Jiang Xiaotong, Gu Mengqi, Xie Yu, Li Yuchen, Lv Qingfeng, Chen Pengzheng, Jing Die, Zhou Yu, Wang Xietong, Li Lei
Department of Obstetrics and Gynecology, Shandong Provincial Hospital, Shandong University, Jinan, 250021, Shandong, China.
Obstetrics and Gynecology, Maternity and Child Health Care of RIZHAO, Rizhao, 276827, Shandong, China.
Stem Cell Rev Rep. 2025 Sep 2. doi: 10.1007/s12015-025-10935-3.
Preeclampsia is amultisystem disorder involving in inflammatory responses and metabolic dysfunction of maternal-fetal circulation. Recently, researchers found it threatens renal health of offspring in adulthood. Growing evidence indicated chronic kidney disease is associated with glomeruli deficiencies during intrauterine development. Our previous study showed placenta-derived exosomes from cord plasma with preeclampsia impede fetal glomerular vascularization, during which we postulate microRNAs may function as epigenetic switches for gene silencing of human glomerular endothelial cells. However, the specific miRNAs in placenta-derived exosomes engaged in glomerular vascularization remain unclear.
Small RNA sequencing of placental-derived exosomes and bioinformatics analysis were applied to identify differentially expressed miRNAs, followed by real-time polymerase chain reaction for verification. Transient expression and inhibition of candidate miRNA were performed by transfection with chemically synthesized miRNA oligonucleotides. Functional assays of HGECs including cell proliferation assays, EDU assays, migration assays, tube formation assays and monolayer cell barrier permeability assays were performed after transfection. Further, dual luciferase assay was used to explore the target genes of candidate miRNA, followed by RT-qPCR, western blot and rescue assays. Antagomirs transfection of C57BL/6 J fetal mice via Amniotic cavity injection in vivo and C57BL/6 J fetal mice kidney explants culture in vitro were performed to evaluate number of glomeruli, renal development.
Preeclampsia downregulates miR-199a-3p in placenta-derived exosomes from cord plasma. Suppression of endogenous miR-199a-3p in HGECs inhibits angiogenesis, proliferation, migration and permeability. A dual luciferase assay and rescue assays confirmed that miR-199a-3p targets PH domain leucine-rich repeat-containing protein phosphatase 2, regulating the phosphorylation of Akt serine/threonine kinase 1 (S473). C57BL/6 J fetal mice with miR-199a-3p downregulation have low glomerulus counts and relative growth rate.
miR-199a-3p in placenta-derived exosomes from cord plasma controls VEGF-induced glomerular angiogenesis. Moreover, it provides a distinct perspective for the mechanisms underlying the increased risk for renal disease in the offspring of preeclampsia patients via placenta-derived exosomal miRNAs.
子痫前期是一种多系统疾病，涉及母婴循环的炎症反应和代谢功能障碍。最近，研究人员发现它会威胁成年后代的肾脏健康。越来越多的证据表明，慢性肾脏病与宫内发育期间的肾小球缺陷有关。我们之前的研究表明，来自子痫前期脐带血血浆的胎盘来源外泌体阻碍胎儿肾小球血管生成，在此过程中我们推测微小RNA可能作为人类肾小球内皮细胞基因沉默的表观遗传开关。然而，胎盘来源外泌体中参与肾小球血管生成的特定微小RNA仍不清楚。
应用胎盘来源外泌体的小RNA测序和生物信息学分析来鉴定差异表达的微小RNA，随后进行实时聚合酶链反应进行验证。通过化学合成的微小RNA寡核苷酸转染进行候选微小RNA的瞬时表达和抑制。转染后对人肾小球内皮细胞进行功能测定，包括细胞增殖测定、EDU测定、迁移测定、管形成测定和单层细胞屏障通透性测定。此外，使用双荧光素酶测定来探索候选微小RNA的靶基因，随后进行RT-qPCR、蛋白质免疫印迹和拯救实验。通过羊膜腔内注射对C57BL/6 J胎鼠进行抗微小RNA转染，并在体外进行C57BL/6 J胎鼠肾外植体培养，以评估肾小球数量和肾脏发育情况。
子痫前期下调脐带血血浆中胎盘来源外泌体中的miR-199a-3p。抑制人肾小球内皮细胞中的内源性miR-199a-3p会抑制血管生成、增殖、迁移和通透性。双荧光素酶测定和拯救实验证实，miR-199a-3p靶向含PH结构域富亮氨酸重复蛋白磷酸酶2，调节Akt丝氨酸/苏氨酸激酶1（S473）的磷酸化。miR-199a-3p下调的C57BL/6 J胎鼠肾小球计数和相对生长率较低。
脐带血血浆中胎盘来源外泌体中的miR-199a-3p控制血管内皮生长因子诱导的肾小球血管生成。此外，它为子痫前期患者后代患肾病风险增加的潜在机制提供了一个独特的视角，即通过胎盘来源的外泌体微小RNA发挥作用。
