Mechanism of miR-146a Mediated by Withaferin A on Alleviating Hippocampal Neuronal Damage in Rats with Cerebral Ischemia-Reperfusion Injury
LIU Hongqi
DING Feng
YAN Haiqing
Abstract：Objective To obvserve the effect of withaferin A on hippocampal neuronal damage in rats with cerebral ischemia-reperfusion,and explore its mechanism of mediating microribonucleic acid-146a(miR-146a)action.Methods Fifty SD rats were used to establish cerebral ischemia-reperfusion models and randomly divided into 5 groups using a number table.The other 10 SD rats underwent the same modeling procedure as before,but were not ligated with the cerebral artery and were recorded as a sham operation group.Withaferin A group was injected with 50 mg·kg-1 of withaferin A via tail vein,the miR-146a upregulation group received tail vein injection of adenovirus 200 μL carrying miR-146a mimetic substance,and the empty-load group received tail vein injection of adenovirus 200 μL without carrying miR-146a simulant,and the nimodipine group was injected with 1.0 mg·kg-1 nimodipine through the tail vein,while the model group and sham operation group were injected with 1 mL·(100 g)-1physiological saline through the tail vein.After 48 hours,real-time reverse transcription polymerase chain reaction(RT-qPCR)was used to detect the expression of miR-146a in the hippocampus tissues of each group,and the neurological function of each group of rats was evaluated.Enzyme linked immunosorbent assay was used for detection of Toll like receptor 4(TLR4)and nuclear transcription factors in serum-κB p65(NF-κB p65),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)levels.Hematoxylin eosin(HE)staining was used to observe pathological changes in rat hippocampal tissue,and terminal dexynucleotidyl transferase(TdT)-mediated dUTP nick end labeling(TUNEL)was used to detect hippocampal neuronal apoptosis.RT-qPCT detection of TLR4 and NF-κB p65,TNF-α and IL-1β gene expressions,immunoblotting was used to detect the expressions of the aforementioned proteins.Results Compared with the sham operation group,the model group had neurological function scores,hippocampal neuron apoptosis rate,serum inflammatory index levels,miR-146a,TLR4,NF-κB p65,TNF-α and IL-1β expressions of genes and proteins increased(P＜0.05).Compared with the model group,the neural function scores,hippocampal neuron apoptosis rates,serum inflammatory index levels,TLR4,NF-κB p65,TNF-α and IL-1β expressions in the withaferin A group,nimodipine group,and miR-146a upregulation group decreased(P＜0.05).Compared with the nimodipine group,the withaferin A group and the miR-146a upregulation group had neurological function scores,hippocampal neuron apoptosis rates,serum inflammatory index levels,TLR4,NF-κB p65,TNF-α and IL-1βexpressions decreased(P＜0.05).Compared with the miR-146a upregulation group,the above indexes except miR-146a were reduced in the withaferin A group(P＜0.05).The nimodipine group,miR-146a upregulation group,and TLR4,NF-κB p65,TNF-α and IL-1β group can all alleviate pathological changes in the hippocampus,but withaferin A group had the lightest pathological changes.Conclusion Withaferin A can upregulate miR-146a to alleviate pathological changes in hippocampal tissue and reduce neuronal apoptosis in rats with cerebral ischemia-reperfusion,which is related to reducing inflammatory response.
Keywords：cerebral ischemia-reperfusionwithaferin Amicroribonucleic acid-146ahippocampusneuronsinflammatory reaction
Publication Date：2025-05-13
Online Publishing Date：2026-09-12(First online date of this platform, not the publication date of the document)
Pages：8( 1577-1584 )