College of Animal Science and Technology, Hebei Agricultural University, Baoding 071000, China.
College of Animal Science and Technology, China Agricultural University, Beijing 100089, China.
Int J Mol Sci. 2023 Sep 22;24(19):14407. doi: 10.3390/ijms241914407.
High prolactin (PRL) concentration has been shown to induce the apoptosis of ovine ovarian granulosa cells (GCs), but the underlying mechanisms are unclear. This study aimed to investigate the mechanism of apoptosis induced by high PRL concentration in GCs. Trial 1: The optimal concentration of glutathion was determined according to the detected cell proliferation. The results showed that the optimal glutathione concentration was 5 μmol/mL. Trial 2: 500 ng/mL PRL was chosen as the high PRL concentration. The GCs were treated with 0 ng/mL PRL (C group), 500 ng/mL PRL (P group) or 500 ng/mL PRL, and 5 μmol/mL glutathione (P-GSH group). The results indicated that the mitochondrial respiratory chain complex (MRCC) I-V, ATP production, total antioxidant capacity (T-AOC), superoxide dismutase (SOD), and thioredoxin peroxidase (TPx) in the C group were higher than those in the P group ( < 0.05), while they were lower than those in the P-GSH group ( < 0.05). Compared to the C group, the P group exhibited elevated levels of reactive oxygen species (ROS) and apoptosis ( < 0.05) and increased expression of ATG7 and ATG5 ( < 0.05). However, MRCC I-V, ATP, SOD, A-TOC, TPx, ROS, and apoptosis were decreased after the addition of glutathione ( < 0.05). The knockdown of either or in P group GCs resulted in a significant reduction ( < 0.05) in MRCC I-V, ATP, T-AOC, SOD and TPx, while the overexpression of either receptor showed an opposite trend ( < 0.05). Our findings suggest that high PRL concentrations induce apoptotic cell death in ovine ovarian GCs by downregulating and , activating oxidative stress and autophagic pathways.
高催乳素（PRL）浓度已被证明可诱导绵羊卵巢颗粒细胞（GC）凋亡，但具体机制尚不清楚。本研究旨在探讨高 PRL 浓度诱导 GC 凋亡的机制。试验 1：根据细胞增殖检测结果，确定谷胱甘肽的最佳浓度。结果表明，谷胱甘肽的最佳浓度为 5μmol/ml。试验 2：选择 500ng/ml PRL 作为高 PRL 浓度。将 GCs 分别用 0ng/ml PRL（C 组）、500ng/ml PRL（P 组）或 500ng/ml PRL+5μmol/ml 谷胱甘肽（P-GSH 组）处理。结果表明，C 组的线粒体呼吸链复合物（MRCC）I-V、ATP 生成、总抗氧化能力（T-AOC）、超氧化物歧化酶（SOD）和硫氧还蛋白过氧化物酶（TPx）均高于 P 组（<0.05），但低于 P-GSH 组（<0.05）。与 C 组相比，P 组的活性氧（ROS）和细胞凋亡增加（<0.05），ATG7 和 ATG5 表达增加（<0.05）。然而，加入谷胱甘肽后，MRCC I-V、ATP、SOD、T-AOC、TPx、ROS 和细胞凋亡均降低（<0.05）。在 P 组 GC 中敲低 或 后，MRCC I-V、ATP、T-AOC、SOD 和 TPx 均显著降低（<0.05），而过表达两种受体则呈现相反趋势（<0.05）。综上所述，高 PRL 浓度通过下调 和 、激活氧化应激和自噬途径，诱导绵羊卵巢 GC 凋亡。
