ICAR-National Institute of Animal Nutrition and Physiology, Bangalore, India.
ICAR-National Institute of Animal Nutrition and Physiology, Bangalore, India; Veterinary College, Bangalore Campus, Hebbal, Bangalore, India.
Theriogenology. 2024 Jan 15;214:266-272. doi: 10.1016/j.theriogenology.2023.10.023. Epub 2023 Oct 17.
Study was carried out to examine the influence of plant bioactive compounds [Kaempferol (KAE), Querectin (QUE) and Myricetin (MYR)] on the survival and growth parameters of cultured ovine preantral follicles (PFs) granulosa cells (GCs) and expression of some key developmental genes. Ovine PFs were isolated from slaughterhouse derived ovaries and KAE, QUE and MYR were supplemented to the standard culture medium of GCs and PFs at concentrations of 0, 5, 10, 25, 50 and 100 μM and cultured for 5 and 7 days respectively. PFs morphological and functional parameters [follicle and enclosed oocyte growth rate, viability of follicles, antrum formation rate, oocyte maturation rate, estradiol concentration, reactive oxygen species (ROS) production] and GC growth parameters (metabolic activity, viability rate, cell number increment, ROS production) were measured after culture. Significantly higher PF growth, viability rate and estradiol concentration was observed at 10 μM, 25 μM and 10 μM concentration of KAE, MYR and QUE respectively compared to the control. ROS production was significantly decreased in the PF culture media treated with 10 μM KAE or MYR 25 μM or 10 μM QUE compared to those observed in the control group. Likewise, metabolic activity of GCs, viability rate and cell number increment cultured with KAE, MYR and QUE was significantly higher at 10, 25 and 10 μM concentrations respectively compared to those observed in control group. ROS production was significantly lower in the GC cultured with KAE, MYR and QUE at 10, 25 and 10 μM concentrations respectively compared to the control. Based on the results of the growth parameters, gene expression of PFs and GCs were studied by qPCR at selected concentrations (KAE, MYR and QUE at 10, 25 and 10 μM concentrations respectively) in the cultured PFs and GCs. Gene expression of GDF9, FGF2, CYP19A1 was significantly higher and Bax, Bcl2 expression was significantly lower in the PFs and GCs cultured with the KAE or QUE at 10 μM concentration. KAE, MYR and QUE have dose dependant responses on PFs and GCs morphological and functional parameters; however, KAE is more potent amongst the three in augmenting the ovarian functions.
研究旨在探讨植物生物活性化合物[山奈酚（KAE）、槲皮素（QUE）和杨梅素（MYR）]对培养绵羊原始卵泡（PFs）颗粒细胞（GCs）的存活和生长参数以及一些关键发育基因表达的影响。从屠宰场来源的卵巢中分离绵羊 PFs，并将 KAE、QUE 和 MYR 分别添加到 GCs 和 PFs 的标准培养基中，浓度为 0、5、10、25、50 和 100μM，并分别培养 5 和 7 天。在培养后测量 PFs 形态和功能参数[卵泡和包被卵母细胞生长率、卵泡活力、腔形成率、卵母细胞成熟率、雌二醇浓度、活性氧（ROS）产生]和 GC 生长参数（代谢活性、活力率、细胞数量增加、ROS 产生）。与对照组相比，在 10μM、25μM 和 10μM 浓度下，KAE、MYR 和 QUE 分别观察到 PF 生长、活力率和雌二醇浓度显著升高。与对照组相比，用 10μM KAE 或 MYR 25μM 或 10μM QUE 处理的 PF 培养物中的 ROS 产生显著降低。同样，在 KAE、MYR 和 QUE 培养的 GCs 中，代谢活性、活力率和细胞数量增加在 10、25 和 10μM 浓度下均显著高于对照组。与对照组相比，在 10、25 和 10μM 浓度下，KAE、MYR 和 QUE 培养的 GC 中的 ROS 产生显著降低。基于生长参数的结果，在选定浓度（KAE、MYR 和 QUE 分别为 10、25 和 10μM）下通过 qPCR 研究了 PFs 和 GCs 的基因表达在培养的 PFs 和 GCs 中。在 10μM 浓度下，用 KAE 或 QUE 培养的 PFs 和 GCs 中的 GDF9、FGF2、CYP19A1 基因表达显著升高，Bax、Bcl2 表达显著降低。KAE、MYR 和 QUE 对 PFs 和 GCs 的形态和功能参数具有剂量依赖性反应；然而，在增强卵巢功能方面，KAE 比其他三种更为有效。
