Kinetics of increased generation of ·NO in endotoxaemic rats as measured by EPR.

• Authors: Przemyslaw M Plonka , Stefan Chlopicki , Magdalena Wisniewska , >Beata K Plonka
• Languages of publication: EN

Abstracts

EN

Ferrous-diethyldithiocarbamate (Fe(DETC)2) chelate is a lipophilic spin trap developed for g·NO detection by electron paramagnetic resonance (EPR) spectroscopy. Using this spin trap we investigated the kinetics of ·NO production in endotoxaemia in rats induced by lipopolysaccharide (Escherichia coli, 10 mg/kg). The NO-Fe(DETC)2 complex was found to give a characteristic EPR signal, and the amplitude of the 3rd (high-field) component of its hyperfine splitting was used to monitor the level of ·NO. We found that in blood, kindey, liver, heart and lung ·NO production starts to increase as early as 2 h after LPS injection, reaches the maximum 6 h after LPS injection and then returns to basal level within further 12-18 h. Interestingly, in the eye bulb the maximum of ·NO production was detected 12 h after LPS, and the signal was still pronounced 24 h after LPS. In brief, the highly lipophilic exogenous spin trap, Fe(DETC)2 is well suited for assessment of ·NO production in endotoxaemia. We demonstrated that the kinetics of increased production of ·NO in endotoxaemic organs, with the notable exception of the eye, do not follow the known pattern of NOS-2 induction under those conditions. Accordingly, only in early endotoxaemia a high level of ·NO is detected, while in late endotoxaemia ·NO detectability is diminished most probably due to concomitant oxidant stress.

Keywords

EN

NO-metry; DETC; NOS; eye; septic shock; oxidative stress

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2003
• Volume: 50
• Issue: 3
• Pages: 807-813

Dates

published

2003

received

2003-05-30

revised

2003-08-23

accepted

2003-09-04

Contributors

author

Przemyslaw M Plonka

• Department of Biophysics, Faculty of Biotechnology, Jagiellonian University, Kraków, Poland

author

Stefan Chlopicki

• Department of Pharmacology, Collegium Medicum of the Jagiellonian University, Kraków, Poland

author

Magdalena Wisniewska

• Department of Biophysics, Faculty of Biotechnology, Jagiellonian University, Kraków, Poland

author

>Beata K Plonka

• Department of Biophysics, Faculty of Biotechnology, Jagiellonian University, Kraków, Poland

References

• Andriambeloson E, Kleschyov AL, Muller B, Beretz A, Stoclet JC, Andriantsitohaina R. (1997) Nitric oxide production and endothelium-dependent vasorelaxation induced by wine polyphenols in rat aorta. Br J Pharmacol.; 120: 1053-8.
• Brandes RP, Koddenberg G, Gwinner W, Kim D, Kruse HJ, Busse R, Mugge A. (1999) Role of increased production of superoxide anions by NAD(P)H oxidase and xanthine oxidase in prolonged endotoxemia. Hypertension.; 33: 1243-9.
• Chlopicki S, Bartus JB, Walski M, Wolkow PP, Gryglewski RJ. (2001) Nitric oxide - a safeguard of pulmonary microcirculation in early endotoxaemia. In Nitric oxide. Basic research and clinical applications. Gryglewski RJ, Minuz P. eds, pp 137-48, IOS Press, Amsterdam.
• Cieslik K, Zhu Y, Wu KK. (2002) Salicylate suppresses macrophage nitric-oxide synthase-2 and cyclo-oxygenase-2 expression by inhibiting CCAAT/enhancer-binding protein-beta binding via a common signaling pathway. J Biol Chem.; 277: 49304-10.
• Kirkeboen KA, Strand OA. (1999) The role of nitric oxide in sepsis - an overview. Acta Anaesthesiol Scand.; 43: 275-88.
• Kubes P. (2000) Inducible nitric oxide synthase: a little bit of good in all of us. Gut.; 47: 6-9.
• Kubrina LN, Caldwell WS, Mordvintcev PI, Malenkova IV, Vanin AF. (1992). EPR evidence for nitric oxide production from guanidino nitrogens of L-arginine in animal tissues in vivo. Biochim Biophys Acta.; 1099: 233-7.
• Laszlo F, Whittle BJ, Moncada S. (1995) Attenuation by nitrosothiol ·NO donors of acute intestinal microvascular dysfunction in the rat. Br J Pharmacol.; 115: 498-502.
• Mülsch A, Vanin A, Mordvintcev P, Hauschildt S, Busse R. (1992) ·NO accounts completely for the oxygenated nitrogen species generated by enzymic L-arginine oxygenation. Biochem J.; 288: 597-603.
• Plonka PM, Chlopicki S, Plonka BK, Jawien J, Gryglewski RJ. (1999) Endotoxaemia in rats: detection of nitrosyl-haemoglobin in blood and lung by EPR. Curr Topics Biophys.; 23(1): 47-53.
• Plonka PM, Wisniewska M, Chlopicki S, Elas M, Rosen JM. (2003) X-band and S-band EPR detection of nitric oxide in murine endotoxemia using spin trapping by ferro-di(N-(dithiocarboxy)sarcosine). Acta Biochim Polon., 50: 799-806.
• Rocha G, Baines MG, Deschenes J, Duclos A, Antecka E, Di Silvio M. (1997) Nitric oxide and transforming growth factor-beta levels during experimental uveitis in the rabbit. Can J Ophthalmol.; 32: 1-3.
• Salvemini D, Riley DP, Lennon PJ, Wang ZQ, Currie MG, Macarthur H, Misko TP. (1999) Protective effects of a superoxide dismutase mimetic and peroxynitrite decomposition catalysts in endotoxin-induced intestinal damage. Br J Pharmacol.; 127: 685-92.
• Shultz PJ, Raij L. (1992) Endogenously synthesized nitric oxide prevents endotoxin-induced glomerular thrombosis. J Clin Invest.; 90: 1718-25.
• Spain DA, Wilson MA, Garrison RN. (1994) Nitric oxide synthase inhibition exacerbates sepsis-induced renal hypoperfusion. Surgery.; 116: 322-30.
• Streilein JW, Stein-Streilein J. (2000) Does innate immune privilege exist? J Leukoc Biol.; 67: 479-87.
• Taylor AW, Yee DG, Streilein JW. (1998) Suppression of nitric oxide generated by inflammatory macrophages by calcitonin gene-related peptide in aqueous humor. Invest Ophthalmol Vis Sci.; 39: 1372-8.
• Thiemermann C. (1997) Nitric oxide and septic shock. Gen Pharmacol.; 29: 159-66.
• Vanin AN, Kleschyov AL. (1998) EPR studies and biological implications of nitrosyl nonheme iron complexes. In Nitric oxide in transplant rejection and anti-tumor defense. Lukiewicz SJ, Zweier JL. eds, pp 49-82. Kluwer Academic Publishers, Boston- Dordrecht-London.
• Vanin AF, Kubrina LN, Kurbanov IS, Mordvintcev PI, Khrapova NV, Galagan ME, Matkhanov EI. (1989) Iron as an inducer of nitric oxide formation in animal organisms. Biokhimiia.; 54: 1974-9 (in Russian).
• Westenberger U, Thanner S, Ruf HH, Gesonde K, Suter G, Trentz O. (1990) Formation of free radicals and nitric oxide derivative of hemoglobin in rats during shock syndrome. Free Radic Res Commun.; 11: 167-78.
• Yamashita T, Kawashima S, Ohashi Y, Ozaki M, Ueyama T, Ishida T, Inoue N, Hirata K, Akita H, Yokoyama M. (2000) Resistance to endotoxin shock in transgenic mice overexpressing endothelial nitric oxide synthase. Circulation.; 101: 931-7.