Authors
Department of physiology, Isfahan university of medical sciences, Isfahan, Iran
Abstract
Background: Nitric oxide (NO) is an important molecule in maintaining endothelial survival and normal function. It is a unique mediator, which may promote or suppress both inflammation and apoptosis. Endothelial cell (EC) injury, dysfunction, and death in response to cytokines, especially interferon gamma (IFN-γ), represent the critical event for the initiation of several inflammatory diseases.
Objective(s): EC injury or death result in endothelial dysfunction that precedes the development of atherosclerosis and its subsequent vascular events. We examine the effect of different concentrations of IFN-γ on human umbilical vein ECs (HUVECs) NO production and apoptosis.
Materials and Methods: HUVECs were cultured at 37°C for 24 h in the absence (control) or presence of 10, 100, and 1000 μg IFN-γ, respectively. The apoptotic cells were determined as annexin V-positive propidium iodide (PI)-negative cells by flow cytometry. Total nitrite concentration was measured in cell cultures supernatant by Griess method.
Results: A comparison of the effect of IFN-γ on EC NO production with untreated cells showed that pretreatment of HUVEC with IFN-γ failed to have a significant effect on NO production by these cells at 10 and 100 U/mL, whereas it led to a significant decreased NO production at 1000 U/mL ( P < 0.05). The cells stimulated with IFN-γ showed significantly higher apoptotic cells (PI negative and annexin V-positive cells) after 24 h, compared with cells with no stimulations ( P < 0.05).
Conclusion: IFN-γ has detrimental effects on ECs in high doses. This might be due to inducible NO synthase activation.
Keywords
1. | Cotran RS, Pober JS. Endothelial activation and inflammation. ProgImmunol 1989;8:747. |
2. | Pober JS, Cotran RS. The role of endothelial cells in inflammation. Transplantation 1990;50:537-44. [PUBMED] |
3. | Pober JS, Min W, Bradley JR. Mechanisms of endothelial dysfunction, injury, and death. Annu Rev Pathol 2009;4:71-95. [PUBMED] |
4. | Hansson GK, Libby P. Schönbeck U, Yan ZQ. Innate and Adaptive Immunity in the Pathogenesis of Atherosclerosis. Circ Res 2002;91:281-.-91. |
5. | Steinberg D. Atherogenesis in perspective: Hypercholesterolemia and inflammation as partners in crime. Nat Med 2002;8:1211-7. |
6. | Aird WC. Endothelial cell heterogeneity and Atherosclerosis. Curr Atheroscler Rep 2006;8:69-75. |
7. | Flammer AJ, Lüscher TF. Human endothelial dysfunction: EDRFs. Pflugers Arch 2010;459:1005-13. |
8. | Cooke JP. The pivotal role of nitric oxide for vascular health. Can J Cardiol 2004;20 Suppl B:7B-15B. |
9. | Tellides G, Pober JS. Interferon-gamma axis in graft arteriosclerosis. Circ Res 2007;100:622-32. |
10. | Hansson GK, Jonasson L, Holm J, Clowes MM, Clowes AW. Gamma-interferon regulates vascular smooth muscle proliferation and Ia antigen expression in vivo and in vitro. Circ Res 1988;63:712-9. |
11. | Mach B, Steimle V, Martinez-Soria E, Reith W. Regulation of MHC class II genes: Lessons from a disease. Annu Rev Immunol 1996;14:301-31. |
12. | Miller CH. Maher SG. Young HA. Clinical Use of Interferon-gamma. Ann N Y Acad Sci 2009;1182:69-79. |
13. | Billiau A, Matthys P. Interferon-gamma: A historical perspective. Cytokine Growth Factor Rev 2009;20:97-113. |
14. | McLaren JE, Ramji DP. Interferon gamma: A master regulator of atherosclerosis. Cytokine Growth Factor Rev 2009;20:125-35. |
15. | Schoenborn JR, Wilson CB. Regulation of interferon-gamma during innate and adaptive immune responses. Adv Immunol 2007;96:41-101. |
16. | Boehm U, Klamp T, Groot M, Howard JC. Cellular responses to interferon-gamma. Annu Rev Immunol 1997;15:749-95. |
17. | Zohlnhofer D, Richte T, Neumann F, Nuhrenberg T, Wessely R, Brandl R, et al. Transcriptome analysis reveals a role of interferon-gamma in human neointima formation. Mol Cell 2001;7:1059-69. |
18. | Der SD, Zhou A, Williams BR, Silverman RH. Identification of genes differentially regulated by interferon alpha, beta, or gamma using oligonucleotide arrays. Proc Natl Acad Sci U S A 1998;95:15623-8. |
19. | Roselaar SE, Schonfeld G, Daugherty A. Enhanced development of atherosclerosis in cholesterol-fed rabbits by suppression of cell-mediated immunity. J Clin Invest 1995;96:1389-94. |
20. | Roselaar SE, Kakkanathu PX, Daugherty A. Lymphocyte populations in atherosclerotic lesions of apoE-/- and LDL receptor-/- mice: Decreasing density with disease progression. Arterioscler Thromb Vasc Biol 1996;16:1013-8. |
21. | Zhou XH, Stemme S, Hansson GK. Evidence for a local immune response in atherosclerosis: CD4(+) T cells infiltrate lesions of apolipoprotein-E-deficient mice. Am J Pathol 1996;149:359-66. |
22. | Emeson EE, Shen ML, Bell CG, Qureshi A. Inhibition of atherosclerosis in CD4 T-cell-ablated and nude (nu/nu) C57BL/6 hyperlipidemic mice. Am J Pathol 1996;149:675-85. |
23. | Zhou XH, Paulsson G, Stemme S, Hansson GK. Hypercholesterolemia is associated with a T helper (Th) 1/Th2 switch of the autoimmune response in atherosclerotic apo E-knockout mice. J Clin Invest 1998;101:1717-25. |
24. | Fong LG, Fong TA, Cooper AD. Inhibition of mouse macrophage degradation of acetyl low density lipoprotein by interferon-γ. J Biol Chem 1990;265:11751-60. |
25. | Li JH, Kluger MS, Madge LA, Zheng L, Bothwell AL, Pober JS. Interferon-γ augments CD95(APO-1/Fas) and procaspase-8 expression and sensitizes human vascular endothelial cells to CD95-mediated apoptosis. Am J Pathol 2002;161:1485-95. |
26. | Laroux FS, Pavlick KP, Hines IN, Kawachi S, Harada H, Bharwani S, et al. Role of nitric oxide in inflammation. Acta Physiol Scand 2001;173:113-8. |
27. | Kim YM, Talanian RV, Billiar TR. Nitric oxide inhibits apoptosis by preventing increases in caspase-3-like activity via two distinct mechanisms. J Biol Chem 1997;272:31138-48. |
28. | Javanmard SH, Nematbakhsh M, Mahmoodi F, Mohajeri MR. l-Arginine supplementation enhances eNOS expression in experimental model of hypercholesterolemic rabbits aorta. Pathophysiology 2009;16:9-13. |
29. | Marumo T, Nakaki T, Adachi H, Esumi H, Suzuki H, Saruta T, et al. Nitric oxide synthase mRNA in endothelial cells: Synergistic induction by interferon-gamma, tumor necrosis factor-alpha and lipopolysaccharide and inhibition by dexamethasone. Jpn J Pharmacol 1993;63:327-34. |
30. | Cornicelli JA, Butteiger D, Rateri DL, Welch K, Daugherty A. Interleukin-4 augments acetylated LDL induced cholesterol esterification in macrophages. J Lipid Res 2000;41:376-83. |
31. | Lamas S, Michel T, Collins T, Brenner BM, Marsden PA. Effects of interferon-gamma on nitric oxide synthase activity and endothelin-1 production by vascular endothelial cells. J Clin Invest 1992;90:879-87. |
32. | Morikawa A, Koide N, Kato Y, Sugiyama T, Chakravortty D, Yoshida T, et al. Augmentation of nitric oxide production by gamma interferon in a mouse vascular endothelial cell line and its modulation by tumor necrosis factor alpha and lipopolysaccharide. Infect Immun 2000;68:6209-14. |
33. | Kubes P, Suzuki M, Granger DN. Nitric oxide: An endogenous modulator of leukocyte adhesion. Proc Natl Acad Sci USA 1991;88:4651-5. |
34. | Yamaoka J, Kabashima K, Kawanishi M, Toda K, Miyachi Y. Cytotoxicity of IFN-gamma and TNF-alpha for vascular endothelial cell is mediated by nitric oxide. Biochem Biophys Res Commun 2002;291:780-6. |