In vitro Effects of Berberine on the Vitality and Oxidative Profile of Bovine Spermatozoa
The aim of this study was to evaluate the dose- and time-dependent in vitro effects of berberine (BER), a natural alkaloid with numerous biological properties on bovine spermatozoa during three time periods (0 h, 2 h, 24 h). Bovine semen samples were diluted and cultivated in physiological saline solution containing 0.5% DMSO together with 200, 100, 50, 10, 5, and 1 μmol/L BER. Spermatozoa motility was assessed using the computer assisted semen analyzer. The viability of spermatozoa was assessed by the metabolic (MTT) assay, production of superoxide radicals was quantified using the nitroblue tetrazolium (NBT) test, and chemiluminescence was used to evaluate the generation of reactive oxygen species (ROS). Cell lysates were prepared and the extent of lipid peroxidation (LPO) was evaluated using the TBARS assay. The results of the movement activity showed a significant increase in the motility during long term cultivation in case of concentrations ranging between 1 and 10 μmol/L BER (P < 0.01; P < 0.001; 24 h). At the same time, supplementation of 1, 5 and 10 μmol/L BER led to a significant preservation of the cell viability (P < 0.001; 24 h). BER addition at a range of 1-50 μmol/L also provided a significantly higher protection against superoxide (P < 0.05) and ROS (P < 0.001; P < 0.01) overgeneration as well as LPO (P < 0.01; P<0.05) after a 24 h cultivation. We may suggest that supplementation of BER to bovine spermatozoa, particularly at concentrations ranging between 1 and 50 μmol/L, may offer protection to the motility, viability and oxidative status of the spermatozoa, particularly notable at 24 h.
Digital Object Identifier (DOI): doi.org/10.5281/zenodo.Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 47
 W. M. Maxwell, and T. Stohjanov, “Liquid storage of ram semen in the absence or presence of some antioxidants,” Reproduction, Fertility and Development, vol. 8, no. 6, pp. 1013-1020, 1996.
 H. Funahashi, and T. Sano, “Select antioxidants improve the function of extended boar semen stored at 10 °C,” Theriogenology, vol. 63, no. 6, 1 pp. 1605-1616, 2005.
 J. C. Calamera, P. J. Fernandez, M. G. Buffone, A. A. Acosta, and G. F. Doncel, “Effects of long-term in vitro incubation of human spermatozoa: functional parameters and catalase effect,” Andrologia, vol. 33, pp. 79-86, 2001.
 H. J. Chi, J. H. Kim, C. S. Ryu, J. Y. Lee, J. S. Park, D. Y. Chung, S. Y. Choi, M. H. Kim, E. K. Chun, and S. I. Roh, “Protective effect of antioxidant supplementation in sperm-preparation medium against oxidative stress in human spermatozoa,” Human Reproduction, vol. 23, no.5 pp. 1023-1028, 2008.
 R. J. Aitken, J. K. Wingate, G. N. De Iuliis, A. J. Koppers, and E. A. McLaughlin, “Cis-unsaturated fatty acids stimulate reactive oxygen species generation and lipid peroxidation in human spermatozoa,” The Journal of Clinical Endocrinology & Metabolism, vol. 91, no. 10, pp. 4154-4163, 2006.
 M. N. Bucak, P. B. Tuncer, S. Sarıözkan, N. Başpınar, M. Taşpınar, K. Coyan, A. Bilgili, P. P. Akalın, S. Büyükleblebici, S. Aydos, S. Ilgaz, A. Sunguroğlu, and D. Oztuna, “Effects of antioxidants on post-thawed bovine sperm and oxidative stress parameters: antioxidants protect DNA integrity against cryodamage,” Cryobiology, vol. 61, no. 3, pp. 248-253, 2010.
 F. J. Peña, H. Rodríguez Martínez, J. A. Tapia, C. Ortega Ferrusola, L. González Fernández, and B. Macías García, “Mitochondria in mammalian sperm physiology and pathology: a review,” Reproduction in Domestic Animals, vol 44, no. 2, pp. 345-349, 2009.
 S. S. du Plessis, K. Makker, N. R. Desai, and A. Agarwal, “Impact of oxidative stress on IVF,” Expert Review of Obstetrics and Gynecology, vol. 3, no. 4, pp. 539-554, 2008.
 E. Tvrdá, Z. Kňažická, L. Bárdos, P. Massányi, and N. Lukáč, “Impact of oxidative stress on male fertility - a review,” Acta Veterinaria Hungarica, vol. 59, no. 4, pp. 465-484, 2011.
 B. A. Fatma, Ch. F. Nozha, D. Ines, A. Hamadi, H. Basma, and A. K. Leila, “Sperm quality improvement after date seed oil in vitro supplementation in spontaneous and induced oxidative stress,” Asian Journal of Andrology, vol. 11, pp. 393-398, 2009.
 S. K. Kulkarni, and A. Dhir, “Berberine: a plant alkaloid with therapeutic potential for central nervous system disorders,” Phytotherapy Research, vol. 24, no. 3, pp. 317-324, 2010.
 C. B. N. Timothy, S. Gregory, and N. D. Kelly, “Berberine: therapeutic potential of an alkaloid found in several medicinal plants,” Alternative Medicine Review, vol. 13, pp. 94-103, 1997.
 C. L. Kuo, C. W. Chi, and T. Y. Liu, “The anti-inflammatory potential of berberine in vitro and in vivo,” Cancer Letters, vol. 2003, pp. 127-137, 2004.
 R. Khosrokhavar, and A. S. F. Ahmadiani, “Antihistaminic and anticholinergic activity of methanolic extract of barberry fruit (Berberis vulgaris) in the guinea- pig ileum,” Journal of Medicinal Plants, vol. 13, pp. 99-105, 2010.
 A. E. A. El-Wahab, D. A. Ghareeb, E. E. M. Sarhan, M. M. Abu-Serie, and M. A. El Demellawy, “In vitro biological assessment of berberis vulgaris and its active constituent, berberine: antioxidants, anti-acetylcholinesterase, anti-diabetic and anticancer effects,” BMC Complementary and Alternative Medicine, vol. 13, pp. 218-230, 2013.
 S. Schaffer, and M. Heinrich, “Understanding local Mediterranean diets: a multidisciplinary pharmacological and ethnobotanical approach,” Pharmacological Research, vol. 52, pp. 353-366, 2005.
 R. N. Gacche, and N. A. Dhole, “Antioxidant and possible anti-inflammatory potential of selected medicinal plants prescribed in the Indian traditional system of medicine,” Pharmaceutical Biology, vol. 44, pp. 389-395, 2006.
 E. Tvrdá, M. Halenár, H. Greifová, A. Mackovich, F. Hashim, and N. Lukáč, “The effect of curcumin on cryopreserved bovine semen,” International Journal of Animal and Veterinary Sciences, vol. 10, no. 11, pp. 707-711, 2016a.
 E. Tvrdá, A. Kováčik, E. Tušimová, D. Paál, A. Mackovich, J. Alimov, and N. Lukáč, “Antioxidant efficiency of lycopene on oxidative stress - induced damage in bovine spermatozoa,” Journal of Animal Science and Biotechnology, vol. 7, no. 1, pp. 50-63, 2016b.
 F. Amidi, A. Pazhohan, M. Shabani Nashtaei, M. Khodarahmian, and S. Nekoonam, “The role of antioxidants in sperm freezing: a review,” Cell Tissue Banking, vol. 17, no. 4, pp. 745-756, 2016.
 L. Chen, T. Wang, and J. Liu, “Effect of berberine on human sperm parameters in vitro,” Translational Andrology and Urology, vol. 5, pp. AB299, 2016.
 N. Adel, H. El Maghraby, D. Ghareeb, and M. Elmahdy, “Vitamin E and berberine counteract the adverse effects of ROS on sperm and seminal parameters,” Poster published at the ESHRE Meeting 2015, doi: 10.13140/RG.2.2.19568.46089, 2015.
 H. D. Guthrie, and G. R. Welch, “Effects of reactive oxygen species on sperm function,” Theriogenology, vol. 78, no. 8, pp. 1700-1708, 2012.
 A. J. Koppers, G. N. De Iuliis, J. M. Finnie, E. A. McLaughlin, and R. J. Aitken, “Significance of mitochondrial reactive oxygen species in the generation of oxidative stress in spermatozoa,” Journal of Clinical Endocrinology & Metabolism, vol. 93, no. 8, pp. 3199-3207, 2008.
 Q. Li, T. Zhou, Ch. Liu, X. Y. Wang, J. Q. Zhang, F. Wu, G. Lin, Y. M. Ma, B. L. Ma, “Mitochondrial membrane potential played crucial roles in the accumulation of berberine in HepG2 cells,” Bioscience Reports, vol. 39, pp. 1-13, 2019.
 J. Ye, J. Le, and Y. Sun, “Berberine improves mitochondrial function in colon epithelial cells to protect L-cells from necrosis in preservation of GLP-1 secretion,” Diabetes, vol. 67, pp. A637, 2018.
 M. Zovko Koncić, D. Kremer, K. Karlović, and I. Kosalec, “Evaluation of antioxidant activities and phenolic content of Berberis vulgaris L. and Berberis croatica Horvat,” Food and Chemical Toxicology, vol. 48, pp. 2176-2180, 2010.
 S. R. Saleh, R. Attia, and D. A. Ghareeb, “The ameliorating effect of berberine-rich fraction against gossypol-induced testicular inflammation and oxidative stress,” Oxidative Medicine and Cellular Longevity, vol. 2018, doi: 10.1155/2018/1056173, 2018.