Amaranthus hybridus (Amaranthaceae) prevents the detrimental effects of cyclophosphamide on ovarian function in Wistar rats: An experimental study
Background: Cyclophosphamide (CP) is an anticancer agent, but its chronic administration induces ovarian toxicity
Objective: We evaluated the effects of aqueous extract (AE) and methanol extract (ME) of Amaranthus hybridus (A. hybridus) on CP-induced ovarian toxicity in rats.
Materials and Methods: 40 female Wistar rats (10 wk, 170-200 gr) were distributed into 8 groups (n = 5/each) as follows: 1) healthy control; 2) CP+distilled water (10 ml/kg/d); 3) CP+3%-tween 80 (10 mL/kg/d); 4) CP+clomiphene citrate (2 mg/kg/d); 5, 6) CP+AE of A. hybridus (55 and 110 mg/kg/d); and 7, 8) CP+ME of A. hybridus (55 and 110 mg/kg/d). After 28 days of treatment, estrus cyclicity, ovarian and uterine weights as well as estradiol levels and ovarian histology were determined.
Results: CP induced ovarian toxicity after 28 days of exposure. More specifically, CP disturbed the estrus cycle, decreased ovary and uterus weights (p = 0.04), and the 17-β estradiol level (p = 0.04), and induced severe ovarian damages. Remarkably, A. hybridus significantly increased (p = 0.03) the ovarian weight (AE and ME at all doses) and uterus weight (ME at 110 mg/kg/d), compared with the CP-treated rats. Moreover, the 17-β estradiol level was significantly elevated (p = 0.02) in rats given clomiphene citrate and A. hybridus (AE 110 mg/kg/d; ME 55 mg/kg/d). Finally, the ovaries of rats given plant extracts had many corpus luteum and normal follicles, and no cystic follicles.
Conclusion: A. hybridus prevented the detrimental effects of CP on ovarian function, which could support its traditional use as a fertility enhancer.
Key words: Cyclophosphamide, Amaranthus hybridus, Toxicity, Estradiol, Rat.
 Zhao YR, Song HM, Ni L. Cyclophosphamide for the treatment of acute lymphoblastic leukemia: A protocol for systematic review. Medicine (Baltimore) 2019; 98: e14293.
 Codacci-Pisanelli G, Del Pup L, Del Grande M, Peccatori FA. Mechanisms of chemotherapy-induced ovarian damage in breast cancer patients. Crit Rev Oncol Hematol 2017; 113: 90–96.
 Kocahan S, Dogan Z, Erdemli E, Taskin E. Protective effect of quercetin against oxidative stress induced toxicity associated with doxorubicin and cyclophosphamide in rat kidney and liver tissue. Iran J Kidney Dis 2017; 11: 124–131.
 Kalich-Philosoph L, Roness H, Carmely A, Fishel-Bartal M, Ligumsky H, Shoshana P, et al. Cyclophosphamide triggers follicle activation and “Burnout”; AS101 prevents follicle loss and preserves fertility. Sci Transl Med 2013; 5: 185ra62.
 Khedr NF. Protective effect of mirtazapine and hesperidin on cyclophosphamide-induced oxidative damage and infertility in rat ovaries. Exp Biol Med 2015; 240: 1682–1689.
 Meirow D, Biederman H, Anderson RA, Wallace WH. Toxicity of chemotherapy and radiation on female reproduction. Clin Obstet Gynecol 2010; 53: 727– 739.
 Yilmaz S, Yilmaz N. Safety of clomiphene citrate: A literature review. Cytotechnology 2018; 70: 489–495.
 Yener NA, Sinanoglu O, Ilter E, Celik A, Serzin G, Midi A, et al. Effects of spirulina on cyclophosphamideinduced ovarian toxicity in rat: Biochemical and histomorphometric evaluation of the ovary. Biochem Res Int 2013; 2013: 764262.
 Kamarzaman S, Shaban M, Abdul S. The prophylactic effect of Nigella sativa against cyclophosphamide in the ovarian follicles of matured adult mice: A preliminary study. J Anim Plant Sci 2014; 24: 81–88.
 Melekoglu R, Ciftci O, Eraslan S, Cetin A, Basak. N. Beneficial effects of curcumin and capsaicin on cyclophosphamide-induced premature ovarian failure in a rat model. J Ovarian Res 2018; 11: 33.
 Huang C, Song K, Ma W, Ding J, Chen Z, Zhang M. Immunomodulatory mechanism of Bushen Huoxue Recipe alleviates cyclophosphamide induced diminished ovarian reserve in mouse model. J. Ethnopharmacol 2017; 208: 44–56.
 Noumedem JA, Mihasan M, Lacmata ST, Stefan M, Kuiate JR, Kuete V. Antibacterial activities of the methanol extracts of ten Cameroonian vegetables against gram-negative multidrug-resistant bacteria. BMC Complement Altern Med 2013; 13: 26.
 Balasubramanian T, Karthikeyan M. Therapeutic effect of amaranthus hybridus on diabetic nephropathy. J Dev Drugs 2016; 5: 1.
 Al-Mamun MA, Husna J, Khatun M, Hasan R, Kamruzzaman M, Hoque KM, et al. Assessment of antioxidant, anticancer and antimicrobial activity of two vegetables species of Amaranthus in Bangladesh. BMC Complement Altern Med 2016; 16: 157.
 Medoua GN, Oldewage-Theron WH. Effect of drying and cooking on nutritional value and antioxidant capacity of morogo (Amaranthus hybridus) a traditional leafy vegetable grown in South Africa. J Food Sci Technol 2014; 51: 736–742.
 Ngadjui E, Nkeng-Efouet PA, Nguelefack TB, Kamanyi A, Watcho P. High fat diet-induced estrus cycle disruption: Effects of Ficus asperifolia. J Complement Integr Med 2015; 12: 205–215.
 Ndeingang ECh, Deeh PBD, Watcho P, Kamanyi A. Phyllanthus muellerianus (Euphorbiaceae) restores ovarian functions in letrozole-induced polycystic ovarian syndrome in rats. Evid Based Complement Alternat Med 2019; 2019: 2965821.
 Hamzeh M, Hosseinimehr SJ, Mohammadi HR, Yaghubi Beklar S, Dashti A, Talebpour Amiri F. Atorvastatin attenuates the ovarian damage induced. by cyclophosphamide in rat: An experimental study. Int J Reprod Biomed 2018; 16: 323–334.
 Council Directive of 24 November 1986, on the approximation of laws, regulations and administrative provisions of the Member States regarding the protection of animals used for experimental and other scientific purposes (86/609/EEC). Official J Eur Communities 1986; 358: 1–29.
 Horicks F, Van Den Steen G, Houben S, Englert Y, Demeestere I. Folliculogenesis is not fully inhibited during GnRH analogues treatment in mice challenging their efficiency to preserve the ovarian reserve during chemotherapy in this model. PLoS One 2015; 10: e0137164.
 Firas AA, Hasan HF. The ameliorative effects of Persia americana pulps alcoholic extract on fertility of female rats treated with cyclophosphoamide. J Nat Sci Res 2016; 6: 93–98.
 Roness H, Kalich-Philosoph L, Meirow D. Prevention of chemotherapy-induced ovarian damage: Possible roles for hormonal and non-hormonal attenuating agents. Hum Reprod Update 2014; 20: 759–774.
 Sun J, Zhang X, Cao Y, Zhao Q, Bao E, Lv Y. Ovarian toxicity in female rats after oral administration of melamine or melamine and cyanuric acid. PLoS One 2016; 11: e0149063.
 Akunna GG, Nwafor J, Egwu OA, Ezemagu UK, Obaje G, Adepoju LH, et al. Cisplatin-induced ovarian. cytotoxicity and the modulating role of aqueous zest. extract of Citrus limonium (AZECL) in rat models. J. Tradit Med Clin Natur 2017; 6: 3.
 Hassanpour A, Yousefian S, Askaripour M, Sharififar F, Ezzatabadipour M. Ovarian protection in cyclophosphamide-treated mice by fennel. Toxicol Rep 2017; 4: 160–164.
 Saleh DO, Mansour DF. Ovario-protective effects of genistein against cyclophosphamide toxicity in rats: Role of anti-müllerian hormone and estradiol. Eur J Pharmacol 2016; 789: 163–171.
 Nana FW, Hilou A, Millogo JF, Nacoulma OG. Phytochemical composition, antioxidantand xanthine oxidase inhibitory activities of Amaranthus cruentus L. and Amaranthus hybridus L. extracts. Pharmaceuticals 2012; 5: 613–628.
 Butchi-Akondi R, Kumar P, Annapurna A, Pujari M. Protective effect of rutin and naringin on sperm quality in streptozotocin (STZ) induced type 1 diabetic rats. Iran J Pharm Res 2011; 10: 585–596.
 Adetutu A, Awe E. Modulatory of effect of fresh Amaranthus caudatus and Amaranthus hybridus aqueous leaf extracts on detoxify enzymes and micronuclei formation after exposure to sodium arsenite. Pharmacognosy Res 2013; 5: 300–305.
 Comish PB, Drumond AL, Kinnell HL, Anderson RA, Matin A, Meistrich ML, et al. Fetal cyclophosphamide exposure induces testicular cancer and reduced. spermatogenesis and ovarian follicle numbers in. mice. PLoS One 2014; 9: e93311.