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Determination of 8-OHdG and 4-HNE Expressions in Sheep with Hepatic Lipidosis by Immunohistochemical Method

Year 2025, Volume: 15 Issue: 1, 9 - 15, 26.06.2025

Özhan Karataş , Gökhan Akçakavak , Murat Eren , Zeynep Çelik , Ayşenur Tural , Osman Dağar , Osman Doğan , Bahadır Kılınç , Mehmet Tuzcu

https://doi.org/10.53518/mjavl.1572721

Abstract

Lipidosis is generally defined as the accumulation of triglycerides in limited droplets within the cytoplasm of parenchymal cells. Lipidosis occurs due to toxic, chemical, infectious, and metabolic causes. This study aimed to reveal the local 8-hydroxy-2'-deoxyguanosine (8-OHdG) and 4-hydroxynonenal (4-HNE) expressions immunohistochemically according to the severity of the disease in sheep with hepatic lipidosis. The study material consisted of a total of 30 male sheep livers, including 6 healthy and 24 with hepatic lipidosis. After the liver samples were fixed in 10% formaldehyde solution, they underwent routine tissue processing to obtain paraffin blocks. Sections taken from the paraffin blocks were then subjected to Hematoxylin-Eosin (H-E) and immunohistochemical staining. Microscopically, control group liver samples showed normal histology. In livers with hepatic lipidosis, sharp-edged vacuoles of various sizes were detected in hepatocytes, and cell nuclei were pushed to the periphery. Additionally, focal hemorrhage and congestion, inflammatory cell infiltration in the portal area, bile duct proliferation, and connective tissue cells were observed. Microscopically, hepatic lipidosis cases were divided into two groups as moderate and severe based on the distribution of vacuoles in the section. In the immunohistochemical examination, 8-OHdG and 4-HNE expressions significantly increased in hepatic lipidosis cases compared to the control group (p<0.001). More intense immunoreactivity was detected especially in cases where disease severity increased (p<0.001). These results indicate that 8-OHdG and 4-HNE proteins play an important role in the pathogenesis of hepatic lipidosis and may be effective in increasing the severity of the disease.

Keywords

8-hydroxy-2 4-Hydroxynonenal hepatic lipidosis sheep

References

  • Akcakavak, G., Kazak, F., Karatas, O., Alakus, H., Alakus, I., Kirgiz, O., Celik, Z., Deveci MZY., Ozdemir, O., & Tuzcu, M. (2024). Eucalyptol regulates Nrf2 and NF-kB signaling and alleviates gentamicin-induced kidney injury in rats by downregulating oxidative stress, oxidative DNA damage, inflammation, and apoptosis. Toxicol. Mech. Methods, 34(4), 413-422.
  • Akcakavak, G., Kazak, F., & Deveci, MZY (2023). Eucalyptol Protects against Cisplatin-Induced Liver Injury in Rats. Biol. Bull. 50(5), 987-994.
  • Al-Habsi, K., Johnson, EH., Kadim, IT., Srikandakumar, A., Annamalai, K., Al- Busaidy, R., & Mahgoub, O. (2007). Effects of low concentrations of dietary cobalt on liveweight gains, haematology, serum vitamin B12 and biochemistry of Omani goats. Vet. J. 173(1), 131-137.
  • Altun, S., & Sağlam, Y. (2014). Erzurum ilinde kesimi yapılan sığırlarda karaciğer lezyonları üzerinde patolojik incelemeler. Atatürk Üni. Vet. Bilim. Derg. 9(1), 7-15.
  • Anderson, N., & Borlak, J. (2008). Molecular mechanisms and therapeutic targets in steatosis and steatohepatitis. Pharm. Rev. 60(3), 311-357.
  • Araujo, FB., Barosa, DS., Hsin, CY., Maranhão, RC., & Abdalla, DS. (1995). Evaluation of oxidative stress in patients with hyperlipidemia. Atherosclerosis, 117(1), 61-71.
  • Ayala, A., Muñoz, MF., & Argüelles, S. (2014). Lipid peroxidation: production, metabolism, and signaling mechanisms of malondialdehyde and 4‐hydroxy‐2‐nonenal. Oxid. Med. Cell Longev. 2014(1), 360438.
  • Boden, G. (1997). Role of fatty acids in the pathogenesis of insulin resistance and NIDDM. Diabetes, 46(1), 3-10.
  • Dinçel, GÇ., Yıldırım, S., & Kul, O. (2018). Role of nitric oxide and oxidative stress in pathophysiology of liver injury in streptozotocin-induced type 1 diabetic rats. Ankara Univ. Vet. Fak. Derg. 65, 39-50.
  • El-Khodery, S., Hussein, H., Metwally, N., Nassif, M., El-Boshy, M., & Galbat, S. (1994). Ultrasonographic and histopathological evaluation of hepatic lipidosis in sheep with cobalt deficiency. OJVR, 12, 23-32.
  • Elshafey, BG., Elfadadny, A., Metwally, S., Saleh, AG., Ragab, RF., Hamada, R., Mandour, AS., Hendawy, A. O., Alkazmi, L., & Ogaly, H. A. (2023). Association between biochemical parameters and ultrasonographic measurement for the assessment of hepatic lipidosis in dairy cows. Italian J. Anim. Sci. 22(1), 136-147.
  • Erer, H., Kıran, M., & Ciftci, M. (2009). Veterinary Genel Patoloji. In: Bahcivanlar Press, Konya, Türkiye, pp. 236.
  • Fu, A., Li, J., Ding, Q., Guo, R., Pi, A., Yang, W., Chen, Y., Dou, X., Song, Z., & Li, S. (2022). Upregulation of 4-hydroxynonenal contributes to the negative effect of n-6 polyunsaturated fatty acid on alcohol-induced liver injury and hepatic steatosis. J Agric Food Chem. 70(21), 6418-6428.
  • Goff, J., & Horst, R. (1997). Physiological changes at parturition and their relationship to metabolic disorders. J dairy sci. 80(7), 1260-1268.
  • Gözün, H., & Kıran, M. (1999). Konya mezbahalarında kesime alınan koyunların karaciğerinde patolojik incelemeler. Veterinarium, 10, 1-19.
  • Herndon, A., Breshears, M., & McFarlane, D. (2014). Oxidative modification, inflammation and amyloid in the normal and diabetic cat pancreas. J. Comp. Pathol. 151(4), 352-362.
  • Hiramatsu, K., Saigusa, Y., & Nakazawa, H. (1991). Low superoxide scavenging activity associated with enhanced superoxide generation by monocytes from male hypertriglyceridemia with and without diabetes. Atherosclerosis, 90(1), 39-47.
  • Iqbal, R., Beigh, S., Mir, A., Shaheen, M., Hussain, S., Nisar, M., & Dar, A. (2022). Evaluation of metabolic and oxidative profile in ovine pregnancy toxemia and to determine their association with diagnosis and prognosis of disease. Trop. Anim. Health Product. 54(6), 338.
  • Ji, X., Liu, N., Wang, Y., Ding, K., Huang, S., & Zhang, C. (2023). Pregnancy Toxemia in ewes: a review of Molecular Metabolic mechanisms and Management Strategies. Metabolites, 13(2), 149.
  • Johnson, E., Muirhead, D., Annamalai, K., King, G., Al-Busaidy, R., & Shahul Hameed, M. (1999). Hepatic lipidosis associated with cobalt deficiency in Omani goats. Vet. Res. Commun. 23, 215-221.
  • Johnson, E. H., Al-Habsi, K., Kaplan, E., Srikandakumar, A., Kadim, I. T., Annamalai, K., Al-Busaidy, R., & Mahgoub, O. (2004). Caprine hepatic lipidosis induced through the intake of low levels of dietary cobalt. Vet. J. 168(2), 174-179.
  • Jubb, KVF., Kennedy, PC., & Palmer, N. (2012). Pathology of domestic animals. Academic press. p. 701.
  • Kazak, F., Akcakavak, G., Alakus, I., Alakus, H., Kirgiz, O., Karatas, O., Deveci, MZY., & Coskun, P. (2024). Proanthocyanidin alleviates testicular torsion/detorsion-induced ischemia/reperfusion injury in rats. Tissue and Cell, 89, 102459.
  • Menzies, P., Langs, L., Boermans, H., Martin, J., & McNally, J. (2004). Myopathy and hepatic lipidosis in weaned lambs due to vitamin E deficiency. Canadian Vet. J. 45(3), 244.
  • Miyamoto, M., Kotani, K., Ishibashi, S., & Taniguchi, N. (2011). The relationship between urinary 8-hydroxydeoxyguanosine and metabolic risk factors in asymptomatic subjects. Med. Princ. and Pract. 20(2), 187- 190.
  • Oruc, E. (2009). Mezbahada kesilen sığırlarda karaciğer lezyonları üzerine histopatolojik bir çalışma. Atatürk Üniv. Vet. Bilim. Derg. 4(2), 97- 104.
  • Seki, S., Kitada, T., Yamada, T., Sakaguchi, H., Nakatani, K., & Wakasa, K. (2002). In situ detection of lipid peroxidation and oxidative DNA damage in non-alcoholic fatty liver diseases. J. Hepatol. 37(1), 56-62.
  • Sumida, Y., Yoneda, M., Seko, Y., Takahashi, H., Hara, N., Fujii, H., Itoh, Y., Yoneda, M., Nakajima, A., & Okanoue, T. (2021). Role of vitamin E in the treatment of non-alcoholic steatohepatitis. Free Radic. Biol. Med. 177, 391-403.
  • Thapa, B., & Walia, A. (2007). Liver function tests and their interpretation. Indian J. Pediatr. 74, 663-671.
  • Ulvund, M. J. (1990). Ovine white-liver disease (OWLD). Changes in blood chemistry. Acta Vet. Scand. 31(3), 277.
  • Varghese, J., Bhat, V., Chianeh, Y. R., Kamath, V., Al-Haj Husain, N., & Özcan, M. (2020). Salivary 8-hydroxyguanosine levels in smokers and non-smokers with chronic periodontitis. Odontology, 108, 569-577.
  • Webb, C., & Twedt, D. (2008). Oxidative stress and liver disease. Veterinary Clinics of North America: Small Anim. Pract. 38(1), 125-135.
  • Yang, Y., Sharma, R, Sharma, A., Awasthi, S., & Awasthi, Y. (2003). Lipid peroxidation and cell cycle signaling: 4-hydroxynonenal, a key molecule in stress mediated signaling. Acta Biochim. Pol. 50(2), 319-336.
  • Yeh, M. M., & Brunt, E. M. (2014). Pathological features of fatty liver disease. Gastroenterology, 147(4), 754-764.
  • Yoshino, K., Katoh, N., Takahashi, K., & Yuasa, A. (1992). Purification of a protein from serum of cattle with hepatic lipidosis, and identification of the protein as haptoglobin. Am. J. Vet. Res. 53(6), 951-956.

Year 2025, Volume: 15 Issue: 1, 9 - 15, 26.06.2025

Özhan Karataş , Gökhan Akçakavak , Murat Eren , Zeynep Çelik , Ayşenur Tural , Osman Dağar , Osman Doğan , Bahadır Kılınç , Mehmet Tuzcu

https://doi.org/10.53518/mjavl.1572721

Abstract

References

  • Akcakavak, G., Kazak, F., Karatas, O., Alakus, H., Alakus, I., Kirgiz, O., Celik, Z., Deveci MZY., Ozdemir, O., & Tuzcu, M. (2024). Eucalyptol regulates Nrf2 and NF-kB signaling and alleviates gentamicin-induced kidney injury in rats by downregulating oxidative stress, oxidative DNA damage, inflammation, and apoptosis. Toxicol. Mech. Methods, 34(4), 413-422.
  • Akcakavak, G., Kazak, F., & Deveci, MZY (2023). Eucalyptol Protects against Cisplatin-Induced Liver Injury in Rats. Biol. Bull. 50(5), 987-994.
  • Al-Habsi, K., Johnson, EH., Kadim, IT., Srikandakumar, A., Annamalai, K., Al- Busaidy, R., & Mahgoub, O. (2007). Effects of low concentrations of dietary cobalt on liveweight gains, haematology, serum vitamin B12 and biochemistry of Omani goats. Vet. J. 173(1), 131-137.
  • Altun, S., & Sağlam, Y. (2014). Erzurum ilinde kesimi yapılan sığırlarda karaciğer lezyonları üzerinde patolojik incelemeler. Atatürk Üni. Vet. Bilim. Derg. 9(1), 7-15.
  • Anderson, N., & Borlak, J. (2008). Molecular mechanisms and therapeutic targets in steatosis and steatohepatitis. Pharm. Rev. 60(3), 311-357.
  • Araujo, FB., Barosa, DS., Hsin, CY., Maranhão, RC., & Abdalla, DS. (1995). Evaluation of oxidative stress in patients with hyperlipidemia. Atherosclerosis, 117(1), 61-71.
  • Ayala, A., Muñoz, MF., & Argüelles, S. (2014). Lipid peroxidation: production, metabolism, and signaling mechanisms of malondialdehyde and 4‐hydroxy‐2‐nonenal. Oxid. Med. Cell Longev. 2014(1), 360438.
  • Boden, G. (1997). Role of fatty acids in the pathogenesis of insulin resistance and NIDDM. Diabetes, 46(1), 3-10.
  • Dinçel, GÇ., Yıldırım, S., & Kul, O. (2018). Role of nitric oxide and oxidative stress in pathophysiology of liver injury in streptozotocin-induced type 1 diabetic rats. Ankara Univ. Vet. Fak. Derg. 65, 39-50.
  • El-Khodery, S., Hussein, H., Metwally, N., Nassif, M., El-Boshy, M., & Galbat, S. (1994). Ultrasonographic and histopathological evaluation of hepatic lipidosis in sheep with cobalt deficiency. OJVR, 12, 23-32.
  • Elshafey, BG., Elfadadny, A., Metwally, S., Saleh, AG., Ragab, RF., Hamada, R., Mandour, AS., Hendawy, A. O., Alkazmi, L., & Ogaly, H. A. (2023). Association between biochemical parameters and ultrasonographic measurement for the assessment of hepatic lipidosis in dairy cows. Italian J. Anim. Sci. 22(1), 136-147.
  • Erer, H., Kıran, M., & Ciftci, M. (2009). Veterinary Genel Patoloji. In: Bahcivanlar Press, Konya, Türkiye, pp. 236.
  • Fu, A., Li, J., Ding, Q., Guo, R., Pi, A., Yang, W., Chen, Y., Dou, X., Song, Z., & Li, S. (2022). Upregulation of 4-hydroxynonenal contributes to the negative effect of n-6 polyunsaturated fatty acid on alcohol-induced liver injury and hepatic steatosis. J Agric Food Chem. 70(21), 6418-6428.
  • Goff, J., & Horst, R. (1997). Physiological changes at parturition and their relationship to metabolic disorders. J dairy sci. 80(7), 1260-1268.
  • Gözün, H., & Kıran, M. (1999). Konya mezbahalarında kesime alınan koyunların karaciğerinde patolojik incelemeler. Veterinarium, 10, 1-19.
  • Herndon, A., Breshears, M., & McFarlane, D. (2014). Oxidative modification, inflammation and amyloid in the normal and diabetic cat pancreas. J. Comp. Pathol. 151(4), 352-362.
  • Hiramatsu, K., Saigusa, Y., & Nakazawa, H. (1991). Low superoxide scavenging activity associated with enhanced superoxide generation by monocytes from male hypertriglyceridemia with and without diabetes. Atherosclerosis, 90(1), 39-47.
  • Iqbal, R., Beigh, S., Mir, A., Shaheen, M., Hussain, S., Nisar, M., & Dar, A. (2022). Evaluation of metabolic and oxidative profile in ovine pregnancy toxemia and to determine their association with diagnosis and prognosis of disease. Trop. Anim. Health Product. 54(6), 338.
  • Ji, X., Liu, N., Wang, Y., Ding, K., Huang, S., & Zhang, C. (2023). Pregnancy Toxemia in ewes: a review of Molecular Metabolic mechanisms and Management Strategies. Metabolites, 13(2), 149.
  • Johnson, E., Muirhead, D., Annamalai, K., King, G., Al-Busaidy, R., & Shahul Hameed, M. (1999). Hepatic lipidosis associated with cobalt deficiency in Omani goats. Vet. Res. Commun. 23, 215-221.
  • Johnson, E. H., Al-Habsi, K., Kaplan, E., Srikandakumar, A., Kadim, I. T., Annamalai, K., Al-Busaidy, R., & Mahgoub, O. (2004). Caprine hepatic lipidosis induced through the intake of low levels of dietary cobalt. Vet. J. 168(2), 174-179.
  • Jubb, KVF., Kennedy, PC., & Palmer, N. (2012). Pathology of domestic animals. Academic press. p. 701.
  • Kazak, F., Akcakavak, G., Alakus, I., Alakus, H., Kirgiz, O., Karatas, O., Deveci, MZY., & Coskun, P. (2024). Proanthocyanidin alleviates testicular torsion/detorsion-induced ischemia/reperfusion injury in rats. Tissue and Cell, 89, 102459.
  • Menzies, P., Langs, L., Boermans, H., Martin, J., & McNally, J. (2004). Myopathy and hepatic lipidosis in weaned lambs due to vitamin E deficiency. Canadian Vet. J. 45(3), 244.
  • Miyamoto, M., Kotani, K., Ishibashi, S., & Taniguchi, N. (2011). The relationship between urinary 8-hydroxydeoxyguanosine and metabolic risk factors in asymptomatic subjects. Med. Princ. and Pract. 20(2), 187- 190.
  • Oruc, E. (2009). Mezbahada kesilen sığırlarda karaciğer lezyonları üzerine histopatolojik bir çalışma. Atatürk Üniv. Vet. Bilim. Derg. 4(2), 97- 104.
  • Seki, S., Kitada, T., Yamada, T., Sakaguchi, H., Nakatani, K., & Wakasa, K. (2002). In situ detection of lipid peroxidation and oxidative DNA damage in non-alcoholic fatty liver diseases. J. Hepatol. 37(1), 56-62.
  • Sumida, Y., Yoneda, M., Seko, Y., Takahashi, H., Hara, N., Fujii, H., Itoh, Y., Yoneda, M., Nakajima, A., & Okanoue, T. (2021). Role of vitamin E in the treatment of non-alcoholic steatohepatitis. Free Radic. Biol. Med. 177, 391-403.
  • Thapa, B., & Walia, A. (2007). Liver function tests and their interpretation. Indian J. Pediatr. 74, 663-671.
  • Ulvund, M. J. (1990). Ovine white-liver disease (OWLD). Changes in blood chemistry. Acta Vet. Scand. 31(3), 277.
  • Varghese, J., Bhat, V., Chianeh, Y. R., Kamath, V., Al-Haj Husain, N., & Özcan, M. (2020). Salivary 8-hydroxyguanosine levels in smokers and non-smokers with chronic periodontitis. Odontology, 108, 569-577.
  • Webb, C., & Twedt, D. (2008). Oxidative stress and liver disease. Veterinary Clinics of North America: Small Anim. Pract. 38(1), 125-135.
  • Yang, Y., Sharma, R, Sharma, A., Awasthi, S., & Awasthi, Y. (2003). Lipid peroxidation and cell cycle signaling: 4-hydroxynonenal, a key molecule in stress mediated signaling. Acta Biochim. Pol. 50(2), 319-336.
  • Yeh, M. M., & Brunt, E. M. (2014). Pathological features of fatty liver disease. Gastroenterology, 147(4), 754-764.
  • Yoshino, K., Katoh, N., Takahashi, K., & Yuasa, A. (1992). Purification of a protein from serum of cattle with hepatic lipidosis, and identification of the protein as haptoglobin. Am. J. Vet. Res. 53(6), 951-956.
There are 35 citations in total.

Details

Primary Language English
Subjects Veterinary Pathology
Journal Section Research Article
Authors

Özhan Karataş 0000-0002-2778-8059

Gökhan Akçakavak 0000-0001-5949-4752

Murat Eren 0000-0002-8544-7340

Zeynep Çelik 0000-0002-9667-5728

Ayşenur Tural 0000-0003-1585-3359

Osman Dağar 0000-0003-2209-7512

Osman Doğan 0000-0001-8579-3203

Bahadır Kılınç 0000-0003-3426-2116

Mehmet Tuzcu 0000-0003-3118-1054

Publication Date June 26, 2025
Submission Date October 23, 2024
Acceptance Date December 9, 2024
Published in Issue Year 2025 Volume: 15 Issue: 1

Cite

APA Karataş, Ö., Akçakavak, G., Eren, M., Çelik, Z., et al. (2025). Determination of 8-OHdG and 4-HNE Expressions in Sheep with Hepatic Lipidosis by Immunohistochemical Method. Manas Journal of Agriculture Veterinary and Life Sciences, 15(1), 9-15. https://doi.org/10.53518/mjavl.1572721
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