Protective effects of hydroalcoholic extract of Rosa canina L. fruit on cyclophosphamide-induced testicular toxicity in mice

Document Type : Short communication


1 Department of Biology, Payame Noor University, Tehran, Iran

2 Department of Biology, Faculty of Sciences, Razi University, Kermanshah, Iran

3 Department of Forensic Medicine, Judicature, Kermanshah, Iran


Objective: Cyclophosphamide (CP)-induced testicular toxicity has been reported in recipient patients. The current study was designed to evaluate protective effects of hydroalcoholic extract of Rosa canina L. fruit (HARF) against CP-induced testicular toxicity in BALB/c mice.
Materials and Methods: Thirty-five mice were divided into five groups as follows: group I (control), group II (CP, received CP 100 mg/kg on days 1, 8, 15, and 22), group III (CP + HARF 250 mg/kg), group IV (CP + HARF 500 mg/kg), and group V (CP + HARF 750 mg/kg). In the groups III, IV, and V that received CP, the HARF was simultaneously administered via intraperitoneal injections for 28 consecutive days starting from day 1. On the 29th day, sperm parameters, stress oxidative biomarkers, and mRNA expression of nuclear factor erythroid-derived-2 (Nrf2) in testis tissue, as well as blood testosterone were evaluated.
Results: The CP exposure decreased sperm parameters, superoxide dismutase (SOD) activity, testosterone, and Nrf2 mRNA expression levels and increased the malondialdehyde (MDA). HARF at the dose of 500 mg/kg improved sperm count and viability and increased SOD and catalase activities, glutathione peroxidase (GPx) activity, testosterone level, and Nrf2 expression and reduced MDA. Also, HARF at the dose of 750 mg/kg improved sperm parameters and increased SOD, catalase, and GPx activities, total testosterone level, and Nrf2 expression, and reduced MDA in comparison with the CP group.
Conclusion: According to our findings, HARF at the doses of 500 and 750 mg/kg inhibited the ruinous effects of CP on the reproductive system in mice.


Alahmar AT. 2018. The effects of oral
antioxidants on the semen of men with
idiopathic oligoasthenoteratozoospermia.
Clin Exp Reprod Med, 45: 57-66.
Almeer RS, Soliman D, Kassab RB, AlBasher
GI, Alarifi S, Alkahtani S, Ali D,
Metwally D, Abdel Moneim AE. 2018.
Royal jelly abrogates cadmium-induced
oxidative challenge in mouse testes:
involvement of the Nrf2 pathway. Int J
Mol Sci, 19: 3979.
Aprioku JS. 2013. Pharmacology of free
radicals and the impact of reactive oxygen
species on the testis. J Reprod Infertil, 14:
Attia AA, ElMazoudy RH, El-Shenawy NS.
2012. Antioxidant role of propolis extract
against oxidative damage of testicular
tissue induced by insecticide chlorpyrifos
in rats. Pestic Biochem Physiol, 103: 87-
Bisht S, Faiq M, Tolahunase M, Dada R. 2017.
Oxidative stress and male infertility. Nat
Rev Urol, 14: 470-485.
Bussmann RW, Swartzinsky P, Worede A,
Evangelista P. 2011. Plant use in odo-bulu
and demaro, Bale region, Ethiopia. J
Ethnobiol Ethnomed, 7: 28.
Darbandi M, Darbandi S, Agarwal A,
Baskaran S, Sengupta P, Dutta S,
Mokarram P, Saliminejad K, Sadeghi MR.
2019. Oxidative stress‐induced alterations
in seminal plasma antioxidants: Is there
any association with keap1 gene
methylation in human spermatozoa?
Andrologia, 51: e13159.
Elangovan N, Chiou T-J, Tzeng W-F, Chu ST. 2006. Cyclophosphamide treatment
causes impairment of sperm and its
fertilizing ability in mice. Toxicology,
222: 60-70.
Elmallah MI, Elkhadragy MF, Al-Olayan EM,
Abdel Moneim AE. 2017. Protective effect
of Fragaria ananassa crude extract on
cadmium-induced lipid peroxidation,
antioxidant enzymes suppression, and
apoptosis in rat testes. Int J Mol Sci, 18:
Fusco R, Salinaro AT, Siracusa R, D’Amico
R, Impellizzeri D, Scuto M, Ontario ML,
Crea R, Cordaro M, Cuzzocrea S. 2021.
Hidrox® counteracts cyclophosphamideinduced male infertility through NRF2
pathways in a mouse model. Antioxidants,
10: 778.
Ghobadi E, Moloudizargari M, Asghari MH,
Abdollahi M. 2017. The mechanisms of
cyclophosphamide-induced testicular
toxicity and the protective agents. Expert
Opin Drug Metab Toxicol, 13: 525-536.
Gholampour F, Javadifar TS, Karimi S,
Eslam-Zadeh T, Owji SM. 2012. The
effects of the hydroalcohol extract of Rosa
canina L. fruit on ischemic acute renal
failure in Wistar rats. Comp Clin Path, 21:
Hamzeh M, Hosseinimehr SJ, Karimpour A,
Mohammadi HR, Khalatbary AR, Amiri
FT. 2019. Cerium oxide nanoparticles
protect cyclophosphamide-induced
testicular toxicity in mice. Int J Prev Med,
10: 5.
Hosseini A, Zare S, Borzouei Z, Pakdel FG.
2018. Cyclophosphamide-induced
testicular toxicity ameliorate by American
ginseng treatment: An experimental study.
Int J Reprod Med, 16: 711-718.
Imamovic Kumalic S, Pinter B. 2014. Review
of clinical trials on effects of oral
antioxidants on basic semen and other
parameters in idiopathic
oligoasthenoteratozoospermia. Biomed
Res Int, 2014: 426951.
Kim J-K, Jang H-D. 2014. Nrf2-mediated HO1 induction coupled with the ERK
signaling pathway contributes to indirect
antioxidant capacity of caffeic acid
phenethyl ester in HepG2 cells. Int J Mol
Sci, 15: 12149-12165.
Kim S, Viswanath ANI, Park J-H, Lee HE,
Park AY, Choi JW, Kim HJ, Londhe AM,
Jang BK, Lee J. 2020. Nrf2 activator via
interference of Nrf2-Keap1 interaction has
antioxidant and anti-inflammatory
properties in Parkinson's disease animal
model. Neuropharmacology, 167: 107989.
Korkmaz A, Topal T, Oter S. 2007.
Parandin et al.
AJP, Vol. 13, No. 1, Jan-Feb 2023 16
Pathophysiological aspects of
cyclophosphamide and ifosfamide induced
hemorrhagic cystitis; implication of
reactive oxygen and nitrogen species as
well as PARP activation. Cell Biol
Toxicol, 23: 303-312.
Ku BM, Joo Y, Mun J, Roh GS, Kang SS, Cho
GJ, Choi WS, Kim HJ. 2006. Heme
oxygenase protects hippocampal neurons
from ethanol-induced neurotoxicity.
Neurosci Lett, 405: 168-171.
Liu J, Simon LM, Phillips JR, Robin ED.
1977. Superoxide dismutase (SOD)
activity in hypoxic mammalian systems. J
Appl Physiol, 42: 107-110.
Luo T, Zou Q-x, He Y-q, Wang H-f, Li N,
Zeng X-h. 2015. Matrine inhibits mouse
sperm function by reducing sperm [Ca2+]
i and phospho-ERK1/2. Cell Physiol
Biochem, 35: 374-385.
Maremanda KP, Khan S, Jena G. 2014. Zinc
protects cyclophosphamide-induced
testicular damage in rat: Involvement of
metallothionein, tesmin and Nrf2.
Biochem Biophys Res Commun, 445:
NaĎpal JD, Lesjak MM, Šibul FS, Anačkov
GT, Četojević-Simin DD, Mimica-Dukić
NM, Beara IN. 2016. Comparative study
of biological activities and phytochemical
composition of two rose hips and their
preserves: Rosa canina L. and Rosa
arvensis Huds. Food chem, 192: 907-914.
Naziroğlu M. 2003. Enhanced testicular
antioxidant capacity in streptozotocininduced diabetic rats. Biol Trace Elem
Res, 94: 61-71.
Novin MG, Golmohammadi MG, Sagha M,
Ziai SA, Abdollahifar MA, Nazarian H.
2020. Protective effect of gallic acid on
testicular tissue, sperm parameters, and
dna fragmentation against toxicity induced
by cyclophosphamide in adult NMRI
mice. Urol J, 17: 78-85.
Nowicka-Bauer K, Nixon B. 2020. Molecular
changes induced by oxidative stress that
impair human sperm motility.
Antioxidants, 9: 134.
Nowrouzi F, Azadbakht M, Kalehoei E,
Modarresi M. 2019. Protective effect of
Rosa Canina extract against doxorubicininduced testicular toxicity in mice. Braz
Arch Biol Technol, 62: e19180017.
Roman I, Stănilă A, Stănilă S. 2013. Bioactive
compounds and antioxidant activity of
Rosa canina L. biotypes from spontaneous
flora of Transylvania. Chem Cent J, 7: 73.
Sadeghi H, Hosseinzadeh S, Touri MA,
Ghavamzadeh M, Barmak MJ. 2016.
Hepatoprotective effect of Rosa canina
fruit extract against carbon tetrachloride
induced hepatotoxicity in rat. Avicenna J
Phytomed, 6: 181-188.
Sadeghi H, Karimizadeh E, Sadeghi H,
Mansourian M, Abbaszadeh-Goudarzi K,
Shokripour M, Asfaram A, Doustimotlagh
AH. 2021. Protective effects of
hydroalcoholic extract of Rosa canina fruit
on vancomycin-induced nephrotoxicity in
rats. J Toxicol, 2021: 5525714.
Sakr SA, Mahran HA, Abo-El-Yazid SM.
2012. Effect of fenugreek seeds extract on
histomorphometrical, ultrastructural and
biochemical changes in testes of albino
mice. Toxicol Ind Health, 28: 276-288.
Salimnejad R, Rad JS, Nejad DM. 2018.
Protective effect of ghrelin on oxidative
stress and tissue damages of mice testes
followed by chemotherapy with
cyclophosphamide. Crescent J Med Biol
Sci, 5: 138-143.
Schmittgen TD, Livak KJ. 2008. Analyzing
real-time PCR data by the comparative CT
method. Nat protoc, 3: 1101-1108.
Shah NA, Khan MR. 2017. Increase of
glutathione, testosterone and antioxidant
effects of Jurenia dolomiaea on CCl 4
induced testicular toxicity in rat. BMC
Complement. Altern Med, 17: 206.
Shanmugam G, Narasimhan M, Tamowski S,
Darley-Usmar V, Rajasekaran NS. 2017.
Constitutive activation of Nrf2 induces a
stable reductive state in the mouse
myocardium. Redox Biol, 12: 937-945.
Tayefi‐Nasrabadi H, Sadigh‐Eteghad S,
Aghdam Z. 2012. The effects of the
hydroalcohol extract of Rosa canina L.
fruit on experimentally nephrolithiasic
Wistar rats. Phytother Res, 26: 78-85.
Winther K, Hansen ASV, Campbell-Tofte J.
2016. Bioactive ingredients of rose hips
(Rosa canina L) with special reference to
antioxidative and anti-inflammatory
properties: in vitro studies. Botanics, 6:
Wyrobek AJ, Gordon LA, Burkhart JG,
Francis MW, Kapp Jr RW, Letz G,
Malling HV, Topham JC, Whorton MD.
1983. An evaluation of the mouse sperm
Rosa canina and cyclophosphamide toxicity
AJP, Vol. 13, No. 1, Jan-Feb 2023 17
morphology test and other sperm tests in
nonhuman mammals: A report of the US
Environmental Protection Agency GeneTox Program. Mutat Res, 115: 1-72.
Young I, Trimble E. 1991. Measurement of
malondialdehyde in plasma by high
performance liquid chromatography with
fluorimetric detection. Ann clin biochem,
28: 504-508.