility of low density lipoproteins. BMC Clin Pathol. 2001;1:1.

  29. 29.

    Lynch SM, Frei B. Reduction of copper, but not iron, by human low density lipoprotein (LDL). Implications for metal ion-dependent oxidative modification of LDL. J Biol Chem. 1995;270:5158–63.

  30. 30.

    Dikalov S, Griendling KK, Harrison DG. Measurement of reactive oxygen species in cardiovascular studies. Hypertension. 2007;49:717–27.

  31. 31.

    Chen J, Liu Y, Liu H, et al. Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) transcriptional regulation by Oct-1 in human endothelial cells: implications for atherosclerosis. Biochem J. 2006;393:255–65.

  32. 32.

    Simon F, Stutzin A. Protein kinase C-mediated phosphorylation of p47phox modulates platelet-derived growth factor-induced H2O2 generation and cell proliferation in human umbilical vein endothelial cells. Endothelium. 2008;15:175–88.

  33. 33.

    Simon F, Fernández R. Early lipopolysaccharide-induced reactive oxygen species production evokes necrotic cell death in human umbilical vein endothelial cells. J Hypertens. 2009;27:1202–16.

  34. 34.

    Hwang J, Kleinhenz DJ, Lassègue B, et al. Peroxisome proliferator-activated receptor-gamma ligands regulate endothelial membrane superoxide production. Am J Physiol Cell Physiol. 2005;288:C899–905.

  35. 35.

    Hofnagel O. Proinflammatory cytokines regulate LOX-1 expression in vascular smooth muscle cells. Arterioscler Thromb Vasc Biol. 2004;24:1789–95.

  36. 36.

    Aird WC. The role of the endothelium in severe sepsis and multiple organ dysfunction syndrome. Blood. 2003;101:3765–77.

  37. 37.

    Pirillo A, Catapano AL, Norata GD. HDL in infectious diseases and sepsis. Handb Exp Pharmacol. 2015;224:483–508.

  38. 38.

    Moriwaki H, Kume N, Sawamura T, et al. Ligand specificity of LOX-1, a novel endothelial receptor for oxidized low density lipoprotein. Arterioscler Thromb Vasc Biol. 1998;18:1541–7.

  39. 39.

    Sawamura T, Kume N, Aoyama T, et al. An endothelial receptor for oxidized low-density lipoprotein. Nature. 1997;386:73–77.

  40. 40.

    Pirillo A, Reduzzi A, Ferri N, et al. Upregulation of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) by 15-lipoxygenase-modified LDL in endothelial cells. Atherosclerosis. 2011;214:331–7.

  41. 41.

    Lubrano V, Balzan S. LOX-1 and ROS, inseparable factors in the process of endothelial damage. Free Radic Res. 2014;48:841–8.

  42. 42.

    Zhao W, Ma G, Chen X. Lipopolysaccharide induced LOX-1 expression via TLR4/MyD88/ROS activated p38MAPK-NF-κB pathway. Vasc Pharmacol. 2014;63:162–72.

  43. 43.

    Son JN, Lho Y, Shin S, et al. Carbamylated low-density lipoprotein increases reactive oxygen species (ROS) and apoptosis via lectin-like oxidized LDL receptor (LOX-1) mediated pathway in human umbilical vein endothelial cells. Int J Cardiol. 2011;146:428–30.

  44. 44.

    Elks CM, Francis J. Central adiposity, systemic inflammation, and the metabolic syndrome. Curr Protoc Cytom. 2010;12:99–104.

  45. 45.

    Echeverría C, Montorfano I, Tapia P, et al. Endotoxin-induced endothelial fibrosis is dependent on expression of transforming growth factors β1 and β2. Infect Immun. 2014;82:3678–86.

  46. 46.

    Becerra A, Rojas M, Vallejos A, et al. Endothelial fibrosis induced by suppressed STAT3 expression mediated by signaling involving the TGF-β1/ALK5/Smad pathway. Lab Invest. 2017;97:1033–46.

  47. 47.

    Montorfano I, Becerra A, Cerro R, et al. Oxidative stress mediates the conversion of endothelial cells into myofibroblasts via a TGF-β1 and TGF-β2-dependent pathway. Lab Invest. 2014;94:1068–82.

  48. 48.

    Echeverría C, Montorfano I, Sarmiento D, et al. Lipopolysaccharide induces a fibrotic-like phenotype in endothelial cells. J Cell Mol Med. 2013;17:800–14.

  49. 49.

    Cutuli L, Pirillo A, Uboldi P, et al. 15-lipoxygenase-mediated modification of HDL3 impairs eNOS activation in human endothelial cells. Lipids. 2014;49:317–26.

  50. 50.

    Chen M, Masaki T, Sawamura T. LOX-1, the receptor for oxidized low-density lipoprotein identified from endothelial cells: implications in endothelial dysfunction and atherosclerosis. Pharmacol Ther. 2002;95:89–100.

  51. 51.

    Spallarossa P, Fabbi P, Manca V, et al. Doxorubicin-induced expression of LOX-1 in H9c2 cardiac muscle cells and its role in apoptosis. Biochem Biophys Res Commun. 2005;335:188–96.

  52. 52.

    Xu S, Ogura S, Chen J, et al. LOX-1 in atherosclerosis: biological functions and pharmacological modifiers. Cell Mol Life Sci. 2013;70:2859–72.

  53. 53.

    Zuniga FA, Ormazabal V, Gutierrez N, et al. Role of lectin-like oxidized low density lipoprotein-1 in fetoplacental vascular dysfunction in preeclampsia. Biomed Res Int. 2014;2014:353616.

  54. 54.

    Prauchner CA. Oxidative stress in sepsis: pathophysiological implications justifying antioxidant co-therapy. Burns. 2017;43:471–85.

  55. 55.

    Weidenbusch M, Anders H-J. Tissue microenvironments define and get reinforced by macrophage phenotypes in homeostasis or during inflammation, repair and fibrosis. J Innate Immun. 2012;4:463–77.

  56. 56.

    Dominguez JH, Mehta JL, Li D, et al. Anti-LOX-1 therapy in rats with diabetes and dyslipidemia: ablation of renal vascular and epithelial manifestations. Am J Physiol Ren Physiol. 2008;294:F110–9.

  57. 57.

    Xu X, Gao X, Potter BJ, et al. Anti-LOX-1 rescues endothelial function in coronary arterioles in atherosclerotic ApoE knockout mice. Arterioscler Thromb Vasc Biol. 2007;27:871–7.

  58. 58.

    Ishino S, Mukai T, Kuge Y, et al. Targeting of lectinlike oxidized low-density lipoprotein receptor 1 (LOX-1) with 99mTc-labeled anti-LOX-1 antibody: potential agent for imaging of vulnerable plaque. J Nucl Med. 2008;49:1677–85.

  59. 59.

    Iwamoto S, Fujita Y, Kakino A, et al. An alternative protein standard to measure activity of LOX-1 ligand containing apoB (LAB)—utilization of anti-LOX-1 single- chain antibody fused to apoB fragment. J Atheroscler Thromb. 2011;18:818–28.

  60. 60.

    Hu W, Xie Q, Liu L, Xiang H. Enhanced bioactivity of the anti-LOX-1 scFv engineered by multimerization strategy. Appl Biochem Biotechnol. 2018;185:233–47.

  61. 61.

    Hu W, Xie Q, Xiang H. Improved scFv Anti-LOX-1 binding activity by fusion with LOX-1-binding peptides. Biomed Res Int. 2017;2017:8946935.

  62. 62.

    Dröge W. Free radicals in the physiological control of cell function. Physiol Rev. 2002;82:47–95.

  63. 63.

    Kunsch C, Medford RM. Oxidative stress as a regulator of gene expression in the vasculature. Circ Res. 1999;85:753–66.

  64. 64.

    Kiritoshi S, Nishikawa T, Sonoda K, et al. Reactive oxygen species from mitochondria induce cyclooxygenase-2 gene expression in human mesangial cells: potential role in diabetic nephropathy. Diabetes. 2003;52:2570–7.

  65. 65.

    Li DW, Spector A. Hydrogen peroxide-induced expression of the proto-oncogenes, c-jun, c-fos and c-myc in rabbit lens epithelial cells. Mol Cell Biochem. 1997;173:59–69.

  66. 66.

    Marshall JC. Inflammation, coagulopathy, and the pathogenesis of multiple organ dysfunction syndrome. Crit Care Med. 2001;29(7 Suppl):S99–106.

  67. 67.

    Ziesmann MT, Marshall JC. Multiple organ dysfunction: the defining syndrome of sepsis. Surg Infect. 2018;19:184–90.

  68. 68.

    Englert JA, Fink MP. The multiple organ dysfunction syndrome and late-phase mortality in sepsis. Curr Infect Dis Rep. 2005;7:335–41.

  69. 69.

    Baue AE. Sepsis, systemic inflammatory response syndrome, multiple organ dysfunction syndrome, and multiple organ failure: are trauma surgeons lumpers or splitters? J Trauma. 2003;55:997–8.

  70. 70.

    Fink MP. Nitric oxide synthase and vascular dysfunction in sepsis. Crit Care Med. 2014;42:1572–5.

  71. 71.

    Yaghi A, Paterson NA, McCormack DG. Vascular reactivity in sepsis: importance of controls and role of nitric oxide. Am J Respir Crit Care Med. 1995;151:706–12.

  72. 72.

    Hoover DB, Brown TC, Miller MK, et al. Loss of sympathetic nerves in spleens from patients with end stage sepsis. Front Immunol. 2017;8:1712.

  73. 73.

    Tracey KJ, Fong Y, Hesse DG, et al. Anti-cachectin/TNF monoclonal antibodies prevent septic shock during lethal bacteraemia. Nature. 1987;330:662–4.

  74. 74.

    Douzinas EE, Tsidemiadou PD, Pitaridis MT, et al. The regional production of cytokines and lactate in sepsis-related multiple organ failure. Am J Respir Crit Care Med. 1997;155:53–59.

  75. 75.

    Pinsky MR. Clinical studies on cytokines in sepsis: role of serum cytokines in the development of multiple-systems organ failure. Nephrol Dial Transplant. 1994;9(Suppl 4):94–98.

Download references


This work was supported by research grants from Fondo Nacional de Desarrollo Científico y Tecnológico—Fondecyt 1161288, 1160900, 11170840, and 1161646. Millennium Institute on Immunology and Immunotherapy P09-016-F. The Millennium Nucleus of Ion Channels-Associated Diseases (MiNICAD) is a Millennium Nucleus supported by the Iniciativa Científica Milenio of the Ministry of Economy, Development and Tourism (Chile). UNAB DI-741-15/N.

Author information

Correspondence to Felipe Simon.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Supplemental Material

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Further reading

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7