Mustafa KAVUTÇU, Recep Onur KARABACAK, Rabia TURAL, Cengiz KARAKAYA, Mehmet ERDEM, Zeynep AYKOL

Investigation of oxidative stress status in cumulus cells in patıents with in vitro fertilization

Background/aim: The negative impact of oxidative stress on oocytes obtained from in vitro fertilization (IVF) patients is a challenge for the optimization of live birth rates. In this study, it is aimed to investigate whether oxidant/antioxidant parameters have a predictive value in terms of determining the count and quality of oocytes. Materials and methods: Catalase (CAT), glutathione-S-transferase (GST), arylesterase (ARE) enzyme activities, and malondialdehyde (MDA) levels were analysed in cumulus cells of poor responder (n = 28, oocyte count ≤ 4), normo responder (n = 48, 5 ≤ oocyte count ≤ 14), and high responder (n = 26, oocyte count ≥ 15) patient groups continuing IVF treatment. Results: The cumulus cell GST enzyme activity were statistically significantly increased in the high responders group compared to the poor responder and the normo responder’s groups (p < 0.001 and p = 0.002, respectively). The cumulus cell MDA levels were significantly decreased in the high responder group compared to the poor responder group (p = 0.008). The cumulus cell CAT (p = 0.175) and ARE (p = 0.124) enzyme activities were examined but no statistically significant difference found between the groups. Conclusion: The significant increase in GST enzyme activity and significant decrease in MDA levels in the high responder group indicate that oxidative stress has an effect oocyte status and quality.Key words: In vitro fertilization, cumulus cell, catalase, glutathione-S-transferase, arylesterase, malondialdehyde

PDF

___

1. Ibrahimi R, Musavi M, Shojaee M, Moossavi M, Moossavi SZ et al. The polymorphism of XRCC1 Arg399Gln (rs25487) and male infertility risk: a meta-analysis of 1,407 cases and 974 control studies. Bratislava Medical Journal 2019; 120 (5): 349- 355. doi: 10.4149/BLL_2019_057
2. Badger-Emeka L. In-vitro Fertilisation. In: Ogbonna, JC, Ubi, BE, and Enweani, IB (editors). Fundamentals, Industrial and Medical Biotechnology. Beijing: Universal Academic Services; 2013. 180-195.
3. Rani K, Paliwal S. A brief reviewon in vitro fertilization (IVF): an advanced and miraculous gateway for infertility treatments. World Journal of Pharmaceutical Sciences 2014; 3: 647-658.
4. Mengden Lv, Klamt F, Smitz J. Redox biology of human cumulus cells: basic concepts, impact on oocyte quality, and potential clinical use. Antioxidants Redox Signaling 2020; 32 (8): 522-535. doi: 10.1089/ars.2019.7984
5. Tatemato H, Sakurai N, Muto N. Protection of porcine oocytes against apaptotic cell death caused by oxidative stress during in vitro maturation: role of cumulus cells. Biology of Reproduction 2000; 63 (3): 805-810. doi: 10.1095/biolreprod63.3.805
6. Tamura H, Takasaki A, Miwa I, Taniguchi K, Maekawa R et al. Oxidative stress impairs oocyte quality and melatonin protects oocytes from free radical damage and improves fertilization rate. Journal of Pineal Research 2008; 44 (3): 280-287. doi: 10.1111/j.1600-079X.2007.00524.x
7. Da Broi MG, Giorgi VSI, Wang F, Keefe DL, Albertini D et al. Influence of follicular fluid and cumulus cells on oocyte quality: clinical implications. Journal of Assisted Reproduction and Genetics 2018; 35 (5): 735-751. doi: 10.1007/s10815-018- 1143-3
8. Scibior D, Czeczot H. Catalase: structure, properties, functions. Postepy Higieny i Medycyny Doswiadczalnej (Online) 2006; 60: 170-180.
9. Narayanan SN, Kumar RS, Kedage V, Nalini K, Nayak S et al. Evaluation of oxidant stress and antioxidant defense in discrete brain regions of rats exposed to 900 MHz radiation. Bratislava Lekarske Listy 2014; 115 (5): 260-266. doi: 10.4149/ BLL_2014_054
10. Sarandol E, Tas S, Serdar Z, Dirican M. Effects of thiamine treatment on oxidative stress in experimental diabetes. Bratislava Medical Journal 2020; 121 (3): 235-241. doi: 10.4149/ BLL_2020_036
11. Smith GS, Walter GL, Walker RM. Clinical Pathology in NonClinical Toxicology Testing. In: Wanda M, Rousseaux CG, Walling MA (editors). Haschek and Rousseaux’s Handbook of Toxicologic Pathology. 3thed. Boston: Academic Press; 2013. pp. 565-594.
12. Başkol G, Köse K. Paraoxanase: biochemical features, functions and clinical importance. Erciyes Medical Journal 2004; 26 (2): 75-80.
13. Hahn M, Subbiah MT. Significant association of lipid peroxidation products with high density lipoproteins. Biochemistry and Molecular Biology International 1994; 33 (4): 699-704.
14. Okyay AG, Aslan M, Taşkın A, Selek S, Cengiz Altın G et al. Oxidative stress markers in ivf patients and their association with pregnancy results. Turkiye Klinikleri Journal of Medical Sciences 2014; 34 (2): 204-209. doi: 10.5336/medsci.2013-36880
15. Rincón JAA, Madeira EM, Campos FT, Mion B, Silva JF et al. Exogenous paraoxonase-1 during oocyte maturation improves bovine embryo development in vitro. Reproduction in Domestic Animals 2016; 51 (5): 827-830. doi: 10.1111/ rda.12730
16. Sezer K, Keskin M. Serbest oksijen radikallerinin hastalıkların patogenezisindeki rolü. Fırat Üniversitesi Sağlık Bilimleri Veteriner Dergisi 2014; 28 (1): 49-56. (in Turkish).
17. Kellogg EW, Fridowich I. Liposome oxidation and erythrocyte lysis by enzymically generated superoxide and hydrogen peroxide. Journal of Biological Chemistry 1977; 252 (19): 6721-6728. doi: 10.1016/S0021-9258(17)39909-X
18. Dasgupta A, Klein K. Chapter 2-Methods for Measuring Oxidative Stress in the Laboratory. pp. 19-40. Antioxidants in Food, Vitamins and Supplements, Prevention and Treatment of Disease, 2014. doi: 10.1016/B978-0-12-405872-9.00002-1
19. Aebi H. Catalase. In: Bergmeyer HU. (editor). Methods of Enzymatic Analysis. New York and London: Academic Press; 1974. pp.673-677.
20. Habig WH, Pabst MJ, Jakoby WB. Glutathione S-transferases: the first enzymatic step in mercapturic acid formation. Journal of Biological Chemistry 1974; 249 (22): 7130-7139. doi: 10.1016/S0021-9258(19)42083-8
21. Furlong CE, Richter RJ, Seidel SL, Costa LG, Motulsky AG. Spectrophotometric assays for the enzymatic hydrolysis of the active metabolites of chlorpyrifos and parathion by plasma paraoxonase/arylesterase: Analytical Biochemistry 1989; 180 (2): 242-247. doi: 10.1016/0003-2697(89)90424-7
22. Van Ye TM, Roza AM, Pieper GM, Henderson JJr, Johnson CP et al. Inhibition of intestinal lipid peroxidation does not minimize morphological damage. Journal of Surgical Research 1993; 55 (5): 553-558. doi: 10.1006/jsre.1993.1183
23. Nuñez-Calonge R, Cortés S, Gutierrez Gonzalez LM, Kireev R, Vara E et al. Oxidative stress in follicular fluid of young women with low response compared with fertile oocyte donors. Reproductive BioMedicine Online 2016; 32 (4): 446-456. doi: 10.1016/j.rbmo.2015.12.010
24. Attaran M, Pasqualotto E, Falcone T, Goldberg JM, Miller KF et al. The effect of follicular fluid reactive oxygen species on the outcome of in vitro fertilization. International Journal of Fertility and Women’s Medicine 2000; 45 (5): 314-320.
25. Wang S, He G, Chen M, Zuo T, Xu W et al. The role of antioxidant enzymes in the ovaries. Oxidative Medicine and Cellular Longevity 2017; 2017: 4371714. doi: 10.1155/2017/4371714
26. Gupta S, Choi A, Yu HY, Czerniak SM, Holick EA et al. Fluctuations in total antioxidant capacity, catalase activity, and hydrogen peroxide levels of follicular fluid during bovine folliculogenesis. Reproduction, Fertility and Development 2011; 23 (5): 673-680. doi: 10.1071/RD10270
27. Elsharkawy SS, Abdrabo MS, Khalil MH, Mannaa HF. Evaluation of oxidative stress markers in serum and follicular fluid of women with infertility related to endometriosis. Journal of Clinical Obstetrics, Gynecology & Infertility 2020; 4 (1): 1045.
28. Kala M, Shaikh MV, Nivsarkar M. Equilibrium between antioxidants and reactive oxygen species: a requisite for oocyte development and maturation. Reproductive Medicine and Biology 2017; 16 (1): 28-35. doi: 10.1002/rmb2.12013
29. Carbone MC, Tatone C, Delle Monache S, Marci R, Caserta D et al. Antioxidant enzymatic defences in human follicular fluid: characterization and age-dependent changes. Molecular Human Reproduction 2003; 9 (11): 639-643. doi: 10.1093/ molehr/gag090
30. Ito M, Muraki M, Takahashi Y, Imai M, Tsukui T et al. Glutathione S-transferase theta 1 expressed in granulosa cells as a biomarker for oocyte quality in age-related infertility. Fertility and Sterility 2008; 90 (4): 1026-1035. doi: 10.1016/j. fertnstert.2007.07.1389
31. Mihalas BP, Redgrove KA, McLaughlin EA, Nixon B. Molecular mechanisms responsible for increased vulnerability of the ageing oocyte to oxidative damage. Oxidative Medicine and Cellular Longevity 2017; 2017: 4015874. doi: 10.1155/2017/4015874
32. Michiels C, Raes M, Toussaint O, Remacle J. Importance of Se-glutathione peroxidase, catalase, and Cu/ZnSOD for cell survival against oxidative stress. Free Radical Biology & Medicine 1994; 17 (3): 235-248. doi: 10.1016/0891- 5849(94)90079-5
33. El-Shahat KH, Kandil M. Antioxidant capacity of follicular fluid in relation to follicular size and stage of estrous cycle in buffaloes. Theriogenology 2012; 77 (8): 1513-1518. doi: 10.1016/j.theriogenology.2011.11.018
34. Nuñez-Calonge R, Rancan L, Cortés S, Elena V, Carolina A et al. Apoptotic markers and antioxidant enzymes have altered expression in cumulus and granulosa cells of young women with poor response to ovarian stimulation. Journal of Reproductive Biology and Endocrinology 2019; 3 (1): 1-6.
35. Behl R, Pandey RS. FSH induced stimulation of catalase activity in goat granulosa cells in vitro. Animal Reproduction Science 2002; 70 (3-4): 215-221. doi: 10.1016/s0378-4320(02)00006-4
36. Meijide S, Hernández ML, Navarro R, Larreategui Z, Ferrando M et al. Glutathione S-transferase activity in follicular fluid from women undergoing ovarian stimulation: role in maturation. Free Radical Biology & Medicine 2014; 75 (Suppl 1): S41. doi: 10.1016/j.freeradbiomed.2014.10.792
37. Agarwal A, Said TM, Bedaiwy MA, Banerjee J, Alvarez JG. Oxidative stress in an assisted reproductive techniques setting. Fertility and Sterility 2006; 86 (3): 503-512. doi: 10.1016/j. fertnstert.2006.02.088
38. Oral O, Kutlu T, Aksoy E, Fıçıcıoğlu C, Uslu H et al. The effects of oxidative stress on outcomes of assisted reproductive techniques. Journal of Assisted Reproduction and Genetics 2006; 23 (2): 81-85. doi: 10.1007/s10815-005-9010-4
39. Kumar S, Mishra V, Thaker R, Gor M, Perumal S et al. Role of environmental factors & oxidative stress with respect to in vitro fertilization outcome. Indian Journal of Medical Research 2018; 148 (Suppl 1): 125-133. doi: 10.4103/ijmr.IJMR_1864_17
40. Karakaya C, Kavutcu M, Gumuslu S, Oktem M, Erdem A et al. Evaluation of follicular fluid antioxidant-oxidant status in high, normo, and poor responder patients undergoing assisted reproductive techniques. Fertility and Sterility 2011; 96 (3): S80. doi: 10.1016/j.fertnstert.2011.07.309
41. Angelucci S, Ciavardelli D, Di Giuseppe F, Eleuterio E, Sulpizio M et al. Proteome analysis of human follicular fluid. Biochimica et Biophysica Acta 2006; 1764 (11): 1775-1785. doi: 10.1016/j. bbapap.2006.09.001
42. Meijide S, Pérez-Ruiz I, Hernández ML, Navarro R, Ferrando M et al. Paraoxonase activities in human follicular fluid: role in follicular maturation. Reproductive BioMedicine Online 2017; 35 (4): 351-362. doi: 10.1016/j.rbmo.2017.06.008