Abstract: We studied cytotoxic effects (CTE) induced in confluent cultures of human umbilical vein endothelial cells (HUVEC) by initiators of free-radical reactions (FRR): H2O2 (10(-6)-10(-9) M), recombinant human tumor necrosis factor-[symbol; see text] (TNF-alpha, 0.05-100 ng/ml), and a combination of TNF-alpha with low-density lipoproteins (LDL, 100 microgram/ml). HUVEC were incubated with these substances for 6 or 24 h in parallel tests performed under aerobic (CO2-incubator) and ischemic conditions (a mixture of 95% N2 + 5% CO2 in RPMI-1640 medium containing no substrate additives, growth factor or protein). HUVEC viability was determined by counting cells adherent to the bottom of wells after 24 h of reincubation under aerobic conditions in the growth medium (Plating Efficiency Index). The data showed that: 1) CTE of these compounds were dose-dependent (H2O2 and TNF-alpha) and time-dependent (TNF-alpha); 2) CTE of FRR initiators and CTE of ischemia were synergistic, that is, their combination produced a greater decrease HUVEC viability than any substance examined or ischemia alone; 3) CTE of TNF-alpha observed in experiments in substrate-deficient, protein-free medium was considerably stronger than in the growth medium; 4) a combination of TNF-a and LDL caused a stronger CTE on HUVEC than either factor alone, and this synergism was more pronounced during incubation under ischemic conditions. Thus, the data indicate that FRR initiators and TNF-alpha + LDL particularly increase the severity of ischemic injuries of EC and therefore they can be factors which in hypercholesterolemic patiens predispose vascular wall to atherosclerosis.
Download PDF:
Reference: Bilenko M.V., Ladygina V.G., Fedosova S.V., Comparative evaluation of cytotoxic effect of hydrogen peroxide and tumor necrosis factor alpha on nonischemic and ischemic endothelial cells, Voprosy meditsinskoi khimii, 1998, vol: 44(3), 229-240.
References
1. Markey В.A., Phan S. H., Varani J. et al. (1990), Free Rad. Biol. Med., 9, 307-314. Scholar google search
2. Britigan B. E., Roeder T. L. and Shasby D. M. (1992), Blood, 79, 699-707. Scholar google search
3. Shatos M.A., Doherty JM. and Hoak J.C. (1991), Arteriosclerosis and Thrombosis, 11, 594-601. Scholar google search
4. Steinbrecher U. P., Parthasarathy S., Leake D. S. and Witztum J. L. (1984), Proc.Natl. Acad. Sci. USA, 81, 3883-3887. Scholar google search
5. Bilenko M.V., Fedosova S.V. and Ladygina V.G. (1996), Eur. Sec. Meet. ISHR Abstracts. J. Mol. Cell. Cardiol., 28, A31. Scholar google search
6. Биленко М. В., Ладыгина В. Г. и Федосова С.В. БЭБиМ, в печати. Scholar google search
7. Schuger L, Varani J., Marks R. M. et al. (1989), Lab. Invest. 61, 62-68. Scholar google search
8. Murphy H. S., Shayman J. A., Till G.O. et al. (1992), Am. J. Physiol., 263, 1, Ptl, L51-L59. Scholar google search
9. Adamson G. M. and Billings R. (1992), Arch. Biochem. Biophys., 294, 223-229. Scholar google search
10. Персидский Ю.В., Кудрявей Ю.И., Барштейн Ю.В. (1991), БЭБиМ, III, 3, 294-297. Scholar google search
11. Pohlman T.H. and Harlan J.M. Cell Immunology, (1989), 119, 41-52. Scholar google search
12. Yoshizumi M., Perrella M. A., Burnett J. С and Lee M. E. (1993), Circ. Res., 73, 205-209. Scholar google search
13. Seidel S. L. and Strong R (1986), Hypertension, 8, 103-108. Scholar google search
14. Crawford D. W. and Kramsch D. M. (1988), Exp. Mol. Pathology, 49, 215-233. Scholar google search
15. Schinetti M.L., Sbarbati R and Scarlattini M. (1989), Cardiovasc. Res., 23, 1, 76-80. Scholar google search
16. Babbs С. F, Cregor M. D. and Badylak S. F. (1992), Cardiovasc. Res. 26, 6, 593-602. Scholar google search
17. Dauber I. M., Parsons P. K, Welsh С. М. et al. (1993),Circulation, 88, 2, 726-735. Scholar google search
18. Squadhto F, Altavilla D., Zingarelli B. et al. (1993), Eur. J. Pharmacol., 237, 2- 3, 223-230. Scholar google search
19. Strenbergh W.С. 3rd, Tuttle T. M, Makhoul R G. et al. (1994), J. Vase. Surgery, 20, 3, 474481. Scholar google search
20. LangelerE. G., Fiers W. and van-Hinsbergh V. W. (1991), Arteriosclerosis and Thrombosis, 11, 4, 872-881. Scholar google search
21. Bilenko M.V., Sevanian A. and Hochstein P. (1993), XIV Eur. Sec. Meet. ISHR. Abstracts. J. Mol. Cell. Cardiol., 25, Suppl. 1, 85. Scholar google search
22. Hodis H.N., Kramsch D.M., Sevanian A. et al. (1994), J. Lipid Res., 35,669-677. Scholar google search
23. Havel R. G, Eder H. and Bragdon J. J. Clin. Invest., (1955), 34, 1345-1353. Scholar google search
24. Ushiyama M. and Michara M. (1978), Anal. Biochem., 86, 271-278. Scholar google search
25. Franzini E., Sellak H., Marquetty С et al. (1996), Free Rad. Biol. Med., 21, 1, 15-23. Scholar google search
26. Nawroth P. P., Bank I., Handley D. et al. (1986), J. Exp. Med., 163, 6, 1363-1375. Scholar google search
27. Spyhdopoulos I, Brogi E., Kearney M. et al. (1997), J. Mol. Cell. Cardiol., 29, 1321-1330. Scholar google search
28. Delius R. E. and Hinshaw D. B. J. (1991), Surg. Res., 50, 4, 314-322. Scholar google search
29. Биленко М.В. (1989).Ишемические и реперфузионные повреждения органов. Медицина, Москва. Scholar google search
30. Bhat G. В., Tinsley S. В., Tolson J. К. et al. (1992), J. Cell. Physiol., 151, 2, 228-238. Scholar google search
31. Li R. K., Shaikh N., Weisel R. et al. (1992), J. Mol. Cell. Cardiol., 24, 6, 595-604. Scholar google search
32. Varani J., Phan S. H., Gibbs D. F. et al. (1990), Lab. Invest., 63, 5, 683-689. Scholar google search
33. Whorton A.R., Montgomery M.E. and Kent R.S. (1985), J. Clin. Invest., 76, 295-302. Scholar google search
34. Polunovsky V.A., Wendt C.H., Ingbar D.H. et al. (1994), Exp. Cell. Res., 214, 584-594. Scholar google search
35. Hiraishi H., Ferano A., Razandi M. et al. (1992), J. Biol. Chem., 267, 14812-14817. Scholar google search
36. Lennon S. V., Martin S. J. and Cotter T. G. (1991), Cell. Prolif, 24, 203-214. Scholar google search
37. Соловьев А.С., Коробко В.Г., Шингарова Л.Н. и др. (1995), Биохимия, 60, 1283-1292. Scholar google search
38. Биленко М.В., Вахрушева Т.В. и Федосова СВ. БЭБиМ, в печати. Scholar google search
39. Frolkis I, Gurevitch J., Yuhas Y. (1997), XVIII Eur. Section Meeting of the ISHR. Abstracts. J. Mol. Cell. Cardiol., 29, 5, Sal, A107. Scholar google search
40. Nowak T.S., Fried R.L., Lust W.D. and Passonneau J. V. (1985), J. Neurochem., 44, 487-494. Scholar google search