Glutathione-dependent enzymes in the follicular fluid of the first-retrieved oocyte and their impact on oocyte and embryos in polycystic ovary syndrome: A cross-sectional study
Background: Oxidative stress and GSH-dependent antioxidant system plays a key role in the pathogenesis of polycystic ovary syndrome (PCOS).
Objective: We compared glutathione peroxidase (GPx) and glutathione reductase activities and reduced glutathione (GSH) levels in serum and follicular fluid (FF) of the first-retrieved follicle and their impact on quality of oocyte and embryo in PCOS women undergoing IVF.
Materials and Methods: This cross-sectional study was conducted on 80 pairs of blood samples and FF of the first-retrieved follicle from PCOS women, at the Infertility center of Ghadir Mother and Child Hospital. The mean activity of GPx and GR, also GSH levels in the serum and FF were compared to the quality of the first follicle and resultant embryo.
Results: Retrieved oocytes included 53 (66.25%) MII, 17 (21.25%) MI, and 10 (12.5%) germinal vesicles; after IVF 42 (52.50%) embryos with grade I and 11 (13.75%) with grade II were produced. The mean values for all three antioxidants were higher in the FF compared to serum (p < 0.001). Also all of the mean measured levels were significantly higher in the FF of the MII oocytes compared to that of oocytes with lower grades (p = 0.012, 0.006 and 0.012, respectively). The mean GPX activity and GSH levels were significantly higher in the serum (p = 0.016 and 0.012, respectively) and FF (p = 0.001 for both) of the high-quality grade I embryos.
Conclusion: GSH-dependent antioxidant system functions more efficiently in the FF of oocytes and embryos with higher quality.
Key words: In vitro fertilization, Glutathione, Antioxidant, Oocyte, Embryo.
 Rotterdam ESHRE/ASRM-Sponsored PCOS Consensus Workshop Group. Revised 2003 consensus on diagnostic criteria and long-term health risks related to polycystic ovary syndrome. Fertil Steril 2004; 81: 19–25.
 Wei D, Xie J, Yin B, Hao H, Song X, Liu Q, et al. Significantly lengthened telomere in granulosa cells from women with polycystic ovarian syndrome (PCOS). J Assist Reprod Genet 2017; 34: 861–866.
 Papalou O, Victor VM, Diamanti-Kandarakis E. Oxidative stress in polycystic ovary syndrome. Curr Pharm Des 2016; 22: 2709–2722.
 Al-Gubory KH, Fowler PA, Garrel C. The roles of cellular reactive oxygen species, oxidative stress and antioxidants in pregnancy outcomes. Int J Biochem Cell Biol 2010; 42: 1634–1650.
 Banuls C, Rovira-LIopis S, Martinez de Maranon A, Veses S, Jover A, Gomez M, et al. Metabolic syndrome enhances endoplasmic reticulum, oxidative stress and leukocyte-endothelium interactions in PCOS. Metabolism 2017; 71: 153–162.
 de Melo AS, Rodrigues JK, Junior AA, Ferriani RA, Navarro PA. Oxidative stress and polycystic ovary syndrome: an evaluation during ovarian stimulation for intracytoplasmic sperm injection. Reproduction 2017; 153: 97–105.
 Rani V, Deep G, Singh RK, Palle K, Yadav UC. Oxidative stress and metabolic disorders: Pathogenesis and therapeutic strategies. Life Sci 2016; 148: 183–193.
 Ashibe S, Miyamoto R, Kato Y, Nagao Y. Detrimental effects of oxidative stress in bovine oocytes during intracytoplasmic sperm injection (ICSI). Theriogenology 2019; 133: 71–78.
 Revelli A, Delle Piane L, Casano S, Molinari E, Massobrio M, Rinaudo P. Follicular fluid content and oocyte quality: From single biochemical markers to metabolomics. Reprod Biol Endocrinol 2009; 7: 40– 52.
 Ambekar AS, Kelkar DS, Pinto SM, Sharma R, Hinduja I, Zaveri K, et al. Proteomics of follicular fluid from women with polycystic ovary syndrome suggests molecular defects in follicular development. J Clin Endocrinol Metab 2015; 100: 744–753.
 Seleem AK, El Refaeey AA, Shaalan D, Sherbiny Y, Badway A. Superoxide dismutase in polycystic ovary syndrome patients undergoing intracytoplasmic sperm injection. J Assist Reprod Genet 2014; 31: 499–504.
 Nishihara T, Matsumoto K, Hosoi Y, Morimoto Y. Evaluation of antioxidant status and oxidative stress markers in follicular fluid for human in vitro fertilization outcome. Reprod Med Biol 2018; 17: 481–486.
 Leroy JL, Vanholder T, Delanghe JR, Opsomer G, Van Soom A, Bols PE, et al. Metabolic changes in follicular fluid of the dominant follicle in high-yielding dairy cows early post partum. Theriogenology 2004; 62: 1131–1143.
 Jana SK, Narendra Babu K, Chattopadhyay R, Chakravarty B, Chaudhury K. Upper control limit of reactive oxygen species in follicular fluid beyond which viable embryo formation is not favorable. Reprod Toxicol 2010; 29: 447–451.
 Agarwal A, Aponte-Mellado A, Premkumar BJ, Shaman A, Gupta S. The effects of oxidative stress on female reproduction: A review. Reprod Biol Endocrinol 2012; 10: 49–79.
 Vignini A, Turi A, Giannubilo SR, Pescosolido D, Scognamiglio P, Zanconi S, et al. Follicular fluid nitric oxide (NO) concentrations in stimulated cycles: the relationship to embryo grading. Arch Gynecol Obstet 2008; 277: 229–232.
 Vàrnagy A, Koszegi T, Gyorgyi E, Szegedi S, Sulyok E, Prèmusz V, et al. Levels of total antioxidant capacity and 8-hydroxy-2’-deoxyguanosine of serum and follicular fluid in women undergoing in vitro fertilization: focusing on endometriosis. Hum Fertil 2018; 13: 1–9.
 Lu JC, Huang YF, Lu NQ. [WHO laboratory manual for the examination and processing of human semen: Its applicability to andrology laboratories in china]. Zhonghua Nan Ke Xue 2010; 16: 867–871. (in China)
 Veeck LL. Oocyte assessment and biological performance. Ann N Y Acad Sci 1988; 541: 259–274.
 Veeck LL. An atlas of human gametes and conceptuses: All illustrated reference for assisted reproductive technology (the encyclopedia of visual medicine series). 1st Ed. CRC Press, USA; 1999.
 Mostafavi-Pour Z, Khademi F, Zal F, Sardarian AR, Amini F. In vitro analysis of csa-induced hepatotoxicity in hepg2 cell line: Oxidative stress and α2 and β1 integrin subunits expression. Hepat Mon 2013; 13: e11447.
 Mashhoody T, Rastegar K, Zal F. Perindopril may improve the hippocampal reduced glutathione content in rats. Adv Pharm Bull 2014; 4: 155–159.
 Agarwal A, Gupta S, Sharma RK. Role of oxidative stress in female reproduction. Reprod Biol Endocrinol 2005; 3: 28–48.
 O’Gorman A, Wallace M, Cottell E, Gibney MJ, McAuliffe FM, Wingfield M, et al. Metabolic profiling of human follicular fluid identifies potential biomarkers of oocyte developmental competence. Reproduction 2013; 146: 389–395.
 Luddi A, Capaldo A, Focarelli R, Gori M, Morgante G, Piomboni P, et al. Antioxidants reduce oxidative stress in follicular fluid of aged women undergoing IVF. Reprod Biol Endocrinol 2016; 14: 57–63.
 Kish I, Ohishi M, Akiba Y, Asada H, Konishi Y, Nakano M, et al. Thioredoxin, an antioxidant redox protein, in ovarian follicles of women undergoing in vitro fertilization. Endocr J 2016; 63: 9–20.
 Zuelke KA, Jeffay SC, Zucker RM, Perreault SD. Glutathione (GSH) concentrations vary with the cell cycle in maturing hamster oocytes, zygotes, and preimplantation stage embryos. Mol Reprod Dev 2003; 64: 106–112.
 Pasqualotto EB, Lara LV, Salvador M, Sobreiro BP, Borges E, Pasqualotto FF. The role of enzymatic antioxidants detected in the follicular fluid and semen of infertile couples undergoing assisted reproduction. Hum Fertil 2009; 12: 166–171.
 Agarwal A, Majzoub A. Role of antioxidants in assisted reproductive techniques. World J Mens Health 2017; 35: 77–93.