子痫前期miR-4531/CX3CL1信号轴参与血管损伤的体外研究  

Effects of miR-4531/CX3CL1 signaling pathway on the vascular injury in preeclampsia in vitro

作　　者：王曼[1] 李骏[1] 李航[1] 宋青 刘艳[1] 王海丽[1] 王晓[1] 程群仙 胡峥 徐灵 WANG Man;LI Jun;LI Hang;SONG Qing;LIU Yan;WANG Haili;WANG Xiao;CHENG Qunxian;HU Zheng;XU Ling(Department of Obstetrics and Gynecology,Minhang Hospital,Fudan University,Shanghai 201199,China)

机构地区：[1]复旦大学附属闵行医院妇产科,上海201199

出　　处：《中国临床医学》2024年第6期868-874,共7页Chinese Journal of Clinical Medicine

基　　金：上海市闵行区科学技术委员会项目(2021MHZ043).

摘　　要：目的探讨miR-4531/CX3CL1信号通路在子痫前期血管损伤中的作用,寻找早期诊断和防治子痫前期的潜在靶点。方法选择2021年10月至2022年12月在复旦大学附属闵行医院诊断为晚发型子痫前期的10例患者及同期10例正常妊娠者的胎盘组织样本,用qRT-PCR检测miR-4531和CX3CL1在胎盘组织中的表达水平。通过lipofectamine 2000进行体外细胞转染,用荧光素酶报告基因实验确定miR-4531与CX3CL1之间的结合关系。检测中性粒细胞的凋亡,人脐静脉内皮细胞(HUVECs)的迁移、修复、集落形成、游离DNA(cfDNA)释放等。结果子痫前期患者胎盘组织中miR-4531下调、CX3CL1上调(P<0.05)。荧光素酶报告基因实验显示,miR-4531与CX3CL1之间存在直接结合关系。上调miR-4531促进HUVECs的迁移、增殖和修复,抑制缺氧条件下HUVECs中cfDNA的释放及中性粒细胞的凋亡(P<0.05);下调miR-4531抑制HUVECs的增殖、迁移和修复,增强缺氧条件下HUVECs中cfDNA的释放,促进中性粒细胞的凋亡(P<0.05)。结论子痫前期患者胎盘组织中miR-4531表达下调,可能通过靶向调控CX3CL1通路导致缺氧条件下血管内皮损伤和异常高凝血状态,是PE潜在的治疗靶点。Objective To investigate the effects of miR-4531/CX3CL1 signaling pathway on the vascular injury in preeclampsia,and to search possible targets for early diagnosis and prevention of preeclampsia.Methods Placental tissue samples were obtained from 10 patients with late-onset preeclampsia and 10 normal pregnant women in the Obstetrics and Gynecology Department of Minhang Hospital,Fudan University from October 2021 to December 2022.The levels of miR-4531 and CX3CL1 were evaluated by qRT-PCR.Cell transfection was performed by lipofectamine 2000 in vitro.The binding relationship between miR-4531 and CX3CL1 was determined by luciferase reporter assay.The apoptosis of neutrophils,the migration,repair,colony formation and cell-free DNA(cfDNA)release of human umbilical vein endothelial cells(HUVECs)were detected.Results MiR-4531 was significantly downregulated,and CX3CL1 was upregulated in placental tissues of preeclampsia patients(P<0.05).Luciferase report assay confirmed the direct binding relationship between miR-4531 and CX3CL1.Upregulation of miR-4531significantly promoted the migration,proliferation,repair,and inhibited cfDNA release of HUVECs and neutrophils apoptosis in medium under hypoxic condition(P<0.05).However,downregulation of miR-4531 obviously inhibited the proliferation,migration,repair,and enhanced cfDNA release of HUVECs and neutrophils apoptosis in medium under hypoxic condition(P<0.05).Conclusions The miR-4531 is downregulated in placental tissues of preeclampsia parients and is a potential therapeutic target for preeclampsia.It might cause endothelial damage and an abnormal hypercoagulable state under hypoxic condition by targeting and regulating the CX3CL1 pathway.

关 键 词：子痫前期 miR-4531 CX3CL1 人脐静脉内皮细胞 细胞游离DNA 

分 类 号：R714.244[医药卫生—妇产科学]