The metabolic pathways of chlorpromazine (CPZ) toxicity were tracked by assessing oxidative/nitrosative stress markers. The main objective of the study was to test the hypothesis that agmatine (AGM) prevents oxidative/nitrosative stress in the liver of Wistar rats 15 days after administration of CPZ. All tested substances were administered intraperitoneally (i.p.) for 15 consecutive days. The rats were divided into four groups: the control group (C, 0.9 % saline solution), the CPZ group (CPZ, 38.7 mg/kg b.w.), the CPZ+AGM group (AGM, 75 mg/kg b.w. immediately after CPZ, 38.7 mg/kg b.w. i.p.) and the AGM group (AGM, 75 mg/kg b.w.).
Rats were decapitated 15 days after the appropriate treatment. In the CPZ group, CPZ concentration was significantly increased compared to C values (p<0.01), while AGM treatment induced the significant decrease in CPZ concentration in the CPZ+AGM group (p<0.05) and the AGM group (p<0.01). CPZ application to healthy rats did not lead to any changes of lipid peroxidation in the liver compared to the C group, but AGM treatment decreased that parameter compared to the CPZ group (p<0.05). In CPZ liver homogenates, nitrite and nitrate concentrations were increased compared to controls (p<0.001), and AGM treatment diminished that parameter in the CPZ group (p<0.05), as well as in the AGM group (p<0.001). In CPZ animals, glutathione level and catalase activity were decreased in comparison with C values (p<0.01 respectively), but AGM treatment increased the activity of catalase in comparison with CPZ animals (p<0.05 respectively). Western blot analysis supported biochemical findings in all groups. Our results showed that treatment with AGM significantly supressed the oxidative/nitrosative stress parameters and restored antioxidant defense in rat liver.
1. Balijepalli S, Kenchappa RS, Boyd MR et al. Protein thiol oxidation by haloperidol results in inhibition of mitochondrial complex I in brain regions: comparison with atypical antipsychotics. Neurochem Int 2001; 38: 425-35. http://dx.doi.org/10.1016/S0197-0186(00)00108-X
2. Parikh V, Khan MM, Mahadik SP et al. Differential effects of antipsychotics on expression of antioxidant enzymes and membrane lipid peroxidation in rat brain. J Psychiatr Res 2003; 37: 43-51. http://dx.doi.org/10.1016/S0022-3956(02)00048-1
4. Sulaiman A, Al-Shawi N, Jwaied A et al. Protective effect of melatonin against chlorpromazine-induced liver disease in rats. Saudi Med J 2006; 27: 1477-82.
5. Tukov FF, Maddox JF, Amacher DE et al. Modeling inflammation-drug interactions in vitro: a rat Kupffer cell-hepatocyte coculture system. Toxicol In Vitro 2006; 20(8): 1488-99. http://dx.doi.org/10.1016/j.tiv.2006.04.005
6. Hassan HA, Yousef MI. Ameliorating effect of chicory (Cichorium intybus L.)-supplemented diet against nitrosamine precursors-induced liver injury and oxidative stress in male rats. Food Chem Toxicol 2010; 48: 2163-9. http://dx.doi.org/10.1016/j.fct.2010.05.023
7. Stankovic MN, Mladenovic D, Ninkovic M et al. The effects of α-lipoic acid on liver oxidative stress and free fatty acid composition in methionine-choline deficient diet-induced NAFLD. J Med Food 2014; 17: 254-61. http://dx.doi.org/10.1089/jmf.2013.0111
8. Maslak E, Zabielski P, Kochan K et al. The liverselective NO donor, V-PYRRO/NO, protects against liver steatosis and improves postprandial glucose tolerance in mice fed high fat diet. Biochem Pharmacol 2015; 93: 389-400. http://dx.doi.org/10.1016/j.bcp.2014.12.004
9. Sharipov RR, Kotsiuruba AV, Kop"iak BS et al. Induction of nitrosative stress in mitochondria of rats hearts in experimental ischemia-reperfusion of the brain and its correction by ecdysterone. Fiziol Zh 2014; 60: 3-13.
10. Akyol O, Herken H, Uz E et al. The indices of endogenous oxidative and antioxidative processes in plasma from schizophrenic patients. The possible role of oxidant/antioxidant imbalance. Prog Neuropsychopharmacol Biol Psychiatry 2002; 26: 995-1005. http://dx.doi.org/10.1016/S0278-5846(02)00220-8
11. Dejanovic B, Stevanovic I, Ninkovic M. et al. Protective effect of agmatine in acute chlorpromazine hepatotoxicity in rats. Acta Vet Brno 2014; 83: 305-12. http://dx.doi.org/10.2754/avb201483040305
12. Ostojic JN, Mladenovic D, Ninkovic M et al. The effects of cold-induced stress on liver oxidative injury during binge drinking. Hum Exp Toxicol 2012; 31: 387-96. http://dx.doi.org/10.1177/0960327111433899
13. Regunathan S, Youngson C, Raasch W et al. Imidazoline receptors and agmatine in blood vessels: a novel system inhibiting vascular smooth muscle proliferation. J Pharmacol Exp Ther 1996; 276: 1272-82.
14. Bhutada A, Ismail-Beigi F. Serum and growth factor induction of Na(+)-K(+)-ATPase subunit mRNAs in Clone 9 cells: role of protein kinase C. Am J Physiol 1991; 261: C699-707.
15. Li X, Fan X, Zheng ZH et al. Protective effects of agmatine on lipopolysaccharide-induced acute hepatic injury in mice. Zhonghua Wei Zhong Bing Ji Jiu Yi Xue 2013; 25: 720-4.
16. El-Agamy DS, Makled MN, Gamil NM. Protective effects of agmatine against Dgalactosamine and lipopolysaccharide-induced fulminant hepatic failure in mice. Inflammopharmacology 2014; 22: 187-94. http://dx.doi.org/10.1007/s10787-013-0188-2
17. Heitzman RJ. Veterinary drug residues. Commission of the European Communities, 2rd ed., 1994.
18. Gurd JW, Jones LR, Mahler HR et al. Isolation and partial characterization of rat brain synaptic membrane. J Neurochem 1974; 22: 281-90. http://dx.doi.org/10.1111/j.1471-4159.1974.tb11591.x
19. Lowry OH, Rosenbrongh NJ, Farr AL et al. Protein measurement with the folin phenol reagent. J Biol Chem 1951; 193: 265-75.
20. Girotti M, Khan N, Mc Lellan B. Early measurement of systemic lipid peroxidation products in the plasma of major blunt trauma patients. J Trauma 1991; 31: 32-5. http://dx.doi.org/10.1097/00005373-199101000-00007
21. Navarro-Gonzalez JA, Garcia-Benayas C, Arenas J. Semiautomated measurement of nitrate in biological fluids. Clin Chem 1998; 44: 679-81.
22. Anderson ME. The DTNB-GSSG reductase recycling assay for total glutathione (GSH + 1/2GSSG). In: Greenwald RA (ed), Tissue glutathione, Florida, CRC Press, 1986: 31723.
23. Góth L. A simple method for determination of serum catalase activity and revision of reference range. Clin Chim Acta 1991; 196: 143-51. http://dx.doi.org/10.1016/0009-8981(91)90067-M
24. Harry GJ, Schmitt TJ, Gong Z et al. Lead-induced alterations of glial fibrillary acidic protein (GFAP) in the developing rat brain. Toxicol Appl Pharmacol 1996; 139: 84-93. http://dx.doi.org/10.1006/taap.1996.0145
25. Couée I, Tipton KF. The inhibition of glutamate dehydrogenase by some antipsychotic drugs. Biochem Pharmacol 1990; 39: 827-32. http://dx.doi.org/10.1016/0006-2952(90)90196-R
26. Dejanovic B, Stevanovic I, Ninkovic M et al. Agmatine protection against chlorpromazineinduced forebrain cortex injury in rats. J Vet Sci 2015.
27. Di Ciero Miranda M, de Bruin VM, Vale MR et al. Lipid peroxidation and nitrite plus nitrate levels in brain tissue from patients with AD. Gerontology 2000; 46: 179-84. http://dx.doi.org/10.1159/000022156
28. Cardillo S, Iuliis AD, Battaglia V, Toninello A, Stevanato R, Vianello F. Novel copper amine oxidase activity from rat liver mitochondria matrix. Arch Biochem Biophys 2009; 485(2): 97-101. http://dx.doi.org/10.1016/j.abb.2009.03.006
29. Agostinelli E, Marques MPM, Calheiros R. Polyamines: fundamental characters in chemistry and biology. Amino Acids 2010; 38: 393-403. http://dx.doi.org/10.1007/s00726-009-0396-7
30. Lores-Arnaiz S, D'Amico G, Czerniczyniec A et al. Brain mitochondrial nitric oxide synthase: in vitro and in vivo inhibition by chlorpromazine. Arch Biochem Biophys 2004; 430: 170-7. http://dx.doi.org/10.1016/j.abb.2004.07.012
31. Lilic A, Dencic S, Pavlovic SZ et al. Activity of antioxidative defense enzymes in the blood of patients with liver echinococcosis. Vojnosanit Pregl 2007; 64: 235-40. http://dx.doi.org/10.2298/VSP0704235L
32. Bannai S, Sato H, Ishii T et al. Enhancement of glutathione levels in mouse peritoneal macrophages by sodium arsenite, cadmium chloride and glucose/glucose oxidase. Biochim Biophys Acta 1991; 1092: 175-9. http://dx.doi.org/10.1016/0167-4889(91)90153-O
33. Lopert P, Patel M. Brain mitochondria from DJ-1 knockout mice show increased respirationdependent hydrogen peroxide consumption. Redox Biol 2014; 2: 667-72. http://dx.doi.org/10.1016/j.redox.2014.04.010
34. Li T, Zhou Q, Zhang N et al. Toxic effects of chlorpromazine on Carassius auratus and its oxidative stress. J Environ Sci Health B 2008; 43: 638-43. http://dx.doi.org/10.1080/03601230802352674
35. Can B, Demirci A, Uysal YD et al. Contradictory effects of chlorpromazine on endothelial cells in a rat model of endotoxic shock in association with its actions on serum TNF-alpha levels and antioxidant enzyme activities. Pharmacol Res 2003; 48: 223-30. http://dx.doi.org/10.1016/S1043-6618(03)00093-8
36. Yabuki M, Tani N, Yoshioka T et al. Local thrombus formation in the site of intravenous injection of chlorpromazine: possible colloidosmotic lysis of the local endothelial cells. Biol Pharm Bull 2000; 23: 957-61. http://dx.doi.org/10.1248/bpb.23.957
37. Kelder PP, Fischer MJ, de Mol NJ et al. Oxidation of chlorpromazine by methemoglobin in the presence of hydrogen peroxide. Formation of chlorpromazine radical cation and its covalent binding to methemoglobin. Arch Biochem Biophys 1991; 284: 313-9. http://dx.doi.org/10.1016/0003-9861(91)90301-X
38. Koek GH, Liedorp PR, Bast A. The role of oxidative stress in non-alcoholic steatohepatitis. Clin Chim Acta 2011; 412: 1297-305. http://dx.doi.org/10.1016/j.cca.2011.04.013
39. Qi XM, Miao LL, Cai Y et al. ROS generated by CYP450, especially CYP2E1, mediate mitochondrial dysfunction induced by tetrandrine in rat hepatocytes. Acta Pharmacol Sin 2013; 34: 1229-36. http://dx.doi.org/10.1038/aps.2013.62