Shen Wei, Wang Qingfu, Shen Guofang, Gu Meiling, Shen Qifeng, Zhang Ailan, Zhu Xiaohong
Department of Obstetrics and Gynecology, Affiliated Xiaoshan Hospital, Hangzhou Normal University, Hangzhou, Zhejiang, 311200, China.
Department of Anesthesia, Affiliated Xiaoshan Hospital, Hangzhou Normal University, Hangzhou, Zhejiang, 311200, China.
Placenta. 2025 Feb;160:89-99. doi: 10.1016/j.placenta.2025.01.002. Epub 2025 Jan 3.
Pre-eclampsia (PE) is a pregnancy complication featuring hypertension and proteinuria. Metformin exerts clinically preventive effects on PE with an unspecified mechanism.
Placental tissues from PE patients and normal pregnant (NP) women were collected. Twenty-four pregnant mice were divided into control, PE (40 μg/kg lipopolysaccharides (LPS)-induced modeling), aspirin, and metformin groups. After acquisition of bone marrow-derived macrophages (BMDM) and THP-1 cells, cells were categorized into control, LPS (100 ng/mL), metformin, and metformin + toll-like receptor 4 (TLR4) agonist RS 09 groups. Inflammatory factors and macrophage polarization were detected by ELISA, flow cytometry, immunofluorescence, immunohistochemistry, and qRT-PCR methods. TLR4/Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway protein expression was examined using Western blot.
Both PE patients and PE-like mice enhanced inducible nitric oxide synthase (iNOS), TLR4, p-NF-κB/NF-κB, and p-inhibitor of NF-κB (IκBα)/IκBα expression, and lower arginase 1 (Arg-1) expression. Moreover, metformin treatment in PE-like mice increased fetal number and weight and reduced hypertension, proteinuria, insulin resistance, tumor necrosis factor-α (TNF-α), IL-6, IL-1β, chemokine ligand 4 (CCL4), C-X-C motif chemokine ligand 2 (CXCL2) expression and M1 macrophage polarization, with similar inhibition to aspirin. In LPS-induced cells, metformin had the same effects mentioned above. Decreased TLR4, p-NF-κB/NF-κB, and p-IκBα/IκBα protein expression was caused by metformin both in vivo and in vitro. In vitro, RS 09 intervention inhibited anti-inflammatory and pro-M2 polarizing effects of metformin, activating TLR4/NF-κB pathway.
Metformin may ameliorate PE by promoting M2 macrophage polarization through up-regulating TLR4/NF-κB pathway, laying theoretical basis for metformin clinical application in PE.
子痫前期（PE）是一种以高血压和蛋白尿为特征的妊娠并发症。二甲双胍对PE具有临床预防作用，但其机制尚不明确。
收集PE患者和正常妊娠（NP）妇女的胎盘组织。将24只妊娠小鼠分为对照组、PE组（40 μg/kg脂多糖（LPS）诱导建模）、阿司匹林组和二甲双胍组。获取骨髓来源的巨噬细胞（BMDM）和THP-1细胞后，将细胞分为对照组、LPS组（100 ng/mL）、二甲双胍组和二甲双胍 + toll样受体4（TLR4）激动剂RS 09组。采用ELISA、流式细胞术、免疫荧光、免疫组织化学和qRT-PCR方法检测炎症因子和巨噬细胞极化情况。使用蛋白质印迹法检测TLR4/活化B细胞核因子κB（NF-κB）通路蛋白表达。
PE患者和类PE小鼠的诱导型一氧化氮合酶（iNOS）、TLR4、p-NF-κB/NF-κB和p-核因子κB抑制蛋白（IκBα）/IκBα表达均增强，精氨酸酶1（Arg-1）表达降低。此外，在类PE小鼠中，二甲双胍治疗可增加胎儿数量和体重，降低高血压、蛋白尿、胰岛素抵抗、肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）、趋化因子配体4（CCL4）、C-X-C基序趋化因子配体2（CXCL2）表达以及M1巨噬细胞极化，其抑制作用与阿司匹林相似。在LPS诱导的细胞中，二甲双胍具有上述相同作用。二甲双胍在体内和体外均导致TLR4、p-NF-κB/NF-κB和p-IκBα/IκBα蛋白表达降低。在体外，RS 09干预抑制了二甲双胍的抗炎和促M2极化作用，激活了TLR4/NF-κB通路。
二甲双胍可能通过上调TLR4/NF-κB通路促进M2巨噬细胞极化来改善PE，为二甲双胍在PE中的临床应用奠定理论基础。
