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Improvement of sperm function, chromatin damage, and oxidative damage by N-Acetyl cysteine in varicocelized rats model

Marziyeh Tavalaee, Maurizio Dattilo, Parisa Mohammadi, Mohammad Hossein Nasr-Esfahani




Introduction: N-Acetylcysteine (NAC), an acetylated form of the amino acid cysteine and precursor of reduced glutathione, plays important roles in a multitude of cellular processes, such as oxidative damage and detoxification of many electrophiles. Considering the pathophysiology of oxidative stress induced infertility in varicocele, we aimed to investigate the effect of NAC on semen analysis parameters (light microscopy), chromatin structure (aniline blue and acridine orange staining), and lipid peroxidation (BODIPY probe) in varicocelized rats.

Methods: In this experimental study, varicocelizing surgery was carried out on 30 Wistar rats. Ten of them were sacrificed after two months (one round spermatogenesis), together with control rats (n=10) and sham operated rats (n=10), to verify the varicocele model. Out of the remaining twenty varicocelized rats, ten received NAC while ten were treated with water (control group) for two months.

Results: All the investigational parameters (sperm parameters, chromatin integrity, and lipid peroxidation) severely worsened 2 and 4 months after surgical varicocele. The administration of NAC for two months significantly improved all the investigational parameters as compared to control rats at four months (p<0.05).

Conclusion: The supplementation of varicocelized rats with NAC was effective in antagonizing the damage as well as in preserving testicular structure and spermatogenetic function. These effects are likely to occur also in clinical varicocele.


Samuni Y, Goldstein S, Dean OM, Berk M. The chemistry and biological activities of N-acetylcysteine. Biochimica et Biophysica Acta (BBA)-General Subjects. 2013;1830(8):4117-29.

Berk M, Malhi GS, Gray LJ, Dean OM. The promise of N-acetylcysteine in neuropsychiatry. Trends in pharmacological sciences. 2013;34(3):167-77.

Manjula Ramen T. Depletion of glutathione during oxidative stress and efficacy of N-acetyl cysteine: an old drug with new approaches. Med Chem. 2015;5(1):037-9.

Mytilineou C, Kramer BC, Yabut JA. Glutathione depletion and oxidative stress. Parkinsonism & related disorders. 2002;8(6):385-7.

Ballatori N, Krance SM, Marchan R, Hammond CL. Plasma membrane glutathione transporters and their roles in cell physiology and pathophysiology. Molecular aspects of medicine. 2009;30(1-2):13-28.

Skvarc DR, Dean OM, Byrne LK, Gray L, Lane S, Lewis M, et al. The effect of N-acetylcysteine (NAC) on human cognition–A systematic review. Neuroscience & Biobehavioral Reviews. 2017;78:44-56.

Ciftci H, Verit A, Savas M, Yeni E, Erel O. Effects of N-acetylcysteine on semen parameters and oxidative/antioxidant status. Urology. 2009;74(1):73-6.

Alsaikhan B, Alrabeeah K, Delouya G, Zini A. Epidemiology of varicocele. Asian journal of andrology. 2016;18(2):179.

Cho C-L, Esteves SC, Agarwal A. Novel insights into the pathophysiology of varicocele and its association with reactive oxygen species and sperm DNA fragmentation. Asian Journal of Andrology. 2016;18(2):186.

Garg H, Kumar R. An update on the role of medical treatment including antioxidant therapy in varicocele. Asian journal of andrology. 2016;18(2):222.

Barekat F, Tavalaee M, Deemeh MR, Bahreinian M, Azadi L, Abbasi H, et al. A preliminary study: N-acetyl-L-cysteine improves semen quality following varicocelectomy. International journal of fertility & sterility. 2016;10(1):120.

Ko KW, Chae JY, Kim SW, Moon DG, Kim JJ, Yoon DK, et al. The effect of the partial obstruction site of the renal vein on testis and kidney in rats: is the traditional animal model suitable for varicocele research? Korean journal of urology. 2010;51(8):565-71.

Freireich EJ, Gehan E, Rall D, Schmidt L, Skipper H. Quantitative comparison of toxicity of anticancer agents in mouse, rat, hamster, dog, monkey, and man. Cancer chemotherapy reports. 1966;50(4):219.

Afiyani AA, Deemeh MR, Tavalaee M, Razi M, Bahadorani M, Shokrollahi B, et al. Evaluation of heat‐shock protein A2 (HSPA2) in male rats before and after varicocele induction. Molecular reproduction and development. 2014;81(8):766-76.

Aitken RJ, Wingate JK, De Iuliis GN, McLaughlin EA. Analysis of lipid peroxidation in human spermatozoa using BODIPY C11. MHR: Basic science of reproductive medicine. 2007;13(4):203-11.

Agarwal A, Prabakaran S, Allamaneni SS. Relationship between oxidative stress, varicocele and infertility: a meta-analysis. Reproductive BioMedicine Online. 2006;12(5):630-3.

Atkuri KR, Mantovani JJ, Herzenberg LA, Herzenberg LA. N-Acetylcysteine—a safe antidote for cysteine/glutathione deficiency. Current opinion in pharmacology. 2007;7(4):355-9.

Dhouib IE, Jallouli M, Annabi A, Gharbi N, Elfazaa S, Lasram MM. A minireview on N-acetylcysteine: an old drug with new approaches. Life sciences. 2016;151:359-63.

Banerjee R. Redox outside the box: linking extracellular redox remodeling with intracellular redox metabolism. Journal of biological chemistry. 2012;287(7):4397-402.

Ji L, Liu R, Zhang XD, Chen HL, Bai H, Wang X, et al. N-acetylcysteine attenuates phosgene-induced acute lung injury via up-regulation of Nrf2 expression. Inhalation toxicology. 2010;22(7):535-42.

Cai Z, Lou Q, Wang F, Li E, Sun J, Fang H, et al. N-acetylcysteine protects against liver injure induced by carbon tetrachloride via activation of the Nrf2/HO-1 pathway. International journal of clinical and experimental pathology. 2015;8(7):8655.

Duarte F, Blaya R, Telöken PE, Becker D, Fernandes M, Rhoden EL. The effects of N-acetylcysteine on spermatogenesis and degree of testicular germ cell apoptosis in an experimental model of varicocele in rats. International urology and nephrology. 2010;42(3):603-8.

Bahreinian M, Tavalaee M, Abbasi H, Kiani-Esfahani A, Shiravi AH, Nasr-Esfahani MH. DNA hypomethylation predisposes sperm to DNA damage in individuals with varicocele. Systems biology in reproductive medicine. 2015;61(4):179-86.

Pallotti F, Paoli D, Carlini T, Vestri A, Martino G, Lenzi A, et al. Varicocele and semen quality: a retrospective case–control study of 4230 patients from a single centre. Journal of endocrinological investigation. 2018;41(2):185-92.

Angelopoulou R, Plastira K, Msaouel P. Spermatozoal sensitive biomarkers to defective protaminosis and fragmented DNA. Reproductive Biology and Endocrinology. 2007;5(1):36.

Yassine S, Escoffier J, Martinez G, Coutton C, Karaouzene T, Zouari R, et al. Dpy19l2-deficient globozoospermic sperm display altered genome packaging and DNA damage that compromises the initiation of embryo development. Mhr: Basic science of reproductive medicine. 2015;21(2):169-85.

Agarwal A, Sharma R, Durairajanayagam D, Cui Z, Ayaz A, Gupta S, et al. Differential proteomic profiling of spermatozoal proteins of infertile men with unilateral or bilateral varicocele. Urology. 2015;85(3):580-8.

Hackney A. Endurance exercise training and reproductive endocrine dysfunction in men alterations in the hypothalamic-pituitary-testicular axis. Current pharmaceutical design. 2001;7(4):261-73.

Hayden RP, Tanrikut C. Testosterone and varicocele. Urologic Clinics. 2016;43(2):223-32.

Jana K, Dutta A, Chakraborty P, Manna I, Firdaus SB, Bandyopadhyay D, et al. Alpha‐lipoic acid and N‐acetylcysteine protects intensive swimming exercise‐mediated germ‐cell depletion, pro‐oxidant generation, and alteration of steroidogenesis in rat testis. Molecular reproduction and development. 2014;81(9):833-50.

Kalmar B, Greensmith L. Induction of heat shock proteins for protection against oxidative stress. Advanced drug delivery reviews. 2009;61(4):310-8.

Skakkebaek NE, Rajpert-De Meyts E, Buck Louis GM, Toppari J, Andersson A-M, Eisenberg ML, et al. Male reproductive disorders and fertility trends: influences of environment and genetic susceptibility. Physiological reviews. 2016;96(1):55-97.

Rao AK, Shaha C. N‐acetylcysteine prevents MAA induced male germ cell apoptosis: role of glutathione and cytochrome c. FEBS letters. 2002;527(1-3):133-7.

Lachance C, Goupil S, Tremblay R, Leclerc P. The immobilization of human spermatozoa by STAT 3 inhibitory compound V results from an excessive intracellular amount of reactive oxygen species. Andrology. 2016;4(1):133-42.

Safarinejad MR, Safarinejad S. Efficacy of selenium and/or N-acetyl-cysteine for improving semen parameters in infertile men: a double-blind, placebo controlled, randomized study. The Journal of urology. 2009;181(2):741-51.

Chen X, Jhee K-H, Kruger WD. Production of the neuromodulator H2S by cystathionine β-synthase via the condensation of cysteine and homocysteine. Journal of Biological Chemistry. 2004;279(50):52082-6.

Xie Z-Z, Liu Y, Bian J-S. Hydrogen sulfide and cellular redox homeostasis. Oxidative medicine and cellular longevity. 2016;2016.

Singh S, Padovani D, Leslie RA, Chiku T, Banerjee R. Relative contributions of cystathionine β-synthase and γ-cystathionase to H2S biogenesis via alternative trans-sulfuration reactions. Journal of Biological Chemistry. 2009;284(33):22457-66.

Toohey JI. Possible involvement of hydrosulfide in B12-dependent methyl group transfer. Molecules. 2017;22(4):582.

Ge S-Q, Lin S-L, Zhao Z-H, Sun Q-Y. Epigenetic dynamics and interplay during spermatogenesis and embryogenesis: implications for male fertility and offspring health. Oncotarget. 2017;8(32):53804.


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