Amaral Ana, Sadowska Agnieszka, Cerveira-Pinto Marta, Kordowitzki Pawel, Skarzynski Dariusz, Ferreira-Dias Graça, Szóstek-Mioduchowska Anna
CIISA-Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Avenida da Universidade Técnica, Lisbon, 1300-477, Portugal.
Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Lisbon, 1300-477, Portugal.
Sci Rep. 2025 Jan 2;15(1):538. doi: 10.1038/s41598-024-84250-1.
Equine endometrosis is a major cause of infertility in mares and is characterized by degenerative, functional and fibrotic changes in the endometrium with increased collagen (COL) deposition. Transforming growth factor (TGF)-β1 is one of the major pro-fibrotic factors involved in the excessive deposition of extracellular matrix (ECM) components in the equine endometrium. It has been demonstrated that ovarian steroids, specifically 17β-estradiol (E2) and progesterone (P4), not only regulate the cyclicity of the estrous cycle, but also have been implicated as anti- or pro-fibrotic factors. This study aimed to evaluate (i) the effect of E2 and P4 on the expression of ECM-associated genes including COL1A1, COL3A1, matrix metalloproteases (MMPs): MMP-1, MMP-2, MMP-3, MMP-9, and MMP-13, and tissue inhibitors of MMPs (TIMPs): TIMP-1 and TIMP-2 in equine endometrial fibroblasts, and (ii) the effect of ovarian steroids on TGF-β1-induced COL1 expression in equine endometrial explants from the follicular and mid-luteal phases of the estrous cycle. The mRNA expression of ECM-associated genes in endometrial fibroblasts and TGF-β1-induced COL1 expression in endometrial explants was modulated by ovarian steroids, with variations depending on the type of steroid and the duration of treatment. Moreover, P4 decreased TGF-β1-induced COL1 protein abundance in the mid-luteal phase of the estrous cycle after 48 h (p < 0.05). The results of our study indicate that during the estrous cycle, the ovarian steroids E2 or P4 may act directly on endometrial fibroblasts, thereby affecting the expression of genes involved in tissue remodeling, namely MMPs and TIMPs. Furthermore, P4 appears to affect not only the ECM-associated genes in endometrial fibroblasts, but also to attenuate the pro-fibrotic action of TGF-β1 in the mid-luteal stage of the estrous cycle.
马子宫内膜异位症是母马不育的主要原因，其特征是子宫内膜出现退行性、功能性和纤维化变化，胶原蛋白（COL）沉积增加。转化生长因子（TGF）-β1是参与马子宫内膜细胞外基质（ECM）成分过度沉积的主要促纤维化因子之一。已经证明，卵巢类固醇，特别是17β-雌二醇（E2）和孕酮（P4），不仅调节发情周期的循环，还被认为是抗纤维化或促纤维化因子。本研究旨在评估（i）E2和P4对马子宫内膜成纤维细胞中包括COL1A1、COL3A1、基质金属蛋白酶（MMPs）：MMP-1、MMP-2、MMP-3、MMP-9和MMP-13，以及MMP组织抑制剂（TIMPs）：TIMP-1和TIMP-2在内的ECM相关基因表达的影响，以及（ii）卵巢类固醇对发情周期卵泡期和黄体中期马子宫内膜外植体中TGF-β1诱导的COL1表达的影响。卵巢类固醇调节子宫内膜成纤维细胞中ECM相关基因的mRNA表达以及子宫内膜外植体中TGF-β1诱导的COL1表达，其变化取决于类固醇类型和处理持续时间。此外，在发情周期的黄体中期，P4在48小时后降低了TGF-β1诱导的COL1蛋白丰度（p<0.05）。我们的研究结果表明，在发情周期中，卵巢类固醇E2或P4可能直接作用于子宫内膜成纤维细胞，从而影响参与组织重塑的基因即MMPs和TIMPs的表达。此外，P4似乎不仅影响子宫内膜成纤维细胞中与ECM相关的基因，还能减弱发情周期黄体中期TGF-β1的促纤维化作用。
