Ratlarda Karbon Tetraklorür ile Oluşturulan Nefrotoksisitede Resveratrol’ün Antiapoptotik ve Antioksidan Rolü
Abstract
Bu çalışma, karbon tetraklorür (CCl4) kaynaklı nefrotoksisite modelinde histopatoloji, immünohistokimya ve immünofloresan metotlarını kullanarak güçlü bir antioksidan olan Resveratrol'ün böbrek dokuları üzerindeki koruyucu etkisini incelemeyi amaçlamaktadır. Rastgele seçilen erkek Wistar sıçanları (n=32) dört gruba ayrıldı. Kontrol grubuna herhangi bir işlem uygulanmazken, ikinci gruba CCl4 (2 mg/kg), ve üçüncü gruba Resveratrol (30 mg/kg/gün) verildi ve dördüncü gruba CCl4 ve Resveratrol birlikte verildi. Deneyden sonra sıçanlara anestezi altında ötenazi uygulandı ve böbrek örnekleri toplandı. CCl4 grubunda histopatolojik olarak, tübüler epitelyumda belirgin dejenerasyon ve nekroz, hem glomerüler hem de interstisyel bölgede yer alan arterlerde belirgin steatoz ve hiperemi görüldü. CCl4+Resveratrol grubunda böbrek dokuları tübüler epitelde hafif bozulma ve glomerüler ve interstisyel alanların arterlerinde hiperemi gösterdi. Böbrek dokusunda DNA hasarını belirlemek için 8-OHdG antikoru ile immünohistokimyasal boyama yapıldı. İmmünohistokimya sonuçları hem kontrol hem de resveratrol gruplarında 8-OHdG antikoruna karşı bir reaksiyon oluşmadığını, ancak CCl4 grubuna ait tübüler epitelde sitoplazmik 8-OHdG ekspresyonunda istatistiksel olarak anlamlı bir artış görüldüğünü ortaya koydu (p<0.05). CCl4+Resveratrol grubunda hafif intrasitoplazmik tutulum tespit edildi. Böbrek dokusunda apoptozu belirlemek için kaspaz 3 antikoru ile immünofloresan boyaması yapıldı. Sonuçlar, CCl4 grubunun tübüler epitelde diğer gruplara kıyasla intrasitoplazmik kaspaz 3 ekspresyonunda istatistiksel olarak anlamlı bir artış gösterdiğini (p<0.05) ortaya koyarken, CCl4+Resveratrol grubundakaspaz 3'e karşı oluşan yanıt minör ve intrasitoplazmik özellikteydi. Sonuç olarak, resveratrol, CCl-4 kaynaklı nefrotoksisite sıçan modelinde güçlü antioksidan, antiapoptotik ve DNA koruyucu etkiler göstermiştir ve CCl-4 kaynaklı nefrotoksisiteye karşı daha iyi böbrek sağlığı için günlük diyete eklenebilir.
Keywords
References
- Andritoiu CV, Ochiuz L, Andritoiu V, Popa M (2014). Effect of apitherapy formulations against carbon tetrachloride-induced toxicity in Wistar rats after three weeks of treatment. Molecules, 19, 13374-13391.
- Atmaca E, Aksoy A (2009). Oksidatif DNA hasarı ve kromatografik yöntemlerle tespit edilmesi. Van Vet J, 20, 79-83.
- Bolat İ, Yıldırım S, Sağlam YS et al. (2024). Investigation of the effects of pulmonary adenomatosis on oxidative DNA damage, inflammation, apoptosis, and autophagy in lung tissues in sheep. Small Rumin Res, 230, 107171.
- Cooksey C (2012). Health concerns of heavy metals and metalloids. Sci Pro, 95, 73-88.
- Darwish MA, Abo-Youssef AM, Khalaf AA, Abo-Saif IG (2018). Resveratrol influences platinum pharmacokinetics: A novel mechanism in protection against cisplatin -induced nephrotoxicity. Toxicol Lett, 290, 73-82.
- Emam NM, Anjum S, Okail HA, Ibrahim MAR, Ahmad T (2020). Pomegranate peel extract protects against carbon tetrachloride‑induced nephrotoxicity in mice through increasing antioxidants status. Biomed Rep, 13, 1-9.
- Fahmy MA, Diab KA, Abdel-Samie NS, Omara EA, Hassan ZM (2018). Carbon tetrachloride induced hepato/renal toxicity in experimental mice: antioxidant potential of Egyptian Salvia officinalis L essential oil. Environ Sci Pollut Res, 25, 27858-27876.
- Green DR (2005). Apoptotic pathways: ten minutes to dead. Cell, 121, 671-674.
- Gülçin İ (2010). Antioxidant properties of resveratrol: A structure–activity insight. IFSET, 11, 210-218.
- Iuga C, Alvarez-Idaboy JRl, Russo N (2012). Antioxidant activity of trans-resveratrol toward hydroxyl and hydroperoxyl radicals: a quantum chemical and computational kinetics study. J Org Chem, 77, 3868-3877.
- Jan S, Khan MR (2016). Protective effects of Monotheca buxifolia fruit on renal toxicity induced by CCl 4 in rats. BMC Complement Altern Med. 16, 1-15.
- Kieffer DA, Martin RJ, Adams SH (2016). Impact of dietary fibers on nutrient management and detoxification organs: gut, liver, and kidneys. Adv Nutr, 7, 1111-1121.
- Konyalioglu S, Armagan G, Yalcin A, Atalayin C, Dagci T (2013). Effects of resveratrol on hydrogen peroxide-induced oxidative stress in embryonic neural stem cells. Neural Regen Res, 8, 485-495.
- Lash LH (1994). Role of renal metabolism in risk to toxic chemicals. Environ Health Perspect, 102, 75-79.
- Ma J-Q, Ding J, Xiao Z-H, Liu C-M (2014). Ursolic acid ameliorates carbon tetrachloride-induced oxidative DNA damage and inflammation in mouse kidney by inhibiting the STAT3 and NF-κB activities. Int Immunopharmacol, 21, 389-395.
- Ma J-Q, Liu C-M, Yang W (2018). Protective effect of rutin against carbon tetrachloride-induced oxidative stress, inflammation and apoptosis in mouse kidney associated with the ceramide, MAPKs, p53 and calpain activities. Chem Biol Interact, 286, 26-33.
- Marino G, Niso-Santano M, Baehrecke EH, Kroemer G (2014). Self-consumption: the interplay of autophagy and apoptosis. Nat Rev Mol Cell Biol, 15, 81-94.
- Marques FZ, Markus MA, Morris BJ (2009). Resveratrol: cellular actions of a potent natural chemical that confers a diversity of health benefits. Int J Biochem Cell Biol, 41, 2125-2128.
- Means JC, Gerdes BC, Koulen P (2017). Distinct mechanisms underlying resveratrol-mediated protection from types of cellular stress in C6 glioma cells. Int J Mol Sci, 18, 1521.
- Nowsheen S, Yang E (2012). The intersection between DNA damage response and cell death pathways. Exp onc. 34, 243.
- Ozdemir A, Tumkaya L, Kalcan S et al. (2022). The effects of TNF-α inhibitors on carbon tetrachloride-induced nephrotoxicity. Clin Exp Hypertens, 44, 291-296.
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- Pellegrino D, La Russa D, Marrone A (2019). Oxidative imbalance and kidney damage: New study perspectives from animal models to hospitalized patients. Antioxidants, 8, 594.
- Porter AG, Jänicke RU (1999). Emerging roles of caspase-3 in apoptosis. Cell Death Differ, 6, 99-104.
- Prakash J, Sen D, Sarat Kumar N et al. (2003). Acute renal failure due to intrinsic renal diseases: review of 1122 cases. Ren Fail, 25, 225-233.
- Safhi MM (2018). Nephroprotective effect of zingerone against CCl4‐induced renal toxicity in Swiss albino mice: Molecular mechanism. Oxid Med Cell Longev, 2018, 2474831.
- Saif MS, Waqas M, Hussain R et al (2024). Potential of CME ZIF-8 MOF Nanoformulation: Smart Delivery of Silymarin for Enhanced Performance and Mechanism in Albino Rats. ACS Applied Bio Mat. 7: 6919-6931.
- Shaban NZ, El-Kot SM, Awad OM, Hafez AM, Fouad GM (2021). The antioxidant and anti-inflammatory effects of Carica Papaya Linn. seeds extract on CCl 4-induced liver injury in male rats. BMC Comp Med and Ther, 21, 1-15.
- Tekeli H, Bildik A (2016) Karbon Tetraklorür İle Oluşturulan Karaciğer Hasarında Glutatyon (Gsh) Ve Glutatyon S-Transferaz (Gst) Aktivitesi Üzerine N-Asetil Sisteinin Etkisi.BAUN Health Sci J, 5 (2), 83-87.
- Unsal V, Cicek M, Sabancilar İ (2021). Toxicity of carbon tetrachloride, free radicals and role of antioxidants. REVEH, 36, 279-295.
- Wang Y, Ye D (2021). A caspase-3 activatable photoacoustic probe for in vivo imaging of tumor apoptosis. In: Methods Enzymol, Volume 657, edn Elsevier: 21-57.
- Wu GJ, Chen JT, Tsai HC, Chen TL, Liu SH, Chen RM (2017). Protection of dexmedetomidine against ischemia/reperfusion‐induced apoptotic insults to neuronal cells occurs via an intrinsic mitochondria‐dependent pathway. J Cell Biochem, 118, 2635-2644.
- Xu B, Zheng J, Tian X et al. (2022). Protective mechanism of traditional Chinese medicine guizhi fuling pills against carbon tetrachloride-induced kidney damage is through inhibiting oxidative stress, inflammation and regulating the intestinal flora. Phytomed, 101, 154129.
Resveratrol’s Antiapoptotic and Antioxidant Role in Carbon Tetrachloride-Induced Nephrotoxicity of Rats
Abstract
This study investigates the protective effect of Resveratrol on renal tissues utilizing histopathology, immunohistochemistry, and immunofluorescence methods in a carbon tetrachloride (CCl4)-induced nephrotoxicity model. Wistar albinos (male, n=32) were randomly selected and divided into groups. The initial control group, followed by the second group, received CCl4 (2mg/kg), the third group was administered Resveratrol (30 mg/kg/day), and the fourth group was co-administered with CCl4 and Resveratrol. After the experiment, rats were euthanized under anesthesia, and their kidneys were collected. Results revealed that in the CCl4 group, notable degeneration and necrosis of tubular epithelium, significant steatosis, and arterial hyperemia wereobserved. In the CCl4+Resveratrol group, renal tissues exhibited slight deterioration of tubular epithelium and hyperemia in the glomerular arteries. The immunohistochemistry approach employing 8- Hydroxy-deoxyguanosine (8-OHdG) was utilized to identify DNA damage in kidney tissue. Immunohistochemistry results indicated that both the control and resveratrol groups had a negative reaction for 8-OHdG, but the CCl4 group demonstrated a statistically significant increase in 8-OHdG expression in the tubular epithelium located cytoplasmically (p<0.05). Modest cytoplasmic presence was detected in the CCl4+Resveratrol group. The immunofluorescence approach utilizing caspase 3 was employed to identify apoptosis in kidney tissue. The results indicated that the CCl4 group demonstrated a statistically significant increase in intracytoplasmic caspase 3 expression in the tubular epithelium compared to other groups (p<0.05) while a minor intracytoplasmic response for caspase 3 was observed in the CCl4+Resveratrol group. As a conclusion, resveratrol demonstrated strong antioxidant, antiapoptotic, and DNA-protective effects in a CCl-4-induced nephrotoxicity rat model and can be added to the daily diet for better kidney health against CCl-4-induced nephrotoxicity.
Keywords
Ethical Statement
Animal Welfare Act, US Public Health Service Policy on the Humane Care and Use of Laboratory Animals, National Research Council’s Guide for the Care and Use of Laboratory Animals ve Guide for the Care and Use of Agricultural Animals in Research and Teaching was performed for the study.
References
- Andritoiu CV, Ochiuz L, Andritoiu V, Popa M (2014). Effect of apitherapy formulations against carbon tetrachloride-induced toxicity in Wistar rats after three weeks of treatment. Molecules, 19, 13374-13391.
- Atmaca E, Aksoy A (2009). Oksidatif DNA hasarı ve kromatografik yöntemlerle tespit edilmesi. Van Vet J, 20, 79-83.
- Bolat İ, Yıldırım S, Sağlam YS et al. (2024). Investigation of the effects of pulmonary adenomatosis on oxidative DNA damage, inflammation, apoptosis, and autophagy in lung tissues in sheep. Small Rumin Res, 230, 107171.
- Cooksey C (2012). Health concerns of heavy metals and metalloids. Sci Pro, 95, 73-88.
- Darwish MA, Abo-Youssef AM, Khalaf AA, Abo-Saif IG (2018). Resveratrol influences platinum pharmacokinetics: A novel mechanism in protection against cisplatin -induced nephrotoxicity. Toxicol Lett, 290, 73-82.
- Emam NM, Anjum S, Okail HA, Ibrahim MAR, Ahmad T (2020). Pomegranate peel extract protects against carbon tetrachloride‑induced nephrotoxicity in mice through increasing antioxidants status. Biomed Rep, 13, 1-9.
- Fahmy MA, Diab KA, Abdel-Samie NS, Omara EA, Hassan ZM (2018). Carbon tetrachloride induced hepato/renal toxicity in experimental mice: antioxidant potential of Egyptian Salvia officinalis L essential oil. Environ Sci Pollut Res, 25, 27858-27876.
- Green DR (2005). Apoptotic pathways: ten minutes to dead. Cell, 121, 671-674.
- Gülçin İ (2010). Antioxidant properties of resveratrol: A structure–activity insight. IFSET, 11, 210-218.
- Iuga C, Alvarez-Idaboy JRl, Russo N (2012). Antioxidant activity of trans-resveratrol toward hydroxyl and hydroperoxyl radicals: a quantum chemical and computational kinetics study. J Org Chem, 77, 3868-3877.
- Jan S, Khan MR (2016). Protective effects of Monotheca buxifolia fruit on renal toxicity induced by CCl 4 in rats. BMC Complement Altern Med. 16, 1-15.
- Kieffer DA, Martin RJ, Adams SH (2016). Impact of dietary fibers on nutrient management and detoxification organs: gut, liver, and kidneys. Adv Nutr, 7, 1111-1121.
- Konyalioglu S, Armagan G, Yalcin A, Atalayin C, Dagci T (2013). Effects of resveratrol on hydrogen peroxide-induced oxidative stress in embryonic neural stem cells. Neural Regen Res, 8, 485-495.
- Lash LH (1994). Role of renal metabolism in risk to toxic chemicals. Environ Health Perspect, 102, 75-79.
- Ma J-Q, Ding J, Xiao Z-H, Liu C-M (2014). Ursolic acid ameliorates carbon tetrachloride-induced oxidative DNA damage and inflammation in mouse kidney by inhibiting the STAT3 and NF-κB activities. Int Immunopharmacol, 21, 389-395.
- Ma J-Q, Liu C-M, Yang W (2018). Protective effect of rutin against carbon tetrachloride-induced oxidative stress, inflammation and apoptosis in mouse kidney associated with the ceramide, MAPKs, p53 and calpain activities. Chem Biol Interact, 286, 26-33.
- Marino G, Niso-Santano M, Baehrecke EH, Kroemer G (2014). Self-consumption: the interplay of autophagy and apoptosis. Nat Rev Mol Cell Biol, 15, 81-94.
- Marques FZ, Markus MA, Morris BJ (2009). Resveratrol: cellular actions of a potent natural chemical that confers a diversity of health benefits. Int J Biochem Cell Biol, 41, 2125-2128.
- Means JC, Gerdes BC, Koulen P (2017). Distinct mechanisms underlying resveratrol-mediated protection from types of cellular stress in C6 glioma cells. Int J Mol Sci, 18, 1521.
- Nowsheen S, Yang E (2012). The intersection between DNA damage response and cell death pathways. Exp onc. 34, 243.
- Ozdemir A, Tumkaya L, Kalcan S et al. (2022). The effects of TNF-α inhibitors on carbon tetrachloride-induced nephrotoxicity. Clin Exp Hypertens, 44, 291-296.
- Pegoraro CMR, Nai GA, Garcia LA et al. (2021). Protective effects of Bidens pilosa on hepatoxicity and nephrotoxicity induced by carbon tetrachloride in rats. Drug Chem Toxicol, 44, 64-74.
- Pellegrino D, La Russa D, Marrone A (2019). Oxidative imbalance and kidney damage: New study perspectives from animal models to hospitalized patients. Antioxidants, 8, 594.
- Porter AG, Jänicke RU (1999). Emerging roles of caspase-3 in apoptosis. Cell Death Differ, 6, 99-104.
- Prakash J, Sen D, Sarat Kumar N et al. (2003). Acute renal failure due to intrinsic renal diseases: review of 1122 cases. Ren Fail, 25, 225-233.
- Safhi MM (2018). Nephroprotective effect of zingerone against CCl4‐induced renal toxicity in Swiss albino mice: Molecular mechanism. Oxid Med Cell Longev, 2018, 2474831.
- Saif MS, Waqas M, Hussain R et al (2024). Potential of CME ZIF-8 MOF Nanoformulation: Smart Delivery of Silymarin for Enhanced Performance and Mechanism in Albino Rats. ACS Applied Bio Mat. 7: 6919-6931.
- Shaban NZ, El-Kot SM, Awad OM, Hafez AM, Fouad GM (2021). The antioxidant and anti-inflammatory effects of Carica Papaya Linn. seeds extract on CCl 4-induced liver injury in male rats. BMC Comp Med and Ther, 21, 1-15.
- Tekeli H, Bildik A (2016) Karbon Tetraklorür İle Oluşturulan Karaciğer Hasarında Glutatyon (Gsh) Ve Glutatyon S-Transferaz (Gst) Aktivitesi Üzerine N-Asetil Sisteinin Etkisi.BAUN Health Sci J, 5 (2), 83-87.
- Unsal V, Cicek M, Sabancilar İ (2021). Toxicity of carbon tetrachloride, free radicals and role of antioxidants. REVEH, 36, 279-295.
- Wang Y, Ye D (2021). A caspase-3 activatable photoacoustic probe for in vivo imaging of tumor apoptosis. In: Methods Enzymol, Volume 657, edn Elsevier: 21-57.
- Wu GJ, Chen JT, Tsai HC, Chen TL, Liu SH, Chen RM (2017). Protection of dexmedetomidine against ischemia/reperfusion‐induced apoptotic insults to neuronal cells occurs via an intrinsic mitochondria‐dependent pathway. J Cell Biochem, 118, 2635-2644.
- Xu B, Zheng J, Tian X et al. (2022). Protective mechanism of traditional Chinese medicine guizhi fuling pills against carbon tetrachloride-induced kidney damage is through inhibiting oxidative stress, inflammation and regulating the intestinal flora. Phytomed, 101, 154129.
Details
| Primary Language | English |
|---|---|
| Subjects | Veterinary Histology and Embryology, Veterinary Pathology |
| Journal Section | Araştırma Makaleleri |
| Authors | |
| Publication Date | July 27, 2025 |
| Submission Date | March 20, 2025 |
| Acceptance Date | June 19, 2025 |
| Published in Issue | Year 2025 Volume: 36 Issue: 2 |
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Accepted papers are licensed under Creative Commons Attribution-NonCommercial 4.0 International License