Neuroprotective effects of microRNA-140-5p on ischemic stroke in mice via regulation of the TLR4/NF-κB axis

Abstract

Background and aimIschemic stroke is one of the main causes of death worldwide and permanent global disability. On the basis of existing literature data, the study was carried out in an effort to explore how miR-140-5p affects ischemic stroke and whether the mechanism relates to toll-like receptor-4 (TLR4) and nuclear factor-kappa B (NF-κB). MethodsFirstly, middle cerebral artery occlusion (MCAO) was performed to establish mouse models of ischemic stroke in vivo, while primary neurons were exposed to oxygen-glucose deprivation (OGD) to set up an ischemic stroke model in vitro. RT-qPCR was then applied to detect the miR-140-5p expression patterns, whereas Western blot was adopted to detect the expression patterns of TLR4, NF-κB, and apoptosis-related factors. In addition, based gain-function of experiments using miR-140-5p mimic and TLR4 over-expression plasmid, neurological function score, TTC staining, TUNEL staining, as well as flow cytometry were carried out to evaluate the effects of miR-140-5p and TLR4 on MCAO mice and OGD neurons. Moreover, dual-luciferase reporter assay was applied to validate the targeting relationship between miR-140-5p and TLR4. ResultsInitial findings revealed that miR-140-5p was poorly-expressed, while TLR4 was highly-expressed in ischemic stroke. It was verified that miR-140-5p targeted TLR4 and downregulated its expression. MiR-140-5p over-expression was observed to inhibit the apoptosis of neurons under OGD exposure and restrain the progression of ischemic stroke, while TLR4 over-expression promoted the apoptosis and disease progression. Besides, miR-140-5p over-expression led to a decrease in NF-κB protein levels, which were increased by TLR4 over-expression. ConclusionIn conclusion, our data indicates that miR-140-5p over-expression may be instrumental for the therapeutic targeting of ischemic stroke by alleviating neuron injury with the involvement of the TLR4/NF-κB axis.