Acute ischemic stroke (AIS) is one of the leading causes of death and disabilities, and as such, it is of utmost importance to identify novel treatment options. Current acute treatments for AIS are limited to either thrombectomy or thrombolysis, both of which must be initiated within 4.5-6 hours of symptom onset. Remote ischemic conditioning (RIC) is a promising non-invasive treatment that is thought to activate the body’s own protective mechanisms against damaging ischemia through circulating microRNAs (miRNAs). Here, we investigate the transcriptional changes in human brain microvascular endothelial cells (HBMECs) transfected with four selected RIC-upregulated miRNAs (RIC-miRNAs), miR-16-5p, miR-144-3p, miR-182-5p, and miR-451a, under oxygen and glucose deprivation (OGD) - mimicking the initial stages of AIS. Pronounced transcriptional changes were present after RIC-miRNA transfection, with 149 unique downregulated and 212 upregulated differentially expressed genes in HBMECs after OGD and RIC-miRNA transfection compared to all other conditions. These genes were involved in cell cycle regulation, DNA replication and pathways of energy metabolism. However, we saw no direct effect on cell viability after RIC-miRNA transfection and OGD. In conclusion, our study suggests that the selected RIC-miRNAs regulate pathways that may facilitate endothelial cell recovery and remodeling events from ischemic damage, offering new therapeutic avenues for AIS.