Department of Reproductive Medicine, West China Second University Hospital of Sichuan University, Chengdu, 610041, Sichuan, China.
Key Laboratory of Birth Defects and Related Diseases of Women and Children of Ministry of Education, Chengdu, 610041, Sichuan, China.
Reprod Sci. 2023 Dec;30(12):3578-3589. doi: 10.1007/s43032-023-01315-8. Epub 2023 Aug 2.
Endometriosis is a hormone-dependent disease associated with impaired immunoregulation. In our recent study, we have characterized the trascriptomic transformation of eutopic endometrium from patients with minimal/mild endometriosis and controls across the menstrual cycle. However, the regulatory mechanism of altered immune microenvironment in eutopic endometrial stromal cells (ESCs) remains unclear. Here, we want to explore the regulation of immune cell to progesterone resistance and endometrial receptivity in the eutopic ESCs by cytokine (TGF-β1), and to understand the effect of TGF-β1 on the decidualization of the eutopic ESCs. Primary culture of eutopic ESCs was performed to explore the effects of TGF-β1 on the expression of Smad and progesterone receptor (PR) and the in vitro decidualization. Additionally, co-immunoprecipitation (Co-IP) was used to explore the direct interaction between Smad and PR. We found an attenuate expression of PRB protein (p=0.026) after using TGF-β1 in eutopic ESCs, although the difference of PRA before and after treatment was not significant (p=0.678). Similarly, the results of qRT-PCR showed that the mRNA level of PR (p<0.001), PRB (p=0.003) and HOXA10 (p<0.001) decreased significantly after TGF-β1 treatment, but that increased (p<0.023, for all) after SB431542 treatment in the eutopic ESCs. Moreover, TGF-β1 has a negative effect on the in vitro decidualization of eutopic ESCs (p=0.003). And the group with treatment of both TGF-β1 and SB435142 in eutopic ESCs showed significant decidual-like changes with increased prolactin level (p=0.01). We did not observe any physical interaction between the PR and p-Smad3/Smad3 proteins by using Co-IP. By activating TGF-β/Smad signaling in eutopic ESCs, elevated TGF-β1 from CD45+ immune cells could attenuate expression of PR, and further decrease endometrial receptivity.
子宫内膜异位症是一种与免疫调节受损相关的激素依赖性疾病。在我们最近的研究中，我们描述了患有轻度子宫内膜异位症的患者和对照组在整个月经周期中，其在位子宫内膜的转录组转化。然而，在位子宫内膜基质细胞（ESC）中免疫微环境改变的调控机制尚不清楚。在这里，我们希望通过细胞因子（TGF-β1）来探索免疫细胞对孕激素抵抗和在位 ESC 子宫内膜容受性的调节作用，并了解 TGF-β1 对在位 ESC 蜕膜化的影响。我们进行了在位 ESC 的原代培养，以探讨 TGF-β1 对 Smad 和孕激素受体（PR）表达的影响及其对在位 ESC 蜕膜化的影响。此外，还使用了免疫共沉淀（Co-IP）来探讨 Smad 和 PR 之间的直接相互作用。我们发现，在使用 TGF-β1 处理后，在位 ESC 中 PRB 蛋白的表达减弱（p=0.026），尽管治疗前后 PRA 的差异不显著（p=0.678）。同样，qRT-PCR 的结果显示，PR（p<0.001）、PRB（p=0.003）和 HOXA10（p<0.001）的 mRNA 水平在 TGF-β1 处理后显著降低，但在 SB431542 处理后增加（p<0.023，均为）。此外，TGF-β1 对在位 ESC 的体外蜕膜化有负面影响（p=0.003）。而且，在在位 ESC 中同时使用 TGF-β1 和 SB435142 的组表现出明显的蜕膜样变化，催乳素水平升高（p=0.01）。我们通过 Co-IP 没有观察到 PR 和 p-Smad3/Smad3 蛋白之间的任何物理相互作用。通过激活在位 ESC 中的 TGF-β/Smad 信号通路，CD45+免疫细胞产生的 TGF-β1 可降低 PR 的表达，进一步降低子宫内膜的容受性。
