The effect of interferon γ on endothelial cell nitric oxide production and apoptosis

Authors
Shaghayegh Haghjooy Javanmard
Nasim Dana
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.  Back to cited text no. 1
    
2. Pober JS, Cotran RS. The role of endothelial cells in inflammation. Transplantation 1990;50:537-44.  Back to cited text no. 2
[PUBMED]    
3. Pober JS, Min W, Bradley JR. Mechanisms of endothelial dysfunction, injury, and death. Annu Rev Pathol 2009;4:71-95.  Back to cited text no. 3
[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.  Back to cited text no. 4
    
5. Steinberg D. Atherogenesis in perspective: Hypercholesterolemia and inflammation as partners in crime. Nat Med 2002;8:1211-7.  Back to cited text no. 5
    
6. Aird WC. Endothelial cell heterogeneity and Atherosclerosis. Curr Atheroscler Rep 2006;8:69-75.  Back to cited text no. 6
    
7. Flammer AJ, Lüscher TF. Human endothelial dysfunction: EDRFs. Pflugers Arch 2010;459:1005-13.  Back to cited text no. 7
    
8. Cooke JP. The pivotal role of nitric oxide for vascular health. Can J Cardiol 2004;20 Suppl B:7B-15B.  Back to cited text no. 8
    
9. Tellides G, Pober JS. Interferon-gamma axis in graft arteriosclerosis. Circ Res 2007;100:622-32.  Back to cited text no. 9
    
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.  Back to cited text no. 10
    
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.  Back to cited text no. 11
    
12. Miller CH. Maher SG. Young HA. Clinical Use of Interferon-gamma. Ann N Y Acad Sci 2009;1182:69-79.  Back to cited text no. 12
    
13. Billiau A, Matthys P. Interferon-gamma: A historical perspective. Cytokine Growth Factor Rev 2009;20:97-113.  Back to cited text no. 13
    
14. McLaren JE, Ramji DP. Interferon gamma: A master regulator of atherosclerosis. Cytokine Growth Factor Rev 2009;20:125-35.  Back to cited text no. 14
    
15. Schoenborn JR, Wilson CB. Regulation of interferon-gamma during innate and adaptive immune responses. Adv Immunol 2007;96:41-101.  Back to cited text no. 15
    
16. Boehm U, Klamp T, Groot M, Howard JC. Cellular responses to interferon-gamma. Annu Rev Immunol 1997;15:749-95.  Back to cited text no. 16
    
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.  Back to cited text no. 17
    
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.  Back to cited text no. 18
    
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.  Back to cited text no. 19
    
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.  Back to cited text no. 20
    
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.  Back to cited text no. 21
    
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.  Back to cited text no. 22
    
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.  Back to cited text no. 23
    
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.  Back to cited text no. 24
    
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.  Back to cited text no. 25
    
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.  Back to cited text no. 26
    
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.  Back to cited text no. 27
    
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.  Back to cited text no. 28
    
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.  Back to cited text no. 29
    
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.  Back to cited text no. 30
    
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.  Back to cited text no. 31
    
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.  Back to cited text no. 32
    
33. Kubes P, Suzuki M, Granger DN. Nitric oxide: An endogenous modulator of leukocyte adhesion. Proc Natl Acad Sci USA 1991;88:4651-5.  Back to cited text no. 33
    
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.  Back to cited text no. 34
    
Advanced Biomedical Research
Volume 2012, October
October 2012
Pages 1-5