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Eugenol Prevents Sodium Nitrate-induced Altered Antioxidant Status of Spermatozoa: An In Vitro Study

Rajini S.V., Chaithra B., Shiva basavaiah

Abstract


Background: High level of nitrate (NO3) induces oxidative stress which affects the male and female reproductive system. In this regard, there is a great concern for the prevention of NO3-induced oxidative stress in the reproductive system by using herbal products. Eugenol is one such natural product found in clove extracted from cloves (Syzygium aromaticum) and is known for its antioxidant, anti-nociceptive, anti-inflammatory, and anti-apoptotic properties. Aim and Objectives: The present study was undertaken to estimate the protective effect of eugenol in preventing sodium nitrate (NaNO3) induced oxidative stress in spermatozoa under in vitro conditions. Methods: Epididymal sperm suspension was treated with 10 mg/ml of eugenol along with NaNO3 (100 mg/ml) for 15 min of a time interval. In addition, different experimental groups viz., control (without eugenol and NaNO3), vehicle control (30% Dimethyl sulfoxide), and NaNO3 (100 mg/ml) alone were maintained. After the treatment period, sperm motility was analyzed. Further, different oxidative stress markers viz., activities of superoxide dismutase (SOD) and catalase (CAT) and concentrations of malondialdehyde (MDA) and nitric oxide (NO) were evaluated. Results and Discussion: There was a complete loss of sperm motility in NaNO3-treated groups compared to control and vehicle control groups. In addition, treatment of NaNO3 caused a significant increase in the activities of SOD and CAT, and MDA levels compared to control and vehicle control groups. Further, there was a significant increase in the concentration of NO in the NaNO3-treated group compared to that of the control and vehicle control groups. These results established the fact that NO3 is reduced to NO and induced oxidative stress in spermatozoa. In addition, a significant increase in NO concentration revealed the negative correlation between NO concentration and sperm motility. In the present study, treatment of eugenol prevented NaNO3-induced alterations in the antioxidant system as the activities of SOD and CAT and concentrations of MDA and NO in eugenol + NaNO3 treated groups were similar to that of control and vehicle control groups. In contrast, the sperm motility in the eugenol + NaNO3 treated group was significantly higher than NaNO3 alone treated group and lower than that of the control and vehicle control groups. These findings suggested the protective effect of eugenol against NaNO3-induced altered antioxidant status of spermatozoa. Conclusion: Exposure to a high level of NO3 causes the overproduction of free radicals and subsequent induction of oxidative stress which will affect sperm motility and thereby reduce the capacity of spermatozoa in fertilization. Eugenol is effectively preventing NO3-induced oxidative stress in spermatozoa.

Keywords


Eugenol, nitric oxide, motility, oxidative stress, sodium nitrate, spermatozoa

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References


Doshi SB, Khullar K, Sharma RK, Agarwal A. Role of reactive nitrogen species in male infertility. Reprod Biol Endocrinol. 2012;10(1):109. doi: 10.1186/1477–7827–10–109.

Pizzino G, Irrera N, Cucinotta M, Pallio G, Mannino F, Arcoraci V, et al. Oxidative stress: harms and benefits for human health. Oxid Med Cell Longev. 2017;8416763. doi: 10.1155/2017/8416763.

Ergün Y, Kurutaş EB, Ozdil B, Güneşaçar R, Ergün Y. Evaluation of nitrite/nitrate levels in relation to oxidative stress parameters in liver cirrhosis. Clin Res Hepatol Gastroenterol. 2011;35(4):303–8. doi: 10.1016/j.clinre.2010.12.009.

Pacher P, Beckman JS, Liaudet L. Nitric oxide and peroxynitrite in health and disease. Physiol Rev. 2007;87(1):315–424. doi: 10.1152/physrev.00029.2006.

Aitken RJ, Baker MA, O’Bryan M. Shedding light on chemiluminescence: the application of chemiluminescence in diagnostic andrology. J Androl. 2004;25(4):455–65. doi: 10.1002/j.1939–4640.2004.tb02815.x.

Spalding RF, Exner ME. Occurrence of nitrate in groundwater—a review. J Environ Qual. 1993;22(3):392–402. doi: 10.2134/jeq1993.00472425002200030002x.

Sleight SD, Atallah OA. Reproduction in the guinea pig as affected by chronic administration of potassium nitrate and potassium nitrite. Toxicol Appl Pharmacol. 1968;12(2):179–85. doi: 10.1016/0041–008x(68)90029-x.

Aly HAA, Mansour AM, Abo-Salem OM, Abd-Ellah HF, Abdel-Naim AB. Potential testicular toxicity of sodium nitrate in adult rats. Food Chem Toxicol. 2010;48(2):572–8. doi: 10.1016/j.fct.2009.11.034.

Adelakun SA, Ukwenya VO, Ogunlade BS, Aniah AJ, Ibiayo GA. Nitrite-induced testicular toxicity in rats: therapeutic potential of walnut oil. JBRA Assist Reprod. 2019;23(1):15–23. doi: 10.5935/1518–0557.20180062.

Clausen HS, Ebdrup NH, Barsøe IM, Lyngsø J, Schullehner J, Ramlau-Hansen CH, et al. Association between drinking water nitrate and adverse reproductive outcomes: a systematic PRISMA review. Water. 2020;12(8):2287. doi: 10.3390/w12082287.

Hassan HA, El-Agmy SM, Gaur RL, Fernando A, Raj MH, Ouhtit A. In vivo evidence of hepato- and reno-protective effect of garlic oil against sodium nitrite-induced oxidative stress. Int J Biol Sci. 2009;5(3):249–55. doi: 10.7150/ijbs.5.249.

Rouag M, Berrouague S, Djaber N, Khaldi T, Boumendjel M, Taibi F, et al. Pumpkin seed oil alleviates oxidative stress and liver damage induced by sodium nitrate in adult rats: biochemical and histological approach. Afr Health Sci. 2020;20(1):413–25. doi: 10.4314/ahs.v20i1.48.

Bouaziz-Ketata H, Salah GB, Salah HB, Marrekchi R, Jamoussi K, Boudawara T, et al. Nitrate-induced biochemical and histopathological changes in the liver of rats: ameliorative effect of Hyparrhenia hirta. Biomed Environ Sci. 2014;27(9):695–706. doi: 10.3967/bes2014.105.

Adewale OO, Samuel ES, Manubolu M, Pathakoti K. Curcumin protects sodium nitrite-induced hepatotoxicity in Wistar rats. Toxicol Rep. 2019;6:1006–11. doi: 10.1016/j.toxrep.2019.09.003.

Azza M, et al. Use of turmeric and curcumin to alleviate adverse reproductive outcomes of water: nitrate pollution in male rats. Nat Sci. 2011;9:229–39.

Tiku AB, Abraham SK, Kale RK. Eugenol as an in vivo radioprotective agent. J Radiat Res. 2004;45(3):435–40. doi: 10.1269/jrr.45.435.

Barceloux DG. Medical Toxicology of Natural Substances: Foods Fungi Medicinal Herbs Plants and Venomous Animals. Hoboken N.J: Wiley; 2008.

Park SH, Sim YB, Lee JK, Kim SM, Kang YJ, Jung JS, et al. The analgesic effects and mechanisms of orally administered eugenol. Arch Pharm Res. 2011;34(3):501–7. doi: 10.1007/s12272–011–0320-z.

Mohammadi Nejad S, Özgüneş H, Başaran N. Pharmacological and Toxicological Properties of Eugenol. Turk J Pharm Sci. 2017;14(2):201–6. doi: 10.4274/tjps.62207.

Said MM. The protective effect of eugenol against gentamicin-induced nephrotoxicity and oxidative damage in rat kidney. Fundam Clin Pharmacol. 2011;25(6):708–16. doi: 10.1111/j.1472–8206.2010.00900.x.

Nagababu E, Lakshmaiah N. Inhibitory effect of eugenol on non-enzymatic lipid peroxidation in rat liver mitochondria. Biochem Pharmacol. 1992;43(11):2393–400. doi: 10.1016/0006–2952(92)90318-d.

Reddy, A. Ch Pulla, and B. R. Lokesh. Effect of curcumin and eugenol on iron-induced hepatic toxicity in rats. Toxicology 107.1 1996: 39–45.

Saalu LC, et al. The dose-dependent testiculoprotective and testiculotoxic potentials of Telfairia occidentalis Hook. f. leaves extract in rat." International Journal of Applied Research in Natural Products 3.3 2010: 27–38.

Marklund S, Marklund G. Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem. 1974;47(3):469–74. doi: 10.1111/j.1432–1033.1974.tb03714.x.

Hugo A, Lester P. Catalase in vitro. Methods Enzymol. 1984;105:121–6. doi: 10.1016/s0076–6879(84)05016–3.

Ohkawa H, Ohishi W, Yagi K. Colorimetric method for determination of MDA activity. Biochemistry. 1979;95:351.

Avdagić N, Zaćiragić A, Babić N, Hukić M, Seremet M, Lepara O, et al. Nitric oxide as a potential biomarker in inflammatory bowel disease. Bosnian J Basic Med Sci. 2013;13(1):5–9. doi: 10.17305/bjbms.2013.2402.

Guillette Jr LJ, Edwards TM. Is nitrate an ecologically relevant endocrine disruptor in vertebrates? Integr Comp Biol. 2005;45(1):19–27. doi: 10.1093/icb/45.1.19.

Kuchakulla M, Masterson T, Arora H, Kulandavelu S, Ramasamy R. Effect of nitroso-redox imbalance on male reproduction. Transl Androl Urol. 2018;7(6):968–77. doi: 10.21037/tau.2018.08.14.

Weinberg JB, Doty E, Bonaventura J, Haney AF. Nitric oxide inhibition of human sperm motility. Fertil Steril. 1995;64(2):408–13. doi: 10.1016/s0015–0282(16)57743–7.

Li W, Tsubouchi R, Qiao S, Haneda M, Murakami K, Yoshino M. Inhibitory action of eugenol compounds on the production of nitric oxide in RAW264.7 macrophages. Biomed Res. 2006;27(2):69–74. doi: 10.2220/biomedres.27.69.

Nagababu E, et al. Assessment of antioxidant activity of eugenol; 2010. p. 165–80.

Gülçin İ. Antioxidant activity of eugenol: A structure-activity relationship study. J Med Food. 2011;14(9):975–85. doi: 10.1089/jmf.2010.0197.

Oroojan AA, Chenani N, An’aam M. Antioxidant effects of eugenol on oxidative stress induced by hydrogen peroxide in islets of Langerhans isolated from male mouse. Int J Hepatol. 2020;2020:5890378. doi: 10.1155/2020/5890378.


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