AntimiR-223-5p ameliorates ischemic damage and improves neurological function by preventing NCKX2 downregulation after ischemia in rats

Abstract It has been demonstrated that the K+-dependent Na+/Ca2+ exchanger, NCKX2, is a new promising stroke neuroprotective target. However, since no pharmacological activator of NCKX2 is still available, microRNA may represent an alternative method to modulate NCKX2 expression. In particular, by bioinformatics analysis, miR-223-5p emerged as a possible modulator of NCKX2 expression. In the light of these premises, the aims of the present study were: (1) to evaluate miRNA223-5p and NCKX2 expression in temporoparietal cortex and striatum of rats subjected to transient middle cerebral artery occlusion; (2) To evaluate whether miR223-5p targets 3’UTR of NCKX2 transcript; (3) to evaluate the effect of miR-223-5p modulation on brain ischemic volume and neurological deficits. Our results showed that miR-223-5p expression increased in a time-dependent manner in the striatum of ischemic rats in parallel with NCKX2 downregulation and the transfection of cortical neurons with miR-223-5p induced a reduction of NCKX2 expression. Moreover, luciferase assay showed that miR-223-5p specifically interacts with NCKX2 3’UTR subregion (+7037 to +8697) thus repressing NCKX2 translation. More interestingly, intracerebroventricular infusion of antimiR-223-5p prevented NCKX2 downregulation after ischemia thus promoting neuroprotection. The present findings support the idea that blocking miR-223-5p by antimiRNA is a reasonable strategy to reduce the neurodetrimental effect induced by NCKX2 downregulation during brain ischemia.