Faculty of Natural Sciences, Constantine the Philosopher University in Nitra, Nitra, Slovakia.
Department of Zoology and Anthropology, Constantine the Philosopher University, Tr. A. Hlinku 1, 949 74, Nitra, Slovakia.
Reprod Sci. 2024 Aug;31(8):2293-2308. doi: 10.1007/s43032-024-01554-3. Epub 2024 Apr 17.
The objective of this study was to elucidate the intricate interplay among miR-105-1, kisspeptin, and their synergistic influence on basic ovarian granulosa cell functions. The effects of miR-105-1 mimics or miR-105-1 inhibitor, kisspeptin (0, 1, and 10 ng/ml), and its combinations with miR-105-1 mimics on porcine granulosa cells were assessed. The expression levels of miR-105-1, viability, proliferation (accumulation of PCNA, cyclin B1, XTT-, and BrdU-positive cells), apoptosis (accumulation of bcl-2, bax, caspase 3, p53, TUNEL-positive cells), proportion of kisspeptin-positive cells, and the release of steroid hormones and IGF-I were analyzed. Transfection of cells with miR-105-1 mimics promoted cell viability and proliferation, the occurrence of kisspeptin, and the release of progesterone and IGF-I; in contrast, miR-105-1 mimics inhibited apoptosis and estradiol output. MiR-105-1 inhibitor had the opposite effect. Kisspeptin amplified the expression of miR-105-1, cell viability, proliferation, steroid hormones, and IGF-I release and reduced apoptosis. Furthermore, the collaborative action of miR-105-1 mimics and kisspeptin revealed a synergistic relationship wherein miR-105-1 mimics predominantly supported the actions of kisspeptin, while kisspeptin exhibited a dual role in modulating the effects of miR-105-1 mimics. These findings not only affirm the pivotal role of kisspeptin in regulating basic ovarian cell functions but also represent the inaugural evidence underscoring the significance of miR-105-1 in this regulatory framework. Additionally, our results show the ability of kisspeptin to promote miR-105-1 expression and the ability of miR-105-1 to promote the occurrence and effects of kisspeptin and, therefore, indicate the existence of the self-stimulating kisspeptin-miR-105-1 axis.
本研究旨在阐明 miR-105-1、kisspeptin 之间的复杂相互作用及其对基本卵巢颗粒细胞功能的协同影响。评估了 miR-105-1 模拟物或 miR-105-1 抑制剂、kisspeptin(0、1 和 10ng/ml)及其与 miR-105-1 模拟物的组合对猪颗粒细胞的影响。分析了 miR-105-1 的表达水平、活力、增殖(PCNA、cyclin B1、XTT-和 BrdU 阳性细胞的积累)、凋亡(bcl-2、bax、caspase 3、p53、TUNEL 阳性细胞的积累)、kisspeptin 阳性细胞的比例以及类固醇激素和 IGF-I 的释放。用 miR-105-1 模拟物转染细胞促进了细胞活力和增殖、kisspeptin 的发生以及孕酮和 IGF-I 的释放;相反,miR-105-1 模拟物抑制了细胞凋亡和雌二醇的产生。miR-105-1 抑制剂则产生相反的效果。Kisspeptin 增强了 miR-105-1 的表达、细胞活力、增殖、类固醇激素和 IGF-I 的释放,并减少了凋亡。此外,miR-105-1 模拟物和 kisspeptin 的协同作用表明它们之间存在协同关系,其中 miR-105-1 模拟物主要支持 kisspeptin 的作用,而 kisspeptin 在调节 miR-105-1 模拟物的作用方面具有双重作用。这些发现不仅证实了 kisspeptin 在调节基本卵巢细胞功能中的关键作用,而且还首次证明了 miR-105-1 在这个调控框架中的重要性。此外,我们的结果显示了 kisspeptin 促进 miR-105-1 表达的能力以及 miR-105-1 促进 kisspeptin 的发生和作用的能力,因此表明自我刺激的 kisspeptin-miR-105-1 轴的存在。