[Effects of human placental mesenchymal stem cell (hPDMSC)-derived exosome (PdE) shuttling miR-144-3p on vascular endothelial cell dysfunction in gestational diabetes mellitus (GDM)].

Abstract

Objective Gestational diabetes mellitus (GDM) is associated with long-term endothelial dysfunction of maternal-fetal vessels, and human placental mesenchymal stem cell (hPDMSC)-derived exosomes (PdE) from GDM patients are closely related to the progression of the disease. This study aims to investigate the expression of miR-144-3p in PdE and its effects and the underlying mechanism on vascular endothelial cell dysfunction in GDM. Methods PdE from normal pregnant patients and GDM patients (the Nor-PdE group and the GDM-PdE group) were isolated and collected. Anti-miR144-3p and negative control were transfected into hPDMSC using the Lipofectamine. Then, the PdE were collected and co-cultured with human umbilical vein endothelial cell (HUVEC). Real-time quantitative PCR (qRT-PCR) was performed to detect miR-144-3p expression, while the CCK-8 assay was used to assess HUVEC viability. Transwell^TM assay was performed to analyze cell migration. Matrix gel tube formation assay was carried out to determine tube formation ability. Bioinformatics and dual-luciferase reporter analysis were used to evaluate the targeting relationship between miR-144-3p and suppressors of cytokine signaling 6 (SOCS6). The protein expression of SOCS6 was analyzed by Western blot. Results The PdE from normal pregnant patients and GDM patients were successfully isolated. Compared with Nor-PdE group, the expression levels of miR-144-3p were increased in GDM-PdE group. Moreover, the expression of miR-144-3p was obviously decreased after anti-miR-144-3p treatment. Compared with Nor-PdE group, GDM-PdE treatment dramatically suppressed HUVEC viability, migration and tube formation, these inhibitory effects were markedly attenuated upon miR-144-3p knockdown. Compared with Nor-PdE group, the transcriptional levels of SOCS6 were decreased in HUVECs after treatment with GDM-PdE. Bioinformatics and dual-luciferase reporter analysis confirmed that SOCS6 was a potential target of miR-144-3p. Down-regulation of miR-144-3p obviously increased HUVEC viability, migration and tube formation under GDM-PdE condition, which was abrogated after SOCS6 down-regulation. Conclusion Abnormal high expression of miR-144-3p in GDM-PdE may regulate the dysfunction of vascular endothelial cells by targeting SOCS6, supporting it as a new potential target for PdE in the prevention and treatment of GDM.