Senescence Human Endothelial Cells Produce ExtracellularVesicles with Dis tinct miRNAs Cargo

Objective: Extracellular vesicles (EVs) derived from senescenceendothelial cells (ECs) contribute to endothelium dysfunction.We hypothesized whether EVs from human umbilicalvein cells (HUVECs) contain dis tinct miRNAs cargo in a modelof senescence in vitro.Materials and Methods: Cellular senescent was induced byH۲O۲-treatment of HUVECs. The protein levels of P۲۱, P۵۳,and P۱۶ were measured by wes tern blotting analysis, while theexpression of miRNAs -۲۱ and miRNAs -۱۲۶ were inves tigatedby real-time PCR (q-PCR) in cells. Small EVs (sEVs) wereisolated and characterized using AChE assay, wes tern blotting,and transmission electron microscopy (TEM). The expressionof miR-۲۱ and miR-۱۲۶ was measured by q-PCR. An in vitrowound healing assay was used to evaluate ECs migration rateupon incubation with sEVs.Results: Data showed that the protein levels of P۲۱, P۵۳, andP۱۶ were increased, however the expression of miRNAs -۲۱and miRNAs -۱۲۶ was up-regulated and down-regulated intreated cells respectively (P value <۰.۰۵). Isolated sEVs wereconfirmed by wes tern blotting and TEM. H۲O۲ increased sEVsproduction by treated cells (P value <۰.۰۵). A high level ofmiRNAs -۲۱ concurred with a decrease in miRNAs -۱۲۶ levelsin sEVs of treated cells (P value <۰.۰۵). The wound healing rateof ECs incubated with sEVs of H۲O۲-induced cells was reduced(P value <۰.۰۵).Conclusion: Our results showed that H۲O۲-induced HUVECsproduced sEVs with dis tinct miRNAs cargo and function, proposingbiomarker application in age-associated vascular disease

کلیدواژه ها:

Cellular Senescence ، Endothelial Cells ، ExtracellularVesicles ، H۲O۲