Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Anhui Medical University, No 678 Furong Road, Hefei, 230601, Anhui, China.
Department of Obstetrics and Gynecology, Bengbu Hospital of Shanghai General Hospital, Bengbu, 233040, Anhui, China.
BMC Med. 2024 Sep 2;22(1):351. doi: 10.1186/s12916-024-03571-0.
Endometriosis is well known as a chronic inflammatory disease. The development of endometriosis is heavily influenced by the estrogen receptor β (ERβ), while NOD-like receptors (NLRs) family CARD domain-containing 5 (NLRC5) exhibits anti-inflammatory properties during endometriosis. However, whether NLRC5-mediated anti-inflammation is involved in the ERβ-mediated endometriosis is still uncertain. This study aimed to assess that relation.
Nine cases of eutopic endometrial tissue and ten cases of ectopic endometrial tissue were collected from patients with endometriosis, and endometrial samples from ten healthy fertile women were analyzed, and the expression levels of ERβ were quantified using immunohistochemistry (IHC). Subsequently, we constructed mouse model of endometriosis by intraperitoneal injection. We detected the expression of ERβ, NLRC5, tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6, and IL-10 and measured the volume of ectopic lesions in mice with endometriosis. In vitro, human endometrial stromal cells (hESCs) were transfected respectively with ERβ-overexpressing and NLRC5-overexpressing plasmids. We then assessed the expression of ERβ and NLRC5 using quantitative real-time PCR (qRT-PCR) and western blot analysis. Furthermore, we measured the concentrations of TNF-α, IL-6, and IL-10 in the cell culture supernatant through enzyme-linked immunosorbent assay (ELISA). Additionally, we evaluated the migration and invasion ability of hESCs using transwell and wound healing assays.
Inhibition of NLRC5 expression promotes the development of ectopic lesions in mice with endometriosis, upregulates the expression of pro-inflammatory factors TNF-α and IL-6, and downregulates the expression of anti-inflammatory factor IL-10. The high expression of NLRC5 in endometriosis depended on the ERβ overexpression. And ERβ promoted the migration of hESCs partially depend on inflammatory microenvironment. Lastly, NLRC5 overexpression inhibited ERβ-mediated development and inflammatory response of endometriosis.
Our results suggest that the innate immune molecule NLRC5-mediated anti-inflammation participates in ERβ-mediated endometriosis development, and partly clarifies the pathological mechanism of endometriosis, expanding our knowledge of the specific molecules related to the inflammatory response involved in endometriosis and potentially providing a new therapeutic target for endometriosis.
子宫内膜异位症是一种众所周知的慢性炎症性疾病。子宫内膜异位症的发展受雌激素受体β（ERβ）的影响很大，而核苷酸结合寡聚化结构域样受体（NLRs）家族 CARD 结构域包含 5（NLRC5）在子宫内膜异位症中表现出抗炎特性。然而，NLRC5 介导的抗炎作用是否参与 ERβ 介导的子宫内膜异位症仍然不确定。本研究旨在评估这种关系。
收集 9 例在位子宫内膜组织和 10 例异位子宫内膜组织，10 例健康生育期妇女的子宫内膜组织进行分析，采用免疫组织化学（IHC）方法定量检测 ERβ 的表达。随后，通过腹腔注射构建子宫内膜异位症小鼠模型。检测 ERβ、NLRC5、肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-6 和 IL-10 的表达，并测量子宫内膜异位症小鼠异位病变的体积。在体外，人子宫内膜基质细胞（hESCs）分别转染 ERβ 过表达和 NLRC5 过表达质粒。通过实时定量 PCR（qRT-PCR）和 Western blot 分析检测 ERβ 和 NLRC5 的表达。此外，通过酶联免疫吸附试验（ELISA）检测细胞培养上清液中 TNF-α、IL-6 和 IL-10 的浓度。另外，通过 Transwell 和划痕愈合实验评估 hESCs 的迁移和侵袭能力。
抑制 NLRC5 的表达促进子宫内膜异位症小鼠异位病变的发展，上调促炎因子 TNF-α和 IL-6 的表达，下调抗炎因子 IL-10 的表达。子宫内膜异位症中 NLRC5 的高表达依赖于 ERβ 的过表达。并且 ERβ 部分依赖于炎症微环境促进 hESCs 的迁移。最后，NLRC5 过表达抑制 ERβ 介导的子宫内膜异位症的发生和炎症反应。
我们的研究结果表明，先天免疫分子 NLRC5 介导的抗炎作用参与 ERβ 介导的子宫内膜异位症的发生发展，部分阐明了子宫内膜异位症的病理机制，扩大了我们对涉及子宫内膜异位症的炎症反应的特定分子的认识，并可能为子宫内膜异位症提供新的治疗靶点。
