Toxicology: Open Access
Open Access

Abstract

In the current assisted reproductive practice in cows the IVF technique is used in increasing proportion. However, the used methods are not always standardized and are needed to be improved. The first challenge in optimization of IVF techniques is obtaining mature oocytes by growing them in culture media and by this to preserve their high fertilization quality. A crucial factor in improving IVF results is the prevention of oocytes from in vitro cultivation stress effects. In the presented study the beneficial effect of antioxidant supplementation in maturation culture media of cow oocytes was evaluated by apoptotic genes expression quantification. The oocytes were cultivated for 24 hours on conventional (control variant), supplemented with rosmarinic acid (RA variant) and ascorbic acid (C variant), maturation media. The oocytes were classified in three quality classes by morphological observation from which the total RNA was isolated. Quantitative PCR technique was used for quantification of BAX and BCL2 apoptotic genes expression. Results of qPCR were interpreted by �?�? (�?�?Ct) method. The ratio BCL2/BAX was considered as an indicator of maturated oocytes homeostasis. Antioxidants culture media supplementation resulted in a better expansion of cumulus cells. The level of expression of the BAX gene has an increasing trend in all COC��?s, inversely proportional to oocyte quality, indicating the overcoming of cell adaptation process for the inferior class. Regarding the BCL2 gene, significantly higher expression levels can be observed in class I oocytes supplemented with antioxidants. The level of maintenance of cell homeostasis, as reflected by the ratio of BAX/ BCL-2, with a value above 7, indicates that apoptotic processes have been installed in all class III oocytes. Supplementation with antioxidants exerts a beneficial effect on inferior class cells, which have a high stress level, to some extent assuring their protection, indicating the effectiveness of administering this supplement. References 1. Mbemya G T (2017) Reports on in vivo and in vitro contribution of medicinal plants to improve the female reproductive function. Reprod Clim.; 32(2): 109-119. 2. Roychoudhury S, Agarwal A, Virk G, Cho C-L (2017) Potential role of green tea catechins in the management of oxidative stress-associated infertility. Reproductive BioMedicine Online; 34(5): 487-498. 3. Kang J T, Moon J H, Choi J I, Park S J, Kim S J, Saadeldin I M, Lee B C (2016) Effect of Antioxidant Flavonoids (Quercetin and Taxifolin) on In vitro Maturation of Porcine Oocytes, Asian Australias. J. Anim. Sci.; 29(3): 352-358. 4. Luno V, Gil L, Olaciregui M, Gonzalez N, Jerez R A, Blas I (2014) Rosmarinic acid improves function and in vitro fertilising ability of boar sperm after cryopreservation, Cryobiology; 69(1): 157-162. 5. Sadeesh E, Shah F, Balhara A K, Thirumaran S M K, Yadav S, Yadav P S (2014) Effect of growth factor and antioxidant on in vitro maturation of oocytes and cleavage rates of in vitro produced Indian buffalo (Bubalus bubalis) embryos. Veterinarski Arhiv; 84(5): 459-474.

Biography

Camelia Tulcan is an Assistant Professor at Biochemistry Department, Faculty of Veterinary Medicine, Timisoara and Coordinator of Antioxidant Research LabHoria Cernescu Research Unit. She has expertise in oxidative stress evaluation in different physiological or pathological condition and was involved in management team of research infrastructure project and in implementation of quality management systems.
Email:cameliatulcan@usab-tm.ro, tulcancamelia@gmail.com