Effects of Dietary Pomegranate Peel on Antioxidant Gene Expression and DJ-1 Protein Abundance in Ram Testes

Nikfarjam, Mahdieh; Rashki Ghaleno, Leila; Shahverdi, Abdolhossein; Mirshahvaladi, Shahab; Ghoreishi, Seyed Mehdi; Alizadeh, AliReza

doi:10.22074/ijfs.2021.141725.1052

Effects of Dietary Pomegranate Peel on Antioxidant Gene Expression and DJ-1 Protein Abundance in Ram Testes

Document Type : Original Article

Authors

¹ Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

² Department of Cellular and Molecular Biology, Faculty of Basic Sciences and Advanced Technologies in Biology, University of Science and Culture, Tehran, Iran

³ Reproductive Epidemiology Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

⁴ Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran Department of Clinical Medicine, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, Australia

⁵ Department of Clinical Medicine, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, Australia

⁶ Department of Animal Science, School of Agriculture, Shiraz University, Shiraz, Iran

10.22074/ijfs.2021.141725.1052

Abstract

Background: Pomegranate is an ancient fruit containing Punicalagin, which has known as an effective antioxidant.
Pomegranate peel was recognized as a phenol and tannin source, and pomegranate seed contains unique fatty acid
(Punicic acid). Limited information exists about the influences of pomegranate peel and seed on antioxidant enzymes
and proteins in the male reproduction system. This study was performed to determine the pomegranate peel and seed
effects on the expression of antioxidant genes and DJ-1 protein in ram’s testis.
Materials and Methods: In this experimental study, twenty-one mature Iranian rams were randomly divided into
three groups (n=7 in each group), and fed experimental diets consisted of a control diet (C), a diet containing dry
pomegranate seed pulp (S), and a diet containing pomegranate peel (P) for 80 days. All rams were offered isoenergetic
and isonitrogenous rations. Testicular tissue samples were collected, and expression of Gpx1, Gpx4, Prdx4, Prdx5, and
Sod2 genes was quantified by real-time polymerase chain reaction (RT-PCR). In addition, western blotting was used
to evaluate DJ-1 expression at the protein level.
Results: Gpx1 and Sod2 mRNA levels in the peel group were significantly (P<0.05) higher than control. Prdx5 mRNA
level was increased (P<0.05) in the seeds group than in the control group. Gpx4 and Prdx4 expression were statistically not affected significantly by the experimental diet. Data analysis showed a significant (P<0.05) increase (1.5-fold) in the expression level of DJ-1 in peel groups than in control.
Conclusion: The expression of antioxidant genes and DJ-1 protein in ram testes are more influenced by pomegranate
peel than seed.

Keywords

20.1001.1.2008076.2021.15.4.4.4

References

Agarwal A, Mulgund A, Hamada A, Chyatte MR. A unique view on male infertility around the globe. Reprod Biol Endocrinol. 2015; 13(1): 37.
Moretti E, Collodel G, Fiaschi AI, Micheli L, Iacoponi F, Cerretani D. Nitric oxide, malondialdheyde and non-enzymatic antioxidants assessed in viable spermatozoa from selected infertile men. Reprod Biol. 2017; 17(4): 370-375.
Darbandi M, Darbandi S, Agarwal A, Baskaran S, Dutta S, Sengupta P, et al. Reactive oxygen species-induced alterations in H19-Igf2 methylation patterns, seminal plasma metabolites, and semen quality. J Assist Reprod Genet. 2019; 36(2): 241-253.
Darbandi M, Darbandi S, Agarwal A, Baskaran S, Sengupta P, Dutta S, et al. Oxidative stress-induced alterations in seminal plasma antioxidants: is there any association with keap1 gene methylation in human spermatozoa? Andrologia. 2019; 51(1): e13159.
Meseguer M, Martinez-Conejero JA, Muriel L, Pellicer A, Remohi J, Garrido N. The human sperm glutathione system: a key role in male fertility and successful cryopreservation. Drug Metab Lett. 2007; 1(2): 121-126.
Barati E, Nikzad H, Karimian M. Oxidative stress and male infertility: current knowledge of pathophysiology and role of antioxidant therapy in disease management. Cell Mol Life Sci. 2020; 77(1): 93-113.
Badr YAA, Sepehran E, Del Azar A, Sadeghi H, Nouri M. The effect of saffron on semen analysis in infertile men with clinical varicocele after varicocelectomy. Nephro-Urol Mon. 2017; 9(5): e59939.
Heidary M, Vahhabi S, Reza Nejadi J, Delfan B, Birjandi M, Kaviani H, et al. Effect of saffron on semen parameters of infertile men. Urol J. 2008; 5(4): 255-259.
Rao F, Tian H, Li W, Hung H, Sun F. Potential role of punicalagin against oxidative stress induced testicular damage. Asian J Androl. 2016; 18(4): 627-632.
Turk G, Sonmez M, Aydin M, Yuce A, Gur S, Yuksel M, et al. Effects of pomegranate juice consumption on sperm quality, spermatogenic cell density, antioxidant activity and testosterone level in male rats. Clin Nutr. 2008; 27(2): 289-296.
Al-Olayan EM, El-Khadragy MF, Metwally DM, Abdel Moneim AE. Protective effects of pomegranate (Punica granatum) juice on testes against carbon tetrachloride intoxication in rats. BMC Complement Altern Med. 2014; 14: 164.
Zarepourfard H, Riasi A, Frouzanfar M, Hajian M, Nasr Esfahani MH. Pomegranate seed in diet, affects sperm parameters of cloned goats following freezing-thawing. Theriogenology. 2019; 125: 203-209.
Makkar HP. Measurement of total phenolics and tannins using Folin-Ciocalteu method. Quantification of tannins in tree and shrub foliage. Netherlands: Springer Science & Business Media Dordrecht; 2003; 49-51.
Gil MI, Tomás-Barberán FA, Hess-Pierce B, Holcroft DM, Kader AA. Antioxidant Activity of pomegranate juice and its relationship with phenolic composition and processing. J Agric Food Chem. 2000; 48(10): 4581-4589.
Dkhil M, Al-Quraishy S, Abdel Moneim A. Effect of pomegranate (Punica granatum L.) juice and methanolic peel extract on testis of male rats. Pakistan J Zool. 2013; 45(5): 1343-1349.
Kang SJ, Choi BR, Lee EK, Kim SH, Yi HY, Park HR, et al. Inhibitory effect of dried pomegranate concentration powder on melanogenesis in B16F10 melanoma cells; involvement of p38 and PKA signaling pathways. Int J Mol Sci. 2015; 16(10): 24219-24242.
Im JY, Lee KW, Woo JM, Junn E, Mouradian MM. DJ-1 induces thioredoxin 1 expression through the Nrf2 pathway. Hum Mol Genet. 2012; 21(13): 3013-3024.
Shi H, Liu J, Zhu P, Wang H, Zhao Z, Sun G, et al. Expression of peroxiredoxins in the human testis, epididymis and spermatozoa and their role in preventing H2O2-induced damage to spermatozoa. Folia Histochem Cytobiol. 2018; 56(3): 141-150.