Sıçanlarda Karbon Tetraklorür İle Uyarılmış Beyin Hasarında Morus nigra'nın Koruyucu Mekanizması

Morus nigra (MN) tüm dünyada yaygın olarak kullanılan bir meyvedir. Hepatik ensefalopati (HE) hem akut hem de kronik karaciğer yetmezliğinde tehlikeli bir nöropsikiyatrik komplikasyondur ve son dönem karaciğer hastalığı olan hastalarda en sık görülen ölüm nedenidir. Bilgimize göre bu, karbon tetraklorür (CCI4) ile indüklenen sirozlu sıçan karaciğeri modeli üzerinde HE tedavisinde MN’nin etkilerinin araştırıldığı ilk çalışmadır. Bu çalışmada, karaciğer dokusunda aspartat aminotransferaz (AST) ve alanin aminotransferaz (ALT) seviyelerinin yanısıra beyin dokusunda 8-hidroksideoksiguanozin (8-OHdG) imünohistokimya ve Hematoksilen-eozin boyaması, superoksid dismutaz (SOD) ve katalaz (CAT) aktivite testleri kullanıldı. Bu amaçla 42 Spraque-Dawley erkek sıçan 6 gruba ayrıldı: kontrol, HE kontrol (1 mL/kg CCI4/haftada 2, intraperitonal), 125 mg/kg MN, 250 mg/kg MN, CCI4+125 mg/kg MN ve CCI4+250 mg/kg MN grupları. CCI4 sıçan karaciğerlerinde kontrol ile karşılaştırıldığında serum enzim seviyelerinde belirgin bir artışa sebep oldu. Diğer taraftan MN uygulaması biyokimyasal parametreleri doza bağımlı olarak önemli ölçüde düzeltti. 8-OHdG aktivitesi CCI4 grubunda belirgin olarak arttı. Ancak MN tedavisi ile bu artışlar önemli ölçüde azaldı. MN'nin intraperitoneal olarak uygulanması, kontrol grubuna kıyasla oksidatif stresi önemli ölçüde azaltırken, apoptozisi önledi ve beyindeki antioksidan savunma mekanizması aktivitesinde artışa neden oldu. MN ile tedavi edilen sıçan beyinleri CCI4 ile tedavi edilen sıçanlara göre daha az DNA hasarı gösterdi. Bu çalışmanın sonucu MN'nin anti-inflamatuar, antiapoptotik, antioksidan özelliklerinden dolayı nöroprotektif etkiye sahip olduğunu gösterdi.

Anahtar Kelimeler:

Morus nigra, hepatik ensephalopati, Karbon tetraklorit

Protective Mechanism of Morus nigra on Carbon Tetrachloride Induced Brain Damage in Rats

Morus nigra (MN) is a common fruit used all over the world. Hepatic encephalopathy (HE) is a dangerous neuropsychiatric complication of both acute and chronic liver failure, and is the most common cause of death in patients with end-stage liver disease. To our knowledge, this is the first study investigated the efficiencies of MN on the treatment of HE on cirrhotic rat liver model induced by carbon tetrachloride (CCI4). In this study it has been used superoxide dismutase (SOD) and catalase (CAT) activities assays, 8-hydroxydeoxyguanosine (8-OHdG) immunohistochemistry and Haematoxylin-Eosin (H&E) staining in the brain, as well as aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels in liver tissue With this aim, 42 Sprague-Dawley male rats were divided into six groups: control, HE control (1 mL/kg CCI4/twice a week, intraperitoneal), 125 mg/kg MN, 250 mg/kg MN, CCI4+125 mg/kg MN and CCI4+250 mg/kg MN groups. CCl4 caused a significant increase in serum enzyme levels in rat livers, compared with control. On the other hand, MN treatment restored the biochemical parameters significantly in a dose-dependent manner. 8-OHdG activity was significantly increased in CCI4 group. However, these increases were significantly decreased by MN treatment. While intraperitoneal administration of MN significantly reduced oxidative stress, prevented apoptosis and caused an increase in the antioxidant defence mechanism activity in brain compared to the control group. Brains of rats treated with MN showed less DNA damage than treated groups with CCI4. As a result of this study showed that MN has a neuroprotective effect because of its anti-inflammatory, anti-apoptotic, antioxidant properties.

Keywords:

Morus nigra hepatic encephalopathy, Carbon tetrachloride, Antioxidant enzymes,

PDF

___

1. Butterworth RF. Hepatic encephalopathy: a neuropsychiatric disorder involving multiple neurotransmitter systems. Cur Op Neurol. 2000;13:721–727.
2. Jalan R, Olde Damink SW, Hayes PC, Deutz NE, Lee A. Pathogenesis of intracranial hypertension in acute liver failure: inflammation, ammonia and cerebral blood flow. J Hepatol. 2004;41:613–20.
3. Rama Rao KV, Norenberg MD. Brain energy metabolism and mitochondrial dysfunction in acute and chronic hepatic encephalopathy. Neurochem Int. 2012;60:697–706.
4. Bosoi CR, Rose CF. Oxidative stress: a systemic factor implicated in the pathogenesis of hepatic encephalopathy. Metab Brain Dis. 2013;28:175–8.
5. Basu, S., Carbon tetrachloride-induced lipid peroxidation: eicosanoid formation and their regulation by antioxidant nutrients. Toxicology. 2003; 189, 113– 127.
6. Dani, C., Pasquali, M.A.B., Oliveira, M.R., Umezu, F.M., Salvador, M., Henriques, J.A.P., Moreira, J.C.F., Protective effects of purple grape juice on carbon tetrachloride-induced oxidative stress in brains of adult Wistar rats. J. Med. Food 2008; 11, 55–61.
7. Botsoglou, N.A., Taitzoglou, I.A., Botsoglou, E., Lavrentiadou, S., Kokoli, A.N., Roubies, N., Effect of long-term dietary administration of oregano on the alleviation of carbon tetrachloride -induced oxidative stress in rats. J. Agric. Food Chem. 2008; 56, 6287–6293.
8. Nomura, T. and Hano, Y., Isoprenoid-substituted phenolic compounds of moraceous plants. Natural Product Reports, 1994; 11, 2, 205-218. PMid:15209130. http://dx.doi.org/10.1039/np9941100205.
9. J. R. G. D. S. Almeida, A. L. Guimarães, A. P. de Oliveira et al., “Evaluation of hypoglycemic potential and pre-clinical toxicology of Morus nigra l. (Moraceae),” Latin American Journal of Pharmacy, 2011;30, 1, 96–100.
10. G. R. Souza, R. G. Oliveira-Junior, T. C. Diniz , A. Branco, S. R. G. Lima-Saraiva et al., Braz. J. Biol., ahead of print Epub Aug 17, 2017: Assessment of the antibacterial, cytotoxic and antioxidant activities of Morus nigra L. (Moraceae). Brazilian Journal of Biology. 1678-4375. doi.org/10.1590/1519-6984.05316
11. Mecocci P, Polidori MC. Antioxidant clinical trials in mild cognitive impairment and Alzheimer's disease. Biophys. Acta. 2012;1822:631–638. 12. J. R. G. D. S. Almeida, A. L. Guimarães, A. P. de Oliveira et al., “Evaluation of hypoglycemic potential and pre-clinical toxicology of Morus nigra l. (Moraceae),” Latin American Journal of Pharmacy, 2011;30, 1, 96–100
13. Abd El-Mawla A. M. A., Mohamed K. M., Mostafa A. M. Induction of biologically active flavonoids in cell cultures of Morus nigra and testing their hypoglycemic efficacy. Scientia Pharmaceutica. 2011;79(4):951–961. doi: 10.3797/scipharm.1101-1106.
14. Dalmagro AP1,2, Camargo A2, Zeni ALB3,4. Morus nigra and its major phenolic, syringic acid, have antidepressant-like and neuroprotective effects in mice. Metab Brain Dis. 2017 Aug 18. doi: 10.1007/s11011-017-0089-y.
15. Bémeur C, Butterworth RF (2013) Liver-brain proinflammatory signalling in acute liver failure: role in the pathogenesis of hepatic encephalopathy and brain edema. Metab Brain Dis 28:145–150. doi:10.1007/s11011-012-9361-3
16. Coballase-Urrutia E., Pedraza-Chaverri J., Cárdenas-Rodríguez N., Huerta-Gertrudis B., García-Cruz M.E., Ramírez-Morales A., Sánchez-González D.J., Martínez-Martínez C.M., Camacho-Carranza R., Espinosa-Aguirre J.J. Hepatoprotective effect of acetonic and methanolic extracts of Heterothecainuloides against CCl (4)-induced toxicity in rats. Exp. Toxicol. Pathol. 2011;63:363–370.
17. M. Bagchi, S. Ghosh, D. Bagchi, E. Hassoun, S.J. Stohs, Protective effects of lazaroid U74389F (16-desmethyl tirilazad) on endrin-induced lipid peroxidation and DNA damage in brain and liver and regional distribution of catalase activity in rat brain, Free Radic Biol Me. 19 (6) (1995) 867–872.
18. Khan RA. Protective effect of Launaea procumbens (L.) on lungs against CCl4-induced pulmonary damages in rat. BMC Complement Altern Med 2012; 12:133.
19. Ahmad B, Khan MR, Shah NA. Amelioration of carbon tetrachloride-induced pulmonary toxicity with Oxalis corniculata. Toxicol Ind Health 2015; 31:1243-1251.
20. Korhonen R, Lahti A, Kankaanranta H, Moilanen E. Nitric oxide production and signaling in inflammation. Curr Drug Targets Inflamm Allergy. 2005;4(4):471–479.
21. Boots A.W., Kubben N., Haenen G.R., Bast A. Oxidized quercetin reacts with thiols rather than with ascorbate: implication for quercetin supplementation. Biochem. Biophys. Res. Commun. 2003;308:560–565.
22. Lotito S.B., Frei B. Consumption of flavonoid-rich foods and increased plasma antioxidant capacity in humans: cause, consequence, or epiphenomenon? Free Radical Biol. Med. 2006;41:1727–1746.
23. A.R.T.Silva,A.C.F.Santos,J.M.Farfeletal.,“Repairofoxidative DNA damage, cell-cycle regulation and neuronal death may influence the clinical manifestation of Alzheimer’s disease,” PLoSONE,vol.9,no.6,ArticleIDe99897,2014.
24. Acharya C, Wade JB, Fagan A, White M, Gavis E, Ganapathy D, Gilles H, Heuman DM, Bajaj JS. Overt Hepatic Encephalopathy impairs learning on the Encephalapp Stroop which is Reversible after Liver Transplant. Liver Transpl. 2017. doi: 10.1002/lt.24864.