Microvesicles‐mediated communication between endothelial cells modulates, endothelial survival, and angiogenic function via transferring of miR‐125a‐5p

Abstract

Endothelial cells (ECs) released microvesicles (EMVs) could modulate the functions of target cells by transferring their microRNAs (miRs). We have reported that miR-125a-5p protected EC function. In this study, we determined whether EMVs provided beneficial effects on ECs by transferring miR-125a-5p. Human brain microvessel ECs were transfected with miR-125a-5p mimic or miR-125a-5p short hairpin RNA to obtain miR-125a-5p overexpressing ECs and miR-125a-5p knockdown ECs, and their derived EMVs. For the functional study, ECs or hypoxia/reoxygenation injured ECs were coincubated with various EMVs. The survival and angiogenic function of ECs were measured. Western blot and quantitative real time polymerase chain reaction (qRT-PCR) were used for measuring the levels of phosphoinositide 3-kinase (PI3K), phosphorylation-Akt (p-Akt)/Akt, p-endothelial nitric oxide synthase (p-eNOS), cleaved caspase-3, and miR-125a-5p. PI3K inhibitor was used for pathway analysis. EMVs promoted the proliferation, migration, and tube formation ability of ECs, and alleviated the apoptotic rate of ECs. These effects were associated by an increase in p-Akt/Akt and p-eNOS, and a decrease in cleaved caspase-3 could be abolished by LY294002. Overexpression or downregulation of miR-125a-5p in EMVs promoted or inhibited those effects of EMVs. EMVs could enhance the survival and angiogenic function of ECs via delivering miR-125a-5p to modulate the expression of PI3K/Akt/eNOS pathway and caspase-3. K E Y W O R D S brain endothelial cells (ECs), cell function, hypoxia and reperfusion, microvesicles (MVs), miR-125a-5p J Cell Biochem. 2019;120:3160-3172. wileyonlinelibrary.com/journal/jcb 3160 |

Exosomes Derived from Mesenchymal Stem Cells Ameliorate Hypoxia/Reoxygenation-Injured ECs via Transferring MicroRNA-126

“…The number and size of MSC-EXs were detected by the NanoSight NS300 instrument as we previously described [21, 24]. In this study, diluted suspensions containing MSC-EXs were loaded into the sample chamber and the camera level was maintained at 9 for light scatter mode.…”

“…Cells from 3 wells were counted at each time point, and the experiment was repeated 6 times. For pathway exploration, cells were preincubated with PI3K inhibitor (LY294002; 20 μ M; Selleckchem) for 2 h [21] and then incubated with MSC-EXs miR-126 .…”

“…The migration of ECs was measured by scratch assay as we previously described [21, 24]. Briefly, after H/R added with a culture medium (vehicle), MSC-EXs, or MSC-EXs miR-126 , a scratch was made through the cultured ECs.…”

“…The PI3K/Akt/eNOS signal pathway and caspase-3 activation have been shown to be involved in EC survival, angiogenesis, and apoptosis processes [21]. Studies showed that miR-126 can regulate the PI3K/Akt/eNOS signal pathway in human cardiac microvascular ECs and human coronary artery ECs [14, 15].…”

An Insight into the microRNAs Associated with Arteriovenous and Cavernous Malformations of the Brain

“…Furthermore, the overexpression of miR-125a-5p resulted in a reduction of seizures and lessened hippocampal inflammation via the suppression of its target gene, calmodulindependent protein kinase IV (CAMK4) [139]. Pan et al showed that microvesicles carrying miR-125a-5p fostered the proliferation, migration, the ability to form tubes of transfected microvessel BECs, improved their apoptotic rate [141], while an earlier study discovered the role of this microRNA in protecting BECs from the injurious effects of oxidized low-density lipoproteins such as, senescence, decrease of nitric oxide (NO), and upsurge of reactive oxygen species (ROS) [142]. It is therefore likely that this microRNA plays a distinctive role in CCM patients presenting with epilepsy, or it could hypothetically act as a biomarker in identifying the cases liable to develop CCM-stimulated seizures.…”