Lipopolisakkarit ile İndüklenen Sıçan Sepsis Modelinde S-Allil Sisteinin İnflamatuar Kaskat Üzerine Etkileri
Yıl 2021,
Cilt: 43 Sayı: 2, 94 - 105, 30.03.2021
Neslihan Tekin
,
Umut Kolaç
,
Cihan Tanrıkut
Mehmet Cengiz Üstüner
,
Emre Entok
,
Fahrettin Akyüz
,
İ. Özkan Alataş
Öz
Lipopolysaccharide (LPS) is a main constituent of Gram-negative bacterial cell walls and is considered a leading cause of sepsis. S-allyl cysteine (SAC) is a water-soluble organosulfur component present in garlic which has a potent antioxidant and free radical scavenger activity. The purpose of this study is to examine the antioxidant and anti-inflammatory potential of SAC on endotoxin LPS-induced sepsis. Female Wistar albino rats were divided into 6 groups. LPS (5 mg/kg) was applied to rats in sepsis and treatment groups intraperitoneally. After 24 hours from LPS injection 50 mg/kg and 100 mg/kg SAC was orally administrated to treatment groups. Lung and liver 18F-fluoro-deoxy-D-glucose (18F-FDG) uptake was measured by 18FDG-PET scan. Serum levels of nuclear factor-kappa B (NF-κβ), tumor necrosis factor-alpha (TNF-α), matrix metalloproteinase-9 (MMP-9), plasma levels of interleukin-1β (IL-1β), IL-6 and tissue levels of oxidative stress markers catalase (CAT), superoxide dismutase (SOD), malondialdehyde (MDA) and nitric oxide (NO) were determined. As a result of the study, MDA and NO levels of sepsis group were significantly higher than treatment groups in lung tissue. SOD activities of sepsis group was determined to significantly lower in the liver and lung tissues than the groups which were treated with SAC. Likewise, it was concluded that serum MMP-9, TNF-α and NF-κβ levels of sepsis group was significantly higher compared to levels of treatment groups. It was determined that SAC administration reduced 18F-FDG uptake in septic rats. In conclusion, SAC was observed to diminish effects of the acute toxicity and oxidative stress formed with LPS.
Kaynakça
- [1] Thimmulappa RK, Lee H, Rangasamy T, Reddy SP, Yamamoto M, Kensler TW, Biswal S. Nrf2 is a critic,al regulator of the innate immune response and survival during experimental sepsis. J Clin Invest. 2016;116:984-995.
- [2] Kolac UK, Ustuner MC, Tekin N, Ustuner D, Colak E, Entok E. The Anti-Inflammatory and Antioxidant Effects of Salvia officinalis on Lipopolysaccharide-Induced Inflammation in Rats. J Med Food. 2017;20:1193-1200.
- [3] Caroff M, Karibian D. Structure of bacterial lipopolysaccharides. Carbohydrate Res. 2003;338:2431-47.
- [4] Entok E, Ustuner MC, Ozbayer C, Tekin N, Akyuz F, Yangi B, Kurt H, Degirmenci I, Gunes HV. Anti-inflammatuar and anti-oxidative effects of Nigella sativa L.: (18)FDG-PET imaging of inflammation. Mol Biol Rep. 2014;41:2827-2834.
- [5] Armstrong L, Jordan N, Millar A. Interleukin 10 (IL-10) regulation of tumour necrosis factor alpha (TNF-alpha) from human alveolar macrophages and peripheral blood monocytes. Thorax. 1996;51:143-9.
- [6] Libby P. Inflammatory mechanisms: the molecular basis of inflammation and disease. Nutr Rev. 2007;65:S140-6.
- [7] Dinarello CA. The IL-1 family and inflammatory diseases. Clin Exp Rheumatol. 2002;20:S1-13.
- [8] de Souza P, Schulz R, da Silva-Santos JE. Matrix metalloproteinase inhibitors prevent sepsis-induced refractoriness to vasoconstrictors in the cecal ligation and puncture model in rats. Eur J Pharmacol. 2015;765:164-170.
- [9] Halliwell B, Gutteridge JM. Oxygen toxicity, oxygen radicals, transition metals and disease. Biochem J. 1984;219:1-14.
- [10] Devasagayam TP, Tilak JC, Boloor KK, Sane KS, Ghaskadbi SS, Lele RD. Free radicals and antioxidants in human health: current status and future prospects. J Assoc Physicians India. 2004;52:794-804.
- [11] Arreola R, Quintero-Fabián S, López-Roa RI, Flores-Gutiérrez EO, Reyes-Grajeda JP, Carrera-Quintanar L, Ortuño-Sahagún D. Ortuño-Sahagún, Immunomodulation and anti-inflammatory effects of garlic compounds. J Immunol Res. 2015;2015:401630.
- [12] You S, Nakanishi E, Kuwata H, Chen J, Nakasone Y, He X, He J, Liu X, Zhang S, Zhang B, Hou DX. Inhibitory effects and molecular mechanisms of garlic organosulfur compounds on the production of inflammatory mediator. Mol Nutr Food Res. 2013;57:2049-2060.
- [13] Bayraktar O, Tekin N, Aydın O, Akyuz F, Musmul A, Burukoglu D. Effects of S-allyl cysteine on lung and liver tissue in a rat model of lipopolysaccharide-induced sepsis. Naunyn Schmiedebergs Arch Pharmacol. 2015;388:327-335.
- [14] Brady Z, Taylor ML, Haynes M, Whitaker M, Mullen A, Clews L, Partridge M, Hicks RJ, Trapp JV. The clinical application of PET/CT: a contemporary review. Australas Phys Eng S. 2008;31:90-109.
- [15] Krause BJ, Beyer T, Bockisch A, Delbeke D, Kotzerke J, Minkov V, Reiser M, Willich N. FDG-PET/CT in oncology. Nuklearmedizin. 2007;46:291-301.
- [16] Cortas NK, Wakid NW. Determination of inorganic nitrate in serum and urine by a kinetic cadmium-reduction method. Clin Chem. 1990;36:1440-1443.
- [17] Lowe VJ, Naunheim KS. Current role of positron emission tomography in thoracic oncology. Thorax. 1998;53:703-712.
- [18] Seydanoğlu A. Dose-Dependent Effects of Octreotide on Oxidant-Antioxidant Status and Lung Histopathology During Experimental Sepsis, Selçuk Üniversitesi Tıp Fakültesi. Dissertation, Selcuk University, 2006.
- [19] Tsukioka T, Takemura S, Minamiyama Y, Mizuguchi S, Toda M, Okada S. Attenuation of Bleomycin-Induced Pulmonary Fibrosis in Rats with S-Allyl Cysteine. Molecules. 2017;22:543.
- [20] Kim KM, Chun SB, Koo MS, Choi WJ, Kim TW, Kwon YG, Chung HT, Billiar TR, Kim YM. Differential regulation of NO availability from macrophages and endothelial cells by the garlic component S-allyl cysteine. Free Radical Bio Med. 2001;30:747-756.
- [21] Numagami Y, Ohnishi ST. S-allylcysteine inhibits free radical production, lipid peroxidation and neuronal damage in rat brain ischemia. J Nutr. 2001;131:1100S-1105S.
- [22] Goetz FW, Planas JV, MacKenzie S. Tumor necrosis factors. Dev Comp Immunol. 2004;28:487-497.
- [23] Michie HR, Manogue KR, Spriggs DR, Revhaug A, O'Dwyer S, Dinarello CA, Cerami A, Wolff SM, Wilmore DW. Detection of circulating tumor necrosis factor after endotoxin administration. N Engl J Med. 1988;318:1481-1486.
- [24] Kabe Y, Ando K, Hirao S, Yoshida M, Handa H. Redox regulation of NF-κB activation: distinct redox regulation between the cytoplasm and the nucleus. Antioxid Redox Sign. 2005;7:395-403.
- [25] Fitzgerald DC1, Meade KG, McEvoy AN, Lillis L, Murphy EP, MacHugh DE, Baird AW. Tumour necrosis factor-α (TNF-α) increases nuclear factor κB (NFκB) activity in and interleukin-8 (IL-8) release from bovine mammary epithelial cells. Vet Immunol Immunopathol. 2007;116:59-68.
[26] Qin H, Wilson CA, Lee SJ, Zhao X, Benveniste EN. LPS induces CD40 gene expression through the activation of NF-κB and STAT-1α in macrophages and microglia. Blood. 2005;106:3114-3122.
- [27] Chandel NS, Trzyna WC, McClintock DS, Schumacker PT. Role of oxidants in NF-κB activation and TNF-α gene transcription induced by hypoxia and endotoxin. J Immunol. 2000;165:1013-1021.
- [28] Wang T,Zhang X, Li JJ. The role of NF-κB in the regulation of cell stress responses. Int Immunopharmacol. 2002;2:1509-1520.
- [29] Geng Z, Rong Y, Lau BH. S-allyl cysteine inhibits activation of nuclear factor kappa B in human T cells. Free Radical Bio Med. 1997;23:345-350.
- [30] Mong MC, Yin MC. Nuclear factor κB-dependent anti-inflammatory effects of s-allyl cysteine and s-propyl cysteine in kidney of diabetic mice. J Agr Food Chem. 2012;60:3158-3165.
- [31] Soyocak A, Kurt H, Özgen M, Coşan DT, Değirmenci İ, Çolak E, Güneş HV. Nuclear factor kappa B (NF-ΚB) activity according to grade of knee osteoarthritis patients. Osmangazi J Med. 2016;38:49-54.
- [32] Liu MW, Liu R, Wu HY, Zhang W, Xia J, Dong MN, Yu W, Wang Q, Xie FM, Wang R1, Huang YQ, Qian CY. Protective effect of Xuebijing injection on D-galactosamine-and lipopolysaccharide-induced acute liver injury in rats through the regulation of p38 MAPK MMP-9 and HO-1 expression by increasing TIPE2 expression. Int J Mol Med. 2016;38:1419-1432.
- [33] Trofimov A, Strekalova T, Mortimer N, Zubareva O, Schwarz A, Svirin E, Umriukhin A, Svistunov A, Lesch KP, Klimenko V. Klimenko, Postnatal LPS Challenge Impacts Escape Learning and Expression of Plasticity Factors Mmp9 and Timp1 in Rats: Effects of Repeated Training. Neurotox Res. 2017;32:175-186.
- [34] Kurt AN, Aygun AD, Godekmerdan A, Kurt A, Dogan Y, Yilmaz E. Serum IL-1β, IL-6, IL-8, and TNF-α levels in early diagnosis and management of neonatal sepsis. Mediators Inflamm. 2007;2007:31397.
- [35] Li R, Wang Y, Ma Z, Ma M, Wang D, Xie G, Yin Y1 Zhang P, Tao K. Maresin 1 Mitigates Inflammatory Response and Protects Mice from Sepsis. Mediators Inflamm. 2016;2016:3798465.
- [36] Çalış U, Turgut Cosan D, Saydam F, Kolac UK, Soyocak A, Kurt H, Gunes HV, Sahinturk V, Sahin Mutlu F, Ozdemir Koroglu Z, Degirmenci I.The Effects of Monosodium Glutamate and Tannic Acid on Adult Rats. Iran Red Crescent Med J. 2016;18:e37912.
- [37] Crimi E, Sica V, Williams-Ignarro S, Zhang H, Slutsky AS, Ignarro LJ, Napoli C. The role of oxidative stress in adult critical care. Free Radical Bio Med. 2006;40:398-406.
- [38] Zarezadeh M, Baluchnejadmojarad T, Kiasalari Z, Afshin-Majd S, Roghani M. Garlic active constituent s-allyl cysteine protects against lipopolysaccharide-induced cognitive deficits in the rat: possible involved mechanisms. Eur J Pharmacol. 2017;795:13-21.
- [39] Baluchnejadmojarad T, Kiasalari Z, Afshin-Majd S, Ghasemi Z, Roghani M. S-allyl cysteine ameliorates cognitive deficits in streptozotocin-diabetic rats via suppression of oxidative stress, inflammation, and acetylcholinesterase. Eur J Pharmacol. 2017;794:69-76.
- [40] Bergamini S, Rota C, Canali R, Staffieri M, Daneri F, Bini A, Giovannini F, Tomasi A, Iannone A. N-acetylcysteine inhibits in vivo nitric oxide production by inducible nitric oxide synthase. Nitric Oxide. 2001;5:349-360.
- [41] Kirkebøen KA, Strand ØA; The role of nitric oxide in sepsis–an overview. Acta Anaesthesiol Scand. 1999;43:275-288.
- [42] Yoshimura T, Yokoyama H, Fujii S, Takayama F, Oikawa K, Kamada H. In vivo EPR detection and imaging of endogenous nitric oxide in lipopolysaccharide-treated mice. Nat Biotechnol. 1996;14:992-4.
The Effects of S-Allyl Cysteıne on Inflammatory Cascade in Lipopolysaccharide Induced Rat Sepsis Model
Yıl 2021,
Cilt: 43 Sayı: 2, 94 - 105, 30.03.2021
Neslihan Tekin
,
Umut Kolaç
,
Cihan Tanrıkut
Mehmet Cengiz Üstüner
,
Emre Entok
,
Fahrettin Akyüz
,
İ. Özkan Alataş
Öz
Lipopolysaccharide (LPS) is a main constituent of Gram-negative bacterial cell walls and is considered a leading cause of sepsis. S-allyl cysteine (SAC) is a water-soluble organosulfur component present in garlic which has a potent antioxidant and free radical scavenger activity. The purpose of this study is to examine the antioxidant and anti-inflammatory potential of SAC on endotoxin LPS-induced sepsis. Female Wistar albino rats were divided into 6 groups. LPS (5 mg/kg) was applied to rats in sepsis and treatment groups intraperitoneally. After 24 hours from LPS injection 50 mg/kg and 100 mg/kg SAC was orally administrated to treatment groups. Lung and liver 18F-fluoro-deoxy-D-glucose (18F-FDG) uptake was measured by 18FDG-PET scan. Serum levels of nuclear factor-kappa B (NF-κβ), tumor necrosis factor-alpha (TNF-α), matrix metalloproteinase-9 (MMP-9), plasma levels of interleukin-1β (IL-1β), IL-6 and tissue levels of oxidative stress markers catalase (CAT), superoxide dismutase (SOD), malondialdehyde (MDA) and nitric oxide (NO) were determined. As a result of the study, MDA and NO levels of sepsis group were significantly higher than treatment groups in lung tissue. SOD activities of sepsis group was determined to significantly lower in the liver and lung tissues than the groups which were treated with SAC. Likewise, it was concluded that serum MMP-9, TNF-α and NF-κβ levels of sepsis group was significantly higher compared to levels of treatment groups. It was determined that SAC administration reduced 18F-FDG uptake in septic rats. In conclusion, SAC was observed to diminish effects of the acute toxicity and oxidative stress formed with LPS.
Destekleyen Kurum
Eskisehir Osmangazi Üniversitesi
Teşekkür
We are grateful to Eskisehir Osmangazi University for the use of the Medical Biology Laboratory and Medical. This study was funded by Eskisehir Osmangazi University Scientific Research Commission. Contract grant number: 201611D29
Kaynakça
- [1] Thimmulappa RK, Lee H, Rangasamy T, Reddy SP, Yamamoto M, Kensler TW, Biswal S. Nrf2 is a critic,al regulator of the innate immune response and survival during experimental sepsis. J Clin Invest. 2016;116:984-995.
- [2] Kolac UK, Ustuner MC, Tekin N, Ustuner D, Colak E, Entok E. The Anti-Inflammatory and Antioxidant Effects of Salvia officinalis on Lipopolysaccharide-Induced Inflammation in Rats. J Med Food. 2017;20:1193-1200.
- [3] Caroff M, Karibian D. Structure of bacterial lipopolysaccharides. Carbohydrate Res. 2003;338:2431-47.
- [4] Entok E, Ustuner MC, Ozbayer C, Tekin N, Akyuz F, Yangi B, Kurt H, Degirmenci I, Gunes HV. Anti-inflammatuar and anti-oxidative effects of Nigella sativa L.: (18)FDG-PET imaging of inflammation. Mol Biol Rep. 2014;41:2827-2834.
- [5] Armstrong L, Jordan N, Millar A. Interleukin 10 (IL-10) regulation of tumour necrosis factor alpha (TNF-alpha) from human alveolar macrophages and peripheral blood monocytes. Thorax. 1996;51:143-9.
- [6] Libby P. Inflammatory mechanisms: the molecular basis of inflammation and disease. Nutr Rev. 2007;65:S140-6.
- [7] Dinarello CA. The IL-1 family and inflammatory diseases. Clin Exp Rheumatol. 2002;20:S1-13.
- [8] de Souza P, Schulz R, da Silva-Santos JE. Matrix metalloproteinase inhibitors prevent sepsis-induced refractoriness to vasoconstrictors in the cecal ligation and puncture model in rats. Eur J Pharmacol. 2015;765:164-170.
- [9] Halliwell B, Gutteridge JM. Oxygen toxicity, oxygen radicals, transition metals and disease. Biochem J. 1984;219:1-14.
- [10] Devasagayam TP, Tilak JC, Boloor KK, Sane KS, Ghaskadbi SS, Lele RD. Free radicals and antioxidants in human health: current status and future prospects. J Assoc Physicians India. 2004;52:794-804.
- [11] Arreola R, Quintero-Fabián S, López-Roa RI, Flores-Gutiérrez EO, Reyes-Grajeda JP, Carrera-Quintanar L, Ortuño-Sahagún D. Ortuño-Sahagún, Immunomodulation and anti-inflammatory effects of garlic compounds. J Immunol Res. 2015;2015:401630.
- [12] You S, Nakanishi E, Kuwata H, Chen J, Nakasone Y, He X, He J, Liu X, Zhang S, Zhang B, Hou DX. Inhibitory effects and molecular mechanisms of garlic organosulfur compounds on the production of inflammatory mediator. Mol Nutr Food Res. 2013;57:2049-2060.
- [13] Bayraktar O, Tekin N, Aydın O, Akyuz F, Musmul A, Burukoglu D. Effects of S-allyl cysteine on lung and liver tissue in a rat model of lipopolysaccharide-induced sepsis. Naunyn Schmiedebergs Arch Pharmacol. 2015;388:327-335.
- [14] Brady Z, Taylor ML, Haynes M, Whitaker M, Mullen A, Clews L, Partridge M, Hicks RJ, Trapp JV. The clinical application of PET/CT: a contemporary review. Australas Phys Eng S. 2008;31:90-109.
- [15] Krause BJ, Beyer T, Bockisch A, Delbeke D, Kotzerke J, Minkov V, Reiser M, Willich N. FDG-PET/CT in oncology. Nuklearmedizin. 2007;46:291-301.
- [16] Cortas NK, Wakid NW. Determination of inorganic nitrate in serum and urine by a kinetic cadmium-reduction method. Clin Chem. 1990;36:1440-1443.
- [17] Lowe VJ, Naunheim KS. Current role of positron emission tomography in thoracic oncology. Thorax. 1998;53:703-712.
- [18] Seydanoğlu A. Dose-Dependent Effects of Octreotide on Oxidant-Antioxidant Status and Lung Histopathology During Experimental Sepsis, Selçuk Üniversitesi Tıp Fakültesi. Dissertation, Selcuk University, 2006.
- [19] Tsukioka T, Takemura S, Minamiyama Y, Mizuguchi S, Toda M, Okada S. Attenuation of Bleomycin-Induced Pulmonary Fibrosis in Rats with S-Allyl Cysteine. Molecules. 2017;22:543.
- [20] Kim KM, Chun SB, Koo MS, Choi WJ, Kim TW, Kwon YG, Chung HT, Billiar TR, Kim YM. Differential regulation of NO availability from macrophages and endothelial cells by the garlic component S-allyl cysteine. Free Radical Bio Med. 2001;30:747-756.
- [21] Numagami Y, Ohnishi ST. S-allylcysteine inhibits free radical production, lipid peroxidation and neuronal damage in rat brain ischemia. J Nutr. 2001;131:1100S-1105S.
- [22] Goetz FW, Planas JV, MacKenzie S. Tumor necrosis factors. Dev Comp Immunol. 2004;28:487-497.
- [23] Michie HR, Manogue KR, Spriggs DR, Revhaug A, O'Dwyer S, Dinarello CA, Cerami A, Wolff SM, Wilmore DW. Detection of circulating tumor necrosis factor after endotoxin administration. N Engl J Med. 1988;318:1481-1486.
- [24] Kabe Y, Ando K, Hirao S, Yoshida M, Handa H. Redox regulation of NF-κB activation: distinct redox regulation between the cytoplasm and the nucleus. Antioxid Redox Sign. 2005;7:395-403.
- [25] Fitzgerald DC1, Meade KG, McEvoy AN, Lillis L, Murphy EP, MacHugh DE, Baird AW. Tumour necrosis factor-α (TNF-α) increases nuclear factor κB (NFκB) activity in and interleukin-8 (IL-8) release from bovine mammary epithelial cells. Vet Immunol Immunopathol. 2007;116:59-68.
[26] Qin H, Wilson CA, Lee SJ, Zhao X, Benveniste EN. LPS induces CD40 gene expression through the activation of NF-κB and STAT-1α in macrophages and microglia. Blood. 2005;106:3114-3122.
- [27] Chandel NS, Trzyna WC, McClintock DS, Schumacker PT. Role of oxidants in NF-κB activation and TNF-α gene transcription induced by hypoxia and endotoxin. J Immunol. 2000;165:1013-1021.
- [28] Wang T,Zhang X, Li JJ. The role of NF-κB in the regulation of cell stress responses. Int Immunopharmacol. 2002;2:1509-1520.
- [29] Geng Z, Rong Y, Lau BH. S-allyl cysteine inhibits activation of nuclear factor kappa B in human T cells. Free Radical Bio Med. 1997;23:345-350.
- [30] Mong MC, Yin MC. Nuclear factor κB-dependent anti-inflammatory effects of s-allyl cysteine and s-propyl cysteine in kidney of diabetic mice. J Agr Food Chem. 2012;60:3158-3165.
- [31] Soyocak A, Kurt H, Özgen M, Coşan DT, Değirmenci İ, Çolak E, Güneş HV. Nuclear factor kappa B (NF-ΚB) activity according to grade of knee osteoarthritis patients. Osmangazi J Med. 2016;38:49-54.
- [32] Liu MW, Liu R, Wu HY, Zhang W, Xia J, Dong MN, Yu W, Wang Q, Xie FM, Wang R1, Huang YQ, Qian CY. Protective effect of Xuebijing injection on D-galactosamine-and lipopolysaccharide-induced acute liver injury in rats through the regulation of p38 MAPK MMP-9 and HO-1 expression by increasing TIPE2 expression. Int J Mol Med. 2016;38:1419-1432.
- [33] Trofimov A, Strekalova T, Mortimer N, Zubareva O, Schwarz A, Svirin E, Umriukhin A, Svistunov A, Lesch KP, Klimenko V. Klimenko, Postnatal LPS Challenge Impacts Escape Learning and Expression of Plasticity Factors Mmp9 and Timp1 in Rats: Effects of Repeated Training. Neurotox Res. 2017;32:175-186.
- [34] Kurt AN, Aygun AD, Godekmerdan A, Kurt A, Dogan Y, Yilmaz E. Serum IL-1β, IL-6, IL-8, and TNF-α levels in early diagnosis and management of neonatal sepsis. Mediators Inflamm. 2007;2007:31397.
- [35] Li R, Wang Y, Ma Z, Ma M, Wang D, Xie G, Yin Y1 Zhang P, Tao K. Maresin 1 Mitigates Inflammatory Response and Protects Mice from Sepsis. Mediators Inflamm. 2016;2016:3798465.
- [36] Çalış U, Turgut Cosan D, Saydam F, Kolac UK, Soyocak A, Kurt H, Gunes HV, Sahinturk V, Sahin Mutlu F, Ozdemir Koroglu Z, Degirmenci I.The Effects of Monosodium Glutamate and Tannic Acid on Adult Rats. Iran Red Crescent Med J. 2016;18:e37912.
- [37] Crimi E, Sica V, Williams-Ignarro S, Zhang H, Slutsky AS, Ignarro LJ, Napoli C. The role of oxidative stress in adult critical care. Free Radical Bio Med. 2006;40:398-406.
- [38] Zarezadeh M, Baluchnejadmojarad T, Kiasalari Z, Afshin-Majd S, Roghani M. Garlic active constituent s-allyl cysteine protects against lipopolysaccharide-induced cognitive deficits in the rat: possible involved mechanisms. Eur J Pharmacol. 2017;795:13-21.
- [39] Baluchnejadmojarad T, Kiasalari Z, Afshin-Majd S, Ghasemi Z, Roghani M. S-allyl cysteine ameliorates cognitive deficits in streptozotocin-diabetic rats via suppression of oxidative stress, inflammation, and acetylcholinesterase. Eur J Pharmacol. 2017;794:69-76.
- [40] Bergamini S, Rota C, Canali R, Staffieri M, Daneri F, Bini A, Giovannini F, Tomasi A, Iannone A. N-acetylcysteine inhibits in vivo nitric oxide production by inducible nitric oxide synthase. Nitric Oxide. 2001;5:349-360.
- [41] Kirkebøen KA, Strand ØA; The role of nitric oxide in sepsis–an overview. Acta Anaesthesiol Scand. 1999;43:275-288.
- [42] Yoshimura T, Yokoyama H, Fujii S, Takayama F, Oikawa K, Kamada H. In vivo EPR detection and imaging of endogenous nitric oxide in lipopolysaccharide-treated mice. Nat Biotechnol. 1996;14:992-4.