Yue Xiaojing, Pang Menglan, Chen Yun, Cheng Zhixing, Zhou Ruisi, Wang Yu, Zha Zhiqiang, Huang Liping
Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.
Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.
Placenta. 2024 Dec;158:145-155. doi: 10.1016/j.placenta.2024.10.013. Epub 2024 Oct 22.
Preeclampsia (PE) is a pregnancy-associated complication characterised by new-onset hypertension and proteinuria. This study explored the therapeutic potential of puerarin (Pue) in PE and investigated the underlying mechanism, with a focus on placental ferroptosis.
Using an N-nitro-L-arginine methyl ester (L-NAME)-induced PE mouse model, we assessed the effects of Pue on PE phenotypes and placental ferroptosis. Antioxidative and anti-ferroptotic effects of Pue were studied in three ferroptotic cell models (hypoxia/reperfusion, cobalt chloride, and erastin). The regulation of Pue on cAMP response element binding protein (CREB) and heme oxygenase-1 (HO-1) was evaluated through gain- and loss-of-function assays. Luciferase assays were used to elucidate the effect of Flag-CREB on Hmox1 promoter fragments. CREB/HO-1 modulation by Pue was validated in mouse placentas with PE.
Pue significantly alleviated maternal hypertension, proteinuria, fetal growth restriction, and placental damage in PE mice. This was associated with an upregulation of the anti-ferroptosis system (glutathione peroxidase 4 [GPX4], cys2/glutamate antiporter [SLC7A11], and glutathione [GSH]) and repression of reactive oxygen species (ROS) and malondialdehyde (MDA) in trophoblasts. Pue reduced HO-1 and CREB, and HO-1 deficiency upregulated GPX4 and SLC7A11. Manipulation of CREB expression led to changes in HO-1/GPX4; whereas, the regulation reversed by Pue administration. Flag-CREB enhanced luciferase activity on the full length Hmox1 promoter (-2000/+78), which contains three CREB1 binding sites (S1-S3). In contrast, no increase in luciferase activity was observed with promoter fragments (-850/+78) and (-550/+78), which contain only the CREB1 binding sites S2 and S3, respectively.
Pue ameliorated PE-like symptoms in mice by repressing trophoblast ferroptosis via inhibition of CREB signalling and affecting the Homx1 promoter.
子痫前期（PE）是一种与妊娠相关的并发症，其特征为新发高血压和蛋白尿。本研究探讨了葛根素（Pue）在子痫前期中的治疗潜力，并研究了其潜在机制，重点关注胎盘铁死亡。
使用N-硝基-L-精氨酸甲酯（L-NAME）诱导的子痫前期小鼠模型，我们评估了葛根素对子痫前期表型和胎盘铁死亡的影响。在三种铁死亡细胞模型（缺氧/复氧、氯化钴和埃拉斯汀）中研究了葛根素的抗氧化和抗铁死亡作用。通过功能获得和功能丧失试验评估了葛根素对环磷酸腺苷反应元件结合蛋白（CREB）和血红素加氧酶-1（HO-1）的调节作用。荧光素酶试验用于阐明Flag-CREB对Hmox1启动子片段的影响。葛根素对CREB/HO-1的调节作用在子痫前期小鼠胎盘中得到验证。
葛根素显著减轻了子痫前期小鼠的母体高血压、蛋白尿、胎儿生长受限和胎盘损伤。这与抗铁死亡系统（谷胱甘肽过氧化物酶4 [GPX4]、胱氨酸/谷氨酸反向转运体[SLC7A11]和谷胱甘肽[GSH]）的上调以及滋养层细胞中活性氧（ROS）和丙二醛（MDA）的抑制有关。葛根素降低了HO-1和CREB，而HO-1缺乏上调了GPX4和SLC7A11。操纵CREB表达导致HO-1/GPX4发生变化；然而，葛根素给药可逆转这种调节。Flag-CREB增强了全长Hmox1启动子（-2000/+78）上的荧光素酶活性，该启动子包含三个CREB1结合位点（S1-S3）。相比之下，仅包含CREB1结合位点S2和S3的启动子片段（-850/+78）和（-550/+78）未观察到荧光素酶活性增加。
葛根素通过抑制CREB信号通路并影响Homx1启动子来抑制滋养层铁死亡，从而改善小鼠子痫前期样症状。
