Zeng Yuhua, Feng Qing
Health Management Medicine Center, The Third Xiangya Hospital of Central South University, Changsha, China.
Department of Obstetrics and Gynecology, The Third Xiangya Hospital of Central South University, Changsha, China.
Kaohsiung J Med Sci. 2025 Jan;41(1):e12915. doi: 10.1002/kjm2.12915. Epub 2024 Dec 4.
Intrauterine adhesion (IUA) is the second most common cause of secondary infertility in women and can also lead to menstrual abnormalities and multiple adverse pregnancy outcomes. Therefore, elucidating the mechanism of its development is crucial for the prevention and treatment of IUA. This study will investigate the function and mechanism of forkhead box P1 (FOXP1)/DNA methyltransferase 1 (DNMT1)/unc-51-like autophagy activating kinase 1 (ULK1) in IUA. Expression levels of key genes were detected using western blot and quantitative - real time reverse transcription polymerase chain reaction. Cell proliferation was detected by CCK-8 and EdU staining. Transcriptional regulation relationships were detected by dual luciferase reporter gene and chromatin immunoprecipitation (ChIP) assay. Methylation station of ULK1 was detected by methylmion specific PCR (MSP). Fibrosis and pathological changes in the uterine cavity tissues were detected by Masson and hematoxylin and eosin staining. It was observed that the expression of FOXP1 and DNMT1 increased in transforming growth factor (TGF)-β1-induced cells, while ULK1 expression decreased. Downregulation of FOXP1 could inhibit human endometrial stromal cells proliferation and autophagy, as well as decrease the expression of fibrogenic factors (collagen type I alpha 1 chain [COL1A1], fibronectin [FN], and alpha-smooth muscle actin [α-SMA]). The results of MSP and ChIP experiments showed that DNMT1 promotes methylation of the ULK1 promoter region and inhibits its transcription. In an animal model, knockdown of FOXP1 alleviated pathological fibrosis and uterine adhesions. Knockdown of FOXP1 can inhibit endometrial fibrosis in IUA rats; FOXP1 could be a potential target for the treatment of IUA.
宫腔粘连（IUA）是女性继发性不孕的第二大常见原因，还可导致月经异常和多种不良妊娠结局。因此，阐明其发病机制对于IUA的防治至关重要。本研究将探讨叉头框蛋白P1（FOXP1）/DNA甲基转移酶1（DNMT1）/unc-51样自噬激活激酶1（ULK1）在IUA中的作用及机制。采用蛋白质免疫印迹法和定量实时逆转录聚合酶链反应检测关键基因的表达水平。通过CCK-8法和EdU染色检测细胞增殖。采用双荧光素酶报告基因检测和染色质免疫沉淀（ChIP）分析检测转录调控关系。通过甲基化特异性PCR（MSP）检测ULK1的甲基化状态。采用Masson染色及苏木精-伊红染色检测宫腔组织的纤维化及病理变化。结果发现，在转化生长因子（TGF）-β1诱导的细胞中，FOXP1和DNMT1的表达增加，而ULK1表达降低。下调FOXP1可抑制人子宫内膜基质细胞增殖和自噬，并降低纤维化因子（Ⅰ型胶原α1链[COL1A1]、纤连蛋白[FN]和α-平滑肌肌动蛋白[α-SMA]）的表达。MSP和ChIP实验结果表明，DNMT1促进ULK1启动子区域的甲基化并抑制其转录。在动物模型中，敲低FOXP1可减轻病理纤维化和宫腔粘连。敲低FOXP1可抑制IUA大鼠子宫内膜纤维化；FOXP1可能是治疗IUA的潜在靶点。
