Constantine the Philosopher University, Nitra, Slovakia.
Constantine the Philosopher University, Nitra, Slovakia.
Reprod Biol. 2022 Mar;22(1):100580. doi: 10.1016/j.repbio.2021.100580. Epub 2021 Nov 26.
The present study aims to examine the role of kisspeptin (KP), FSH, and its receptor (FSHR), and their interrelationships in the control of basic human ovarian granulosa cells functions. We investigated: (1) the ability of granulosa cells to produce KP and FSHR, (2) the role of KP in the control of ovarian functions, and (3) the ability of KP to affect FSHR and to modify the FSH action on ovarian functions. The effects of KP alone (0, 10 and 100 ng/mL); or of KP (10 and 100 ng/mL) in combination with FSH (10 ng/mL) on cultured human granulosa cells were assessed. Viability, markers of proliferation (PCNA and cyclin B1) and apoptosis (bax and caspase 3), as well as accumulation of KP, FSHR, and steroid hormones, IGF-I, oxytocin (OT), and prostaglandin E2 (PGE2) release were analyzed by the Trypan blue exclusion test, quantitative immunocytochemistry, and ELISA. KP given at a low dose (10 ng/mL) stimulated viability, proliferation, inhibited apoptosis, promoted the release of progesterone (P4), estradiol (E2), IGF-I, OT, and PGE2, the accumulation of FSHR, but not testosterone (T) release. KP given at a high dose (100 ng/mL) had the opposite, inhibitory effect. FSH stimulated cell viability, proliferation and inhibited apoptosis, promoted P4, T, E2, IGF-I, and OT, but not PGE2 release. Furthermore, KP at a low dose promoted the stimulatory effect of FSH on viability, proliferation, P4, E2, and OT release, promoted its inhibitory action on apoptosis, but did not modify its action on T, IGF-I, and PGE2 output. KP at a high dose prevented and inverted FSH action. These results suggest an intra-ovarian production and a functional interrelationship between KP and FSH/FSHR in direct regulation of basic ovarian cell functions (viability, proliferation, apoptosis, and hormones release). The capability of KP to stimulate FSHR, the ability of FSH to promote ovarian functions, as well as the similarity of KP (10 ng/mL) and FSH action on granulosa cells' viability, proliferation, apoptosis, steroid hormones, IGF-I, OT, and PGE2 release, suggest that FSH influence these cells could be mediated by KP. Moreover, the capability of KP (100 ng/mL) to decrease FSHR accumulation, basal and FSH-induced ovarian parameters, suggest that KP can suppress some ovarian granulosa cell functions via down-regulation of FSHR. These observations propose the existence of the FSH-KP axis up-regulating human ovarian cell functions.
本研究旨在探讨 kisspeptin (KP)、FSH 及其受体 (FSHR) 在调控人类卵巢颗粒细胞基本功能中的作用及其相互关系。我们研究了：(1) 颗粒细胞产生 KP 和 FSHR 的能力，(2) KP 在卵巢功能调控中的作用，以及 (3) KP 影响 FSHR 的能力及其对 FSH 作用于卵巢功能的修饰作用。检测了 KP 单独（0、10 和 100ng/mL）或与 FSH（10ng/mL）联合作用于培养的人卵巢颗粒细胞的效果。通过台盼蓝排除试验、定量免疫细胞化学和 ELISA 分析细胞活力、增殖标志物（PCNA 和细胞周期蛋白 B1）和凋亡标志物（bax 和 caspase 3）以及 KP、FSHR 和甾体激素、IGF-I、催产素 (OT) 和前列腺素 E2 (PGE2) 的积累。KP 低剂量（10ng/mL）刺激细胞活力、增殖、抑制凋亡、促进孕激素 (P4)、雌二醇 (E2)、IGF-I、OT 和 PGE2 释放，促进 FSHR 积累，但不促进睾酮 (T) 释放。高剂量（100ng/mL）的 KP 则产生相反的抑制作用。FSH 刺激细胞活力、增殖并抑制凋亡，促进 P4、T、E2、IGF-I 和 OT 释放，但不促进 PGE2 释放。此外，低剂量的 KP 促进了 FSH 对细胞活力、增殖、P4、E2 和 OT 释放的刺激作用，促进了其对凋亡的抑制作用，但不改变其对 T、IGF-I 和 PGE2 输出的作用。高剂量的 KP 阻止并反转了 FSH 的作用。这些结果表明，在直接调节基本卵巢细胞功能（活力、增殖、凋亡和激素释放）方面，KP 和 FSH/FSHR 之间存在着卵巢内的产生和功能的相互关系。KP 刺激 FSHR 的能力、FSH 促进卵巢功能的能力，以及 KP（10ng/mL）和 FSH 对颗粒细胞活力、增殖、凋亡、甾体激素、IGF-I、OT 和 PGE2 释放的相似作用，表明 FSH 对这些细胞的影响可能是由 KP 介导的。此外，KP（100ng/mL）降低 FSHR 积累、基础和 FSH 诱导的卵巢参数的能力表明，KP 可以通过下调 FSHR 来抑制一些卵巢颗粒细胞的功能。这些观察结果提出了存在 FSH-KP 轴来上调人卵巢细胞功能的假设。
