Journal of Ethnopharmacology and Toxicology | Volume 4 Issue 1 | Pages: 10-18 | Doi : 10.37446/jet/rsa/4.1.2026.10-18

Research Article

OPEN ACCESS | Published on : 19-Jan-2026

Dose-dependent protective effect of Cucurbita pepo seed extract on induced reproductive toxicity

• Anyanwu C. F.
• Department of Pharmacology, Faculty of Basic Clinical Sciences, University of Port Harcourt, P.M.B. 5323, Nigeria.

Siminialayi I.M.
• Department of Pharmacology, Faculty of Basic Clinical Sciences, University of Port Harcourt, P.M.B. 5323, Nigeria.

Kemebirinemughan T.
• Department of Pharmacology, Faculty of Basic Clinical Sciences, University of Port Harcourt, P.M.B. 5323, Nigeria.

Abstract

In recent times, there has been growing interest in the search for natural products as possible remedies for mitigating reproductive toxicity caused by intake of medications. This study aims to investigate the effect of ethanol extract of Cucurbita pepo (C. pepo) seed on pregabalin-induced reproductive toxicity in male Wistar rats. Group A (normal control) = 0.5ml and 5% Tween 80; B (negative control) = 75mg/kg pregabalin, group C= 500mg/kg of C. pepo extract; D = 1000mg/kg of C. pepo extract, group E = 75 mg/kg of pregabalin and 500mg/kg of C. pepo extract and group F = 75mg/kg of pregabalin and 1000mg/kg of C. pepo extract. There was a Significant (p<0.05) increase in testosterone, viable sperm cell, and decrease in abnormal sperm cell observed in test group C relative to controls. No significant variation in FSH and LH relative to both controls. A Significant (p<0.01) increase in dead sperm cells and significant (p<0.01) decrease in normal and viable sperm cells, in group F compared to controls. Cucurbita pepo, at lower doses may possess beneficial effects on male reproductive parameters. Higher doses in combination with pregabalin demonstrate potential reproductive toxicity.

Keywords

Cucurbita pepo seed, testosterone, luteinizing hormone, pregabalin, reproductive toxicity

References

• Abarikwu, S. O., Oleribe, A. L., Mgbudom-Okah, C. J., Onuah, C. L., Chikwendu, C. S., & Onyeike, E. N. (2022). The protective effect of fluted pumpkin seeds against atrazine-induced testicular injury. Drug and Chemical Toxicology, 45(2), 799–809.

  Aghaei, S., Nikzad, H., Taghizadeh, M., Tameh, A. A., Taherian, A., & Moravveji, A. (2014). Protective effect of pumpkin seed extract on sperm characteristics, biochemical parameters and epididymal histology in adult male rats treated with cyclophosphamide. Andrologia, 46(8), 927–935.

  Akomolafe, S. F. (2021). Effects of roasting on the phenolic phytochemicals and antioxidant activities of pumpkin seed. Vegetos, 34(3), 505–514.

  Akomolafe, S. F., Ogundare, M. A. B., Akinlua, I., Ajayi, O. O., Odesanmi, E. O., & Adewale, O. B. (2025). Comparative effect of dietary fortification of raw and roasted seeds of pumpkin (Cucurbita pepo L.) on fertility biomolecules in cisplatin-induced spermatotoxic rats. Journal of Trace Elements and Minerals, 12, 100236. https://doi.org/10.1016/j.jtemin.2025.100236.

  Almohaimeed, H. M., Hamed, S., Seleem, H. S., Batawi, A. H., Mohammedsaleh, Z. M., Balgoon, M. J., … Ayuob, N. (2021). An ethanolic extract of Cucurbita pepo L. seeds modifies neuroendocrine disruption in chronically stressed rats and adrenal expression of inflammatory markers and HSP70. Frontiers in Pharmacology, 12, 749766. https://doi.org/10.3389/fphar.2021.749766.

  Anyanwu, C. F., Georgewill, O. A., & Georgewill, U. O. (2024). Effect of Cucurbita pepo (Pumpkin) seed extracts on the cyclicity and reproductive hormones of female Wistar rats. Journal of Herbmed Pharmacology, 13(4), 620-629.

  Anyanwu, C. F., Georgewill, O. A., & Obinna, V. C. (2025). Reproductive outcome in female Wistar rats following treatment with Cucurbita pepo (pumpkin) seed extracts. British Journal of Healthcare and Medical Research, 12(02), 01–12.

  Anyanwu, C. F., Georgewill, U. O., Onuama, V. I., & Phil-Ashiri, N. U. (2024). Potential reproductive toxicity of Lacatomtom drink in male Wistar rats. Tropical Journal of Natural Product Research, 8(6).

  Bilinska, B., Hejmej, A., & Kotula-Balak, M. (2018). Preparation of testicular samples for histology and immunohistochemistry. In Sertoli cells: Methods and protocols (pp. 17–36). Springer.

  Carey, J. L., Nader, N., Chai, P. R., Carreiro, S., Griswold, M. K., & Boyle, K. L. (2017). Drugs and medical devices: Adverse events and the impact on women’s health. Clinical Therapeutics, 39(1), 10–22.

  Cheesbrough, M. (2006). District laboratory practice in tropical countries (Part 2, 2nd ed.). Cambridge University Press.

  de Fátima Mestre, V., Martins, C. C. N., de Brito, L. V., Zeffa, A. C., Sestário, C. S., & Salles, M. J. S. (2023). Pregabalin alters reproductive performance in male mice and causes congenital anomalies in offspring. Reproduction, Fertility and Development, 35(18), 750–759.

  ElAkhdar, N., Bawab, R., & Borjac, J. (2020). Effect of Ceratonia siliqua and Cucurbita pepo seed extracts on spermatogenesis in male mice. BAU Journal – Health and Wellbeing, 3(1), 7.

  Elshafie, H. S., Camele, I., & Mohamed, A. A. (2023). A comprehensive review on the biological, agricultural and pharmaceutical properties of plant-based secondary metabolites. International Journal of Molecular Sciences, 24(4), 3266.

  Fawzy, E. I., El Makawy, A. I., El-Bamby, M. M., & Elhamalawy, H. O. (2018). Improved effect of pumpkin seed oil against bisphenol-A adverse effects in male mice. Toxicology Reports, 5, 857–863.

  Hamed, S. A. (2018). Sexual dysfunctions induced by pregabalin. Clinical Neuropharmacology, 41(4), 116–122. https://doi.org/10.1097/WNF.0000000000000286.

  Harborne, A. J. (1998). Phytochemical methods: A guide to modern techniques of plant analysis. Springer Science & Business Media.

  Hashemi, B., & Dadkhah, T. (2014). Properties of Cucurbita pepo L. in Islamic Persian medicine. Health, Spirituality and Medical Ethics, 1(3), 24–28.

  Ivoh, G., Anyanwu, C. F., & Obinna, V. C. (2023). Oestrogenic activity of aqueous ethanol extract of Cucurbita pepo (pumpkin) seed in female Wistar rats. Asian J. Res. Rep. Endocrinol, 6(1), 170-178.

  Krzastek, S. C., Farhi, J., Gray, M., & Smith, R. P. (2021). Impact of environmental toxin exposure on male fertility potential. Translational Andrology and Urology, 9(6), 2797–2813. https://doi.org/10.21037/tau-20-685.

  Kumar, P., Srivastava, N., Pande, M., Prasad, J. K., & Sirohi, A. S. (2017). Evaluating sperm cell viability and membrane integrity. In Protocols in semen biology (comparing assays) (pp. 57–71). Springer.

  Lestari, B., & Meiyanto, E. (2018). The emerging nutraceutical potential of pumpkin seeds: A review. Indonesian Journal of Cancer Chemoprevention, 9(2), 92–101. https://doi.org/10.14499/indonesianjcanchemoprev9iss2pp92-101.

  Malan, D. F., & Neuba, D. F. (2011). Traditional practices and medicinal plants use during pregnancy by Anyi-Ndenye women (Eastern Côte d'Ivoire). African journal of reproductive health, 15(1), 85-93.

  Mascarenhas, C., Sousa, A. C., & Rato, L. (2024). Effects of pharmaceutical substances with obesogenic activity on male reproductive health. International Journal of Molecular Sciences, 25(4), 2324. https://doi.org/10.3390/ijms25042324.

  Matus, Z., Molnár, P., & Szabó, L. G. (1993). Main carotenoids in pressed seeds (Cucurbitae semen) of oil pumpkin (Cucurbita pepo convar. pepo var. styriaca). Acta Pharmaceutica Hungarica, 63(5), 247–256.

  Mehda, S., Laib, I., Diab, F., Attia, R., Benaissa, Y., Bellabidi, M., … Croun, D. (2025). Protective effects of Calligonum comosum against pregabalin-induced toxicity. Food Science & Nutrition, 13(7), e70681.

  Mohammadi, N., Shirian, S., Gorji, A., Roshanpajouh, M., Ahmadi, E., Nazari, H., & Arezoomandan, R. (2023). The potential protective effect of melatonin and N-acetylcysteine alone and in combination on opioid-induced testicular dysfunction and degeneration in rat. Reproductive Toxicology, 120, 108453.

  Mudau, F. N., Chimonyo, V. G. P., Modi, A. T., & Mabhaudhi, T. (2022). Neglected and underutilised crops: a systematic review of their potential as food and herbal medicinal crops in South Africa. Frontiers in Pharmacology, 12, 809866.

  Noh, S., Go, A., Kim, D. B., Park, M., Jeon, H. W., & Kim, B. (2020). Role of antioxidant natural products in management of infertility: a review of their medicinal potential. Antioxidants, 9(10), 957.

  Odigie, J. O., Anyanwu, C. F., & Ameja, C. N. (2025). Protective Effect of Ethanol Extract of Cucurbita pepo (Pumpkin) Seed on Pregabaline Induced Hepatorenal Toxicity. Journal of Complementary and Alternative Medical Research, 26(8), 35-43.

  Ofoego, U. C., Nwachukwu, O., Ekwujuru, E. U., Mbagwu, S., Nwakanma, A., Nweke, E. O., & Ofoego, A. N. (2017). Ethanolic Seed Extract of Cucurbita Pepo (Pumpkin) Protects the Testis from Azadirachta Indica (Neem) Induced Damage. Journal of Natural Sciences Research, 7(24), 120-131.

  Oh, J., Hong, S., Ko, S. H., & Kim, H. S. (2024). Evaluation of antioxidant effects of pumpkin (Cucurbita pepo L.) seed extract on aging-and menopause-related diseases using Saos-2 cells and ovariectomized rats. Antioxidants, 13(2), 241.

  Oyetayo, F. L., Akomolafe, S. F., & Osesanmi, T. J. (2020). Effect of dietary inclusion of pumpkin (Cucurbita pepo L.) seed on cisplatin-induced nephrotoxicity in rats. Journal of Food Biochemistry, 44(10), e13439.

  Oyetayo, F. L., Akomolafe, S. F., & Osesanmi, T. J. (2020). Effect of dietary inclusion of pumpkin (Cucurbita pepo L.) seed on nephrotoxicity occasioned by cisplatin in experimental rats. Journal of Food Biochemistry, 44(10), e13439.

  Rieger, J., Pelckmann, L.-M., & Drewes, B. (2020). Preservation and processing of intestinal tissue for the assessment of histopathology. In Animal models of allergic disease: Methods and protocols (pp. 267–280). Springer.

  Salehi, B., Capanoglu, E., Adrar, N., Catalkaya, G., Shaheen, S., Jaffer, M., ... & Capasso, R. (2019). Cucurbits plants: A key emphasis to its pharmacological potential. Molecules, 24(10), 1854.

  Salem Hareedy, M., Mohamed Tawfik, K., Badary, D. M., Ahmed Mahmoud, W., & Mohamed Mohamed, E. (2020). Pregabalin administration and withdrawal affect testicular structure and functions in rats. Andrologia, 52(11), e13808.

  Shokry, D. A., El Nabrawy, N., Yassa, H. D., Gaber, S. S., Batiha, G. E. S., & Welson, N. N. (2020). Pregabalin induced reproductive toxicity and body weight changes by affecting caspase3 and leptin expression: protective role of wheat germ oil. Life Sciences, 260, 118344.

  Tao, G., Dagher, F., Moballegh, A., & Ghose, R. (2020). Role of oxidative stress in the efficacy and toxicity of herbal supplements. Current Opinion in Toxicology, 20, 36-40.

  Theil, G., Richter, M., Schulze, M., Köttig, T., Patz, B., Heim, S., … Fornara, P. (2022). Extract from Cucurbita pepo improves BPH symptoms without affecting sexual function. World Journal of Urology, 40(7), 1769–1775.

  Wei, L. S., Goh, K. W., Abdul Hamid, N. K., Abdul Kari, Z., Wee, W., & Van Doan, H. (2022). Co-supplementation of probiotics and medicinal herbs in aquaculture: A mini-review. Frontiers in Veterinary Science, 9, 869564.

  World Health Organization. (2019). WHO consolidated guideline on self-care interventions for health: sexual and reproductive health and rights. World Health Organization.