Human Placental Mesenchymal Stem Cells‐Exosomes Alleviate Endothelial Barrier Dysfunction via Cytoskeletal Remodeling through hsa‐miR‐148a‐3p/ROCK1 Pathway

Abstract

Background. Endothelial barrier disruption of human pulmonary vascular endothelial cells (HPVECs) is an important pathogenic factor for acute lung injury (ALI)/acute respiratory distress syndrome (ARDS). Mesenchymal stem cells‐exosome (MSCs‐Exo) represents an ideal carrier for cell‐free therapy. The therapeutic implication and underlying mechanism of human placental MSCs‐Exo (HPMSCs‐Exo) in ALI/ARDS need to be further explored. Materials and Methods. HPMSCs‐Exo was extracted from HPMSCs and characterized. Then, the therapeutic effects of exosomes were evaluated in ALI mice and HPVECs. RNA‐sequencing was applied to reveal the miRNA profile of HPMSCs‐Exo and differentially expressed genes (DEGs) in HPMSCs‐Exo‐pretreated HPVECs. The targets of miRNAs were predicted by bioinformatics methods and correlated to DEGs. Finally, the role of hsa‐miR‐148a‐3p/ROCK1 pathway in HPVECs has been further discussed. Results. The results showed that HPMSCs‐Exo could downregulate Rho‐associated coiled‐coil‐containing protein kinase 1 (ROCK1), upregulate the expression of zonula occludens‐1 (ZO‐1) and F‐actin, promote HPVECs migration and tube formation, reduce cytoskeletal disorders and cell permeability, and thus improve ALI/ARDS. RNA‐sequencing revealed the DEGs were mainly enriched in cell junction, angiogenesis, inflammation, and energy metabolism. HPMSCs‐Exo contains multiple miRNAs which are associated with cytoskeletal function; the expression abundance of hsa‐miR‐148a‐3p is the highest. Bioinformatic analysis identified ROCK1 as a target of hsa‐miR‐148a‐3p. The overexpression of hsa‐miR‐148a‐3p in HPMSCs‐Exo promoted the migration and tube formation of HPVECs and reduced ROCK1 expression. However, the overexpression of ROCK1 on HPVECs reduced the therapeutic effect of HPMSCs‐Exo. Conclusions. HPMSCs‐Exo represents a protective regimen against endothelial barrier disruption of HPVECs in ALI/ARDS, and the hsa‐miR‐148a‐3p/ROCK1 pathway plays an important role in this therapeutics implication.