Abstract
BackgroundPreeclampsia (PE) is a frequently occurring pregnancy disorder in the placenta, which results in various maternal and fetal complications. The current study aims to evaluate the role of extracellular vesicles (EVs)-encapsulated microRNA (miR)-101 in biological processes of trophoblasts in PE and its underlying mechanism.MethodsHuman umbilical cord mesenchymal stem cell (HUCMSC) and HUCMSC-derived EVs were isolated and cultured, after which EV characterization was carried out using PKH67 staining. In silico analyses were adopted to predict the downstream target genes of miR-101, and dual luciferase reporter gene assay was applied to validate the binding affinity. Furthermore, loss- and gain-of-function approaches were adopted to determine the role of miR-101 and bromodomain-containing protein 4 (BRD4) in trophoblast proliferation and invasion using EDU staining and transwell assay. In addition, a rat model of PE was established to verify the function of EV-encapsulated miR-101 in vivo.ResultsPlacental tissues obtained from PE patients presented with downregulated miR-101 expression and upregulated BRD4 and CXCL11 expression. EV-encapsulated miR-101 from HUCMSCs could be delivered into the trophoblast HTR-8/SVneo cells, thus enhancing proliferation and migration of trophoblasts. Mechanically, miR-101 targeted and negatively regulated BRD4 expression. BRD4 knockdown promoted the proliferation and migration of trophoblasts by suppressing NF-ÎșB/CXCL11 axis. EV-encapsulated miR-101 from HUCMSCs also reduced blood pressure and 24 h urine protein in vivo, thereby ameliorating PE.ConclusionIn summary, EV-encapsulated miR-101 promoted proliferation and migration of placental trophoblasts through the inhibition of BRD4 expression via NF-ÎșB/CXCL11 inactivation.
Highlights
Preeclampsia (PE) is a frequently occurring pregnancy disorder in the placenta, which results in various maternal and fetal complications
A prior study noted that miR-101 regulates apoptosis of trophoblast HTR-8/SVneo cells during PE [10], which led to us exploring the function of extracellular vesicles (EVs)-encapsulated miR-101 during the early phase of PE
When GW4869 was added to inhibit EVs released from Human umbilical cord mesenchymal stem cells (HUCMSCs), co-culturing of transfected HUCMSCs and HTR-8/SVneo cells, no red fluorescence was observed from cy3-miR-101 mimic in HTR-8/SVneo cells (Fig. 2c). These results suggested that miR-101 from HUCMSCs could be delivered into HTR-8/SVneo cells and this transportation may rely on EVs
Summary
Preeclampsia (PE) is a frequently occurring pregnancy disorder in the placenta, which results in various maternal and fetal complications. The current study aims to evaluate the role of extracellular vesicles (EVs)encapsulated microRNA (miR)-101 in biological processes of trophoblasts in PE and its underlying mechanism. Elucidating the underlying mechanism by which placental trophoblast proliferation and invasion affect PE is crucial in order to attenuate PE. The pathogenesis of PE is involved in the alteration in protein expression in the human umbilical cord mesenchymal stem cell (HUCMSC)-derived EVs [7]. Altered expression of CXCL11 has been found in PE [18] These findings unveil a possible mechanism underlying the involvement of EV-encapsulated miR-101 and BRD4dependent NF-ÎșB-CXCL11 axis in PE. In the current study, PE rat models and human extravillous trophoblast cell lines HTR-8/SVneo were established to explore the potential regulatory network. Understanding the molecular regulation of EV-encapsulated miR-101 is essential to develop therapeutic methods that can attenuate PE
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