Department of Anesthesiology, Wuxi Maternity and Child Health Care Hospital, Wuxi, China.
Immun Inflamm Dis. 2023 Sep;11(9):e1008. doi: 10.1002/iid3.1008.
Endometrial injury is a common disease in women caused by intrauterine inflammation, infections, and endocrine disorders. Human endometrial stromal cells (hEndoSCs) can maintain endometrial homeostasis and play an important role in repairing endometrial injury. Mifepristone, a steroidal anti-progesterone drug, is widely used in the field of reproductive medicine worldwide. Mifepristone-induced hEndoSC injury has been used to study endometrial injury in vitro. At present, the pathogenesis and potential regulatory mechanisms of oxycodone in endometrial injury remain unknown.
We aimed to evaluate the functions of oxycodone in mifepristone-stimulated hEndoSC injury and analyze its potential molecular mechanism.
MATERIALS & METHODS: hEndoSC viability, cytotoxicity, and apoptosis were analyzed using the methyl thiazolyl tetrazolium assay, the lactate dehydrogenase assay, and flow cytometry, respectively. Furthermore, the levels of cleaved-Caspase3, Keap1, Nrf2, HO-1, and NQO1 were assessed using reverse transcription quantitative polymerase chain reaction and western blot analysis, and the release of inflammatory cytokines was determined using the enzyme-linked immunosorbent assay.
We observed that oxycodone had no adverse effects on hEndoSCs; rather, it protected hEndoSCs against mifepristone-induced endometrial damage, as confirmed by the enhanced cell viability, reduced number of apoptotic cells, decreased Caspase3 activity and inflammatory cytokine secretion, and increased Keap1/Nrf2/HO-1 pathway-related protein expression. In addition, we found that the protective effects of oxycodone on mifepristone-induced hEndoSC injury were inhibited by ML385 (a Keap1/Nrf2/HO-1 inhibitor).
In summary, we confirmed that oxycodone alleviates mifepristone-induced hEndoSC injury by activating the Keap1/Nrf2/HO-1 signaling pathway.
子宫内膜损伤是一种常见的妇科疾病，由宫腔内炎症、感染和内分泌失调引起。人子宫内膜基质细胞（hEndoSCs）可维持子宫内膜稳态，在修复子宫内膜损伤中发挥重要作用。米非司酮是一种甾体类抗孕激素药物，在全球生殖医学领域广泛应用。米非司酮诱导的 hEndoSC 损伤已被用于体外研究子宫内膜损伤。目前，羟考酮在子宫内膜损伤中的发病机制和潜在调控机制尚不清楚。
本研究旨在评估羟考酮在米非司酮刺激的 hEndoSC 损伤中的作用，并分析其潜在的分子机制。
采用噻唑蓝比色法、乳酸脱氢酶法和流式细胞术分别检测 hEndoSC 活力、细胞毒性和细胞凋亡。此外，采用逆转录定量聚合酶链反应和 Western blot 分析检测 cleaved-Caspase3、Keap1、Nrf2、HO-1 和 NQO1 水平，采用酶联免疫吸附试验检测炎症细胞因子的释放。
我们发现羟考酮对 hEndoSCs 无不良影响；相反，它可保护 hEndoSCs 免受米非司酮诱导的子宫内膜损伤，这表现在细胞活力增强、凋亡细胞减少、Caspase3 活性和炎症细胞因子分泌减少以及 Keap1/Nrf2/HO-1 通路相关蛋白表达增加。此外，我们发现羟考酮对米非司酮诱导的 hEndoSC 损伤的保护作用被 ML385（一种 Keap1/Nrf2/HO-1 抑制剂）所抑制。
综上所述，我们证实羟考酮通过激活 Keap1/Nrf2/HO-1 信号通路减轻米非司酮诱导的 hEndoSC 损伤。
