Abstract
Endothelial progenitor cells (EPCs) are multipotential stem cells considered to have immense clinical value for revascularization. However, the clinical application of EPCs has been hampered by their clinical potency in ischemic anoxic environments. This study aimed to explore the effect of microRNA-210 (miR-210) on EPCs under oxygen-glucose deprivation (OGD) conditions. We generated a model of EPCs cultured under OGD conditions to simulate ischemia and explore the expression of miR-210 in vitro. With longer exposure to hypoxia, we found that miR-210-3p expression was highly upregulated in OGD groups compared to that in controls from 4 to 24 h, but not miR-210-5p. We then transfected a miR-210-3p mimic and inhibitor into EPCs, and after 24 h, we exposed them to OGD conditions for 4 h to simulate ischemia. We detected miR-210 by real-time polymerase chain reaction (RT-PCR) and tested the proliferation, migration, and tube formation of normal EPCs and OGD-treated EPCs by CCK-8, transwell chamber, and Matrigel assays, respectively. The direct targets of miR-210-3p were predicted using miRWalk. Compared to that in normal EPCs, higher miR-210-3p expression was found in OGD-treated EPCs (p < 0.05). Moreover, upregulation of miR-210-3p was found to promote proliferation, migration, and tube formation in EPCs under normal and OGD conditions (p < 0.05), whereas down-regulation inhibited these abilities in OGD-treated EPCs (p < 0.05). Repulsive guidance molecule A (RGMA), a negative regulator of angiogenesis, was predicted to be a target of miR-210-3p. Accordingly, upregulation of miR-210-3p was found to inhibit its expression at the protein level in OGD-treated EPCs, whereas downregulation of miR-210-3p inhibited its expression (p < 0.05). A dual-luciferase reporter system confirmed that RGMA is a direct target of miR-210-3p. MicroRNA-210-3p overexpression enhances the angiogenic properties of OGD-treated EPCs by inhibiting RGMA.
Highlights
Characterized by high mortality, morbidity, and disability, ischemic stroke causes immense health and economic burdens to families and society (Benjamin et al, 2017)
We investigated the possible functions of miR210 in Endothelial progenitor cells (EPCs) under the hypoxic condition and further explored the potential underlying molecular mechanism in controlling cellular behavior
We subjected the fourth generation of EPCs to direct staining (Figure 1B) and under the microscope, we could see that nearly half of EPCs were stained for CD34 and CD31, whereas most of the EPCs were stained for KDR and Von Willebrand Factor (vWF)
Summary
Characterized by high mortality, morbidity, and disability, ischemic stroke causes immense health and economic burdens to families and society (Benjamin et al, 2017). Previous research has demonstrated that EPCs can differentiate into mature endothelial cells (ECs) to take part in angiogenesis and vasculogenesis after cerebral ischemic injury (Chong et al, 2016). They have been shown to participate in the repair of dysfunctional endothelia and to suppress endothelial injury through their direct incorporation into newly forming vessels or the secretion of pro-angiogenic growth factors or cytokines (Asahara et al, 2011). Our study began with the key point of improving the ability of EPCs to resist hypoxia and ischemia
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