Deneysel Sepsisli Sıçanlarda Aktif Anyonik Oksijen Uygulamasının Testis Dokusu Üzerindeki Koruyucu Etkisi

Öz

Bu çalışma, sıçanlarda sepsis kaynaklı testis hasarında aktif anyonik oksijen (AOO) uygulamasının oksidatif stres, enflamasyon ve steroidogenez üzerindeki etkilerini araştırmayı amaçlamıştır. Otuz iki erkek Sprague-Dawley sıçanı dört gruba ayrılmıştır: Kontrol, Sepsis (LPS), AOO ve Sepsis+AOO. Oksidatif stres parametreleri (glutatyon [GSH], malondialdehit [MDA], süperoksit dismutaz [SOD], katalaz [CAT]), enflamatuvar sitokinler (interlökin-6 [IL-6], tümör nekroz faktörü-alfa [TNF-α], transforme edici büyüme faktörü-beta1 [TGF-β1], interlökin-1 beta [IL-1β]) ve steroidojenik belirteçlerin (3β-hidroksisteroid dehidrogenaz [3β-HSD], 17β-hidroksisteroid dehidrogenaz [17β-HSD], steroidojenik akut düzenleyici protein [StAR]) biyokimyasal analizi için testis dokuları toplanmıştır. Sepsis grubunda, kontrol grubuna kıyasla GSH düzeyleri (p < .001), SOD aktivitesi (p < .01) ve CAT aktivitesi (p < .001) anlamlı derecede azalırken, MDA düzeyleri (p < .001) belirgin şekilde artmıştır. IL-6 (p < .001), TNF-α (p < .01), TGF-β1 (p < .05) ve IL-1β (p < .001) dahil olmak üzere pro-enflamatuvar sitokinler anlamlı derecede yükselirken, steroidojenik enzimlerden 3β-HSD (p < .01), 17β-HSD (p < .05) ve StAR (p < .05) sepsis grubunda anlamlı derecede azalmıştır. Sepsis+AOO grubunda ise, sepsis grubuna kıyasla AOO tedavisi GSH düzeylerini (p < .001), SOD aktivitesini (p < .05) ve CAT aktivitesini (p < .001) anlamlı derecede artırırken, MDA düzeylerini (p < .001) azaltmıştır. Ek olarak, AOO, IL-6 (p < .01), TNF-α (p < .01), TGF-β1 (p < .001) ve IL-1β düzeylerini anlamlı derecede baskılamış ve 3β-HSD (p < .001), 17β-HSD (p < .05) ve StAR (p < .01) ekspresyonunu anlamlı derecede yukarı regüle etmiştir. Bu çalışma, AOO'nun güçlü antioksidan ve anti-enflamatuvar özellikler sergilediğini, sepsis kaynaklı testis oksidatif stresini ve enflamasyonunu etkili bir şekilde baskılarken testis steroidogenezini koruduğunu ve sepsis kaynaklı testis disfonksiyonu için terapötik bir ajan olarak potansiyelini ortaya koymaktadır.

Anahtar Kelimeler

• Aktif Anyonik Oksijen
• Enflamasyon;
• Oksidatif Stres
• Sepsis
• Steroidogenez

Destekleyen Kurum

Bu çalışma için herhangi bir finansal destek alınmamıştır.

Etik Beyan

Bu çalışma için etik onay, Kafkas Üniversitesi Hayvan Deneyleri Yerel Etik Kurulu'ndan alınmıştır (Onay No: KAÜ-HADYEK/2026-121).

Protective Effect of Active Anionic Oxygen Administration on Testicular Tissue in Rats with Experimental Sepsis

Öz

This study aimed to investigate the effects of active anionic oxygen (AAO) on oxidative stress, inflammation, and steroidogenesis in sepsis-induced testicular injury in rats. Thirty male Wistar Albino rats were randomly assigned to four groups: Control, Sepsis (lipopolysaccharide, LPS), AAO, and Sepsis+AAO. Testicular tissues were analyzed for oxidative stress markers (reduced glutathione [GSH], malondialdehyde [MDA], superoxide dismutase [SOD], and catalase [CAT]), inflammatory cytokines (interleukin-6 [IL-6], tumor necrosis factor-alpha [TNF-α], transforming growth factor-beta 1 [TGF-β1], and interleukin-1 beta [IL-1β]), and steroidogenic markers (3β-hydroxysteroid dehydrogenase [3β-HSD], 17β-hydroxysteroid dehydrogenase [17β-HSD], and steroidogenic acute regulatory protein [StAR]). Sepsis significantly decreased GSH (p < .0001), SOD (p < .01), and CAT (p < .0001) levels, while increasing MDA (p < .0001) levels compared with the control group. In addition, IL-6 (p < .0001), TNF-α (p < .001), TGF-β1 (p < .05), and IL-1β (p < .0001) levels were significantly elevated, whereas the expression of 3β-HSD (p < .01), 17β-HSD (p < .05), and StAR (p < .01) was significantly reduced. Compared with the sepsis group, AAO treatment significantly improved antioxidant status (all p < .05), suppressed inflammatory cytokine levels (all p < .01), and enhanced steroidogenic marker expression (all p < .05). These findings demonstrate that AAO exerts potent antioxidant and anti-inflammatory effects, attenuates sepsis-induced testicular injury, and preserves steroidogenesis, suggesting its potential as a protective agent for sepsis-associated testicular dysfunction.

Anahtar Kelimeler

• Active anionic oxygen
• Inflammation
• Oxidative stress
• Sepsis
• Steroidogenesis

Destekleyen Kurum

No funding was received for this study.

Etik Beyan

Ethical approval for this study was obtained from the Animal Experiments Local Ethics Committee of Kafkas University (Approval No: KAÜ-HADYEK/2026-121).

Kaynakça

1. Aebi, H. (1974). Catalase. Methods of Enzymatic Analysis, 2, 673–684. https://doi.org/10.1016/B978-0-12-091302-2.50032-3
2. Aguilar Diaz de Leon, J., & Borges, C. R. (2020). Evaluation of oxidative stress in biological samples using the thiobarbituric acid reactive substances (TBARS) assay. Journal of Visualized Experiments, (159), e61122. https://doi.org/10.3791/61122
3. Aksak Karamese, S., Gelen, V., Yildiz, G. N., Albayrak, K., Gedikli, S., Kara, A., & Karamese, M. (2025). Investigation of the anti-inflammatory, anti-oxidant and anti-apoptotic activity of 18β- glycyrrhetinic-acid on the model of LPS-induced lung injury in rats. Molecular Immunology, 181, 93–101. https://doi.org/10.1016/j.molımm.2025.03.009
4. Ali, W., Khatyan, U., Sun, J., Alasmari, A., Alshahrani, M. Y., Qazi, I. H., Wang, T., Liu, Z., & Zou, H. (2024). Mitigating effect of pomegranate peel extract against the furan induced testicular injury by apoptosis, steroidogenic enzymes and oxidative stress. Chemosphere, 358, 142086. https://doi.org/10.1016/j.chemosphere.2024.142086
5. Atanassova, N., & Koeva, Y. (2012). Hydrohysteroid dehydrogenases - Biological role and clinical importance - Review. In R. A. Canuto (Eds.), Dehydrogenases. IntechOpen. https://doi.org/10.5772/54149
6. Aydin, S., Bacanli, M., Taner, G., Şahin, T., Başaran, A. A., & Başaran, N. (2013). Protective effects of resveratrol on sepsis-induced DNA damage in the lymphocytes of rats. Human & Experimental Toxicology, 32(10), 1048–1057. https://doi.org/10.1177/0960327112467047
7. Bati, Y. U., Sezer, M., Yilmaz, A., Baser, L., Guraslan, A., Bayram, P., & Karamese, M. (2025). The protective effects of dose-dependent umbelliferone application on clp-induced acute lung injury (ALI) model. Journal of Biochemical and Molecular Toxicology, 39(10), e70549. https://doi.org/10.1002/jbt.70549
8. Beutler, E., Duron, O., & Kelly, B. M. (1963). Improved method for the determination of blood glutathione. The Journal of Laboratory and Clinical Medicine, 61, 882-888.

9. Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72(1–2), 248–254. https://doi.org/10.1016/0003-2697(76)90527-3
10. Cecconi, M., Evans, L., Levy, M., & Rhodes, A. (2018). Sepsis and septic shock. The Lancet, 392(10141), 75–87. https://doi.org/10.1016/S0140-6736(18)30696-2
11. El-Sherbiny, M., Elkattawy, H. A., Hadhoud, S., Nasr, A. N., Ibrahim, A. M., Ameer, O. Z., Alsaleebi, N., Asfari, J., Zakari, M. O., Mojaddidi, M. A., Ali, E. K., Almohaimeed, H. M., Abdeen, A., Ali, S. K., & Eldesoqui, M. (2025). Oxytocin ameliorates lipopolysaccharide-induced acute orchitis model: interplay of oxidative stress and inflammatory pathways. Frontiers in Pharmacology, 15, 1506777. https://doi.org/10.3389/fphar.2024.1506777
12. Galley, H. F. (2011). Oxidative stress and mitochondrial dysfunction in sepsis. British Journal of Anaesthesia, 107(1), 57–64. https://doi.org/10.1093/BJA/AER093
13. Gerek, M., Erdem, O., Aşık, M. B., Demiriz, M., Alkoclar, E. C., Diler, A. S., Kusbaygı, H., & Bursalı, U. (2021). A biocidal active anionic h2o8 oxygen solution with bronchodilative & oxygenizing features for the treatment of coronaviral ınfection related hypoxia - GATA animal trials abstract. Zenodo. https://doi.org/10.5281/zenodo.5501039
14. Guo, M., Zhang, Y., Yang, H., & Liu, Q. (2025). On anxiolytic and antidepressant effects of curcumin in sepsis-induced post-intensive care syndrome: Antioxidant and anti-ınflammatory effects. Neuropsychobiology, 84(5–6), 231–247. https://doi.org/10.1159/000547791
15. Hafez, M. H., Gad, S. B., & El-Sayed, Y. S. (2025). Quercetin-mediated restoration of high-fat diet-induced male reproductive dysfunction through modifying spermatogenesis and unraveling 3β-HSD, 17β-HSD, and StAR pathways. BMC Pharmacology and Toxicology, 26(1), 90. https://doi.org/10.1186/S40360-025-00918-y
16. Hu, G. X., Zhao, B. H., Chu, Y. H., Zhou, H. Y., Akingbemi, B. T., Zheng, Z. Q., & Ge, R. S. (2010). Effects of genistein and equol on human and rat testicular 3β-hydroxysteroid dehydrogenase and 17β-hydroxysteroid dehydrogenase 3 activities. Asian Journal of Andrology, 12(4), 519-526. https://doi.org/10.1038/aja.2010.18
17. Jana, K., Jana, S., & Samanta, P. K. (2006). Effects of chronic exposure to sodium arsenite on hypothalamo-pituitary-testicular activities in adult rats: possible an estrogenic mode of action. Reproductive Biology and Endocrinology, 4(1), 9. https://doi.org/10.1186/1477-7827-4-9
18. Jarczak, D., Kluge, S., & Nierhaus, A. (2021). Sepsis-pathophysiology and therapeutic concepts. Frontiers in Medicine, 8, 628302. https://doi.org/10.3389/fmed.2021.628302
19. Kükürt, A., Gelen, V., Başer, Ö. F., Deveci, H. A., & Karapehlivan, M. (2021). Thiols: role in oxidative stress-related disorders. In P. Atukeren (Eds.), Accenting lipid peroxidation. IntechOpen. https://doi.org/10.5772/intechopen.96682
20. Lin, Y., Alhaskawi, A., Chen, L., & Moqbel, S. A. A. (2025). Recent advances in understanding oxidative stress in sepsis: pathogenic roles and antioxidant therapeutic prospects - a narrative review. Frontiers in Pharmacology, 16, 1695992. https://doi.org/10.3389/fphar.2025.1695992
21. Paul, S. N., Nessel, I., Puthucheary, Z., & Henson, S. M. (2026). Sepsis and the immunometabolic inflammatory response. Npj Metabolic Health and Disease, 4(1), 4. https://doi.org/10.1038/s44324-025-00095-w
22. Raheem, A. K., Abu-Raghif, A. R., Abbas, A. H., Ridha-Salman, H., & Oubaid, E. N. (2025). Quercetin mitigates sepsis-induced renal injury via inhibiting inflammatory and oxidative pathways in mice. Journal of Molecular Histology, 56(3), 184. https://doi.org/10.1007/S10735-025-10442-2
23. Remick, D. G., Newcomb, D. E., Bolgos, G. L., & Call, D. R. (2000). Comparison of the mortality and inflammatory response of two models of sepsis: lipopolysaccharide vs. cecal ligation and puncture. Shock, 13(2), 110–116. https://doi.org/10.1097/00024382-200013020-00004
24. Rudd, K. E., Johnson, S. C., Agesa, K. M., Shackelford, K. A., Tsoi, D., Kievlan, D. R., Colombara, D. V., Ikuta, K. S., Kissoon, N., Finfer, S., Fleischmann-Struzek, C., Machado, F. R., Reinhart, K. K., Rowan, K., Seymour, C. W., Watson, R. S., West, T. E., Marinho, F., Hay, S. I., ... Naghavi, M. (2020). Global, regional, and national sepsis incidence and mortality, 1990–2017: Analysis for the global burden of disease study. Lancet, 395(10219), 200–211. https://doi.org/10.1016/S0140-6736(19)32989-7
25. Sahoo, D. K., Wong, D., Patani, A., Paital, B., Yadav, V. K., Patel, A., & Jergens, A. E. (2024). Exploring the role of antioxidants in sepsis-associated oxidative stress: A comprehensive review. Frontiers in Cellular and Infection Microbiology, 14, 1348713. https://doi.org/10.3389/fcimb.2024.1348713
26. Singer, M., Deutschman, C. S., Seymour, C. W., Shankar-Hari, M., Annane, D., Bauer, M., Bellomo, R., Bernard, G. R., Chiche, J.D., Coopersmith, C. M., Hotchkiss, R. S., Levy, M. M., Marshall, J. C., Martin, G. S., Opal, S. M., Rubenfeld, G. D., van der Poll, T., Vincent, J. L., & Angus, D. C. (2016). The third ınternational consensus definitions for sepsis and septic shock (sepsis-3). JAMA, 315(8), 801–810. https://doi.org/10.1001/jama.2016.0287
27. Sun, Y., Oberley, L. W., & Li, Y. (1988). A simple method for clinical assay of superoxide dismutase. Clinical Chemistry, 34(3), 497–500. https://doi.org/10.1093/clınchem/34.3.497
28. Üstündağ, H., Doğanay, S., Kalındemirtaş, F. D., Demir, Ö., Huyut, M. T., Kurt, N., Özgeriş, F. B., & Akbaba, Ö. (2023). A new treatment approach: Melatonin and ascorbic acid synergy shields against sepsis-induced heart and kidney damage in male rats. Life Sciences, 329, 121875. https://doi.org/10.1016/J.LFS.2023.121875
29. Yildirim, S., Sengul, E., Aksu, E. H., Cinar, İ., Gelen, V., Tekin, S., & Dag, Y. (2024). Selenium reduces acrylamide-induced testicular toxicity in rats by regulating HSD17B1, StAR, and CYP17A1 expression, oxidative stress, inflammation, apoptosis, autophagy, and DNA damage. Environmental Toxicology, 39(3), 1402–1414. https://doi.org/10.1002/tox.23996
30. Yoshioka, T., Kawada, K., Shimada, T., & Mori, M. (1979). Lipid peroxidation in maternal and cord blood and protective mechanism against activated-oxygen toxicity in the blood. American Journal of Obstetrics and Gynecology, 135(3), 372–376. https://doi.org/10.1016/0002-9378(79)90708-7