Mechanism underlying ozonated water intra-articular injection in modulating miR-214-3p to alleviate cartilage damage in knee osteoarthritis rats via the nuclear factor-κB signaling pathway
10.3969/j.issn.1671-7856.2025.06.008
- VernacularTitle:基于NF-κB信号通路探索臭氧水关节腔注射调控miR-214-3p对KOA大鼠软骨损伤的作用机制
- Author:
Min LIU
1
;
Weiqi LI
;
Huiru DOU
;
Youlong ZHOU
Author Information
1. 河南中医药大学,郑州 450002
- Publication Type:Journal Article
- Keywords:
knee osteoarthritis;
ozonated water;
cartilage repair;
articular cavity injection;
miR-214-3p;
NF-κB signaling pathway
- From:
Chinese Journal of Comparative Medicine
2025;35(6):74-83
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the mechanism underlying the effect of intra-articular injection of ozonated water on cartilage repair in knee osteoarthritis(KOA)rats via the miR-214-3p/nuclear factor(NF)-κB signaling pathway,to provide a theoretical basis for the clinical application of ozonated water.Methods Sixty male Sprague-Dawley rats were divided randomly into ozonated and experimental groups.Rats in the experimental group were further divided randomly into model,ozonated water,ozonated water+AAV-NC,and adeno-associated virus with ozonated water+AAV-miR-214-3p inhibitor groups.The articular cartilage repair status was evaluated after the respective treatments,using articular cartilage surface gross scoring,hematoxylin/eosin staining,and modified Mankin's score.Expression levels of interleukin(IL)-1β,tumor necrosis factor(TNF)-α,and matrix metalloproteinase(MMP)-13 were detected by enzyme-linked immunosorbent assay(ELISA).Protein expression levels of IκB kinase(IKK)β,p65,and phospho(p)-IκBα were detected in cartilage tissues by Western blot,and gene expression levels of IKKβ,p65,IκBα,and miR-214-3p were detected by real-time polymerase chain reaction(RT-PCR).Results Rats in all groups showed different degrees of cartilage damage compared with the control group,and the scores were significantly lower in the ozonated water and ozonated water+AAV-NC groups compared with the ozonated water+AAV-miR-214-3p inhibitor group(P＜0.05).IL-1β,TNF-α,and MMP-13 levels detected by ELISA were significantly lower in cartilage tissues from rats in the ozonated water and ozonated water+AAV-NC groups compared with the model and ozonated water+AAV-miR-214-3p inhibitor groups(P＜0.05).According to RT-PCR,IKKβ and p65 mRNA levels were significantly down-regulated while IκBα mRNA was significantly up-regulated in cartilage tissues from rats in the ozonated water and ozonated water+AAV-NC groups compared with the model and ozonated water+AAV-miR-214-3p inhibitor groups(P＜0.05).Similarly,IKKβ,p65,and p-IκBα protein expression levels detected by Western blot were significantly down-regulated while IκBα protein expression levels were up-regulated in cartilage tissues from rats in the ozonated water and ozonated water+AAV-NC groups compared with the model and ozonated water+AAV-miR-214-3p inhibitor groups(P＜0.05).Conclusions Ozone hydrotherapy significantly ameliorated cartilage damage in KOA rats,possibly via inhibiting activation of the NF-κB signaling pathway through up-regulation of miR-214-3p expression and promoting cartilage repair in KOA rats.
