Morin Reduces Ciprofloxacin-Induced Brain Damage by Regulating Oxidative Stress, Inflammation, and Histopathological Changes

Öz

Ciprofloxacin (CPFX), one of the fluoroquinones, is an antibacterial antibiotic that is used in the treatment of urinary tract, respiratory, abdominal and gastrointestinal infections as well as having negative side effects on the central nervous system. Morin, a bioactive flavonoid, is an antioxidant with neuroprotective properties thanks to its many pharmacological properties. The protective effect off Morin against brain damage caused by CPFX, broad spectrum antibiotic, was investigated. Twentyeight female rats were divided into four groups as control, Morin, CPFX, CPFX+Morin. CPFX, Morin was administered once a day for seven days. Oxidative stress and inflammation parameters were analyzed to determine brain tissue damage and histopathological analysis was performed to determine tissue damage and structural changes. According to the findings obtained as a result of the analyses, CPFX increased nuclear factor kappa B (NF-B), malondialdehyde (MDA), tumor necrosis factor-alpha (TNF-α), and interleukin 1β (IL-1β) and decreased glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) in brain. Co-administration of CPFX with Morin increased GSH and GPx, SOD, CAT and decreased NF-B, MDA, TNF-α, IL-1β levels. According to findings, when evaluated together, it was found that CPFX caused damage in brain tissue by causing oxidative stress and inflammation and Morin as a supportive treatment reduced the damage in brain tissue by bringing these markers closer to normal.

Anahtar Kelimeler

• Brain
• Ciprofloxacin
• Inflammation
• Morin
• Oxidative stress

Morin, Oksidatif Stresi, İnflamasyonu ve Histopatolojik Değişiklikleri Düzenleyerek Siprofloksasin Kaynaklı Beyin Hasarını Azaltır

Öz

Florokinonlardan olan siprofloksasin (CPFX), idrar yolu, solunum, abdominal ve gastrointestinal enfeksiyonların tedavisinde kullanılmasının yanı sıra merkezi sinir sistemi üzerinde olumsuz yan etkileri bulunan antibakteriyel antibiyotiktir. Biyoaktif flavonoid olan Morin, birçok farmakolojik özellikleri sayesinde nöroprotektif özelliğe sahip antioksidandır. Geniş spektrumlu antibiyotik olan CPFX’in neden olduğu beyin hasarına karşı Morin’in koruyucu etkisi araştırıldı. Bu amaçla yapılan çalışmada yirmisekiz adet dişi rat kontrol, Morin, CPFX, CPFX+Morin olarak dört gruba ayrıldı. CPFX, Morin yedi gün boyunca günde bir defa uygulandı. Beyin dokusunda hasarı belirlemek amacıyla oksidatif stres ve inflamasyon parametleri ve doku hasarını ve yapısal değişiklikleri belirlemek için histopatolojik analiz yapıldı. Analizler sonucunda elde edilen bulgulara göre CPFX, beyinde nuclear factor kappa B (NF-B), malondialdehit (MDA), tumor necrosis factor-alpha (TNF-α) ve interleukin 1β (IL-1β)’i artırdığı, glutatyon peroksidaz (GPx), süperoksit dismutaz (SOD), katalaz (CAT) ile glutatyon (GSH)’yı azalttığı bulundu. CPFX’in Morin ile birlikte uygulanmasının GSH ile GPx, SOD, CAT’i artırdığı, NF-B, MDA, TNF-α, IL-1β düzeylerini azalttığı tespit edildi. Elde edilen bulgulara göre birlikte değerlendirildiğinde, CPFX’in oksidatif stres ve inflamasyona neden olarak beyin dokusunda hasara neden olduğu ve destekleyici tedavi olarak Morin’in bu belirteçleri normale yaklaştırarak beyin dokusunda hasarı azalttığı bulundu.

Anahtar Kelimeler

• Beyin
• İnflamasyon
• Morin
• Oksidatif stres
• Siprofloksasin

Kaynakça

1. Adedara, I. A., Awogbindin, I. O., Mohammed, K. A., Da-Silva, O. F., & Farombi, E. O. (2021). Abatement of the dysfunctional hypothalamic-pituitary-gonadal axis due to ciprofloxacin administration by selenium in male rats. Journal of Biochemical and Molecular Toxicology, 35(5), e22741. https://doi.org/10.1002/jbt.22741 Aebi, H. (1984). [13] Catalase in vitro. Methods in Enzymology. 105, 121-126. https://doi.org/ 10.1016/S0076-6879(84)05016-3
2. Aksu, E. H., Kandemir, F. M., Küçükler, S., & Mahamadu, A. (2018). Improvement in colistin‐induced reproductive damage, apoptosis, and autophagy in testes via reducing oxidative stress by chrysin. Journal of biochemical and molecular toxicology, 32(11), e22201. https://doi.org/10.1002/jbt.22201
3. Aksu, E. H., Kandemir, F. M., & Küçükler, S. (2021). Ameliorative effect of hesperidin on streptozotocin‐diabetes mellitus‐induced testicular DNA damage and sperm quality degradation in Sprague–Dawley rats. Journal of food biochemistry, 45(10), e13938. https://doi.org/10.1111/jfbc.13938
4. Aktas, M. S., Kandemir, F. M., Kirbas, A., Hanedan, B., & Aydin, M. A. (2017). Evaluation of oxidative stress in sheep infected with Psoroptes ovis using total antioxidant capacity, total oxidant status, and malondialdehyde level. Journal of Veterinary Research, 61(2), 197. https://doi.org/10.1515/jvetres-2017-0025
5. Aktas Senocak, E., Utlu, N., Kurt, S., Kucukler, S., & Kandemir, F. M. (2024). Sodium pentaborate prevents acetaminophen-induced hepatorenal injury by suppressing oxidative stress, lipid peroxidation, apoptosis, and inflammatory cytokines in rats. Biological Trace Element Research, 202(3), 1164-1173. https://doi.org/10.1007/s12011-023-03755-4
6. Alla, N., Palatheeya, S., Challa, S. R., & Kakarla, R. (2024). Morin attenuated the global cerebral ischemia via antioxidant, anti-inflammatory, and antiapoptotic mechanisms in rats. Metabolik Brain Disease, 39(7), 1323-1334. https://doi.org/10.1007/s11011-024-01410-y
7. Al-Naely, A. J., Al-Hamadawi, H. A., & Alumeri, J. K. (2022). An Examination of the Effect of Rutin against Neurotoxicity Induced by Ciprofloxacin Antibiotic in Wistar Rats. Archives of Razi Institute, 77(2), 835-841. https://doi.org/10.22092/ARI.2022.357094.1971
8. Aygörmez, S., Küçükler, S., Gür, C., Akaras, N., Maraşli, Ş., & Kandemir, F. M. (2025). Investigation of the effects of morin on potassium bromate-induced brain damage in rats via different pathways with biochemical and histopathological methods. Food and Chemical Toxicology, 201, 115466. https://doi.org/10.1016/j.fct.2025.115466

9. Badawy, S., Yang, Y. Q., Liu, Y., Marawan, M. A., Ares, I., Martinez, M. A., Martínez-Larrañaga, M. R., Wang, X., Anadón, A., & Martinez, M. (2021). Toxicity induced by ciprofloxacin and enrofloxacin: oxidative stress and metabolism. Critical Reviews in Toxicology, 51(9), 754-787. https://doi.org/10.1080/10408444.2021.2024496
10. Banaeeyeh, S., Razavi, B. M., & Hosseinzadeh, H. (2024). Neuroprotective Effects of Morin Against Cadmium- and Arsenic-Induced Cell Damage in PC12 Neurons. Biological Trace Element Research, 22, https://doi.org/10.1007/s12011-024-04407-x
11. Caglayan, C., Kandemir, F. M., Ayna, A., Gür, C., Küçükler, S., & Darendelioğlu, E. (2022). Neuroprotective effects of 18β-glycyrrhetinic acid against bisphenol A-induced neurotoxicity in rats: involvement of neuronal apoptosis, endoplasmic reticulum stress and JAK1/STAT1 signaling pathway. Metabolic Brain Disease, 37(6), 1931-1940. https://doi.org/10.1007/s11011-022-01027-z
12. Cakmak, F., Kucukler, S., Gur, C., Comakli, S., Ileriturk, M., & Fatih Mehmet Kandemir, F. M. (2023). Morin provides therapeutic effect by attenuating oxidative stress, inflammation, endoplasmic reticulum stress, autophagy, apoptosis, and oxidative DNA damage in testicular toxicity caused by ifosfamide in rats. Iranian Journal of Basic Medical Sciences, 26(10), 1227-1236. https://doi.org/10.22038/IJBMS.2023.71702.15580
13. Çelik, H., Kucukler, S., Çomaklı, S., Caglayan, C., Özdemir, S., Yardım, A., Karaman, M., & Kandemir, F. M. (2020). Neuroprotective effect of chrysin on isoniazid-induced neurotoxicity via suppression of oxidative stress, inflammation and apoptosis in rats. Neurotoxicology, 81, 197-208. https://doi.org/10.1016/j.neuro.2020.10.009
14. Dhouibi, R., Affes, H., Salem, M. B., 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
15. Ekinci Akdemir, F. N., Yildirim, S., Kandemir, F. M., Küçükler, S., Eraslan, E., & Güler, M. C. (2025). Protective Effects of Baicalein and Bergenin Against Gentamicin‐Induced Hepatic and Renal Injuries in Rats: An Immunohistochemical and Biochemical Study. Basic & Clinical Pharmacology & Toxicology, 136(1), e14121. https://doi.org/10.1111/bcpt.14121
16. Genc, M., Kandemir, F. M., & Coban, O. (2019). Effects of in-ovo rutin injection to fertile Japanese quail (Coturnix Coturnix Japonica) egg on hatchability, embryonic death, hatchling weight, and hatchling liver oxidative and nitrosative stress. Brazilian Journal of Poultry Science, 21, eRBCA-2019. https://doi.org/10.1590/1806-9061-2018-0786
17. Gür, C., Kandemir, F. M., Darendelioglu, E., Caglayan, C., Kucukler, S., Kandemir, O., & Ileriturk, M. (2021). Morin protects against acrylamide-induced neurotoxicity in rats: an investigation into different signal pathways. Environmental Science and Pollution Research, 28(36), 49808-49819. https://doi.org/10.1007/s11356-021-14049-4
18. Gür, C., & Kandemir, F. M. (2022). Evaluation of the levels of metalloproteinases as well as markers of oxidative stress and apoptosis in lung tissues after malathion and rutin administrations to rats. Turkish Journal of Nature and Science, 11(3), 51-57. https://doi.org/10.46810/tdfd.1132497
19. Hanedan, B., Ozkaraca, M., Kirbas, A., Kandemir, F. M., Aktas, M. S., Kilic, K., Comakli, S., Kucukler, S., & Bilgili, A. (2018). Investigation of the effects of hesperidin and chrysin on renal injury induced by colistin in rats. Biomedicine & Pharmacotherapy, 108, 1607-1616. https://doi.org/10.1016/j.biopha.2018.10.001
20. Hassan, S. M., Jawad, M. J., & Rasool, M. I. (2023). The potential effect of infliximab, dimethyl fumarate (DMF), and their combination in ciprofloxacin-induced renal toxicity in male rats. Journal of Medicine and Life, 16(3), 477-480. https://doi.org/10.25122/jml-2022-0197
21. Ibitoye, O. B., Aliyu, N. O., & Ajiboye, T. O. (2020). Protective Influence of Phyllanthus Muellarianus on Ciprofloxacin-Induced Neurotoxicity in Male Rats. Journal of Dietary Supplements, 17(3), 321-335. https://doi.org/10.1080/19390211.2019.1586805
22. Ibrahim, N. A., Buabeid, M. A., Elmorshedy, K. E., & Arafa, E. S. A. (2025). Cell protective effects of vitamin C against oxidative stress induced by ciprofloxacin on spermatogenesis: involvement of cellular apoptosis. Frontiers in Cell and Developmental Biology, 13, 1489959. https://doi.org/10.3389/fcell.2025.1489959
23. Ileriturk, M., Kandemir, O., & Kandemir, F. M. (2022). Evaluation of protective effects of quercetin against cypermethrin‐induced lung toxicity in rats via oxidative stress, inflammation, apoptosis, autophagy, and endoplasmic reticulum stress pathway. Environmental toxicology, 37(11), 2639-2650. https://doi.org/10.1002/tox.23624
24. Ishola, I. O., Awogbindin, I. O., Olubodun-Obadun, T. G., Oluwafemi, O. A., Onuelu, J. E., & Adeyemi, O. O. (2022). Morin ameliorates rotenone-induced Parkinson disease in mice through antioxidation and anti-neuroinflammation: gut-brain axis involvement. Brain Research, 1789, 147958. https://doi.org/10.1016/j.brainres.2022.147958
25. Kandemir, F. M., Ozkaraca, M., Küçükler, S., Caglayan, C., & Hanedan, B. (2018). Preventive effects of hesperidin on diabetic nephropathy induced by streptozotocin via modulating TGF-β1 and oxidative DNA damage. Toxin reviews, 37(4), 287-293. https://doi.org/10.1080/15569543.2017.1364268
26. Khalaf, M. M., Mahmoud, H. M., Kandeil, M. A., Mahmoud, H. A., & Salama, A. A. (2024). Fumaric acid protects rats from ciprofloxacin-provoked depression through modulating TLR4, Nrf-2, and p190-rho GTP. Drug and Chemical Toxicology, 47(6), 897-908. https://doi.org/10.1080/01480545.2024.2310641
27. Khamchai, S., Chumboatong, W., Hata, J., Tocharus, C., Suksamrarn, A., & Tocharus, J. (2022). Morin Attenuated Cerebral Ischemia/Reperfusion Injury Through Promoting Angiogenesis Mediated by Angiopoietin-1-Tie-2 Axis and Wnt/β-Catenin Pathway. Neurotoxicity Research, 40(1), 14-25. https://doi.org/10.1007/s12640-021-00470-7
28. Kızıl, H.E., Caglayan, C., Darendelioğlu, E., Ayna, A., Gür, C., Kandemir, F. M., & Küçükler, S.(2023). Morin ameliorates methotrexate-induced hepatotoxicity via targeting Nrf2/HO-1 and Bax/Bcl2/Caspase-3 signaling pathways. Molecular Biology Reports, 50(4), 3479-3488. https://doi.org/10.1007/s11033-023-08286-8
29. Küçükler, S., Kandemir, F. M., Özdemir, S., Çomaklı, S., & Caglayan, C. (2021). Protective effects of rutin against deltamethrin-induced hepatotoxicity and nephrotoxicity in rats via regulation of oxidative stress, inflammation, and apoptosis. Environmental Science and Pollution Research, 28(44), 62975-62990. https://doi.org/10.1007/s11356-021-15190-w
30. Küçükler, S., Caglayan, C., Özdemir, S., Çomaklı, S., & Kandemir, F. M. (2024). Hesperidin counteracts chlorpyrifos-induced neurotoxicity by regulating oxidative stress, inflammation, and apoptosis in rats. Metabolic Brain Disease, 39(4), 509-522. https://doi.org/10.1007/s11011-023-01339-8
31. Lawrence, R. A., & Burk, R. F. (1976). Glutathione peroxidase activity in selenium-deficient rat liver. Biochemical and Biophysical Research Communications, 71(4), 952-958. https://doi.org/10.1016/0006-291x(76)90747-6
32. 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, 402-408. https://doi.org/10.1006/meth.2001.1262.
33. Lowry, O. H., Rosebrough, N. J., Farr, A. L., & Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry, 193(1), 265-275. PMID: 14907713
34. Mojoyinola, A. O., Ishaya, H. B., Makena, W., Jacob, C. B., Jonga, U. M., Anochie, V. C., Denis, E. W., & Gadzama, M. N. (2023). Protective effect of ciprofloxacin-induced oxidative stress, testicular and hepatorenal injury by Citrullus lanatus L. (Watermelon) seeds in adult Wistar rats. South African Journal of Botany. 156, 365-375. https://doi.org/10.1016/j.sajb.2023.03.017
35. 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
36. Paxinos, G., & Watson, C., (2007). The Rat Brain in Stereotaxic Coordinates, Sixth Edn. San Diego, CA: Academic Press
37. Salama, A., Mahmoud, H. A. A., Kandeil, M. A., & Khalaf, M. M. (2021). Neuroprotective role of camphor against ciprofloxacin induced depression in rats: modulation of Nrf-2 and TLR4. Immunopharmacology and Immunotoxicology, 43(3), 309-318. https://doi.org/10.1080/08923973.2021.1905658
38. Sedlak, J., & Lindsay, R. H. (1968). Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with Ellman’s reagent. Analytical Biochemistry, 25, 192-205. https://doi.org/10.1016/0003-2697(68)90092-4
39. Sun, Y., Oberleyi 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/clinchem/34.3.497
40. Şimşek, H., Sefa Küçükler, S., Gür, C., İleritürk, M., Aygörmez, S., & Kandemir, F. M. (2023). Protective effects of zingerone against sodium arsenite-induced lung toxicity: A multi-biomarker approach. Iranian Journal of Basic Medical Sciences, 26(9), 1098-1106. https://doi.org/10.22038/IJBMS.2023.71905.15623
41. Thangarajan, S., Vedagiri, A., Somasundaram, S., Sakthimanogaran, R., & Murugesan, M. (2018). Neuroprotective effect of morin on lead acetate- induced apoptosis by preventing cytochrome c translocation via regulation of Bax/Bcl-2 ratio. Neurotoxicology and Teratology, 66, 35-45. https://doi.org/10.1016/j.ntt.2018.01.006
42. Tuncer, S. Ç., Akarsu, S. A., Küçükler, S., Gür, C., & Kandemir, F. M. (2023). Effects of sinapic acid on lead acetate-induced oxidative stress, apoptosis and inflammation in testicular tissue. Environmental Toxicology, 38(11), 2656-2667. https://doi.org/10.1002/tox.23900
43. Tuncer, S. Ç., Gur, C., Kucukler, S., Akarsu, S. A., & Kandemir, F. M. (2024). Effects of zingerone on rat induced testicular toxicity by sodium arsenite via oxidative stress, endoplasmic reticulum stress, inflammation, apoptosis, and autophagy pathways. Iranian Journal of Basic Medical Sciences, 27(5), 603-610. https://doi.org/10.22038/IJBMS.2024.73342.15934
44. Ubaid, M. M., Kadhim, S. H., & Al-kareem, Z. A. (2022). Protective effect of cinnamon oil against ciprofloxacin toxicity on liver and kidney of male Wistar rats. Journal of Applied and Natural Science, 14(4), 1430-1434. https://doi.org/10.31018/jans.v14i4.3823
45. Varışlı, B., Caglayan, C., Kandemir, F. M., Gür, C., Bayav, İ., & Genç, A. (2022). The impact of Nrf2/HO-1, caspase-3/Bax/Bcl2 and ATF6/IRE1/PERK/GRP78 signaling pathways in the ameliorative effects of morin against methotrexate-induced testicular toxicity in rats. Molecular Biology Reports, 49(10), 9641-9649. https://doi.org/10.1007/s11033-022-07873-5
46. Varışlı, B., Caglayan, C., Kandemir, F. M., Gür, C., Ayna, A., Genç, A., & Taysı, S. (2023). Chrysin mitigates diclofenac-induced hepatotoxicity by modulating oxidative stress, apoptosis, autophagy and endoplasmic reticulum stress in rats. Molecular Biology Reports, 50(1), 433-442. https://doi.org/10.1007/s11033-022-07928-7
47. Yakut, S., Atcalı, T., Çaglayan, C., Ulucan, A., Kandemir, F. M., Kara, A., & Anuk, T. (2024). Therapeutic Potential of Silymarin in Mitigating Paclitaxel-Induced Hepatotoxicity and Nephrotoxicity: Insights into Oxidative Stress, Inflammation, and Apoptosis in Rats. Balkan Medical Journal, 41(3), 193-205. https://doi.org/10.4274/balkanmedj.galenos.2024.2024-1-60
48. Yardim, A., Gur, C., Comakli, S., Ozdemir, S., Kucukler, S., Celik, H., & Kandemir, F. M. (2022). Investigation of the effects of berberine on bortezomib-induced sciatic nerve and spinal cord damage in rats through pathways involved in oxidative stress and neuro-inflammation. Neurotoxicology, 89, 127-139. https://doi.org/10.1016/j.neuro.2022.01.011
49. Yılmaz, S., Küçükler, S., Şimşek, H., Aygörmez, S., & Kandemir, F.M. (2024). Naringin protects against colistin-induced sciatic nerve damage by reducing oxidative stress, apoptosis and inflammation damage. Journal of Experimental and Clinical Medicine, 41(1), 53-59. https://doi.org/10.52142/omujecm.41.1.9