The follicular hierarchy in broiler breeder chicken ovary is often deranged due to excessive ovarian follicular recruitment, resulting in a condition that resembles polycystic ovary syndrome (PCOS) in women. Metformin is widely prescribed to correct PCOS and has been shown to affect granulosa cell functions in humans and rodent models. The objectives of this study are to determine the effects of metformin on signal transduction pathways, gene expression related to steroidogenesis, and progesterone secretion from granulosa cells isolated from the most recently recruited preovulatory and prehierarchical follicles of broiler breeder chickens. Granulosa cells were treated with 0, 1, 10, or 20 mM of metformin in the presence of FSH. The abundance of pAMPK, pACC, pERK, and pAkt was determined by Western blotting. The expression of genes related to progesterone biosynthesis was quantified by qPCR. Progesterone concentrations in culture media were quantified by ELISA. Metformin treatment did not have an effect on the abundance of pAMPK and pACC in prehierarchical follicles but significantly decreased the abundance of pERK and pAkt in a dose-dependent manner in preovulatory and prehierarchical follicles. The expression of genes related to steroidogenesis such as FSHR, STAR, CYP11A1, HSD3B, and progesterone secretion was significantly decreased in response to metformin treatment in a dose-dependent manner. Our data suggest that metformin treatment attenuates progesterone secretion via AMPK-independent pathways in granulosa cells of prehierarchical and preovulatory follicles of broiler breeder hens. Further studies are required to determine if metformin administration could ameliorate ovarian dysfunction in obese broiler breeder hens.

中文翻译：

---

二甲双胍减弱肉种鸡卵巢卵泡中的类固醇生成

由于卵巢卵泡过度募集，肉种鸡卵巢中的卵泡等级经常紊乱，导致女性出现类似于多囊卵巢综合征 (PCOS) 的状况。二甲双胍被广泛用于纠正 PCOS，并已被证明会影响人类和啮齿动物模型的颗粒细胞功能。本研究的目的是确定二甲双胍对信号转导途径、与类固醇生成相关的基因表达以及从最近招募的肉种鸡排卵前和分级前卵泡中分离的颗粒细胞中孕酮分泌的影响。在 FSH 存在下，用 0、1、10 或 20 mM 二甲双胍处理颗粒细胞。通过蛋白质印迹测定 pAMPK、pACC、pERK 和 pAkt 的丰度。通过qPCR定量与孕酮生物合成相关的基因的表达。通过ELISA量化培养基中的孕酮浓度。二甲双胍治疗对等级前卵泡中 pAMPK 和 pACC 的丰度没有影响，但在排卵前和等级前卵泡中以剂量依赖性方式显着降低了 pERK 和 pAkt 的丰度。与类固醇生成相关的基因（如 FSHR、STAR、CYP11A1、HSD3B 和孕酮分泌）的表达在二甲双胍治疗后以剂量依赖性方式显着降低。我们的数据表明，二甲双胍治疗可通过非 AMPK 非依赖性途径减弱肉种鸡等级前和排卵前卵泡的颗粒细胞中的孕酮分泌。