Protective effect of Pistacia lentiscus extract on ethanol-induced testicular tissue damage in male Wistar rats: Histopathological alterations and fertility index
Subject Areas : Journal of Medicinal Herbs, "J. Med Herb" (Formerly known as Journal of Herbal Drugs or J. Herb Drug)Roya Asadinezhad 1 , Elham Moghtadaei Khorasgani 2 , Firoozeh Saghaei 3 , Taghi Taktaz Hafeshjani 4 * , Armin Deyhim 5 , Kambiz Yousefi 6
1 - Department of Veterinary, ShK.C., Islamic Azad University, ShahreKord, Iran;
2 - 2Department of Pathobiology, ShK.C., Islamic Azad University, ShahreKord, Iran;
3 - Department of Veterinary, ShK.C., Islamic Azad University, ShahreKord, Iran;
4 - Department of Animal Clinical Science, ShK.C., Islamic Azad University, ShahreKord, Iran;
5 - Department of Veterinary, ShK.C., Islamic Azad University, ShahreKord, Iran;
6 - Department of Surgery, ShK.C., Islamic Azad University, ShahreKord, Iran;
Keywords: Ethanol, mastic extract, spermatogonia, testis, testosterone ,
Abstract :
Background & Aim: Ethanol exposure is known to inflict significant toxic effects on testicular tissue, leading to reduced testosterone levels, impaired spermatogenesis, and ultimately reduced fertility. Given the well-documented antioxidant and anti-inflammatory properties of Pistacia lentiscus (mastic gum), the aim of the present study was to investigate the protective effects of its hydroalcoholic extract against ethanol-induced testicular damage in male Wistar rats. Experimental: Twenty-four adult male rats (weighing 200–250 g) were randomly assigned into four equal groups. The control group received no treatment, while the diseased group was exposed to ethanol (administered intraperitoneally at a dose of 10 mg/kg body weight). Two treatment groups received ethanol in combination with 100 and 200 mg/kg of the hydroalcoholic extract of P. lentiscus, respectively. Following a 3‑week treatment period, the animals were fasted, weighed, and blood samples were collected from the right ventricle under diethyl ether anesthesia. Serum testosterone levels were quantified using biochemical assays, and testicular tissue samples were processed for histopathological evaluation via hematoxylin–eosin staining. Results: Data clearly demonstrated that ethanol exposure significantly reduced serum testosterone levels and adversely affected sperm parameters including the number of seminiferous tubules, spermatogonia, spermatids, and spermatozoa compared with the control group (P<0.001). Although the co-administration of 100 mg/kg extract showed notable improvements, the 200 mg/kg dose restored most parameters almost to control values (P<0.001). Histopathological evaluations further confirmed a marked recovery in testicular architecture and cellular integrity with the higher dose of the extract. Recommended applications/industries: This research suggests that P. lentiscus extract could be developed as a natural therapeutic agent for reproductive health support in individuals exposed to alcohol-related oxidative stress. The findings may be applicable in the phytopharmaceutical, nutraceutical, and male fertility supplement industries.
Abu-Khudir, R., Almutairi, H.H., Abd El-Rahman, S.S., El-Said, K.S. 2023. The palliative and antioxidant effects of hesperidin against lead-acetate-induced testicular injury in male Wistar rats. Biomedicines, 11(9):2390.
Asadi, N., Bahmani, M., Kheradmand, A., Rafieian-Kopaei, M. 2017. The impact of oxidative stress on testicular function and the role of antioxidants in improving it: A review. Journal of Clinical and Diagnostic Research, 11: Ie01-ie5.
Ayala, A., Muñoz, M.F. and Argüelles, S. 2014. Lipid peroxidation: production, metabolism, and signaling mechanisms of malondialdehyde and 4-hydroxy-2-nonenal. Oxidative Medicine and Cell Longevity, 2014: 360438.
Botsaris, G., Orphanides, A., Yiannakou, E., Gekas, V. and Goulas, V. 2015. Antioxidant and Antimicrobial Effects of Pistacia lentiscus L. Extracts in Pork Sausages. Food Technology and Biotechnology, 53: 472-478.
Chouikh, N., Benguedouar, L., Ayad, R., Medjahed, Z., Bento-Silva, A., Duarte, N. .2025. Phytochemical analysis, antioxidant activity and modulatory effect of aqueous leaf extract of Pistacia lentiscus L on oxaliplatin-induced oxidative stress in isolated rat liver mitochondria Drug and Chemical Toxicology, 48: 219-230.
de Freitas, M.B., Moreira, E.A.M., Wilhelm Filho, D., Faccin, G.L., Trindade, E.M., Batista, S.M. 2014. Ethanol in lactation promotes oxidative stress in different phases of rat offspring. SPEN Journal, 9: e1-e6.
Donnelly, G.P., McClure, N., Kennedy, M.S., Lewis, S.E. 1999. Direct effect of alcohol on the motility and morphology of human spermatozoa. Andrologia, 31: 43-47.
Dosumu, O.O., Akinola, O.B. and Akang, E.N. 2012. Alcohol-induced testicular oxidative stress and cholesterol homeostasis in rats – The therapeutic potential of virgin coconut oil. Middle East Fertility Society Journal, 17: 122-128.
Eid, N.A., Shibata, M.A., Ito, Y., Kusakabe, K., Hammad, H., Otsuki, Y. 2002. Involvement of Fas system and active caspases in apoptotic signalling in testicular germ cells of ethanol-treated rats. International Journal of Andrology, 25: 159-67.
El-Sokkary, G.H. 2001. Quantitative study on the effects of chronic ethanol administration on the testis of adult male rat. Neuro Endocrinol Letters, 22: 93-99.
Emanuele, M.A. and Emanuele, N. 2001. Alcohol and the male reproductive system. Alcohol Research and Health, 25: 282-287.
Finelli, R., Mottola, F., Agarwal, A. 2021. Impact of alcohol consumption on male fertility potential: A narrative review. International Journal of Environment Research and Public Health, 19(1): 328-334.
Gao, Y., Jian, L., Lu, W., Xue, Y., Machaty, Z., Luo, H. 2021. Vitamin E can promote spermatogenesis by regulating the expression of proteins associated with the plasma membranes and protamine biosynthesis. Gene, 773: 145364.
Ghzaiel, I., Zarrouk, A., Nury, T., Libergoli, M., Florio, F., Hammouda, S. Antioxidant properties and cytoprotective effect of Pistacia lentiscus L. seed oil against 7β-Hydroxycholesterol-induced toxicity in C2C12 Myoblasts: reduction in oxidative stress, mitochondrial and peroxisomal dysfunctions and attenuation of cell death. Antioxidants, 10(11): 1772.
Hadaya, O., Bransi-Nicola, R., Shalev, Y., Azaizeh, H., Roth, Z., Muklada, H. 2020. Pistacia lentiscus extract enhances mammary epithelial cells’ productivity by modulating their oxidative status. Scientific Reports, 10: 20985.
Howard, S.R. 2021. Interpretation of reproductive hormones before, during and after the pubertal transition-Identifying health and disordered puberty. Clinical Endocrinology, 95: 702-715.
Komsky-Elbaz, A., Saktsier, M., Biran, D., Argov-Argaman, N., Azaizeh, H., Landau, Y.S. 2019. Atrazine-induced toxicity in goat spermatozoa is alleviated to some extent by polyphenol-enriched feed. Chemosphere, 236: 124858.
Krzastek, S.C., Farhi, J., Gray, M. and Smith, R.P. 2020. Impact of environmental toxin exposure on male fertility potential. Translational Andrology and Urology, 9: 2797-2813.
La Vignera, S., Condorelli, R.A., Balercia, G., Vicari, E., Calogero, A.E. 2013. Does alcohol have any effect on male reproductive function? A review of literature. Asian Journal of Andrology, 15: 221-225.
Lee, H.Y., Naseer, M.I., Lee, S.Y. and Kim, M.O. 2010. Time-dependent effect of ethanol on GnRH and GnRH receptor mRNA expression in hypothalamus and testis of adult and pubertal rats. Neuroscience Letters, 471: 25-29.
Ma, Y., Yu, X., Liu, Y.F., Song, B., Sun, Z. and Zhao, S. 2024. Immunoregulation and male reproductive function: Impacts and mechanistic insights into inflammation. Andrology, 1-20.
Malmir, M., Mehranjani, M.S., Faraji, T., Noreini, S.N. 2021. Antioxidant effect of vitamin E on the male rat reproductive system by a high oral dose of Bisphenol-A. Toxicology Research and Application, 2021:5.
Maneesh, M., Jayalekshmi, H., Dutta, S., Chakrabarti, A. and Vasudevan, D.M. 2005. Role of oxidative stress in ethanol induced germ cell apoptosis - An experimental study in rats. Indian Journal of Clinical Biochemistry, 20: 62-67.
Martinez, M., Macera, S., de Assis, G.F., Pinheiro, P.F., Almeida, CC., Tirapelli, L.F, .2009. Structural evaluation of the effects of chronic ethanol ingestion on the testis of Calomys callosus. Tissue and Cell, 41: 199-205.
Nassab, G., Bohlouli, S., Ghanbari, A. 2018. Therapeutic effect of Ferulago angulata extract on reproductive parameters and serum testosterone levels in diabetic male rats. Journal of Reports in Pharmaceutical Sciences, 7: 1-8.
Rosenblum, E.R., Gavaler, J.S., Van Thiel, D.H. 1989. Lipid peroxidation: a mechanism for alcohol-induced testicular injury Free Radical Biology and Medicine, 7: 569-77.
Roychoudhury, S., Chakraborty, S., Choudhury, A.P., Das, A., Jha, N.K., Slama, P .2021. Environmental factors-induced oxidative stress: Hormonal and molecular pathway disruptions in hypogonadism and erectile Dysfunction antioxidants (Basel). Antioxidants, 10(6):837.
Sawidis, T., Yurukova, L., Askitis, T. 2010. Chios mastic, a natural supplement for zinc to enhance male sexuality and prostate function. Pharmacetical Biology, 48: 48-54.
Shaki, F., arab-nozari, M., Elahi, P., Ghasemi, M., Habibi, E. 2019. Ameliorative effect of Viola odorata. L. ethyl acetate extract against nephrotoxicity induced by chronic ethanol exposure in rats. Journal of Mazandaran University of Medical Sciences, 29: 1-13.
Shipa, S.J., Khandokar, L., Bari, M.S., Qais, N., Rashid, M.A. and Haque, M.A. 2022. An insight into the anti-ulcerogenic potentials of medicinal herbs and their bioactive metabolites. Journal of Ethnopharmacology, 293: 115245.
Suzuki, R., Sakagami, H., Amano, S., Fukuchi, K., Sunaga, K., Kanamoto, T. 2017. Evaluation of biological activity of mastic extracts based on chemotherapeutic indices. In Vivo, 31 591-598.
Yousefi, K., Farid, M., Moghtadaei Khorasgani, E and Yadegari, M. 2025. Therapeutic potential of cymbopogon citratus hydroalcoholic extract and vitamin E coadministration in alleviating complications from testicular ischemia–reperfusion-induced injury in rats. Andrologia, 2025: 4028767.
Zakhari, S. 2006. Overview: how is alcohol metabolized by the body? Alcohol Research and Health, 29: 245-254.
Zirkin, B.R., Papadopoulos, V. 2018. Leydig cells: formation, function, and regulation. Biology Reprot, 99: 101-111.