Zinc deficiency decreased cell viability both in endothelial EA.hy926 cells and mouse aortic culture ex vivo and its implication for anti-atherosclerosis

  • Cho, Young-Eun (Department of Food Science and Nutrition, Andong National University) ;
  • Choi, Jee-Eun (Department of Food Science and Nutrition, Andong National University) ;
  • Alam, Md. Jahangir (Department of Food Science and Nutrition, Andong National University) ;
  • Lee, Man-Hyo (Gyeongbuk Institute for BioIndustry) ;
  • Sohn, Ho-Yong (Department of Food Science and Nutrition, Andong National University) ;
  • Beattie, John H. (Division of Vascular Health, Rowett Research Institute) ;
  • Kwun, In-Sook (Department of Food Science and Nutrition, Andong National University)
  • Published : 2008.06.30

Abstract

Zinc plays a protective role in anti-atherosclerosis but the clear mechanism has not been proposed yet. In the present study, we evaluated whether zinc modulates atherosclerotic markers, VACM-1 and ICAM-1 and cell viability both in endothelial cells in vitro and mouse aortic cell viability ex vivo. In study 1, as in vitro model, endothelial EA.hy926 cells were treated with $TNF{\alpha}$ for 5 hours for inducing oxidative stress, and then treated with Zn-adequacy ($15\;{\mu}M$ Zn) or Zn-deficiency ($0\;{\mu}M$ Zn) for 6 hours. Pro-atherosclerosis factors, VCAM-1 and ICAM-1 mRNA expression and cell viability was measured. In study 2, as ex vivo model, mouse aorta ring was used. Mourse aorta was removed and cut in ring then, cultured in a 96-well plate. Aortic ring was treated with various $TNF{\alpha}$ (0-30 mg/ml) and intracellular zinc chelator, N, N, N', N', -tetrakis (2-pyridylmethyl) ethylenediamine (TPEN, $0-30\;{\mu}M$) for cellular zinc depletion for 2 days and then cell viability was measured. The results showed that in in vitro study, Zn-adequate group induced more VCAM-1 & ICAM-1 mRNA expression than Zn-deficient group during 6-hour zinc treatment post-5 hour TNF-$\alpha$ treatment, unexpectedly. These results might be cautiously interpreted that zinc would biologically induce the early expression of anti-oxidative stress through the increased adhesion molecule expression for reducing atherosclerotic action, particularly under the present 6-hour zinc treatment. In ex vivo, mouse aortic ring cell viability was decreased as TNF-$\alpha$ and TPEN levels increased, which suggests that mouse aortic blood vessel cell viability was decreased, when oxidative stress increases and cellular zinc level decreases. Taken together, it can be suggested that zinc may have a protective role in anti-atherosclerosis by cell viability in endothelial cells and aorta tissue. Further study is needed to clarify how pro-atherosclerosis molecule expression is modulated by zinc.

Keywords

References

  1. Alexander RW (1995). Hypertension and the pathogenesis of atherosclerosis: oxidative stress and the mediation of arterial inflammatory response: a new perspective. Hypertension 25:155-161 https://doi.org/10.1161/01.HYP.25.2.155
  2. Beattie JH & Kwun IS (2004). Is zinc deficiency a risk factor for atherosclerosis? Br J Nutr 91:177-181 https://doi.org/10.1079/BJN20031072
  3. Berger M, Rubinraut E, Barshack I, Roth A, Keren G & George J (2004). Zinc reduces intimal hyperplasia in the rat carotid injury model. Atherosclerosis 175:229-234 https://doi.org/10.1016/j.atherosclerosis.2004.03.022
  4. Choi JS, Choi YJ, Park SH, Kang JS & Kang YH (2004). Flavones mitigate tumor necrosis factor-a-induced adhesion molecular upregulation in cultured human endothelial cells: role of nuclear factor-kB. J Nutr 134:10103-1019
  5. Clair J, Talwalkar R, McClainC J & Hennig B (1995). Selective removal of zinc from cell culture media. Journal of Trace Elements in Experimental Medicine 7:143-151
  6. Clegg MS, Hanna LA, Niles BJ, Momma TY & Keen CL (2005). Zinc deficiency-induced cell death. IUBMB Life 57:661-669 https://doi.org/10.1080/15216540500264554
  7. Collins PW, Macey MG, Cahill MR & Newland AC (1993). Von Willebrand factor release and P-selectin expression is stimulated by thrombin and trypsin but not IL-1 in cultured human endothelial cells. Thromb Haemost 70:346-350 https://doi.org/10.1055/s-0038-1649578
  8. Costarelli L, Muti E, Malavolta M, Giacconi R, Cipriano C, Sartini D, Emanuelli M, Silvestrini M, Provinciali L, Gobbi B & Mocchegiani E (2008). Modulation of genes involved in Zinc homeostasis in old low-grade atherosclerotic patients under effects of HMG-CoA reductase inhibitors. Rejuvenation Res 11:287-291 https://doi.org/10.1089/rej.2008.0665
  9. Desai A, ZhaoY & Warren JS (2008). Development of atherosclerosis in Balb/c apolipoprotein E-deficient mice. Cardiovasc Pathol (Epub ahead of print)
  10. Formigari A, Irato P & Santon A (2007). Zinc, antioxidant systems an dmetallothionein in metal mediated-apoptosis: Biochemical and cytochemical aspects. Com Biochem Physiol C Toxicol Pharmacol (Epub ahead of print)
  11. Hennig B, Wang Y, Ramasamy S & McClain CJ (1992). Zinc deficiency alters barrier function of cultured porcine endothelial cells. J Nutr 122:1242-1247 https://doi.org/10.1093/jn/122.6.1242
  12. Hennig B, Meerarani P, Ramadass P, Toborek M, Malecki N, Slim R & McClain CJ (1999). Zinc nutrition and apoptosis of vascular endothelial cells: implications in atherosclerosis. Nutrition 1:744-748
  13. Henriksen PA & Sallenave JM (2008). Medicine in focus Human neutrophil elastase: Mediator and therapeutic target in atherosclerosis. Int J Biochem Cell Biol 40:1095-1100 https://doi.org/10.1016/j.biocel.2008.01.004
  14. Kharbanda R & MacAllister RJ (2005). The Atherosclerosis time-line and the role of the endothelium. Endocr Metab Agents 5:47-52 https://doi.org/10.2174/1568013053005409
  15. Kunsch C & Medford RM (1999). Oxidative stress as a regulator of gene expression in the vasculature. Circ Res 85 (8):753-766 https://doi.org/10.1161/01.RES.85.8.753
  16. Ling S, Nheu L , Dai A, Guo Z & Komesaroff P ( 2007). Effects of four medicinal herbs on human vascular endothelial cells in culture. Int J Cardiol (Epub ahead of print)
  17. McClain C, Morris P & Hennig B (1995). Zinc and endothelial function. Nutrition 11:117-120
  18. Minqin R, En H, K Beck, R Rajendran, Wu BJ, Halliwell B, Watt F & Stocker R (2007). Nuclear microprobe investigation into the trace elemental contents of carotid artery walls of apolipoprotein E deficient mice. Nucl Instrum Methods Phys Res B 260:240-244 https://doi.org/10.1016/j.nimb.2007.02.028
  19. Meerarani P, Ramadass P, Toborek M, Bauer HC, Bauer H & Hennig B (2000). Zinc protect against apoptosis of endothelial cells induced by linoleic acid and tumor necrosis factor alpha. Am J Clin Nutr 71:81-87 https://doi.org/10.1093/ajcn/71.1.81
  20. Papagianni A, Kalovoulos M, Kirmizis D, Vainas A, Belechri A, Alexopoulos E & Memmos D (2003). Carotid atherosclerosis is associated with inflammation and endothelial cell adhesion molecules in chronic haemodialysis patients. Nephrol Dial Transplant 18:113-119 https://doi.org/10.1093/ndt/18.1.113
  21. Reiterer G, Toborek M & Hennig B (2004). Peroxisome proliferator activated receptors and $\gamma$ require zinc for their anti-inflammatory properties in porcine vascular endothelial cells. J Nutr 134:1711-1715
  22. Ren M, Watt F, Huat BTK & Halliwell B (2003). Correlation of iron and zinc levels with lesion depth in newly formed atherosclerotic lesions. Free Radic Biol Med 34:746-752 https://doi.org/10.1016/S0891-5849(02)01427-2
  23. Theilmeier G, Michiels C, Spaepen E, Vreys I, Collen D, Vermylen J & Hoylaerts MF. (2008). Endothelial von Willebrand factor recruits platelets to atherosclerosis-prone sites in response to hypercholesterolemia. Blood 99:4486-4493 https://doi.org/10.1182/blood.V99.12.4486
  24. Watt F, Rajendran R, Ren MQ, Tan BKH & Halliwell B (2006). A nuclear microscopy study of trace elements Ca, Fe, Zn and Cu in atherosclerosis. Nucl Instrum Methods Phys Res 249:646-652 https://doi.org/10.1016/j.nimb.2006.03.073
  25. Yang H, Roberts LJ, Shi MJ, Zhou LC, Ballard BR, Richardson A & Guo ZM (2004). Retardation of atherosclerosis by overexpression of catalase or both Cu/Zn-superoxide dismutase and catalase in mice lacking apolipoprotein. E. Circ Res 95:1075-1081 https://doi.org/10.1161/01.RES.0000149564.49410.0d
  26. Zheng HT, Zhou LN, Huang CJ, Hua X, Jian R, Su BH & Fanga F (2008). Selenium inhibits high glucose and high insulin-induced adhesion molecule expression in vascular endothelial cells. Arch Med Res 39:373-379 https://doi.org/10.1016/j.arcmed.2007.12.007