Anethole reduces oxidative stress and improves in vitro survival and activation of primordial follicles

Primordial follicles, the main source of oocytes in the ovary, are essential for the maintenance of fertility throughout the reproductive lifespan. To the best of our knowledge, there are no reports describing the effect of anethole on this important ovarian follicle population. The aim of the study was to investigate the effect of different anethole concentrations on the in vitro culture of caprine preantral follicles enclosed in ovarian tissue. Randomized ovarian fragments were fixed immediately (non-cultured treatment) or distributed into five treatments: α-MEM+ (cultured control), α-MEM+ supplemented with ascorbic acid at 50 μg/mL (AA), and anethole at 30 (AN30), 300 (AN300), or 2000 µg/mL (AN2000), for 1 or 7 days. After 7 days of culture, a significantly higher percentage of morphologically normal follicles was observed when anethole at 2000 μg/mL was used. For both culture times, a greater percentage of growing follicles was observed with the AN30 treatment compared to AA and AN2000 treatments. Anethole at 30 and 2000 µg/mL concentrations at days 1 and 7 of culture resulted in significantly larger follicular diameter than in the cultured control treatment. Anethole at 30 µg/mL concentration at day 7 showed significantly greater oocyte diameter than the other treatments, except when compared to the AN2000 treatment. At day 7 of culture, levels of reactive oxygen species (ROS) were significantly lower in the AN30 treatment than the other treatments. In conclusion, supplementation of culture medium with anethole improves survival and early follicle development at different concentrations in the caprine species.

原始卵泡（primordial follicle）是卵巢内卵母细胞的主要来源，对维系整个生殖生命周期的生育能力至关重要。据我们所知，目前尚无关于茴香脑（anethole）对这一重要卵巢卵泡群体影响的相关研究报道。本研究旨在探讨不同浓度茴香脑对包裹于卵巢组织内的山羊窦前卵泡（preantral follicle）体外培养的影响。将随机选取的卵巢组织碎片一部分立即固定（设为非培养对照组），其余则随机分配至5种培养体系中，分别培养1天或7天：仅添加α-MEM+的培养体系（培养对照组）、添加50 μg/mL抗坏血酸的α-MEM+培养体系（AA组）、以及分别添加30 μg/mL（AN30组）、300 μg/mL（AN300组）、2000 μg/mL（AN2000组）茴香脑的α-MEM+培养体系。培养7天后，采用2000 μg/mL茴香脑处理的培养体系中，形态正常卵泡的占比显著升高。在两种培养时长下，AN30组的生长卵泡占比均显著高于AA组与AN2000组。在培养第1天与第7天，30 μg/mL及2000 μg/mL茴香脑处理组的卵泡直径均显著大于培养对照组。培养第7天时，30 μg/mL茴香脑处理组的卵母细胞直径显著高于其余各组，仅AN2000组除外。培养第7天时，AN30组的活性氧（reactive oxygen species, ROS）水平显著低于其余各组。综上，在山羊卵巢组织体外培养体系中添加不同浓度的茴香脑，可改善卵泡存活与早期发育状况。