Ratlarda Karbon Tetraklorür ile İndüklenen Karaciğer Toksikasyonu Üzerine Resveratrol’ün Koruyucu Etkisinin Histopatolojik ve TUNEL Metodu ile Araştırılması
Abstract
Bu
çalışmada, deneysel olarak karbon tetraklorür ile hepatotoksisite oluşturulan
ratların karaciğer dokuları üzerine güçlü antioksidan etkisine sahip olan Resveratrol’ün,
histopatoloji ve TUNEL metodu ile koruyucu etkinliğinin araştırılması
amaçlanmıştır. Bu amaçla, 32 adet erkek Wistar cinsi Albino rat kullanıldı.
Denekler rastgele seçilerek, 1. grup kontrol, 2. Grup CCl4, 3. grup Resveratrol.
4. grup ise CCl4+ Resveretrol olmak üzere 4 gruba ayrıldı. Deneme
sonunda hayvanlar anestezi altında sakrifiye edilerek histopatolojik analizler
için karaciğer örnekleri %10 formalin solüsyonuna alındı. Yapılan
histopatolojik incelemede kontrol ve Resveratrol grubunda ratların
karaciğerleri normal histolojik yapıda olduğu tespit edildi. CCI4
grubunda ise karaciğerin periasiner bölgelerindeki hepatositlerde şiddeti
hidropik dejenerasyon, steatozis ve kogulasyon nekrozu tesbit edildi. CCI4+
Resveretrol grubunda ratların karaciğerlerinde periasiner bölgede
hepatositlerde hafif hidropik dejenerasyona rastlanırken nekrotik hepatositlere
hiç rastlanmadı. Apoptozisi belirlemek amacıyla TUNEL metodu kullanıldı. TUNEL
sonuçlarına göre kontrol ve Resveratrol grubunda karaciğerlerde birkaç hücrede
pozitiflik tespit edildi. CCI4 grubunda karaciğerlerin periasiner
bölgelerindeki hepatositlerde çok sayıda pozitif hücre belirlendi. CCl4+Resveretrol
grubunda ratların karaciğerlerinde periasiner bölgede çok az sayıda pozitif
hücreye rastlandı. İstatiski olarak gruplar arasındaki fark önemli (P<0.05)
bulundu. Sonuç olarak, histopatolojik ve immunohistokimyasal bulgulara göre CCI4
neden olduğu oksidatif strese ve lipitperoksidasyonu sonucu oluşturulan
karaciğer hasarına karşı Resveratrolün bu hasarı önemli düzeyde inhibe edici
etkisinin olduğu belirlenmiştir.
References
- 1. Çavusoglu H., 1996. Tibbi Fizyoloji, 9 ed., Yüce Yayinlari, Alemdar Ofset, Istanbul. 2. Junqueira LC., Carneiro J., 2003. Basic Histology 10th ed., McGraw Hill Companies Incorpotated, New York. 3. Çinar A., Yörük M., Meral İ., Kiliçalp D., Koç A., Ertekin A., 1999. The effects of carbon tetrachloride (CCl4) induced experimental acute and chronic intoxication on hystological structure of liver and some hematological values and electrocardiogram in Rabbits. Turk J Vet Anim Sci, 23, 235-242. 4. Hong SW., Jung KH., Zheng HM., Lee HS., Suh JK., Park IS., Hong SS., 2010. The protective effect of Resveratrol on dimethylnitrosamine-induced liver fibrosis in rats. Arch Pharm Res, 33, 601-609. 5. Kasdallah-Grissa A., Mornagui B., Aouani E., Hammami M., Gharbi N., Kamoun A., El-Fazaa S., 2006. Protective effect of Resveratrol on ethanol-induced lipid peroxidation in rats. Alcohol Alcohol, 41, 236-239. 6. Tunalı-Akbay T., Sehirli O., Ercan F., Sener G., 2010. Resveratrol protects against methotrexate-induced hepatic injury in rats. JPPS, 13, 303-310. 7. Tahan G., Tarcin O., Tahan V., Eren F., Gedik N., Sahan E., Yucel O., 2007. The effects of N-acetylcysteine on bile duct ligation–induced liver fibrosis in rats. Dig Dis Sci., 52, 3348-3354. 8. Muriel P., Rivera‐Espinoza Y., 2008. Beneficial drugs for liver diseases. Journal of Applied Toxicology, 28, 93-103. 9. Wang C., Fan RQ., Zhang YX., Nie H., Li K., 2016. Naringenin protects against isoniazid-and rifampicin-induced apoptosis in hepatic injury. World J Gastroenterol, 22, 9775-9783. 10. Karakuş A., Değer Y., Yıldırım S., 2016. Protective effect of Silybum marianum and Taraxacum officinale extracts against oxidative kidney injuries induced by carbon tetrachloride in rats. Ren Fail, 39, 1-6. 11. Mukhopadhyay P., Mukherjee S., Ahsan K., Bagchi A., Pacher P., Das DK., 2010. Restoration of altered microRNA expression in the ischemic heart with Resveratrol. PLoS One, 5, e15705. 12. Bhuvaneswari R., Chidambaranathan N., Jegatheesan K., 2014. Hepatoprotective effect of embilica officinalis and its silver nanoparticles against ccl4 induced hepatotoxicity in wistar albino rats. DJNB, 9, 223-235. 13. MacDonald-Wickks LK., Garg ML., 2003. Vitamin E supplemantation in the mitigation of carbon tetrachloride induced oxidative stres in rats. Nutr Biochem, 14, 211-218. 14. Sotelo-Feliz JI., Martinez-Fong D., Muriel P., Santillan RL., Castillo D., Yahuaca P., 2002. Evaluation of the effectiveness of Rosmarinus officinalis (Lamiaceae) in the alleviation of carbon tetrachloride-induced acte hepatotoxicity in the rat. J Ethnopharmocol, 81, 145-154. 15. Galicia-Moreno M., Rodriguez-Rivera A., Reyes-Gordillo K., Segovia J., Shibayama M., Tsutsumi V., Muriel P., 2009. N-acetylcysteine prevents carbon tetrachloride-induced liver cirrhosis: role of liver transforming growth factor-beta and oxidative stress. Eur J Gastroenterol Hepatol, 21, 908-914. 16. Marchal J., Pifferi F., Aujard F., 2013. Resveratrol in mammals: effects on aging biomarkers, age‐related diseases, and life span. Ann N Y Acad Sci, 1290, 67-73. 17. Yıldırım S., Yener Z., 2016. Immunohistopathological and biochemical study of the effects of Dead Nettle (Urtica Dioica) extract on preventing liver lesions induced by experimental aflatoxicosis in Rats. WIMJ, DOI: 10.7727/wimj.2016.126. 18. Chan CC., Cheng LY., Lin CL., Huang YH., Lin HC., Lee FY., 2011. The protective role of natural phytoalexin Resveratrol on inflammation, fibrosis and regeneration in cholestatic liver injury. Molecular nutrition & food research, 55, 1841-1849. 19. Sinha K., Chaudhary G., Gupta YK., 2002. Protective effect of Resveratrol against oxidative stress in middle cerebral artery occlusion model of stroke in rats. Life sciences, 71, 655-665. 20. Chan CC., Lee KC., Huang YH., Chou CK., Lin HC., Lee FY., 2014. Regulation by Resveratrol of the cellular factors mediating liver damage and regeneration after acute toxic liver injury. JGH, 29, 603-613. 21. Bradamante S., Barenghi L., Villa A., 2004. Cardiovascular protective effects of Resveratrol. Cardiovascular Therapeutics, 22, 169-188. 22. Dinçel GÇ., Atasever A., Yaman D., 2016. Nitric oxide and glial fibrillary acidic protein (GFAP) expression in the liver parenchyma in carbon tetrachloride-induced hepatotoxicity. Kafkas Univ Vet Fak Derg, 22, 671-678. 23. Shi J., Aisaki K., Ikawa Y., Wake K., 1998. Evidence of hepatocyte apoptosis in rat liver after the administration of carbon tetrachloride. AJP, 153, 515-525. 24. Suzek H., Celik I., Dogan A., Yildirim S., 2016. Protective effect and antioxidant role of sweetgum (Liquidambar orientalis) oil against carbon tetrachloride-induced hepatotoxicity and oxidative stress in rats. Pharm Biol., 54, 451-457. 25. Kaneko M., Nagamine T., Nakazato K., Mori M., 2013. The anti-apoptotic effect of fucoxanthin on carbon tetrachloride-induced hepatotoxicity. J Toxicol Sci, 38, 115-126. 26. Öktem S., Özhan MH., Özol D., 2001. Apoptozisin önemi. Toraks Dergisi, 2, 91-95. 27. Karaman M., Özen H., Dağ S., Atakisi O., Cıgsar G., Kaya O., 2017. Ameliorative effect of omega-3 in carbon tetrachloride toxicity. Kafkas Univ Vet Fak Derg, 23(1), 77-85. 28. Khan RA., Khan MR., Sahreen S., Bokhari J., 2010. Prevention of CCl 4-induced nephrotoxicity with Sonchus asper in rat. FCT, 48, 2469-2476. 29. Paolillo R., Carratelli CR., Rizzo A., 2011. Effect of Resveratrol and quercetin in experimental infection by Salmonella enterica serovar Typhimurium. International immunopharmacology, 11, 149-156. 30. Schmatz R., Perreira LB., Stefanello N., Mazzanti C., Spanevello R., Gutierres J., Zanini D., 2012. Effects of Resveratrol on biomarkers of oxidative stress and on the activity of delta aminolevulinic acid dehydratase in liver and kidney of streptozotocin-induced diabetic rats. Biochimie, 94, 374-383.
Investigation by Histopathological and TUNEL Method of the Protective Effect of Resveratrol on Carbon Tetrachloride Induced Liver Toxicity in Rats
Abstract
In this study, it was aimed to investigate the
protective effect of Resveratrol, which has a strong antioxidant effect on
liver tissues, by histopathology and TUNEL method in experimental rats
hepatotoxic with carbon tetrachloride. For this reason 32 male Wistar rats were
used. Rats were randomly chosen and separated into 4 groups. First control,
rats in second group CCl4, 3rd group Resveratrol 4th group CCl4+Resveretrol. At
the end of the experiment, rats were euthanasied under anaesthesia and liver
samples were taken in formalin solution 10%. According to histopathological
examination, livers were in normal histopathological structure in the control
and Resveratrol groups. In CCl4 group, severe hydropic degeneration
in hepatocytes localised in the center regions of the liver, steatosis and
coagulation necrosis were detected. In CCl4+Resveretrol group; while
slight hydropic degeneration were detected in the central regions of livers,
necrotic hepatocytes were not detected. TUNEL method were used in order to
detect apoptosis. According to TUNEL results, positivity was detected in
certain liver cells in control and Resveratrol group. Numerous positive cells
were detected in central regions of livers in CCl4 group. Few
positive cells were detected in central region of rat livers in CCl4+Resveretrol
group. Differences in groups were statistically significant (P<0.05). As a
result of histopathological and immunohistochemical examination, inhibitor effect
of resvetratrol were determined against liver damage resulting from oxidative
stress and lipiperoxidation caused by CCl4.
References
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Resveratrol protects against methotrexate-induced hepatic injury in rats. JPPS, 13, 303-310. 7. Tahan G., Tarcin O., Tahan V., Eren F., Gedik N., Sahan E., Yucel O., 2007. The effects of N-acetylcysteine on bile duct ligation–induced liver fibrosis in rats. Dig Dis Sci., 52, 3348-3354. 8. Muriel P., Rivera‐Espinoza Y., 2008. Beneficial drugs for liver diseases. Journal of Applied Toxicology, 28, 93-103. 9. Wang C., Fan RQ., Zhang YX., Nie H., Li K., 2016. Naringenin protects against isoniazid-and rifampicin-induced apoptosis in hepatic injury. World J Gastroenterol, 22, 9775-9783. 10. Karakuş A., Değer Y., Yıldırım S., 2016. Protective effect of Silybum marianum and Taraxacum officinale extracts against oxidative kidney injuries induced by carbon tetrachloride in rats. Ren Fail, 39, 1-6. 11. Mukhopadhyay P., Mukherjee S., Ahsan K., Bagchi A., Pacher P., Das DK., 2010. Restoration of altered microRNA expression in the ischemic heart with Resveratrol. PLoS One, 5, e15705. 12. Bhuvaneswari R., Chidambaranathan N., Jegatheesan K., 2014. Hepatoprotective effect of embilica officinalis and its silver nanoparticles against ccl4 induced hepatotoxicity in wistar albino rats. DJNB, 9, 223-235. 13. MacDonald-Wickks LK., Garg ML., 2003. Vitamin E supplemantation in the mitigation of carbon tetrachloride induced oxidative stres in rats. Nutr Biochem, 14, 211-218. 14. Sotelo-Feliz JI., Martinez-Fong D., Muriel P., Santillan RL., Castillo D., Yahuaca P., 2002. Evaluation of the effectiveness of Rosmarinus officinalis (Lamiaceae) in the alleviation of carbon tetrachloride-induced acte hepatotoxicity in the rat. J Ethnopharmocol, 81, 145-154. 15. Galicia-Moreno M., Rodriguez-Rivera A., Reyes-Gordillo K., Segovia J., Shibayama M., Tsutsumi V., Muriel P., 2009. N-acetylcysteine prevents carbon tetrachloride-induced liver cirrhosis: role of liver transforming growth factor-beta and oxidative stress. Eur J Gastroenterol Hepatol, 21, 908-914. 16. Marchal J., Pifferi F., Aujard F., 2013. Resveratrol in mammals: effects on aging biomarkers, age‐related diseases, and life span. Ann N Y Acad Sci, 1290, 67-73. 17. Yıldırım S., Yener Z., 2016. Immunohistopathological and biochemical study of the effects of Dead Nettle (Urtica Dioica) extract on preventing liver lesions induced by experimental aflatoxicosis in Rats. WIMJ, DOI: 10.7727/wimj.2016.126. 18. Chan CC., Cheng LY., Lin CL., Huang YH., Lin HC., Lee FY., 2011. The protective role of natural phytoalexin Resveratrol on inflammation, fibrosis and regeneration in cholestatic liver injury. Molecular nutrition & food research, 55, 1841-1849. 19. Sinha K., Chaudhary G., Gupta YK., 2002. Protective effect of Resveratrol against oxidative stress in middle cerebral artery occlusion model of stroke in rats. Life sciences, 71, 655-665. 20. Chan CC., Lee KC., Huang YH., Chou CK., Lin HC., Lee FY., 2014. Regulation by Resveratrol of the cellular factors mediating liver damage and regeneration after acute toxic liver injury. JGH, 29, 603-613. 21. Bradamante S., Barenghi L., Villa A., 2004. Cardiovascular protective effects of Resveratrol. Cardiovascular Therapeutics, 22, 169-188. 22. Dinçel GÇ., Atasever A., Yaman D., 2016. Nitric oxide and glial fibrillary acidic protein (GFAP) expression in the liver parenchyma in carbon tetrachloride-induced hepatotoxicity. Kafkas Univ Vet Fak Derg, 22, 671-678. 23. Shi J., Aisaki K., Ikawa Y., Wake K., 1998. Evidence of hepatocyte apoptosis in rat liver after the administration of carbon tetrachloride. AJP, 153, 515-525. 24. Suzek H., Celik I., Dogan A., Yildirim S., 2016. Protective effect and antioxidant role of sweetgum (Liquidambar orientalis) oil against carbon tetrachloride-induced hepatotoxicity and oxidative stress in rats. Pharm Biol., 54, 451-457. 25. Kaneko M., Nagamine T., Nakazato K., Mori M., 2013. The anti-apoptotic effect of fucoxanthin on carbon tetrachloride-induced hepatotoxicity. J Toxicol Sci, 38, 115-126. 26. Öktem S., Özhan MH., Özol D., 2001. Apoptozisin önemi. Toraks Dergisi, 2, 91-95. 27. Karaman M., Özen H., Dağ S., Atakisi O., Cıgsar G., Kaya O., 2017. Ameliorative effect of omega-3 in carbon tetrachloride toxicity. Kafkas Univ Vet Fak Derg, 23(1), 77-85. 28. Khan RA., Khan MR., Sahreen S., Bokhari J., 2010. Prevention of CCl 4-induced nephrotoxicity with Sonchus asper in rat. FCT, 48, 2469-2476. 29. Paolillo R., Carratelli CR., Rizzo A., 2011. Effect of Resveratrol and quercetin in experimental infection by Salmonella enterica serovar Typhimurium. International immunopharmacology, 11, 149-156. 30. Schmatz R., Perreira LB., Stefanello N., Mazzanti C., Spanevello R., Gutierres J., Zanini D., 2012. Effects of Resveratrol on biomarkers of oxidative stress and on the activity of delta aminolevulinic acid dehydratase in liver and kidney of streptozotocin-induced diabetic rats. Biochimie, 94, 374-383.
Details
| Journal Section | Araştırma Makaleleri |
|---|---|
| Authors | |
| Publication Date | October 30, 2017 |
| Published in Issue | Year 2017 Volume: 12 Issue: 2 |