Department of Gynecology and Obstetrics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, China.
Department of Urology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, China.
Hum Reprod. 2021 Mar 18;36(4):951-964. doi: 10.1093/humrep/deaa363.
Could erastin activate ferroptosis to regress endometriotic lesions?
Erastin could induce ferroptosis to regress endometriotic lesions in endometriosis.
Ectopic endometrial stromal cells (EESCs) are in an iron overloading microenvironment and tend to be more sensitive to oxidative damage. The feature of erastin-induced ferroptosis is iron-dependent accumulation of lethal lipid reactive oxygen species (ROS).
STUDY DESIGN, SIZE, DURATION: Eleven patients without endometriosis and 21 patients with endometriosis were recruited in this study. Primary normal and ectopic endometrial stromal cells were isolated, cultured and subjected to various treatments. The in vivo study involved 10 C57BL/6 female mice to establish the model of endometriosis.
PARTICIPANTS/MATERIALS, SETTING, METHODS: The markers of ferroptosis were assessed by cell viability, lipid peroxidation level and morphological changes. The cell viability was measured by colorimetric method, lipid peroxidation levels were measured by flow cytometry, and morphological changes were observed by transmission electron microscopy. Immunohistochemistry and western blot were used to detect ferroportin (FPN) expression. Prussian blue staining and immunofluorescent microscopy of catalytic ferrous iron were semi-quantified the levels of iron. Adenovirus-mediated overexpression and siRNA-mediated knockdown were used to investigate the role of FPN on erastin-induced ferroptosis in EESCs.
EESCs were more susceptible to erastin treatment, compared to normal endometrial stromal cells (NESCs) (P<0.05). Treatment of cultured EESCs with erastin dramatically increased the total ROS level (P<0.05, versus control), lipid ROS level (P<0.05, versus NESCs) and intracellular iron level (P<0.05, versus NESCs). The cytotoxicity of erastin could be attenuated by iron chelator, deferoxamine (DFO), and ferroptosis inhibitors, ferrostatin-1 and liproxstatin-1, (P<0.05, versus erastin) in EESCs. In EESCs with erastin treatment, shorter and condensed mitochondria were observed by electron microscopy. These findings together suggest that erastin is capable to induce EESC death by ferroptosis. However, the influence of erastin on NESCs was slight. The process of erastin-induced ferroptosis in EESCs accompanied iron accumulation and decreased FPN expression. The overexpression of FPN ablated erastin-induced ferroptosis in EESCs. In addition, knockdown of FPN accelerated erastin-induced ferroptosis in EESCs. In a mouse model of endometriosis, we found ectopic lesions were regressed after erastin administration.
N/A.
LIMITATIONS, REASONS FOR CAUTION: This study was mainly conducted in primary human endometrial stromal cells. Therefore, the function of FPN in vivo need to be further investigated.
Our findings reveal that erastin may serve as a potential therapeutic treatment for endometriosis.
STUDY FUNDING/COMPETING INTEREST(S): This research did not receive any specific grant from funding agencies in the public, commercial or not-for-profit sectors. The authors declare no conflict of interest.
依拉司群能否通过激活铁死亡来消退子宫内膜异位症病灶？
依拉司群可诱导铁死亡来消退子宫内膜异位症中的异位病灶。
异位子宫内膜基质细胞（EESCs）处于铁超载的微环境中，往往更容易受到氧化损伤。依拉司群诱导的铁死亡的特征是铁依赖性累积致命的脂质活性氧（ROS）。
研究设计、大小、持续时间：本研究招募了 11 名无子宫内膜异位症的患者和 21 名子宫内膜异位症患者。分离、培养并对原代正常和异位子宫内膜基质细胞进行了各种处理。体内研究涉及 10 只 C57BL/6 雌性小鼠，建立子宫内膜异位症模型。
参与者/材料、设置、方法：通过细胞活力、脂质过氧化水平和形态变化来评估铁死亡的标志物。通过比色法测量细胞活力，通过流式细胞术测量脂质过氧化水平，通过透射电子显微镜观察形态变化。用免疫组织化学和 Western blot 检测铁转运蛋白（FPN）的表达。普鲁士蓝染色和催化亚铁的免疫荧光显微镜用于半定量检测铁水平。腺病毒介导的过表达和 siRNA 介导的敲低用于研究 FPN 在 EESCs 中对依拉司群诱导的铁死亡中的作用。
与正常子宫内膜基质细胞（NESCs）相比，EESCs 对依拉司群治疗更敏感（P<0.05）。用依拉司群处理培养的 EESCs 可显著增加总 ROS 水平（P<0.05，与对照组相比）、脂质 ROS 水平（P<0.05，与 NESCs 相比）和细胞内铁水平（P<0.05，与 NESCs 相比）。在 EESCs 中，铁螯合剂去铁胺（DFO）和铁死亡抑制剂 ferrostatin-1 和 liproxstatin-1 可减弱依拉司群的细胞毒性（P<0.05，与依拉司群相比）。在依拉司群处理的 EESCs 中，电子显微镜观察到线粒体变短且浓缩。这些发现共同表明，依拉司群能够通过铁死亡诱导 EESC 死亡。然而，依拉司群对 NESCs 的影响较小。EESCs 中依拉司群诱导的铁死亡伴随着铁积累和 FPN 表达减少。FPN 的过表达消除了 EESCs 中依拉司群诱导的铁死亡。此外，FPN 的敲低加速了 EESCs 中依拉司群诱导的铁死亡。在子宫内膜异位症的小鼠模型中，我们发现依拉司群给药后异位病灶消退。
无。
局限性、谨慎的原因：本研究主要在原代人子宫内膜基质细胞中进行。因此，FPN 在体内的功能需要进一步研究。
我们的研究结果表明，依拉司群可能是治疗子宫内膜异位症的一种潜在治疗方法。
研究资金/利益冲突：本研究未从公共、商业或非营利部门的任何特定资助机构获得任何资金。作者声明没有利益冲突。
