Safranal Attenuates Malathion-Induced Oxidative Stress and AChE Inhibition Across Blood, Hepatic, Renal, and Brain Matrices

Öz

Organophosphate pesticides, particularly malathion, are widely used in agriculture but pose severe toxicity risks to non-target organisms through acetylcholinesterase (AChE) enzyme inhibition and the induction of oxidative stress. This study investigated the potential protective effects of safranal, a primary bioactive component of saffron (Crocus sativus L.) known for its antioxidant and anti-inflammatory properties, against malathion-induced systemic and tissue-specific toxicity. Experimental groups were evaluated by measuring malondialdehyde (MDA), nitric oxide (NO), glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), and GPx (glutathione peroxidase) levels, alongside AChE enzyme activity in the blood, liver, kidney, and brain tissues. The findings demonstrated that malathion exposure caused profound toxicity characterized by severe AChE inhibition, accelerated lipid peroxidation, elevated NO levels, and a critical depletion of both enzymatic and non-enzymatic antioxidant defenses across all analyzed matrices. Conversely, safranal administration served as an effective therapeutic intervention. It mitigated oxidative damage, limited the exhaustion of key antioxidant enzymes by reducing reactive oxygen species, and supported the preservation of AChE functionality. Notably, safranal’s protective efficacy exhibited tissue-dependent variations; while it significantly attenuated oxidative stress markers and preserved cellular viability in the plasma, liver, and renal matrices, it demonstrated limited protective capacity in brain tissue under the current experimental conditions. In conclusion, safranal represents a promising bioactive countermeasure against organophosphate-induced systemic injury. However, further research is warranted to elucidate its precise transport mechanisms and potential limitations in crossing the blood-brain barrier.

Anahtar Kelimeler

• Safranal
• Organophosphates
• Malathion
• Oxidative Stress
• AChE

Destekleyen Kurum

This research did not receive any specific support from funders in the public, commercial, or non-profit sectors.

Etik Beyan

The experimental protocol was approved by Animal Experiments Local Ethics Committee of Afyon Kocatepe University (Decision No: B.30.2.AKÜ.0.A2.00.00/249, Date: 09.11.2012).

Teşekkür

The authors would like to express their gratitude to Afyon Kocatepe University and also Çay Vocational School and Faculty of Science and Arts for providing the necessary laboratory facilities and institutional support for this research. We also extend our sincere thanks to our master's students for their valuable technical assistance and dedication during the experimental stages of this study.

Safranal; Kan, Karaciğer, Böbrek ve Beyin Matrislerinde Malation Kaynaklı Oksidatif Stresi ve AChE İnhibisyonunu Hafifletir

Öz

Başta malation olmak üzere organofosfatlı pestisitler tarımda yaygın olarak kullanılmakta, ancak asetilkolinesteraz (AChE) enzim inhibisyonu ve oksidatif stresi tetikleme mekanizmalarıyla hedef olmayan organizmalar üzerinde ciddi toksisite riskleri oluşturmaktadır. Bu çalışmada, safranın (Crocus sativus L.) antioksidan ve anti-enflamatuar özellikleri ile bilinen temel biyoaktif bileşeni safranalın, malation kaynaklı sistemik ve dokuya özgü toksisiteye karşı olası koruyucu etkileri araştırılmıştır. Deneysel gruplar; kan, karaciğer, böbrek ve beyin dokularında malondialdehit (MDA), nitrik oksit (NO), glutatyon (GSH), süperoksit dismütaz (SOD), katalaz (CAT) ve glutatyon peroksidaz (GPx) düzeylerinin yanı sıra AChE enzim aktivitesi ölçülerek değerlendirilmiştir. Bulgular, malation maruziyetinin; şiddetli AChE inhibisyonu, hızlanmış lipid peroksidasyonu, yükselmiş NO seviyeleri ve analiz edilen tüm matrislerde hem enzimatik hem de enzimatik olmayan antioksidan savunma hatlarında kritik bir tükenme ile karakterize derin bir toksisiteye yol açtığını göstermiştir. Buna karşılık, safranal uygulaması etkili bir terapötik müdahale sağlamıştır. Safranal oksidatif hasarı hafifletmiş, reaktif oksijen türlerini azaltarak temel antioksidan enzimlerin tükenmesini sınırlandırmış ve AChE fonksiyonelliğinin korunmasını desteklemiştir. Özellikle, safranalın koruyucu etkinliği dokuya bağlı varyasyonlar göstermiştir; plazma, karaciğer ve böbrek matrislerinde oksidatif stres belirteçlerini önemli ölçüde azaltıp hücresel canlılığı korurken, mevcut deneysel koşullar altında beyin dokusunda sınırlı bir koruyucu kapasite sergilemiştir. Sonuç olarak safranal, organofosfat kaynaklı sistemik hasara karşı gelecek vaat eden biyoaktif bir önlem olarak öne çıkmaktadır. Bununla birlikte, kan-beyin bariyerini geçmedeki kesin taşınma mekanizmalarını ve olası sınırlılıklarını aydınlatmak için daha fazla araştırmaya ihtiyaç duyulmaktadır.

Anahtar Kelimeler

• Safranal
• Organofosfatlar
• Malatyon
• Oksidatif Stres
• AChE

Destekleyen Kurum

Bu araştırma, kamu, ticari veya kar amacı gütmeyen sektörlerdeki fon sağlayıcılardan herhangi bir özel destek almamıştır.

Etik Beyan

Deney protokolü, Afyon Kocatepe Üniversitesi Hayvan Deneyleri Yerel Etik Kurulu tarafından onaylanmıştır (Karar No: B.30.2.AKÜ.0.A2.00.00/249, Tarih: 09.11.2012).

Teşekkür

Bu araştırma için gerekli laboratuvar olanaklarını ve kurumsal desteği sağladıkları için Afyon Kocatepe Üniversitesi Çay Meslek Yüksekokulu'na ve Afyon Kocatepe Üniversitesi Fen ve Edebiyat Fakültesi'ne teşekkürlerimizi ifade etmek istiyoruz. Ayrıca, bu çalışmanın deneysel aşamalarında değerli teknik yardımları ve özverileri için yüksek lisans öğrencilerimize de içten teşekkürlerimizi sunarız.

Kaynakça

1. Aboubakr, H. M., Elzohairy, E. A., Ali, A. A., Rashed, L. A., Elkady, N. K., & Soliman, A. S. A. (2019). Therapeutic effects of N-acetylcysteine against malathion-induced hepatotoxicity. Egyptian Journal of Forensic Sciences, 9(1), 34. https://doi.org/10.1186/s41935-019-0142-6
2. Akbel, E., Arslan-Acaroz, D., Demirel, H. H., Kucukkurt, I., & Ince, S. (2018). The subchronic exposure to malathion, an organophosphate pesticide, causes lipid peroxidation, oxidative stress, and tissue damage in rats: the protective role of resveratrol. Toxicology Research, 7(3), 503–512. https://doi.org/10.1039/c8tx00030aAroniadou-Anderjaska, V., Figueiredo, T. H., de Araujo Furtado, M., Pidoplichko, V. I., & Braga, M. F. M. (2023). Mechanisms of organophosphate toxicity and the role of acetylcholinesterase inhibition. Toxics, 11(10), 866. https://doi.org/10.3390/toxics11100866
3. Barr, D. B., & Angerer, J. (2006). Potential uses of biomonitoring data: a case study using the organophosphorus pesticides chlorpyrifos and malathion. Environmental Health Perspectives, 114(11), 1763– 1769. https://doi.org/10.1289/ehp.9062
4. Basarslan, S. K., Alp, H., Senol, S., Evliyaoglu, O., & Ozkan, U. (2014). Is intralipid fat emulsion a promising therapeutic strategy on neurotoxicity induced by malathion in rats? European Review for Medical and Pharmacological Sciences, 18(4), 471–476.
5. 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.
6. Butnariu, M., Quispe, C., Herrera-Bravo, J., Sharifi-Rad, J., Singh, L., Aborehab, N. M., ... Cho, W. C. (2022). The pharmacological activities of Crocus sativus L.: A review based on the mechanisms and therapeutic opportunities of its phytoconstituents. Oxidative Medicine and Cellular Longevity, 2022(1), 8214821. https://doi.org/10.1155/2022/8214821
7. Buyukokuroglu, M. E., Cemek, M., Yurumez, Y., Yavuz, Y., & Aslan, A. (2008). Antioxidative role of melatonin in organophosphate toxicity in rats. Cell Biology and Toxicology, 24(2), 151–158. https://doi.org/10.1007/s10565-007-9024-z
8. Cemek, M., Büyükben, A., Büyükokuroğlu, M. E., Aymelek, F., & Tür, L. (2010a). Protective roles of vitamin E (α-tocopherol), selenium and vitamin E plus selenium in organophosphate toxicity in vivo: A comparative study. Pesticide Biochemistry and Physiology, 96(3), 113–118. https://doi.org/10.1016/j.pestbp.2009.09.009

9. Cemek, M., Emin Büyükokuroğlu, M., Yürümez, Y., Yavuz, Y., Aslan, A., Büyükben, A., & Aymelek, F. (2010b). Tissue trace and major element levels in organophosphate insecticide fenthion (Lebaycid®) toxicity in rats: Prophylactic and therapeutic effect of exogenous melatonin. Ecotoxicology and Environmental Safety, 73(2), 206– 212. https://doi.org/10.1016/j.ecoenv.2009.09.008
10. Cerdá-Bernad, D., Valero-Cases, E., Pastor, J.-J., & Frutos, M. J. (2022). Saffron bioactives crocin, crocetin and safranal: effect on oxidative stress and mechanisms of action. Critical Reviews in Food Science and Nutrition, 62(12), 3232–3249. https://doi.org/10.1080/10408398.2020.1864279
11. Chen, T.-H., Wang, H.-C., Chang, C.-J., & Lee, S.-Y. (2024). Mitochondrial glutathione in cellular redox homeostasis and disease manifestation. International Journal of Molecular Sciences, 25(2), 1314. https://doi.org/10.3390/ijms25021314
12. Chen, Y., Yang, Z., Nian, B., Yu, C., Maimaiti, D., Chai, M., ... Xu, D. (2024). Mechanisms of neurotoxicity of organophosphate pesticides and their relation to neurological disorders. Neuropsychiatric Disease and Treatment, 20, 2237–2254. https://doi.org/10.2147/NDT.S479757
13. Contreras, H. R., & Bustos-Obregón, E. (1999). Morphological alterations in mouse testis by a single dose of malathion. Journal of Experimental Zoology, 284(3), 355-359. https://doi.org/10.1002/ (SICI)1097-010X(19990801)284:3<355::AID-JEZ13>3.0.CO;2-N
14. Dorri, S. A., Hosseinzadeh, H., Abnous, K., Hasani, F. V., Robati, R. Y., & Razavi, B. M. (2015). Involvement of brain-derived neurotrophic factor (BDNF) on malathion induced depressive-like behavior in subacute exposure and protective effects of crocin. Iranian Journal of Basic Medical Sciences, 18(10), 958–966.
15. Draper, H. H., Dhanakoti, S. N., Hadley, M., & Piché, L. A. (2019). Malondialdehyde in biological systems. In Cellular antioxidant defense mechanisms (pp. 97–109). CRC Press. https://doi.org/10.1201/9780429289309-5
16. Durak, D., Uzun, F. G., Kalender, S., Ogutcu, A., Uzunhisarcikli, M., & Kalender, Y. (2009). Malathion-induced oxidative stress in human erythrocytes and the protective effect of vitamins C and E in vitro. Environmental Toxicology, 24(3), 235–242. https://doi.org/10.1002/tox.20423
17. Han, D., & Wen, L. (2026). Tissue-specific oxidative stress and protective interventions in malathion-induced toxicity in rodent models: A comprehensive review. Ecotoxicology and Environmental Safety, 309, 119607. https://doi.org/10.1016/j.ecoenv.2025.119607
18. Hazman, Ö., & Bozkurt, M. F. (2015). Anti-inflammatory and antioxidative activities of safranal in the reduction of renal dysfunction and damage that occur in diabetic nephropathy. Inflammation, 38(4), 1537–1545. https://doi.org/10.1007/s10753-015-0128-y
19. Ighodaro, O. M., & Akinloye, O. A. (2018). First line defence antioxidants-superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX): Their fundamental role in the entire antioxidant defence grid. Alexandria Journal of Medicine, 54(4), 287–293. https://doi.org/10.1016/j.ajme.2017.09.001
20. Jain, S. K., McVie, R., Duett, J., & Herbst, J. J. (1989). Erythrocyte Membrane Lipid Peroxidation and Glycosylated Hemoglobin in Diabetes. Diabetes, 38(12), 1539–1543. https://doi.org/10.2337/diab.38.12.1539 Janaszak-Jasiecka, A., Płoska, A., Wierońska, J. M., Dobrucki, L. W., & Kalinowski, L. (2023). Endothelial dysfunction due to eNOS uncoupling: molecular mechanisms as potential therapeutic targets. Cellular & Molecular Biology Letters, 28(1), 21. https://doi.org/10.1186/s11658-023-00423-2
21. Karabag-Coban, F., Bulduk, I., Liman, R., Ince, S., Cigerci, I., & Hazman, O. (2016). Oleuropein alleviates malathion-induced oxidative stress and DNA damage in rats. Toxicological & Environmental Chemistry, 98(1), 101–108. https://doi.org/10.1080/02772248.2015.1110156
22. Lasram, M. M., Dhouib, I. B., Bouzid, K., Lamine, A. J., Annabi, A., Belhadjhmida, N., ... Gharbi, N. (2014a). Association of inflammatory response and oxidative injury in the pathogenesis of liver steatosis and insulin resistance following subchronic exposure to malathion in rats. Environmental Toxicology and Pharmacology, 38(2), 542– 553. https://doi.org/10.1016/j.etap.2014.08.007 Lasram, M. M., Bini Douib, I., Bouzid, K., Annabi, A., EL ELJ, N., Dhouib, H., ... Gharbi, N. (2014b). Effects of N-acetyl-L-cysteine, in vivo, against pathological changes induced by malathion. 9 Toxicology Mechanisms and Methods, 24(4), https://doi.org/10.3109/15376516.2014.886003 294–306.
23. Lertnimitphun, P., Jiang, Y., Kim, N., Fu, W., Zheng, C., Tan, H., ... Xu, H. (2019). Safranal alleviates dextran sulfate sodium- induced colitis and suppresses macrophage-mediated inflammation. Frontiers in Pharmacology, 10, 1281. https://doi.org/10.3389/fphar.2019.01281
24. Miranda, K. M., Espey, M. G., & Wink, D. A. (2001). A rapid, simple spectrophotometric method for simultaneous detection of nitrate and nitrite. Nitric Oxide, 5(1), 62–71. https://doi.org/10.1006/niox.2000.0319
25. Mostafalou, S., Eghbal, M. A., Nili-Ahmadabadi, A., Baeeri, M., & Abdollahi, M. (2012). Biochemical evidence on the potential role of organophosphates in hepatic glucose metabolism toward insulin resistance through inflammatory signaling and free radical pathways. Toxicology and Industrial Health, 28(9), 840–851. https://doi.org/10.1177/0748233711425073
26. Ramadan, G., El-Beih, N. M., & Ahmed, R. S. A. (2017). Aged garlic extract ameliorates immunotoxicity, hematotoxicity and impaired burn-healing in malathion- and carbaryl-treated male albino rats. Environmental Toxicology, 32(3), 789–798. https://doi.org/10.1002/tox.22279
27. Saad, H., Elfeky, S. A., El-Gamel, N. E. A., & Abo Dena, A. S. (2025). Organophosphate pesticides: a review on classification, synthesis, toxicity, remediation and analysis. RSC Advances, 15(48), 40802– 40822. https://doi.org/10.1039/D5RA05552K
28. Sadeghnia, H. R., Kamkar, M., Assadpour, E., Boroushaki, M. T., & Ghorbani, A. (2013). Protective Effect of Safranal, a Constituent of Crocus sativus, on Quinolinic Acid-induced Oxidative Damage in Rat Hippocampus. Iranian Journal of Basic Medical Sciences, 16(1), 73- 82. https://doi.org/10.22038/IJBMS.2013.251
29. Samarghandian, S., Borji, A., Delkhosh, M. B., & Samini, F. (2013). Safranal treatment improves hyperglycemia, hyperlipidemia and oxidative stress in streptozotocin-induced diabetic rats. Journal of Pharmacy & Pharmaceutical Sciences, 16(2), 352. https://doi.org/10.18433/J3ZS3Q
30. Samarghandian, S., Azimi-Nezhad, M., & Samini, F. (2015). Preventive effect of safranal against oxidative damage in aged male rat brain. Experimental Animals, 64(1), 65–71. https://doi.org/10.1538/expanim.14-0027
31. Singh, S., Kumar, V., Thakur, S., Banerjee, B. D., Chandna, S., Rautela, R. S., ... Rai, A. (2011). DNA damage and cholinesterase activity in occupational workers exposed to pesticides. Environmental Toxicology and Pharmacology, 31(2), 278–285. https://doi.org/10.1016/j.etap.2010.11.005
32. Tamaddonfard, E., Farshid, A.-A., Eghdami, K., Samadi, F., & Erfanparast, A. (2013). Comparison of the effects of crocin, safranal and diclofenac on local inflammation and inflammatory pain responses induced by carrageenan in rats. Pharmacological Reports, 65(5), 1272–1280. https://doi.org/10.1016/S1734-1140(13)71485-3
33. Ullah, S., Li, Z., Hasan, Z., Khan, S. U., & Fahad, S. (2018). Malathion induced oxidative stress leads to histopathological and biochemical toxicity in the liver of rohu (Labeo rohita, Hamilton) at acute concentration. Ecotoxicology and Environmental Safety, 161, 270– 280. https://doi.org/10.1016/j.ecoenv.2018.06.002