| Abstrakt: |
Simple Summary: The oxidative stress during in vitro culture of ovarian tissues can reduce the production of collagen fibers in the extracellular matrix and impair follicular and stromal cell survival. Consequently, there is a necessity to supplement culture media with antioxidant agents to minimize the damage caused by oxidative stress. α-pinene is a natural monoterpene with a wide range of biological effects, including antioxidant activity. However, it is unclear whether it improves the efficiency of in vitro culture of bovine ovarian tissue. Therefore, we studied the effect of α-pinene on follicular activation, growth, and morphology, on stromal cell density and collagen fibers, as well as on the expression of genes involved in antioxidant defense in bovine ovarian tissues cultured in vitro. The results indicate that the presence of α-pinene in the culture medium improved follicular morphology but did not affect follicular growth. It preserves the density of stromal cells and collagen fibers and increases transcript levels of enzymes involved with antioxidant defense. In conclusion, α-pinene is a promising antioxidant substance, enhancing follicular and stromal cell quality and relieving oxidative stress. Oxidative stress during in vitro of ovarian tissues has adverse effects on follicle survival. α-pinene is a monoterpenoid molecule with antioxidant activity that has great potential to maintain cell survival in vitro. This study investigated the effect of α-pinene (1.25, 2.5, 5.0, 10.0, or 20.0 μg/mL) on primordial follicle growth and morphology, as well as on stromal cells and collagen fibers in bovine ovarian slices cultured for six days. The effect of α-pinene on transcripts of catalase (CAT), superoxide dismutase (SOD), peroxiredoxin 6 (PRDX6), glutathione peroxidase (GPX1), and nuclear factor erythroid 2-related factor 2 (NRF2) was investigated by real-time PCR. The tissues were processed for histological analysis to evaluate follicular growth, morphology, stromal cell density, and collagen fibers. The results showed that 2.5, 5.0, or 10.0 µg/mL α-pinene increased the percentages of normal follicles but did not influence follicular growth. The α-pinene (10.0 µg/mL) kept the stromal cell density and collagen levels in cultured bovine ovarian tissue like uncultured tissues. Ovarian tissues cultured in control medium had reduced expression of mRNA for NRF2, SOD, CAT, GPX1, and PRDX6, but α-pinene (10.0 µg/mL) increased mRNA levels for NRF2 and PRDX6. In conclusion, 10.0 µg/mL α-pinene improves the follicular survival, preserves stromal cell density and collagen levels, and increases transcripts of NRF2 and PRDX6 after in vitro culture of bovine ovarian tissue. [ABSTRACT FROM AUTHOR] |
