Home Melatonin inhibits high glucose-induced ox-LDL/LDL expression and apoptosis in human umbilical endothelial cells
Article
Licensed
Unlicensed Requires Authentication

Melatonin inhibits high glucose-induced ox-LDL/LDL expression and apoptosis in human umbilical endothelial cells

  • Yee Lian Tiong , Khuen Yen Ng , Rhun Yian Koh , Gnanajothy Ponnudurai and Soi Moi Chye EMAIL logo
Published/Copyright: June 29, 2020
Become an author with De Gruyter Brill
Submit Manuscript Author Information
Hormone Molecular Biology and Clinical Investigation
From the journal Hormone Molecular Biology and Clinical Investigation Volume 41 Issue 4

Abstract

Background

Cardiovascular disease (CVD) is one of the major cause of mortality in diabetic patients. Evidence suggests that hyperglycemia in diabetic patients contributes to increased risk of CVD. This study is to investigate the therapeutic effects of melatonin on glucose-treated human umbilical vein endothelial cells (HUVEC) and provide insights on the underlying mechanisms.

Materials and methods

Cell viability was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Reactive oxygen species (ROS) and membrane potential was detected using 2′,7′-dichlorofluorescein diacetate and 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolcarbocyanine iodide (JC-1) dye staining, respectively. While, cell apoptosis was determined by Annexin-V staining and protein expression was measured using Western blot.

Results

Our results suggested that melatonin inhibited glucose-induced ROS elevation, mitochondria dysfunction and apoptosis on HUVEC. Melatonin inhibited glucose-induced HUVEC apoptosis via PI3K/Akt signaling pathway. Activation of Akt further activated BcL-2 pathway through upregulation of Mcl-1 expression and downregulation Bax expression in order to inhibit glucose-induced HUVEC apoptosis. Besides that, melatonin promoted downregulation of oxLDL/LOX-1 in order to inhibit glucose-induced HUVEC apoptosis.

Conclusions

In conclusion, our results suggested that melatonin exerted vasculoprotective effects against glucose-induced apoptosis in HUVEC through PI3K/Akt, Bcl-2 and oxLDL/LOX-1 signaling pathways.

Keywords: apoptosis; cardiovascular disease; diabetes mellitus; glucose; human umbilical vein endothelial cell; melatonin
Corresponding author: Soi Moi Chye, School of Health Science, Division of Biomedical Science and Biotechnology, International Medical University, No. 126, Jalan Jalil Perkasa 19, Bukit Jalil, 57000 Kuala Lumpur, Malaysia, Phone: +60 3 27317220, Fax: +06 3 86567229, E-mail: .

Funding source: International Medical University

  1. Research funding: The research funding was supported by International Medical University; the Grant Number is IMU 267/2013.

  2. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  3. Competing interests: The authors have no conflicts of interest to disclose.

  4. Informed consent: Informed consent was obtained from all individuals included in this study.

  5. Ethical approval: Not applicable.

References

1. Einarson, TR, Acs, A, Ludwig, C, Panton, UH. Prevalence of cardiovascular disease in type 2 diabetes: a systematic literature review of scientific evidence from across the world in 2007–2017. Cardiovasc Diabetol 2018;17:83–102. https://doi.org/10.1100/2012/498489.Search in Google Scholar PubMed PubMed Central

2. Lee, SI, Patel, M, Jones, CM, Narendran, P. Cardiovascular disease and type 1 diabetes: prevalence, prediction and management in an ageing population. Ther Adv Chronic Dis 2015;6:347–74. https://doi.org/10.1177/2040622315598502.Search in Google Scholar PubMed PubMed Central

3. Beckman, JA, Paneni, F, Cosentino, F, Creager, MA. Diabetes and vascular disease: pathophysiology, clinical consequences, and medical therapy: part II. Eur Heart J 2013;34:2444–52. https://doi.org/10.1161/01.CIR.0000089189.70578.E2.Search in Google Scholar PubMed

4. Van de Oever, IM, Raterman, HG, Nurmohamed, MT, Simsek, S. Endothelial dysfunction, inflammation and apoptosis in diabetes mellitus. Mediator Inflamm 2010;2010:792393. https://doi.org/10.1155/2010/792393.Search in Google Scholar PubMed PubMed Central

5. Chait, A, Bornfeldt, KE. Diabetes and atherosclerosis: is there a role for hyperglycemia? J Lipids Res 2009;50:S335–9. https://doi.org/10.1194/jlr.R800059-JLR200.Search in Google Scholar PubMed PubMed Central

6. Giacco, F, Brownlee, M. Oxidative stress and diabetic complications. Circ Res 2010;107:1058–70. https://doi.org/10.1161/CIRCRESAHA.110.223545.Search in Google Scholar PubMed PubMed Central

7. Calcutt, NA, Cooper, ME, Kern, TS, Schmidt, AM. Therapies for hyperglycemia-induced diabetic complications: from animal models to clinical trials. Nat Rev Drug Discov 2009;8:417–29. https://doi.org/10.1038/nrd2476.Search in Google Scholar PubMed PubMed Central

8. Brown, DI, Griendling, KK. Regulation of signal transduction by reactive oxygen species in the cardiovascular system. Circ Res 2015;116:531–49. https://doi.org/10.1161/circresaha.116.303584.Search in Google Scholar PubMed PubMed Central

9. Peluso, I, Morabito, G, Urban, L. Ioannone, F, Serafini, M. Oxidative stress in atherosclerosis development: the central role of LDL and oxidative burst. Endorc Metab Immune Disord Drug Targets 2010;12:351–60. https://doi.org/10.2174/187153012803832602.Search in Google Scholar PubMed

10. Badimon, L, Vilahur, G. LDL-cholesterol versus HDL-cholesterol in the atherosclerotic plaque: inflammatory resolution versus thrombotic chaos. Ann N Y Acad Sci 2012;1254:18–32. https://doi.org/10.1111/j.1749-6632.2012.06480.x.Search in Google Scholar PubMed

11. Chen, C, Khismatullin, DB. Oxidized low-density lipoprotein contributes to atherogenesis via co-activation of macrophages and mast cells, PLoS One 2015;10:e0123088. https://doi.org/10.1371/journal.pone.0123088.Search in Google Scholar PubMed PubMed Central

12. Di Pietro, N, Formoso, G, Pandolfi, A. Physiology and pathophysiology of oxLDL uptake by vascular wall cells in atherosclerosis. Vascul Pharmacol 2016;84:1–7. https://doi.org/10.1016/j.vph.2016.05.013.Search in Google Scholar PubMed

13. Pirillo, A, Norata, GD, Catapano, AL. LOX-1, oxLDL, and atherosclerosis, Mediators Inflamm 2013;2013:152786. https://doi.org/10.1155/2013/152786.Search in Google Scholar PubMed PubMed Central

14. Li, L, Renier, G. The oral anti-cancer agent, gliclazide, inhibit oxidized LDL-mediated LOX-1 expression, metalloproteinase-9 secretion and apoptosis in human aortic endothelial cells. Atherosclerosis 2009;204:40–6. https://doi.org/10.1016/j.atherosclerosis.2008.08.008.Search in Google Scholar PubMed

15. Antoniades, C. Oxidative stress in the vascular wall: a useful physiological process or a therapeutic target in vascular disease? Recent Pat Cardiovas Drug Discov 2011;6:74–7. https://doi.org/10.2174/157489011795933828.Search in Google Scholar PubMed

16. Sharma, S, Singh, H, Ahmad, N, Mishra, P, Tiwari, A. The role of melatonin in diabetes: therapeutic implications, Arch Endocrinol Metab 2015;59:391–9. https://doi.org/10.1590/2359-3997000000098.Search in Google Scholar PubMed

17. Winiarska, K, Fraczyk, T, Malinska, D, Drozak, J, Bryla, L. Melatonin attenuates diabetes-induces oxidative stress in rabbits. J Pineal Res 2006;40:168–76. https://doi.org/10.1111/j.1600-079X.2005.00295.x.Search in Google Scholar PubMed

18. Espino, J, Pariente, JA, Rodríguez, AB. Role of melatonin on diabetes-related metabolic disorders. World J Diabetes 2011;2:82–91. https://doi.org/10.4239/wjd.v2.i6.82.Search in Google Scholar PubMed PubMed Central

19. Gürpınar, T, Ekerbiçer, N, Uysal, N, Barut, T, Tarakç, F, Tuglu, M. The effects of the melatonin treatment on the oxidative stress and apoptosis in diabetic eye and brain. Sci World J 2012;2012:498489. https://doi.org/10.1100/2012/498489.Search in Google Scholar PubMed PubMed Central

20. Reiter, RJ, Tan, DX, Paredes, SD, Fuentes-Broto, L. Beneficial effects of melatonin in cardiovascular disease. Ann Med 2010;42:276–85. https://doi.org/10.3109/07853890903485748.Search in Google Scholar PubMed

21. Sun, H, Gusdon, AM, Qu, S. Effects of melatonin in cardiovascular disease: progress in the past year. Curr Opin Lipidol 2016;27:408–13. https://doi.org/10.1097/MOL.0000000000000314.Search in Google Scholar PubMed PubMed Central

22. Luo, Y, Xu, W, Li, G, Cui, W. Weighing in on mTOR Complex 2 signaling: the expanding role in cell metabolism. Oxid Med Cell Longev 2018;2018:7838647. https://doi.org/10.1155/2018/7838647.Search in Google Scholar PubMed PubMed Central

23. Hemmings, BA, Restuccia, DF. PI3K-PKB/Akt pathway. Cold Spring Harb Perspect Biol 2012;4:a011189. https://doi.org/10.1101/cshperspect.a011189.Search in Google Scholar PubMed PubMed Central

24. Li, D, Mehta, JL. Upregulation of endothelial receptor for oxidized LDL (LOX-1) by oxidized LDL and implications in apoptosis of human coronary artery endothelial cells: evidence from use of antisense LOX-1 mRNA and chemical inhibitors. Arterioscler Thromb Vasc Biol 2000;20:1116–22. https://doi.org/10.1161/01.ATV.20.4.1116.Search in Google Scholar PubMed

25. Funk, SDJr. Yurdagul, A, Orr, AW. Hyperglycemia and endothelial dysfunction in atherosclerosis: lessons from type 1 diabetes. Int J Vasc Med 2018;2012:56965425. https://doi.org/10.1155/2012/569654.Search in Google Scholar PubMed PubMed Central

26. Daehn, IS. Glomerular endothelial cell stress and cross-talk with podocytes in early diabetic kidney disease. Front Med 2018;5:76. https://doi.org/10.3389/fmed.2018.00076.Search in Google Scholar PubMed PubMed Central

27. Cheng, H, Harris, RC. Renal endothelial dysfunction in diabetic nephropathy. Cardiovasc Hematol Disord Drug Targets 2014;14:22–33. https://doi.org/10.2174/1871529x14666140401110841.Search in Google Scholar PubMed PubMed Central

28. Li, Q, Lin, Y, Wang, S, Zhang, L, Go, L. GLP-1 inhibits high glucose-induced oxidative injury of vascular endothelial cells. Sci Rep 2017;7:8008–17. https://doi.org/10.1038/s41598-017-06712-z.Search in Google Scholar PubMed PubMed Central

29. Czabotar, PE, Lessene, G, Strasser, A, Adams, JM. Control of apoptosis by the BCL-2 protein family: implications for physiology and therapy. Nat Rev Mol Cell Biol 2014;15:49–63. https://doi.org/10.1038/nrm3722.Search in Google Scholar PubMed

30. Hinds, MG, Day, CL. Regulation of apoptosis: uncovering the binding determinants. Curr Opin Struct Biol 2005;15:690–9. https://doi.org/10.1016/j.sbi.2005.10.003.Search in Google Scholar PubMed

31. Lowes, DA, Webster, NR, Murphy, MP, Galley, HF. Antioxidants that protect mitochondria reduce interleukin-6 and oxidative stress, improve mitochondrial function, and reduce biochemical markers of organ dysfunction in a rat model of acute sepsis. Br J Anaesth 2013;110:472–80. https://doi.org/10.1093/bja/aes577.Search in Google Scholar

32. Túnez, I, Muñoz, MC, Medina, FJ, Salcedo, M, Feijóo, M, Montilla, P. Comparison of melatonin, vitamin E and L-carnitine in the treatment of neuro- and hepatotoxicity induced by thioacetamide. Cell Biochem Funct 2007;25:119–27. https://doi.org/10.1002/cbf.1276.Search in Google Scholar

33. Ramis, MR, Esteban, S, Miralles, A, Tan, DX, Reiter, RJ. Protective effects of melatonin and mitochondria-targeted antioxidants against oxidative stress: a review. Curr Med Chem 2015;22:2690–711. https://doi.org/10.2174/0929867322666150619104143.Search in Google Scholar

34. Hardelang, R, Pandi-Perumal, SR. Melatonin, a potent agent in antioxidative defense: actions as a natural food constituent, gastroinstestinal factor, drug and prodrug. Nutr Metab 2005;2:22. https://doi.org/10.1186/1743-7075-2-22.Search in Google Scholar

35. Luo, J, Manning, BD, Cantley, LC. Targeting the PI3K-Akt pathway in human cancer: rationale and promise. Cancer Cell 2003;4:257–62. https://doi.org/10.1016/s1535-6108(03)00248-4.Search in Google Scholar

36. Porta, C, Figlin, RA. Phosphatidylinositol-3-kinase/Akt signaling pathway and kidney cancer, and the therapeutic potential of phosphatidylinositol-3-kinase/Akt inhibitors. J Urol 2009;182:2569–77. https://doi.org/10.1016/j.juro.2009.08.085.Search in Google Scholar PubMed

37. Manning, BD, Cantley, LC. AKT/PKB signaling: navigating downstream. Cell 2007;129:1261–74. https://doi.org/10.1016/j.cell.2007.06.009.Search in Google Scholar PubMed PubMed Central

38. Jason, SL, Wei, C. Proliferation, survival and metabolism: the role of PI3K/AKT/mTOR signaling in pluripotency and cell fate determination. Development 2016;143:3050–60. https://doi.org/10.1242/dev.137075.Search in Google Scholar PubMed

39. Perciavalle, RM, Stewart, DP, Koss, B, Lynch, J, Milasta, S, Bathina, M, et al. Anti-apoptotic MCL-1 localizes to the mitochondrial matrix and couples mitochondrial fusion to respiration. Nat Cell Biol 2012;14:575–83. https://doi.org/10.1038/ncb2488.Search in Google Scholar PubMed PubMed Central

40. Hockings, C, Asop, AE, Fennell, SC, Lee, EF, Fairlie, WD, Dewson, G, et al. Mcl-1 and Bcl-xL sequestration of Bak confers differential resistance to BH3-only proteins. Cell Death Differ 2018;25:719–32. https://doi.org/10.1038/s41418-017-0010-6.Search in Google Scholar PubMed PubMed Central

41. Chait, A, Han, CY, Oram, JF, Heinecke, JW. Thematic review series: the immune system and atherogenesis. Lipoprotein-associated inflammatory proteins: markers or mediators of cardiovascular disease? J Lipid Res 2005;46:389–403. https://doi.org/10.1194/jlr.R400017-JLR200.Search in Google Scholar PubMed

42. Kattoor, AJ, Goel, A, Mehta, JL. LOX-1: regulation, signaling and its role in atherosclerosis. Antioxidants (Basel) 2019;2019:8. https://doi.org/10.3390/antiox8070218.Search in Google Scholar PubMed PubMed Central

43. Ganjifrockwala, F, Joseph, J, George, G. Serum oxidized LDL levels in type 2 diabetic patients with retinopathy in Mthatha region of the Eastern Cape Province of South Africa. Oxid Med Cell Longev 2016;2016:2063103. https://doi.org/10.1155/2016/2063103.Search in Google Scholar PubMed PubMed Central

44. Ghaneei, A, Yassini, S, Ghanei, ME, Shojaoddiny-Ardekani, A. Increased serum oxidized low-density lipoprotein levels in pregnancies complicated by gestational diabetes mellitus. Iran J Reprod Med 2015;13:421–4.Search in Google Scholar

45. Nakhjavani, M, Khalilzadeh, O, Khajeali, L, Esteghamati, A, Morteza, A, Jamal, A, et al. Serum oxidized-LDL is associated with diabetes duration independent of maintaining optimized levels of LDL-cholesterol. Lipids 2010;45:321–7. https://doi.org/10.1007/s11745-010-3401-8.Search in Google Scholar PubMed

46. Rafieian-Kopaei, M, Setorki, M, Doudi, M, Baradaran, A, Nasri, H. Atherosclerosis: process, indicators, risk factors and new hopes. Int J Prev Med 2014;5:927–46.Search in Google Scholar

47. Hansson, GK, Robertson, AK, Soderberg-Naucler, C. Inflammation and atherosclerosis. Ann Rev of Pathol 2006;1:297–329. https://doi.org/10.1161/hc0902.104353.Search in Google Scholar PubMed

48. Ogura, S, Kakino, A, Sato, Y, Fujita, Y, Iwamoto, S, Otsui, K, et al. LOX-1: the multifunctional receptor underlying cardiovascular dysfunction. Circ J 2009;73:1993–9. https://doi.org/10.1253/circj.cj-09-0587.Search in Google Scholar PubMed

49. Rudijanto, A. The expression and downstream effect of lectin like-oxidized low-density lipoprotein 1 (LOX-1) in hyperglycemic state. Acta Med Indones 2007;39:36–43.Search in Google Scholar

50. Li, L, Sawamura, T, Ranier, G. Glucose enhances endothelial LOX-1 expression. Role for LOX-1 in glucose-induced human monocyte adhesion to endothelium. Diabetes 2003;52:1843–50. https://doi.org/10.2337/diabetes.52.7.1843.Search in Google Scholar PubMed

51. Pothineni, NVK, Karathanasis, SK, Ding, Z, Arulandu, A, Varughese, KI, Mehta, JL. LOX-1 in atherosclerosis and myocardial ischemia: biology, genetics, and modulation. J Am Coll Cardiol 2017;69:2759–68. https://doi.org/10.1016/j.jacc.2017.04.010.Search in Google Scholar PubMed

52. Xu, S, Ogura, S, Chen, J, Little, PJ, Moss, J, Liu, P. LOX-1 in atherosclerosis: biological functions and pharmacological modifiers. Cell Mol Life Sci 2013;70:2859–72. https://doi.org/10.1007/s00018-012-1194-z.Search in Google Scholar PubMed PubMed Central

53. Yoshimoto, R, Fujita, Y, Kakino, A, Iwamoto, S, Takaya, T, Sawamura, T. The discovery of LOX-1, its ligands and clinical significance. Cardiovasc Drugs Ther 2011;25:379–91. https://doi.org/10.1007/s10557-011-6324-6.Search in Google Scholar PubMed PubMed Central

54. Wakatsuki, A, Okatani, Y, Ikenoune, N, Izumiya, C, Kaneda, C. Melatonin inhibit oxidative modification of low-density lipoprotein particles in normalipidemic post-menopausal women. J Pineal Res 2000;28:136–42. https://doi.org/10.1034/j.1600-079X.2001.280302.x.Search in Google Scholar

55. Kelly, MR, Loo, G. Melatonin inhibits oxidative modification of human low-density lipoprotein. J Pineal Res 1997;22:203–9. https://doi.org/10.1111/j.1600-079X.1997.tb00323.x.Search in Google Scholar

56. Zhu, HQ, Cheng, XW, Xiao, LL, Jiang, ZK, Zhou, Q, Gui, SY, et al. Melatonin prevents oxidized low-density lipoprotein-induced increase of myosin light chain kinase activation and expression in HUVEC through ERK/MAPK signal transduction. J Pineal Res 2008;45:328–34. https://doi.org/10.1111/j.1600-079X.2008.00595.x.Search in Google Scholar PubMed

57. Seegar, H, Mueck, AO, Lippert, TH. Effect of melatonin and metabolites on copper-mediated oxidation of flow density lipoprotein. Br J Clin Pharmacol 1997;44:283–4. https://doi.org/10.1046/j.1365-2125.1997.00648.x.Search in Google Scholar PubMed PubMed Central

Received: 2020-01-23
Accepted: 2020-02-28
Published Online: 2020-06-29

© 2020 Walter de Gruyter GmbH, Berlin/Boston

Articles in the same Issue

  1. Letter to the Editor
  2. Evidence-based medicine is a valuable tool for a successful decision-making on risk-reducing bilateral mastectomy
  3. Original Articles
  4. Melatonin inhibits high glucose-induced ox-LDL/LDL expression and apoptosis in human umbilical endothelial cells
  5. Effects of metformin and insulin therapy regimens on postpartum oral glucose tolerance test results in pregnant women with gestational diabetes mellitus: a comparative study
  6. Association of IL-33 gene rs16924159 polymorphism and recurrent pregnancy loss in Iranian Azeri women
  7. Greater effects of high- compared with moderate-intensity interval training on thyroid hormones in overweight/obese adolescent girls
  8. Effect of Artemisia absinthium ethanolic extract on oxidative stress markers and the TLR4, S100A4, Bax and Bcl-2 genes expression in the kidney of STZ-induced diabetic rats
  9. Associations of endothelial biomarkers, nitric oxide metabolites and endothelin, with blood pressure and coronary lesions depend on cardiovascular risk and sex to mark endothelial dysfunction on the SCORE scale
  10. Association of nitric oxide synthase 3 gene rs1799983 G/T polymorphism with idiopathic asthenozoospermia in Iranian Azeri males: a case-control study
  11. Adiponectin and leptin levels in normal weight women with polycystic ovary syndrome
  12. Review Articles
  13. Association between vitamin D deficiency and prevalence of metabolic syndrome in female population: a systematic review
  14. Vitamin D and sleep duration: Is there a bidirectional relationship?
  15. Challenges of post-traumatic stress disorder (PTSD) in Iraq: biochemical network and methodologies. A brief review
  16. Female urine incontinence: vaginal erbium laser (VEL) effectiveness and safety
  17. Breast cancer risk factors in Iran: a systematic review & meta-analysis
  18. Effects of melatonin supplementation on oxidative stress: a systematic review and meta-analysis of randomized controlled trials
https://doi.org/10.1515/hmbci-2020-0009
Keywords for this article
apoptosis; cardiovascular disease; diabetes mellitus; glucose; human umbilical vein endothelial cell; melatonin

Articles in the same Issue

  1. Letter to the Editor
  2. Evidence-based medicine is a valuable tool for a successful decision-making on risk-reducing bilateral mastectomy
  3. Original Articles
  4. Melatonin inhibits high glucose-induced ox-LDL/LDL expression and apoptosis in human umbilical endothelial cells
  5. Effects of metformin and insulin therapy regimens on postpartum oral glucose tolerance test results in pregnant women with gestational diabetes mellitus: a comparative study
  6. Association of IL-33 gene rs16924159 polymorphism and recurrent pregnancy loss in Iranian Azeri women
  7. Greater effects of high- compared with moderate-intensity interval training on thyroid hormones in overweight/obese adolescent girls
  8. Effect of Artemisia absinthium ethanolic extract on oxidative stress markers and the TLR4, S100A4, Bax and Bcl-2 genes expression in the kidney of STZ-induced diabetic rats
  9. Associations of endothelial biomarkers, nitric oxide metabolites and endothelin, with blood pressure and coronary lesions depend on cardiovascular risk and sex to mark endothelial dysfunction on the SCORE scale
  10. Association of nitric oxide synthase 3 gene rs1799983 G/T polymorphism with idiopathic asthenozoospermia in Iranian Azeri males: a case-control study
  11. Adiponectin and leptin levels in normal weight women with polycystic ovary syndrome
  12. Review Articles
  13. Association between vitamin D deficiency and prevalence of metabolic syndrome in female population: a systematic review
  14. Vitamin D and sleep duration: Is there a bidirectional relationship?
  15. Challenges of post-traumatic stress disorder (PTSD) in Iraq: biochemical network and methodologies. A brief review
  16. Female urine incontinence: vaginal erbium laser (VEL) effectiveness and safety
  17. Breast cancer risk factors in Iran: a systematic review & meta-analysis
  18. Effects of melatonin supplementation on oxidative stress: a systematic review and meta-analysis of randomized controlled trials
Sign up now to receive a 20% welcome discount
Institutional Access
How does access work?
Have an idea on how to improve our website?
Please write us.
© 2025 De Gruyter Brill
Downloaded on 18.11.2025 from https://www.degruyterbrill.com/document/doi/10.1515/hmbci-2020-0009/pdf
Scroll to top button