Protective effects of Moringa oleifera against cyclophosphamide-induced oxidative and reproductive damage in male rabbits
DOI:
https://doi.org/10.55779/nsb17412756Keywords:
antioxidants, cyclophosphamide, Moringa oleifera, Oryctolagus cuniculus, reproductive toxicity, trace elementsAbstract
Cyclophosphamide (CP), a widely used chemotherapeutic agent, induces reproductive toxicity through oxidative stress and associated cellular damage. This study aimed to evaluate the protective effects of Moringa oleifera (MO) leaf extract, a plant rich in antioxidants and micronutrients, against CP-induced testicular and epididymal toxicity in adult male rabbits. Twenty-one rabbits were randomly assigned to three groups (n = 7): Group I (Control): received distilled water orally; Group II (CP): received CP (50 mg/kg/week, intraperitoneally); Group III (MO+CP): received MO extract (300 mg/kg/day, orally) in combination with CP for four weeks. Reproductive assessed included testicular and epididymal weights, semen analysis, DNA fragmentation, testicular and epididymal histopathology, oxidative stress markers (MDA, SOD, GPx), trace elements (Zn, iron), and hormones (testosterone, LH, FSH). CP administration significantly impaired reproductive function, reducing sperm motility decreasing by approximately 43%, and MDA levels increasing by 152%, along with altered trace element levels, and reduced hormone concentrations. MO co-treatment significantly mitigated these effects, reducing MDA levels by approximately 46%, increasing SOD and GPx activities, and partially restoring (70–85%) testosterone and sperm parameters close to control values. These findings suggest that MO leaf extract may serve as a promising natural antioxidant agent to alleviate CP-induced male reproductive toxicity, likely by reducing lipid peroxidation (↓MDA), enhancing antioxidant defenses (↑SOD, GPx), and stabilizing essential trace elements.
Metrics
References
Aitken RJ, De Iuliis GN (2007). Origins and consequences of DNA damage in male germ cells. Reproductive Biomedicine Online 14(6):727-733. https://doi.org/10.1016/S1472-6483(10)60676-1
Ali A, Qamar A, Khan M, Mughal AM, Tanvir S, Iqbal S (2024). Moringa oleifera mitigates the fluoxetine-induced damage to the histological architecture of the seminiferous tubules in adult male rats. Annals of the Pakistan Institute of Medical Sciences 21(1):52-57. https://doi.org/10.48036/apims.v20i1.843
Anan HH, Wahba NS, Abdallah MA, Mohamed DA (2017). Histological and immunohistochemical study of cyclophosphamide effect on adult rat testis. International Journal of Scientific Reports 3(2):39-48.
Bancroft JD, Gamble M (2019). Theory and Practice of Histological Techniques. 8th ed. Elsevier, London.
Bedwal RS, Bahuguna A (1994). Zinc, copper and selenium in reproduction. Experientia 50(7):626-640.
Cengiz M, Sahinturk V, Cetik Yildiz S, Şahin İK, Bilici N, Yaman SO, Altuner Y, Appak-Baskoy S, Ayhanci A (2020). Cyclophosphamide-induced oxidative stress, lipid peroxidation, apoptosis and histopathological changes in rats: Protective role of boron. Biological Trace Element Research 198(2):462-472. https://doi.org/10.1016/j.jtemb.2020.126574
Ceribaşi AO, Türk G, Sönmez M, Sakin F, Ateşşahin A (2010). Toxic effect of cyclophosphamide on sperm morphology, testicular histology and blood oxidant-antioxidant balance, and protective roles of lycopene and ellagic acid. Basic & Clinical Pharmacology & Toxicology 107(3):730-736. https://doi.org/10.1111/j.1742-7843.2010.00571.x
DRG International Inc. (2020). Testosterone, LH, and FSH ELISA kit instruction manual. USA.
El-Desoky NI, Hashem NM, Elkomy AG, Riad EA (2017). Physiological response and semen quality of rabbit bucks supplemented with Moringa oleifera leaves extract during heat stress. Animal 11(7):1129-1136. https://doi.org/10.1017/S1751731117000088
Emadi A, Jones RJ, Brodsky RA (2009). Cyclophosphamide and cancer: golden anniversary. Nature Reviews Clinical Oncology 6(11):638-647. https://www.nature.com/articles/nrclinonc.2009.146
Gameiro S, Boivin J, Vermeulen N, et al. (2015). ESHRE guideline: routine psychosocial care in infertility and medically assisted reproduction—a guide for fertility staff. Human Reproduction 30(11):2476-2485. https://doi.org/10.1093/humrep/dev177
Gopalakrishnan L, Doriya K, Kumar DS (2016). Moringa oleifera: A review on nutritive importance and its medicinal application. Food Science and Human Wellness 5(2):49-56. https://doi.org/10.1016/j.fshw.2016.04.001
Grande G, Barrachina F, Soler-Ventura A, Jodar M, Mancini F, Marana R, Chiloiro S, Pontecorvi A, Oliva R, Milardi D (2022). The role of testosterone in spermatogenesis: lessons from proteome profiling of human spermatozoa in testosterone deficiency. Frontiers in Endocrinology 13:852661. https://doi.org/10.3389/fendo.2022.852661
Hamed NS, Hammad HB, Abdou MI (2024). Moringa seeds mitigate oxidative stress and promote antioxidant activity in aging male rats. Biomedical Research and Therapy 11(11):926-942. https://doi.org/10.15419/bmrat.v11i10.926
Harrer A, Meyron-Holtz EG, Meinhardt A (2025). The role of iron in normal and impaired testicular function. Andrology. https://doi.org/10.1111/andr.70068
Huang L, Yao G, Huang G, Jiang C, Li L, Liao L, Yuan G, Shang L, Xu W (2021). Association of zinc deficiency, oxidative stress and increased double-stranded DNA breaks in globozoospermic infertile patients and its implication for the assisted reproductive technique. Translational Andrology and Urology 10(3):1088-1101. https://doi.org/10.21037/tau-20-1116
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193(1):265-275.
Maremanda KP, Srivalliputturu SB, Jena G (2020). Zinc deficient diet exacerbates the testicular and epididymal damage in type 2 diabetic rat: studies on oxidative stress-related mechanisms. Reproductive Biology 20(2):191-201. https://doi.org/10.1016/j.repbio.2020.03.002
Marklund S, Marklund G (1974). Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. European Journal of Biochemistry 47(3):469-474.
Merker HJ, Günther T (1997). Testis damage induced by zinc deficiency in rats. Journal of Trace Elements in Medicine and Biology 11(1):19-22. https://doi.org/10.1016/S0946-672X(97)80004-1
Miller-Ihli NJ (1988). Accuracy in trace analysis: trace element determinations in biologicals using atomic absorption spectrometry. Journal of Research of the National Bureau of Standards 93(3):247-254. https://doi.org/10.6028/jres.093.073
Najafi T (2017). Chemotherapy‑induced oxidative stress and infertility. International Journal of Women’s Health and Reproduction Sciences 5(2):80-81. https://doi.org/10.15296/ijwhr.2017.15
Nayak G, Rao A, Mullick P, Mutalik S, Kalthur SG, Adiga SK, Kalthur G (2020). Ethanolic extract of Moringa oleifera leaves alleviate cyclophosphamide-induced testicular toxicity by improving endocrine function and modulating cell specific gene expression in mouse testis. Journal of Ethnopharmacology 259:112922. https://doi.org/10.1016/j.jep.2020.112922
Nayak G, Vadinkar A, Nair S, Kalthur SG, D'Souza AS, Shetty PK, Mutalik S, Shetty MM, Kalthur G, Adiga SK (2015). Sperm abnormalities induced by pre-pubertal exposure to cyclophosphamide are effectively mitigated by Moringa oleifera leaf extract. Andrologia 47(6):645-654. https://doi.org/10.1111/and.12422
Obembe OO (2019). Sex hormones and oxidative stress biomarkers of male Wistar rats treated with Moringa oleifera seed fractions. JBRA Assisted Reproduction 23(4):408-413. https://doi.org/10.5935/1518-0557.20190047
Ogunlade BO, Jeje SO, Adelakun SA, Akingbade GT (2022). Moringa oleifera restored semen quality, hormonal profile, and testicular morphology against Highly Active Antiretroviral Therapy-induced toxicity in adult male Wistar rats. JBRA Assisted Reproduction 26(1):3-12. https://doi.org/10.5935/1518-0557.20210032
Ohkawa H, Ohishi N, Yagi K (1979). Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry 95(2):351-358. https://doi.org/10.1016/0003-2697(79)90738-3
Olowe TG, Oyovwi MO, Nwangwa KE, Ohwin EP, Oghenetega OB (2024). Cytotoxic properties of cyclophosphamide: A focus on its mechanistic impacts on male gonadal functions. Journal of Experimental & Reproductive Pharmacology 9(2):106-115. https://doi.org/10.14218/JERP.2023.00025S
Özbilgin MK, Demirören S, Üçöz M, Oztatlici M (2021). Cyclophosphamide suppresses spermatogenesis in the testis of mice through downregulation of miR-34b and miR-34c. Andrologia 53:e14071. https://doi.org/10.1111/and.14071
Peñalver R, Martínez-Zamora L, Lorenzo JM, Ros G, Nieto G (2022). Nutritional and antioxidant properties of Moringa oleifera leaves in functional foods. Foods 11(8):1107. https://doi.org/10.3390/foods11081107
Prasad AS (1995). Trace Elements in Human Health and Disease. 4th ed. Academic Press, San Diego, CA.
Razavi SR, Khadivi F, Hashemi F, Bakhtiari A (2018). Effect of zinc on spermatogenesis and sperm chromatin condensation in bleomycin, etoposide, and cisplatin-treated rats. Cell Journal 20(4):521-526. https://doi.org/10.22074/cellj.2019.5522
Rezvanfar MA, Sadrkhanlou RA, Ahmadi A, Shojaei-Sadee H, Rezvanfar MA, Mohammadirad A, Salehnia A, Abdollahi M (2008). Protection of cyclophosphamide-induced toxicity in reproductive tract histology, sperm characteristics, and DNA damage by an herbal source; evidence for role of free-radical toxic stress. Human & Experimental Toxicology 27(12):953-962. https://doi.org/10.1177/0960327108102046
Rotruck JT, Pope AL, Ganther HE, Swanson AB, Hafeman DG, Hoekstra WG (1973). Selenium: biochemical role as a component of glutathione peroxidase. Science 179(4073):588-590. https://doi.org/10.1126/science.179.4073.588
Salsi G, Greco C, Laudicina VA, Lucia C, Muscarella SM, Greco G, Orlando S, Fascella G, Mammano MM (2025). Preliminary results of Moringa oleifera Lam. grown in a semi-arid Mediterranean environment in a climate change scenario. Frontiers in Sustainable Food Systems 9:1576147. https://doi.org/10.3389/fsufs.2025.1576147
Setiyowati PAI, Yuningtyaswari Y, Primindari RS, Maghfuroh L, Sari NIP, Oktavia NF, Lim V, Hayati A (2025). Moringa oleifera leaf extract modulates oxidative stress and hormonal imbalance to alleviate di-(2-ethylhexyl) phthalate-induced testicular injury in rats. Journal of Animal Health and Production 13(4):1261-1269. https://doi.org/10.17582/journal.jahp/2025/13.4.1261.1269
Sherif IO (2018). Uroprotective mechanism of quercetin against cyclophosphamide-induced urotoxicity: effect on oxidative stress and inflammatory markers. Journal of Cellular Biochemistry 119(9):7441-7448. https://doi.org/10.1002/jcb.27053
Singh S, Mishra AK, Lata S, Tiwari KN (2017). Aggravation of cyclophosphamide-induced reproductive toxicity in mice by aqueous extract of Aegle marmelos (L.). Brazilian Journal of Pharmaceutical Sciences 53(3):e177. https://doi.org/10.1590/s2175-97902017000300177
World Health Organization (WHO) (2021). WHO Laboratory Manual for the Examination and Processing of Human Semen. 6th ed. Geneva: WHO Press.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2025 Lamia RIHANI, Asma SAIHIA

This work is licensed under a Creative Commons Attribution 4.0 International License.
Papers published in Notulae Scientia Biologicae are Open-Access, distributed under the terms and conditions of the Creative Commons Attribution License.
© Articles by the authors; licensee SMTCT, Cluj-Napoca, Romania. The journal allows the author(s) to hold the copyright/to retain publishing rights without restriction.
License:
Open Access Journal - the journal offers free, immediate, and unrestricted access to peer-reviewed research and scholarly work, due SMTCT supports to increase the visibility, accessibility and reputation of the researchers, regardless of geography. Users are allowed to read, download, copy, distribute, print, search, or link to the full texts of the articles, or use them for any other lawful purpose, without asking prior permission from the publisher or the author.







.png)













