Carob (Ceratonia siliqua L.) fruit hydro-alcoholic extract alleviates reproductive toxicity of lead in male mice: Evidence on sperm parameters, sex hormones, oxidative stress biomarkers and expression of Nrf2 and iNOS

Document Type: Original Research Article

Authors

1 Department of Theriogenology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

2 Department of Theriogenology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.

3 Department of Pathobiology, Faculty of Veterinary medicine, Urmia University, Urmia, Iran

Abstract

Objective: Carob (Ceratonia siliqua L.) is an evergreen tree with fruits that have potent antioxidant activity. The aim of this study was to investigate alleviative effects of carob fruit hydro-alcoholic extract (CFHAE) against reproductive toxicity induced by lead (Pb) in male mice.
Material and Methods: Forty-two NMRI adult male mice were randomly categorized into 7 groups (N=6). Group I was the control group and received no treatment. Group II was the sham group and received 0.2 ml distilled water per day. Group III (Pb group) received Pb acetate 1000 ppm/kg/day. Groups IV and V received CFHAE 500 and 1000 mg/kg/day, respectively. Groups VI and VII received both Pb 1000 ppm/kg/day and CFHAE at doses of 500 and 1000 mg/kg/day, respectively at the same time. The groups were treated by gavage. After 35 days, sperm parameters (count, motility, morphology, viability, DNA damage, and teratozoospermia index), total antioxidant capacity (TAC), reduced glutathione content (GSH), antioxidant enzymes (SOD, CAT, and GPx) activity, MDA levels, and sex hormones (FSH, LH, and testosterone) concentrations in serum, testicular expression of Nrf2 and iNOS genes and histopathological alterations were evaluated.
Results: Our findings revealed that co-administration of CFHAE with Pb significantly (p<0.05 to p<0.01) improved sperm parameters, elevated sex hormones, TAC, GSH content, and antioxidant enzymes activity of serum, decreased serum MDA levels, and down-regulated testicular expression of Nrf2 and iNOS genes compared with Pb group. Also, CFHAE ameliorated histopathological alterations in testis tissue caused by Pb.
Conclusion: CFHAE can alleviate reproductive toxicity following Pb exposure in male mice.

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Abdel-Rahman M, Bauomy AA, Salem FEH, Ahmed Khalifa M. 2018. Carob extract attenuates brain and lung injury in rats exposed to waterpipe smoke. Egypt J Basic Appl Sci, 5:31–40.

Aktan F. 2004. iNOS-mediated nitric oxide production and its regulation. Life Sci, 75:639–653.

Al-Olayan EM, El-Khadragy MF, Alajmi RA, Othman MS, Bauomy AA, Ibrahim SR, Moneim AE. 2016. Ceratonia siliqua pod extract ameliorates Schistosoma mansoni-induced liver fibrosis and oxidative stress. BMC Complement Altern Med, 16: no pagination.

Anjum MR, Reddy PS. 2015. Recovery of lead-induced suppressed reproduction in male rats by testosterone. Andrologia, 47:560–567.

Ata A, Yildiz-Gulay O, Güngör S, Balic A, Gulay MS. 2018. The effect of carob (Ceratonia siliqua) bean extract on Male New Zealand white rabbit semen. World Rabbit Sci, 26:209–215.

Ayinde OC, Ogunnowo S, Ogedegbe RA. 2012. Influence of Vitamin C and Vitamin E on testicular zinc content and testicular toxicity in lead exposed albino rats. BMC Pharmacol Toxicol, 13:1–8.

 Bae D-S, Gennings C, Carter Jr WH, Yang RSH, Campain JA. 2001. Toxicological interactions among arsenic, cadmium, chromium, and lead in human keratinocytes. Toxicol Sci, 63:132–142.

Benzie I,Strain J. 1999. Direct measure of total antioxidant activity of biological fluids and modified version for simultaneous measurement of total antioxidant power and ascorbic. Methods Enzymol, 299:15–27.

Beutler E, Duron O, Kelly BM. 1963. Improved method for the determination of blood glutathione. J Lab Clin Med, 61:882–888.

Choudhary R, Chawala VK, Soni ND, Kumar J, Vyas RK. 2010. Oxidative stress and role of antioxidants in male infertility. Pakistan J Physiol, 6:54–59.

Chowdhury AR. 2009. Recent advances in heavy metals induced effect on male reproductive function-A retrospective. Al Ameen J Med Sci, 2:37–42.

Creasy DM. 2001. Pathogenesis of Male Reproductive Toxicity. Toxicol Pathol, 29:64–76.

Custódio L, Fernandes E, Escapa AL, López-Avilés S, Fajardo A, Aligué R, Alberício F, Romano A. 2009. Antioxidant activity and in vitro inhibition of tumor cell growth by leaf extracts from the carob tree (Ceratonia siliqua). Pharm Biol, 47:721–728.

Dakia PA, Wathelet B, Paquot M. 2007. Isolation and chemical evaluation of carob (Ceratonia siliqua L.) seed germ. Food Chem, 102:1368–1374.

Dewanto V, Wu X, Adom KK, Liu RH. 2002. Thermal processing enhances the nutritional value of tomatoes by increasing total antioxidant activity thermal processing enhances the nutritional value of tomatoes by increasing total antioxidant activity. J Agric Food Chem, 50:3010–3014.

Dkhil MA, Moneim AEA, Al-Quraishy S. 2016. Indigofera oblongifolia ameliorates lead acetate-induced testicular oxidative damage and apoptosis in a rat model. Biol Trace Elem Res, 173:354–361.

Dorostghoal M, Seyyednejad SM, Jabari A. 2014. Protective effects of Fumaria parviflora L. on lead-induced testicular toxicity in male rats. Andrologia, 46:437–446.

Elgawish RAR Abdelrazek HMA. 2014. Effects of lead acetate on testicular function and caspase-3 expression with respect to the protective effect of cinnamon in albino rats. Toxicol Reports, 1:795–801.

El-Nekeety AA, El-Kady AA, Soliman MS, Hassan NS, and Abdel-Wahhab MA. 2009. Protective effect of Aquilegia vulgaris (L.) against lead acetate-induced oxidative stress in rats. Food Chem Toxicol, 47:2209–2215.

Gandhi J, Hernandez RJ, Chen A, Smith NL, Sheynkin YR, Joshi G, Khan SA. 2017. Impaired hypothalamic-pituitary-testicular axis activity, spermatogenesis, and sperm function promote infertility in males with lead poisoning. Zygote, 25: 103-110.

Glade MJ and Smith K. 2015. Oxidative stress, nutritional antioxidants, and testosterone secretion in men. Ann Nutr Disord Ther, 2:1019.

 Hamadouche NA, Nesrine S, Abdelkeder A. 2013. Lead toxicity and the hypothalamic-pituitary-testicular axis. Not Sci Biol, 5:1–6.

Hammond PB. 1977. Exposure of humans to lead. Annu Rev Pharmacol Toxicol, 17:197–214.

Harris ED. 1992. Regulation of antioxidant enzymes. FASEB J, 6:2675–2683.

He X, Wu J, Yuan L, Lin F, Yi J, Li J, Yuan H, Shi J, Yuan T, Zhang S, Fan Y. 2017. Lead induces apoptosis in mouse TM3 Leydig cells through the Fas/FasL death receptor pathway. Environ Toxicol Pharmacol, 56:99–105.

He Y, Zou Q, Chen H, Weng S, Luo T. 2016. Lead inhibits human sperm functions by reducing the levels of intracellular calcium, cAMP, and tyrosine phosphorylation. Tohoku J Exp Med, 238:295–303. 

Huang BM, Liu MY. 2004. Inhibitory actions of lead on steroidogenesis in MA-10 mouse Leydig tumor cells. Syst Biol Reprod Med, 50:5–9.

Jung K-A, Kwak M-K. 2010. The Nrf2 system as a potential target for the development of indirect antioxidants. Molecules, 15:7266–7291.

Kanter M. 2010. Protective effects of melatonin on testicular torsion/detorsion-induced ischemia–reperfusion injury in rats. Exp Mol Pathol, 89:314–320.

Krassas E, Papadopoulou F, Tziomalos K, Zeginiadou T, Pontikides N. 2008. Hypothyroidism has an adverse effect on human spermatogenesis: a prospective, controlled study. Thyroid, 18:1255.

Kumazawa S, Taniguchi M, Suzuki Y, Shimura M, Kwon MS, Nakayama T. 2002. Antioxidant activity of polyphenols in carob pods. J Agric Food Chem, 50:373–377.

Lindl KA, Jordan-Sciutto KL. 2008. Examining the endogenous antioxidant response through immunofluorescent analysis of Nrf2 in tissue. Methods Mol Biol, 477:229–243.

Liu MY, Lai HY, Yang BC, Tsai ML, Yang HY, Huang BM. 2001. The inhibitory effects of lead on steroidogenesis in MA-10 mouse Leydig tumor cells. Life Sci, 68:849–859.

Loraine JA, Bell ET. 1971. Hormone assays and their clinical application. pp. 44-91 and 541-553, Edinburgh, UK: E. & S. Livingstone.

Mabrouk A, Ben Cheikh H. 2014. Thymoquinone supplementation ameliorates lead-induced testis function impairment in adult rats. Toxicol Ind Health, 32:1114–1121.

Mishra M, Acharya UR. 2004. Protective action of vitamins on the spermatogenesis in lead-treated Swiss mice. J Trace Elem Med Biol, 18:173–178.

Mokhtari M, Sharifi E, Azadian S. 2012. The effects of hydro alcoholic extract of Ceratonia siliqua L. seeds on pituitary - Testis hormones and spermatogenesis in rat. Adv Environ Biol, 6:2778–2783.

Moneim AEA. 2015. The neuroprotective effect of berberine in mercury-induced neurotoxicity in rats. Metab Brain Dis, 30:935–942.

Moniem AEA, Dkhil MA, Al-quraishy S. 2010. Protective role of flaxseed oil against lead acetate induced oxidative stress in testes of adult rats. Afr J Biotechnol, 9:7216–7223.

Naghmouchi S, Khouja ML, Romero A, Tous J, Boussaid M. 2009. Tunisian carob (Ceratonia siliqua L.) populations: Morphological variability of pods and kernel. Sci Hortic, 121:125–130.

Nishikimi M, Rao NA, Yagi K. 1972. The ocurrence of superoxide anion in the reaction of reduced phenazine methosufate and molecular oxygen. Biochem Biophys Res Commun, 2:849–854.

Owen RW, Haubner R, Hull WE, Erben G, Spiegelhalder B, Bartsch H, Haber B. 2003. Isolation and structure elucidation of the major individual polyphenols in carob fibre. Food Chem Toxicol, 41:1727–1738.

 Özcan MM, Arslan D, Gökçalik H. 2007. Some compositional properties and mineral contents of carob (Ceratonia siliqua) fruit, flour and syrup. Int J Food Sci Nutr, 58:652–658.

Oziyci HR, Tetik N, Turhan I, Yatmaz E, Ucgun K, Akgul H, Gubbuk H, Karhan M.2014. Mineral composition of pods and seeds of wild and grafted carob (Ceratonia siliqua L.) fruits. Sci Hortic, 167:149–152.

Paglia DE, Valentine WN. 1967. Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med, 70:158–169.

Papagiannopoulos M, Wollseifen HR, Mellenthin A, Haber B, Galensa R. 2004. Identification and quantification of polyphenols in carob fruits (Ceratonia siliqua L.) and derived products by HPLC-UV-ESI/MSn. J Agric Food Chem, 52:3784–3791.

Patra RC, Rautray AK, Swarup D. 2011. Oxidative stress in lead and cadmium toxicity and its amelioration. Vet Med Int, 2011.

Pedersen H. 2001. Late-flowering dune populations of Dactylorhiza incarnata (Orchidaceae): Variation patterns and taxonomic inferences. Nord J Bot, 21:177–186.

Placer ZA, Cushman LL, Johnson BC. 1996. Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical systems. Anal Biochem, 16:359–364.

Rahman MA, H. Salem FE, Bauomy AA, Khalifa MA. 2016. Ameliorative effect of carob aqueous extract on water pipe smoke induced-toxicity in adult male albino rats. Int J Pharm Pharm Sci, 9:246.

Reshma Anjum M, Madhu P, Pratap Reddy K, Sreenivasula Reddy P. 2017. The protective effects of zinc in lead-induced testicular and epididymal toxicity in Wistar rats. Toxicol Ind Health, 33:265–276.

Reshma Anjum M, Sainath SB, Suneetha Y, Sreenivasula Reddy P. 2011. Lead acetate induced reproductive and paternal mediated developmental toxicity in rats. Ecotoxicol Environ Saf, 74:793–799.

Rtibi K, Jabri MA, Selmi S, Souli A, Sebai H, El-Benna J, Amri M, Marzouki L. 2015.  Gastroprotective effect of carob (Ceratonia siliqua L.) against ethanol-induced oxidative stress in rat. BMC Complement Altern Med, 15:1–8.

Rtibi K, Selmi S, Jabri MA, El-Benna J, Amri M, Marzouki L, Sebai H. 2016a. Protective effect of Ceratonia siliqua L. against a dextran sulfate sodium-induced alterations in liver and kidney in rat. J Med Food, 19:882–889.

Rtibi K, Selmi S, Jabri MA, Mamadou G, Limas-Nzouzi N, Sebai H, El-Benna J, Marzouki L, Eto B, Amri M. 2016b. Effects of aqueous extracts from: Ceratonia siliqua L. pods on small intestinal motility in rats and jejunal permeability in mice. RSC Adv, 6:44345–44353.

Said AA, Kairy MH, Shalaby AME, Ismail HT. 2017. The effect of carob pods and fig fruits ether extracts against lead induced hematological and biochemical changes in Oreochromis niloticus. Zagazig Vet J, 45:19–28.

Shotyk W, Le Roux G. 2005. Biogeochemistry and cycling of lead. Met Ions Biol Syst, 43:239–275.

Sigman M. 2007. Medications that impair male fertility. SRM, 11–15.

Simmons SO, Fan C-Y, Yeoman K, Wakefield J, Ramabhadran R. 2011. NRF2 Oxidative Stress Induced by Heavy Metals is Cell Type Dependent. Curr Chem Genomics, 5:1–12.

Sinha AK. 1972. Colorimetric assay of catalase. Anal Biochem, 47:389–394.

Solei

manzadeh A, Kian M, Moradi S, Malekifard F. 2018. Protective effects of hydro-alcoholic extract of Quercus brantii against lead-induced oxidative stress in the reproductive system of male mice. Avicenna J Phytomedicine, 8:448–456.

Suvarna, KS, Layton C, Bancroft, J.D. eds., 2018. Bancroft's Theory and Practice of Histological Techniques E-Book, pp. 40-139, Elsevier Health Sciences.

Suzek H, Celik I, Dogan A. 2017. Nephroprotective hepatoprotective potential and antioxidant role of carob pods (Cerotonia siliqua L.) against carbon tetrachloride-induced toxicity in rats. Indian J Pharm Educ Res, 51:312–320.

Tejada RI, Mitchell JC, Norman A, Marik JJ, Friedman S. 1984. A test for the practical evaluation of male fertility by acridine orange (AO) fluorescence. Fertil Steril, 42:87–91.

Tetik N, Turhan I, Oziyci HR, Gubbuk H, Karhan M, Ercisli S. 2011. Physical and chemical characterization of Ceratonia siliqua L. germplasm in Turkey. Sci Hortic, 129:583–589.

Vafaei A, Mohammadi S, Fazel A, Soukhtanloo M, Pour AM, Beheshti F. 2018. Effects of carob (Ceratonia siliqua) on sperm quality, testicular structure, testosterone level and oxidative stress in busulfan-induced infertile mice. Pharm Sci, 24:104-111.

Wang Y, Fang J, Huang S, Chen L, Fan G, Wang C. 2012. The chronic effects of low lead level on the expressions of Nrf2 and Mrp1 of the testes in the rats. Environ Toxicol Pharmacol, 35:109–116.

 Wong EWP and Cheng CY. 2011. Impacts of environmental toxicants on male reproductive dysfunction. Trends Pharmacol Sci, 32:290–299.

World Health Organisation. 2010. WHO laboratory manual for the examination and processing of human semen. World Health Organization.

Yu B, Lin H, Yang L, Chen K, Luo H, Liu J, Gao X, Xia X, Huang Z.2012. Genetic variation in the Nrf2 promoter associates with defective spermatogenesis in humans. J Mol Med, 90:1333–1342.

Zhang H, Wei K, Zhang M, Liu R, Chen Y. 2014. Assessing the mechanism of DNA damage induced by lead through direct and indirect interactions. J Photochem Photobiol B Biol, 136:46–53.