Modulatory Role of Vitamin E on Oxidative Stress, Inflammation, and Autophagy in Oxytetracycline Induced Renal Toxicity

Öz

The present study aimed to investigate the nephrotoxic effects of oxytetracycline (OTC) and to evaluate the protective role of vitamin E (Evit) through oxidative stress, inflammation, and autophagy pathways. A total of 28 rats were randomly assigned to four experimental groups: control, Evit, OTC, and OTC + Evit and treated for seven consecutive days. At the end of the experimental period, kidney tissues were collected for biochemical and molecular analyses. Oxidative stress parameters, including malondialdehyde (MDA), reduced glutathione (GSH), glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT), were determined using spectrophotometric methods. In addition, the gene expression levels of nuclear factor kappa B (NF-κB), interleukin-1 beta (IL-1β), tumor necrosis factor-alpha (TNF α), and the autophagy-related marker Beclin-1 were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). OTC administration significantly increased MDA levels while decreasing GSH content and the activities of antioxidant enzymes, indicating enhanced oxidative stress. Furthermore, the expression levels of NF-κB, IL-1β, TNF-α, and Beclin-1 were markedly elevated, suggesting activation of inflammatory and autophagic pathways. Evit co administration significantly reduced MDA levels, restored antioxidant enzyme activities, and suppressed the expression of inflammatory and autophagy-related markers compared with the OTC group, although these parameters did not fully return to control levels. In conclusion, Evit exerts a protective effect against OTC-induced renal injury by attenuating oxidative stress and modulating inflammation and autophagy pathways, highlighting its potential as a supportive therapeutic strategy against antibiotic-induced nephrotoxicity.

Anahtar Kelimeler

• Inflammation
• Nephrotoxicity
• Oxidative stress
• Oxytetracycline
• Vitamin E

Oksitetrasiklin Kaynaklı Böbrek Toksisitesinde E Vitamininin Oksidatif Stres, Enflamasyon ve Otofaji Üzerindeki Düzenleyici Rolü

Öz

Bu çalışma, oksitetrasiklinin (OTC) nefrotoksik etkilerini araştırmayı ve E vitamininin (Evit) oksidatif stres, inflamasyon ve otofaji yolları üzerindeki koruyucu rolünü değerlendirmeyi amaçlamıştır. Toplam 28 sıçan rastgele dört deney grubuna (kontrol, Evit, OTC ve OTC + Evit) ayrılmış ve yedi gün boyunca ilgili uygulamalara tabi tutulmuştur. Deney süresinin sonunda, biyokimyasal ve moleküler analizler için böbrek dokuları alınmıştır. Malondialdehit (MDA), indirgenmiş glutatyon (GSH), glutatyon peroksidaz (GPx), süperoksit dismutaz (SOD) ve katalaz (CAT) dahil olmak üzere oksidatif stres parametreleri spektrofotometrik yöntemlerle belirlenmiştir. Ayrıca, nükleer faktör kappa B (NF-κB), interlökin-1 beta (IL-1β), tümör nekroz faktörü-alfa (TNF-α) ve otofaji ile ilişkili bir belirteç olan Beclin-1'in gen ekspresyon düzeyleri, kantitatif gerçek zamanlı polimeraz zincir reaksiyonu (qRT-PCR) ile değerlendirilmiştir. OTC uygulaması, artmış oksidatif strese işaret edecek şekilde MDA düzeylerini önemli ölçüde artırırken, GSH içeriğini ve antioksidan enzim aktivitelerini azaltmıştır. Buna ek olarak, NF-κB, IL-1β, TNF-α ve Beclin-1 ekspresyon düzeyleri belirgin şekilde yükselmiş; bu durum, inflamatuar ve otofajik yolların aktive olduğuna işaret etmiştir. Evit'in birlikte uygulanması, OTC grubuna kıyasla MDA düzeylerini önemli ölçüde düşürmüş, antioksidan enzim aktivitelerini eski haline getirmiş ve inflamatuar ve otofaji ile ilişkili belirteçlerin ekspresyonunu baskılamıştır; ancak bu parametreler kontrol düzeylerine tam olarak dönmemiştir. Sonuç olarak Evit, oksidatif stresi hafifletip inflamasyon ve otofaji yollarını modüle ederek OTC kaynaklı böbrek hasarına karşı koruyucu bir etki göstermekte ve antibiyotik kaynaklı nefrotoksisiteye karşı destekleyici bir tedavi stratejisi olarak potansiyelini ortaya koymaktadır.

Anahtar Kelimeler

• E vitamini
• inflamasyon
• nefrotoksisite
• oksidatif stres
• Oksitetrasiklin

Kaynakça

1. Abdelrazik, E., Hassan, H. M., Abdallah, Z., Magdy, A., & Farrag, E. A. (2022). Renoprotective effect of N-acetylcystein and vitamin E in bisphenol A-induced rat nephrotoxicity; Modulators of Nrf2/ NF-κB and ROS signaling pathway. Acta Bio Medica : Atenei Parmensis, 93(6), e2022301. https://doi.org/10.23750/abm.v93i6.13732
2. Aebi, H. (1984). Catalase in vitro. Methods in Enzymology, 105, 121–126. https://doi.org/10.1016/s0076-6879(84)05016-3
3. Akcakavak, G., Kazak, F., Karatas, O., Alakus, H., Alakus, I., Kirgiz, O., Celik, Z., Deveci, M. Z. Y., 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. Toxicology Mechanisms and Methods, 34(4), 413–422. https://doi.org/10.1080/15376516.2023.2297234
4. Brigelius-Flohé, R., & Traber, M. G. (1999). Vitamin E: function and metabolism. FASEB Journal, 13(10), 1145–1155.
5. Bülbül, R., Şimşek, H., Akaras, N., Kandemir, Ö., Kandemir, F. M., Çağlayan, C., & Genç, A. (2026). Catechin hydrate alleviates tramadol-induced ovarian and uterine injury through regulation of oxidative and ER stress, apoptosis, steroidogenesis, and hormonal imbalance. Reproductive Toxicology, 140, 109132. https://doi.org/10.1016/j.reprotox.2025.109132
6. Dhouibi, R., Affes, H., Ben Salem, M., Charfi, S., Marekchi, R., Hammami, S., Zeghal, K., & Ksouda, K. (2021). Protective effect of Urtica dioica in induced neurobehavioral changes, nephrotoxicity and hepatotoxicity after chronic exposure to potassium bromate in rats. Environmental Pollution, 287, 117657. https://doi.org/10.1016/j.envpol.2021.117657
7. Elshopakey, G. E., & Elazab, S. T. (2021). Cinnamon aqueous extract attenuates diclofenac sodium and oxytetracycline mediated hepato-renal toxicity and modulates oxidative stress, cell apoptosis, and inflammation in male albino rats. Veterinary Sciences, 8(1), 9. https://doi.org/10.3390/vetsci8010009
8. Halliwell, B., & Gutteridge, J. M. C. (Ed.). (2015). Free radicals in biology and medicine. Oxford University Press.

9. Jayanthi, R., & Subash, P. (2010). Antioxidant effect of caffeic acid on oxytetracycline induced lipid peroxidation in albino rats. Indian Journal of Clinical Biochemistry, 25(4), 371–375. https://doi.org/10.1007/s12291-010-0052-8
10. Kandemir, F. M., Ozkaraca, M., Yildirim, B. A., Hanedan, B., Kirbas, A., Kilic, K., Aktas, E., & Benzer, F. (2015). Rutin attenuates gentamicin-induced renal damage by reducing oxidative stress, inflammation, apoptosis, and autophagy in rats. Renal Failure, 37(3), 518–525. https://doi.org/10.3109/0886022X.2015.1006100
11. Kounatidis, D., Dalamaga, M., Grivakou, E., Karampela, I., Koufopoulos, P., Dalopoulos, V., Adamidis, N., Mylona, E., Kaziani, A., & Vallianou, N. G. (2024). Third-generation tetracyclines: current knowledge and therapeutic potential. Biomolecules, 14(7), 783. https://doi.org/10.3390/biom14070783
12. Küçükler, S., Çomaklı, S., Özdemir, S., Değirmençay, Ş., Kandemir, F. M., Genç, A., Dalkılınç, E., & Aydın, Ş. (2025). Investigating the effects of carvacrol in isoproterenol-induced myocardial injury: Role of H-FABP and Gal-3/TLR4/MYD88/NF-κB signaling pathway modulation. Bratislava Medical Journal, 126(11), 2954–2970. https://doi.org/10.1007/s44411-025-00315-1
13. Lawrence, T. (2009). The nuclear factor NF-κB pathway in inflammation. Cold Spring Harbor Perspectives in Biology, 1(6), a001651. https://doi.org/10.1101/cshperspect.a001651
14. Livak, K. J., & Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods, 25(4), 402–408. https://doi.org/10.1006/meth.2001.1262
15. Lowry, O. H., Rosebrough, N. J., Farr, A. L., & Randall, R. J. (1951). Protein measurement with the folin phenol reagent. The Journal of Biological Chemistry, 193(1), 265–275.
16. Matkovics, B. (1988). Determination of enzyme activity in lipid peroxidation and glutathione pathways. Laboratoriumi Diagnosztika, 15, 248–250.
17. Meng, L., Liu, S., & Cui, W. (2025). Renal protective effects of vitamin E for drug-induced kidney injury: a meta-analysis. Frontiers in Pharmacology, 16, 1461792. https://doi.org/10.3389/fphar.2025.1461792
18. Mittal, M., Siddiqui, M. R., Tran, K., Reddy, S. P., & Malik, A. B. (2014). Reactive oxygen species in inflammation and tissue injury. Antioxidants & Redox Signaling, 20(7), 1126–1167. https://doi.org/10.1089/ars.2012.5149
19. Ore, A., & Akinloye, O. A. (2019). Oxidative stress and antioxidant biomarkers in clinical and experimental models of non-alcoholic fatty liver disease. Medicina, 55(2), 26. https://doi.org/10.3390/medicina55020026
20. Placer, Z. A., Cushman, L. L., & Johnson, B. C. (1966). Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical systems. Analytical Biochemistry, 16(2), 359–364. https://doi.org/10.1016/0003-2697(66)90167-9
21. Reuter, S., Gupta, S. C., Chaturvedi, M. M., & Aggarwal, B. B. (2010). Oxidative stress, inflammation, and cancer: how are they linked? Free Radical Biology & Medicine, 49(11), 1603–1616. https://doi.org/10.1016/j.freeradbiomed.2010.09.006
22. Samare-Najaf, M., Zal, F., & Safari, S. (2020). Primary and secondary markers of doxorubicin-induced female infertility and the alleviative properties of quercetin and vitamin e in a rat model. Reproductive Toxicology, 96, 316–326. https://doi.org/10.1016/j.reprotox.2020.07.015
23. Sedlak, J., & Lindsay, R. H. (1968). Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with Ellman’s reagent. Analytical Biochemistry, 25(1), 192–205. https://doi.org/10.1016/0003-2697(68)90092-4
24. Sun, Y., Oberley, L. W., & Li, Y. (1988). A simple method for clinical assay of superoxide dismutase. Clinical Chemistry, 34(3), 497–500.
25. Ungurianu, A., Zanfirescu, A., Nițulescu, G., & Margină, D. (2021). Vitamin e beyond its antioxidant label. Antioxidants, 10(5), 634. https://doi.org/10.3390/antiox10050634
26. Varışlı, B., Darendelioğlu, E., Caglayan, C., Kandemir, F. M., Ayna, A., Genç, A., & Kandemir, Ö. (2022). Hesperidin attenuates oxidative stress, inflammation, apoptosis, and cardiac dysfunction in sodium fluoride‐induced cardiotoxicity in rats. Cardiovascular Toxicology, 22(8), 727–735. https://doi.org/10.1007/s12012-022-09751-9
27. Yavuz, H., Kandemir, Ö., Şimşek, H., Akaras, N., & Kandemir, F. M. (2026). Investigation of the effects of carvacrol on oxytetracycline-induced kidney damage. Van Veterinary Journal, 37(1), 66–73. https://doi.org/10.36483/vanvetj.1819779
28. Yun, H. R., Jo, Y. H., Kim, J., Shin, Y., Kim, S. S., & Choi, T. G. (2020). Roles of autophagy in oxidative stress. International Journal of Molecular Sciences, 21(9), 3289. https://doi.org/10.3390/ijms21093289