Morphological and Biochemical Investigation of the Protective Effects of Panax ginseng on Methotrexate-Induced Testicular Damage

Objective: Methotrexate (MTX) is a chemotherapeutic agent that causes testicular toxicity used in the cure of various types of cancer. The anti-oxidant and anti-cancer effects of Panax ginseng (PxG) have been reported in both experimental and clinical studies. This study aims to examine the healing effect of PxG on testicular damage induced by MTX. Materials and Methods: Sprague Dawley male rats (8-week-olds) were used in the study. A single dose of MTX dissolved in saline (20 mg/kg) was given to MTX and MTX+PxG groups by intraperitoneal injection. PxG dissolved in saline (100 mg/kg) was given by orogastric gavage once a day for 5 days to the MTX+PxG group. Saline was given to the control and MTX groups orally during the experiments. After decapitation, the testis samples were obtained. Seminiferous tubules and basement membrane were evaluated histopathologically. Seminiferous tubule diameter and germinal epithelium thickness were measured. Furthermore, oxidative stress parameters such as malondialdehyde, glutathione, superoxide dismutase, and glutathione-S-transferase were measured. Results: MTX treatment caused seminiferous tubule degeneration with a decrease in Johnsen’s score, the seminiferous tubule’s diameter, and the germinal epithelium’s thickness. Parallel with the histopathological results increased testicular oxidative stress with an increase in malondialdehyde level and a decrease of endogenous anti-oxidant activity with a decrease in glutathione level and glutathione-S-transferase and superoxide dismutase activities. PxG treatment improved these histological and biochemical parameters in MTX-induced testis cytotoxicity. Conclusion: MTX treatment causes testicular damage via the oxidative processes. PxG treatment ameliorates MTX-induced testicular damage by inhibiting oxidative stress.

Keywords:

Methotrexate, Panax ginseng, testis, oxidative stress,

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Hamed KM, Dighriri IM, Baomar AF, et al. Overview of Methotrexate Toxicity: A Comprehensive Literature Review. Cureus. 2022;14(9):e29518. doi:10.7759/cureus.29518 google scholar
Bleyer WA. Methotrexate: clinical pharmacology, current status and therapeutic guidelines. Cancer Treat Rev. 1977;4(2):87-101. google scholar
Pinar N, Cakirca G, Ozgur T, Kaplan M. The protective effects of alpha lipoic acid on methotrexate induced testis injury in rats. Biomed Pharmacother. 2018;97:1486-1492. google scholar
Wang Y, Zhao TT, Zhao HY, Wang H. Melatonin protects methotrexate-induced testicular injury in rats. Eur Rev Med Pharmacol Sci. 2018;22(21):7517-7525. google scholar
Abdelhameed RFA, Fattah SA, Mehanna ET, et al. Zygo-Albuside A: New Saponin from Zygophyllum album L. with signif-icant antioxidant, anti-inflammatory and antiapoptotic effects against methotrexate-induced testicular damage. Int J Mol Sci. 2022;23(18):10799. doi:10.3390/yms231810799 google scholar
Kanpalta F, Ozbeyli D, Sen A, Cevik O, Sener G, Ercan F. Bitter melon (Momordica charantia) fruit extract ameliorates methotrexate-induced reproductive toxicity in male rats. Marmara Med J. 2021;34(3):222-228. google scholar
Vardi N, Parlakpinar H, Ates B, Cetin A, Otlu A. Antiapoptotic and antioxidant effects of beta-carotene against methotrexate-induced testicular injury. Fertil Steril. 2009;92(6):2028-2033. google scholar
Armagan A, Uzar E, Uz E, et al. Caffeic acid phenethyl ester modulates methotrexate-induced oxidative stress in testes of rat. Hum Exp Toxicol. 2008;27(7):547-552. google scholar Ganjkhani M, Nourozi S, Bigonah R, Rostami A, Shokri S. Ameliorating impacts of ginseng on the apoptosis of spermato-genic cells and sperm quality in temporal lobe epilepsy rat model treated with valproate. Andrologia. 2019;51(9):e13348. doi: 10.1111/and.13348. google scholar
Padmanabhan S, Tripathi DN, Vikram A, Ramarao P, Jena GB. Methotrexate-induced cytotoxicity and genotoxicity in germ cells of mice: intervention of folic and folinic acid. Mutat Res. 2009;673(1):43-52. google scholar
Ernst E. Panax ginseng: an overview of the clinical evidence. JGinseng Res. 2010;34(4):259-263. google scholar
Ratan ZA, Haidere MF, Hong YH, et al. Pharmacological poten-tial of ginseng and its major component ginsenosides. J Ginseng Res. 2021;45(2):199-210. google scholar
Liu CX, Xiao PG. Recent advances on ginseng research in China. J Ethnopharmacol. 1992;36(1):27-38. google scholar
Lu J-M, Yao Q, Chen C. Ginseng compounds: an update on their molecular mechanisms and medical applications. Curr Vasc Pharmacol. 2009;7(3):293-302. google scholar
Yue PY, Mak NK, Cheng YK, et al. Pharmacogenomics and the Yin/Yang actions of ginseng: anti-tumor, angiomodulating and steroid-like activities of ginsenosides. Chin Med. 2007;2:6. doi: 10.1186/1749-8546-2-6. google scholar
Kim W, Hwang S, Lee H, Song H, Kim S. Panax ginseng protects the testis against 2,3,7, 8-tetrachlorodibenzo-p-dioxin induced tes-ticular damage in guinea pigs. BJU Int. 1999;83(7):842-849. google scholar
Wanderley MI, Saraiva KL, Cesar Vieira JS, Peixoto CA, Udrisar DP. Foetal exposure to Panax ginseng extract reverts the effects of prenatal dexamethasone in the synthesis of testosterone by Leydig cells of the adult rat. Int J Exp Pathol. 2013;94(3):230-240. google scholar
Aslan E, Kumalar K, Güzel H, Demirel HH, Çelik S, Pektaş MB. Effects of Panax ginseng on cisplatin-induced testicular damage of rats. Ant J Bot. 2021;5(1):37-43. google scholar
Uzar E, Sahin O, Koyuncuoglu HR, et al. The activity of adeno-sine deaminase and the level of nitric oxide in spinal cord of methotrexate administered rats: protective effect of caffeic acid phenethyl ester. Toxicology. 2006;218(2-3):125-133. google scholar
Qiao C, Den R, Kudo K, et al. Ginseng enhances contex-tual fear conditioning and neurogenesis in rats. Neurosci Res. 2005;51(1):31-38. google scholar
Koşmaz K, Durhan A, Şenlikçi A, et al. Evaluation of the pro-tective effect of Red Ginseng on lipid profile, endothelial and oxidative damage after splenectomy in rats. Arch Clin Exp Med. 2021;6(2):43-49. google scholar
Johnsen SG. Testicular biopsy score count-a method for regis-tration of spermatogenesis in human testes: normal values and results in 335 hypogonadal males. Hormones. 1970;1(1):2-25. google scholar
Elmas MA, Ozakpinar OB, Kolgazi M, Sener G, Arbak S, Ercan F. Exercise improves testicular morphology and oxidative stress parameters in rats with testicular damage induced by a high-fat diet. Andrologia. 2022;54(11):e14600. doi:10.1111/and.14600. google scholar
Fujii S, Dale GL, Beutler E. Glutathione-dependent protection against oxidative damage of the human red cell membrane. Blood. 1984;63(5):1096-1101. google scholar
Habig WH, Jakoby WB. Assays for differentiation of glutathione S-Transferases. Methods Enzymol. 1981;77:398-405. google scholar
Ledwozyw A, Michalak J, Stepien A, Kadziolka A. The relation-ship between plasma triglycerides, cholesterol, total lipids and lipid peroxidation products during human atherosclerosis. Clin Chim Acta. 1986;155(3):275-283. google scholar
Mylroie AA, Collins H, Umbles C, Kyle J. Erythrocyte superoxide dismutase activity and other parameters of copper status in rats ingesting lead acetate. Toxicol Appl Pharmacol. 1986;82(3):512-520. google scholar
Tracey WR, Linden J, Peach MJ, Johns RA. Comparison of spec-trophotometric and biological assays for nitric oxide (NO) and endothelium-derived relaxing factor (EDRF): nonspecificity of the diazotization reaction for NO and failure to detect EDRF. J Pharmacol Exp Ther. 1990;252(3):922-928. google scholar
Rikans LE, Hornbrook KR. Lipid peroxidation, antioxidant protection and aging. Biochim Biophys Acta. 1997;1362(2-3):116-127. google scholar
Koroglu KM, Cevik O, Sener G, Ercan F. Apocynin alleviates cisplatin-induced testicular cytotoxicity by regulating oxidative stress and apoptosis in rats. Andrologia. 2019;51(4):e13227. doi: 10.1111/and.13227. google scholar
Leung KW, Wong AS. Ginseng and male reproductive function. Spermatogenesis. 2013;3(3):e26391. doi:10.4161/spmg.26391. google scholar
Sanad NH, Abbas HR, Yaseen AA, Habeeb IA, Alsalim HA. Hormonal, histological, and comparative study of the effect of pure ginseng on testicular function in the breeding/non- breeding season of rams in Basrah. Arch Razi Inst. 2021;76(5):1519-1535. google scholar
Thomford NE, Senthebane DA, Rowe A, et al. Natural products for drug discovery in the 21st century: Innovations for novel drug discovery. Int J Mol Sci. 2018;19(6):1578. google scholar
He Y, Yang J, Lv Y, et al. A Review of Ginseng Clini-cal Trials Registered in the WHO international clinical tri-als registry platform. Biomed Res Int. 2018;2018:1843142. doi:10.1155/2018/1843142. google scholar
Wee JJ, Mee Park K, Chung AS. Biological activities of Ginseng and its application to human health. In: Benzie IFF, Wachtel-Galor S, eds. Herbal Medicine: Biomolecular and Clinical Aspects. 2nd ed. 2011. google scholar
Phillips DC, Woollard KJ, Griffiths HR. The anti-inflammatory actions of methotrexate are critically dependent upon the production of reactive oxygen species. Br J Pharmacol. 2003;138(3):501-511. google scholar
Helal MG, Said E. Tranilast attenuates methotrexate-induced renal and hepatic toxicities: Role of apoptosis-induced tissue proliferation. J Biochem Mol Toxicol. 2020;34(5):e22466. doi: 10.1002/jbt.22466. google scholar
Montasser AOS, Saleh H, Ahmed-Farid OA, Saad A, Marie MS. Protective effects of Balanites aegyptiaca extract, mela-tonin and ursodeoxycholic acid against hepatotoxicity induced by methotrexate in male rats. Asian Pac J Trop Med. 2017;10(6):557-565. google scholar
Won YJ, Kim BK, Shin YK, et al. Pectinase-treated Panax ginseng extract (GINST) rescues testicular dysfunction in aged rats via redox-modulating proteins. Exp Gerontol. 2014;53:57-66. google scholar