Study on the Mechanism of miR-381 Ameliorates Ischemic Stroke by Modulating the NF-κB Signaling Pathway
10.11969/j.issn.1673-548X.2025.08.016
- VernacularTitle:miR-381调节NF-κB信号通路改善缺血性脑卒中的机制研究
- Author:
Youtao ZHANG
1
;
Liping DONG
1
;
Shutie LI
1
Author Information
1. 075000 张家口,河北北方学院附属第一医院老年医学科
- Publication Type:Journal Article
- Keywords:
Ischemic stroke;
miR-381;
Inflammatory response;
Apoptosis
- From:
Journal of Medical Research
2025;54(8):94-100
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the role and mechanism of miR-381 in the pathogenesis of ischemic stroke(IS).Methods A total of 90 patients with IS treated in the First Affiliated Hospital of Hebei North University between January 2020 and December 2022 were selected as the IS group,while 110healthy volunteers were selected as the control group.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the difference of serum miR-381 between the two groups,and enzyme-linked immu-nosorbent assay(ELISA)was used to measure the differences of inflammatory markers interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)between the groups.The CCK-8 assay was used to evaluate the effect of miR-381 on the ac-tivity of human umbilical vein endothelial cell(HUVEC).Use TargetScan,Western blot and luciferase reporter assays to validate whether inhibitor of kappa B kinase(IKK)was a target gene regulated by miR-381.An in vitro model of oxygen-glucose deprivation(OGD)was constructed,and the effects of miR-381 over-expression or inhibition on apoptosis-and inflammation-related proteins were de-tected by RT-qPCR and Western blot.The diagnostic value of miR-381 for IS was evaluated using receiver operating characteristic curve(ROC).Results Compared with the control group,the serum miR-381 levels in the IS group were significantly down-regulated(P＜0.05).In comparison to the mild IS group or small infarct volume group,serum miR-381 levels were significantly lower in patients with moderate to severe IS or large infarct volumes(P were＜0.05,＜0.01).Additionally,compared with the control group,the serum levels of IL-1 β,IL-6 and TNF-α in the IS group were significantly higher(P were＜0.01,＜0.05,＜0.01).Furthermore,com-pared to the mild IS group or small infarct volume group,the serum levels of IL-1 β,IL-6,and TNF-α were significantly elevated in patients with moderate to severe IS group or large infarct volume group(P were＜0.01,＜0.05).CCK-8 results indicated that miR-381 promoted the viability of HUVEC.Dual-Luciferase reporter gene assay results demonstrated that IKK was a direct target of miR-381.Western blot results showed that over-expression of miR-381 inhibited the activation of p65 and inhibitor of NF-κB(IKB)pro-teins,promoted the expression of the anti-apoptotic protein B-cell lymphoma 2(Bcl-2),and suppressed the expression of the pro-apoptotic protein Bcl-2 associated X(BAX).RT-qPCR results showed that over-expression miR-381 reduced the levels of NF-κB mRNA and BAX mRNA,and increased the levels of Bcl-2mRNA.ELISA results indicated that over-expression miR-381 decreased the serum levels of TNF-α,IL-6 and IL-1 β(P were＜0.05,＜0.01,＜0.01).ROC curve analysis showed that the sensitivity and specificity of miR-381 for diagnosing IS were 76.23％and 81.25％,respectively.Conclusion miR-381 can inhibit HUVEC apopto-sis and inflammatory responses by targeting IKK,indicating its potential as a therapeutic target for IS.
