目的:多囊卵巢综合征(PCOS)患者下丘脑神经激肽B(NKB)水平升高可能是导致LH脉冲分泌过度活跃的原因之一。然而,神经激肽B-神经激肽3受体(NKB-NK3R)系统在PCOS局部卵巢组织中的表达和作用尚未阐明。我们构建了体内和体外模型来阐明NKB-NK3R通路在PCOS生殖内分泌紊乱中的作用机制。
方法:将颗粒细胞系-KGN细胞置于棕榈酸(PA)和二氢睾酮(DHT)中以模拟PCOS样条件。我们使用高脂/高糖饮食建立PCOS样小鼠模型,并对一半小鼠给予神经激肽3受体拮抗剂(NK3Ra)。NKB-NK3R系统的表达,线粒体功能,激素水平,并评估炎症状态。
结果:PCOS样刺激诱导KGN细胞NKB-NK3R系统和MAPK-ERK通路过表达,以近似的剂量和时间依赖性方式。NKB-NK3R系统过度激活MAPK-ERK通路以增加NNT过表达,干扰NADH/NADPH池,加剧氧化状态,减少ATP的产生。随着NKB-NK3R系统在局部卵巢组织中的过度表达,排卵障碍,孕酮缺乏,和促炎状态在PCOS样小鼠中明显。拮抗受体,NK3R,通过改善线粒体功能障碍逆转不良生殖内分泌表型。
结论:除了中央法规之外,NKB-NK3R系统的局部卵巢过度表达参与不良生殖内分泌表型,支持NK3Ra对PCOS的治疗意义。
The increased hypothalamic neurokinin B (NKB) level may contribute to the hyperactive LH pulse secretion in Polycystic ovary syndrome (PCOS). However, the expression and role of the neurokinin B-neurokinin 3 receptor (NKB-NK3R) system in the local ovarian tissue of PCOS have not been clarified. We constructed in vivo and in vitro models to elucidate the mechanism of the NKB-NK3R pathway in reproductive endocrine disorders of PCOS.
The granulosa cell line-KGN cells were set in palmitic acid (PA) and dihydrotestosterone (DHT) to simulate the PCOS-like conditions. And we used the high-fat/high-glucose diet to build a PCOS-like mice model and neurokinin 3 receptor antagonist (NK3Ra) was administered to half of the mice. The expression of the NKB-NK3R system, mitochondrial functions, hormone levels, and inflammatory state was evaluated.
The PCOS-like stimulations induced the NKB-NK3R system and MAPK-ERK pathway overexpression in KGN cells, in an approximate dose and time-dependent manner. The NKB-NK3R system overactivated the MAPK-ERK pathway to increase NNT overexpression, disturb NADH/NADPH pools, aggravate the oxidation state, and decrease ATP production. With overexpression of the NKB-NK3R system in the local ovarian tissue, ovulatory dysfunction, progesterone deficiency, and pro-inflammatory states were apparent in PCOS-like mice. Antagonizing the receptor, NK3R, reversed the adverse reproductive endocrine phenotypes via improving mitochondrial dysfunction.
In addition to the central regulation, local ovarian overexpression of the NKB-NK3R system participated in the adverse reproductive endocrine phenotypes, supporting the therapeutic implications of NK3Ra for PCOS.