Li Fan, Zhao Xiaofang
Department of Obstetrics and Gynecology, Ankang People's Hospital, No.38 Jiangbei Avenve, Hanbin District, Ankang, 725000, China.
Hereditas. 2025 Jun 13;162(1):104. doi: 10.1186/s41065-025-00465-w.
Gestational diabetes mellitus (GDM) is a common metabolic complication during pregnancy that poses significant risks to both the pregnant woman and her fetus. Astragaloside IV (Ast IV) belongs to the class of triterpenoid saponins and exhibits important physiological roles in various aspects, including antidiabetic, antioxidant, and antiviral effects. The main objective of this study is to investigate the effects of Ast IV on trophoblast damage caused by high glucose (HG) and its underlying mechanism of action.
Cell viability was determined by the CCK8 assay. The levels of oxidative stress in cells were determined by lactate dehydrogenase (LDH), malondialdehyde (MDA), and reactive oxygen species (ROS) kits. Ferroptosis in cells was assessed by the iron content kit. Gene expression levels were detected by real-time quantitative reverse transcription PCR (qRT-PCR) and western blot. The protein stability of Forkhead box protein M1 (FOXM1) was determined by the cycloheximide (CHX) assay. The ubiquitination level of FOXM1 was detected by the immunoprecipitation assay.
Ast IV alleviated the inhibitory effect of HG on the proliferation of HTR-8/SVneo cells and reduced HG-induced oxidative stress and ferroptosis. Ast IV was able to decrease the ubiquitination of FOXM1, thereby ensuring the stability of its expression. The overexpression of FOXM1 significantly mitigated the inhibitory effect of HG on the viability of HTR-8/SVneo cells and concurrently decreased the occurrence of HG-induced oxidative stress and ferroptosis processes. Conversely, knockdown of FOXM1 diminished the protective effect of Ast IV on HTR-8/SVneo cells.
Ast IV ameliorates HG-induced trophoblast injury by modulating deubiquitination of FOXM1, which provides a new insight into the treatment of GDM.
妊娠期糖尿病（GDM）是孕期常见的代谢并发症，对孕妇及其胎儿均构成重大风险。黄芪甲苷IV（Ast IV）属于三萜皂苷类，在包括抗糖尿病、抗氧化和抗病毒作用等多个方面发挥重要生理作用。本研究的主要目的是探讨Ast IV对高糖（HG）所致滋养层细胞损伤的影响及其潜在作用机制。
采用CCK8法测定细胞活力。通过乳酸脱氢酶（LDH）、丙二醛（MDA）和活性氧（ROS）试剂盒测定细胞中的氧化应激水平。用铁含量试剂盒评估细胞中的铁死亡情况。通过实时定量逆转录PCR（qRT-PCR）和蛋白质印迹法检测基因表达水平。用放线菌酮（CHX）试验测定叉头框蛋白M1（FOXM1）的蛋白质稳定性。通过免疫沉淀试验检测FOXM1的泛素化水平。
Ast IV减轻了HG对HTR-8/SVneo细胞增殖的抑制作用，并降低了HG诱导的氧化应激和铁死亡。Ast IV能够降低FOXM1的泛素化，从而确保其表达的稳定性。FOXM1的过表达显著减轻了HG对HTR-8/SVneo细胞活力的抑制作用，同时减少了HG诱导的氧化应激和铁死亡过程的发生。相反，敲低FOXM1减弱了Ast IV对HTR-8/SVneo细胞的保护作用。
Ast IV通过调节FOXM1的去泛素化改善HG诱导的滋养层细胞损伤，这为GDM的治疗提供了新的见解。
