Qingre Huoxue Formula Inhibits Inflammatory Injury in Collagen-Induced Arthritis Rats by Regulating METTL3
LI Lu
ZHANG Xieyu
YANG Yue
NI Qing
CAO Wei
Abstract：Objective To investigate the potential mechanism of Qingre Huoxue formula in treating rheumatoid arthritis(RA).Methods A collagen-induced arthritis(CIA)model was established using 30 SD rats,which were then randomly divided into a model group,a methotrexate group,and a Qingre Huoxue formula group.An additional 10 SD rats were used as a normal control group.The intervention lasted for 4 consecutive weeks.Assessment indicators included measuring joint swelling in rats every 3 days.At the end of the experiment,serum inflammatory markers—anti-cyclic citrullinated peptide antibody(anti-CCP antibody),interleukin(IL)-10,and IL-17—were detected using ELISA.HE staining was used to observe pathological changes in ankle joints and synovial tissues.RT-qPCR and Western blot were employed to detect the gene and protein expression levels of methyltransferase-like protein 3(METTL3).Network pharmacology was used to analyze the effective active components of Qingre Huoxue formula in treating RA and to predict related pathway targets.Results Compared with the model group,Qingre Huoxue formula effectively reduced joint swelling in rats(P<0.01).HE staining showed that the synovial structure in the Qingre Huoxue formula group tended to be more regular,inflammatory infiltration was reduced,and joint damage was significantly alleviated.Inflammatory factor detection revealed downregulation of anti-CCP antibody and IL-17,and upregulation of IL-10 in the Qingre Huoxue formula group(P<0.05).RT-qPCR and Western blot results indicated that Qingre Huoxue formula effectively inhibited the overexpression of METTL3 at both mRNA and protein levels(P<0.05).Network pharmacology analysis showed that the potential core active components of Qingre Huoxue formula in treating RA mainly included quercetin,β-sitosterol,luteolin,stigmasterol,and apigenin.Key targets involved IL6,tumor necrosis factor(TNF),IL1B,AKT serine/threonine kinase 1(AKT1),signal transducer and activator of transcription 3(STAT3),among others.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses identified 177 related pathways,primarily involving inflammation and immune responses(IL-17 signaling pathway,TNF signaling pathway,Toll-like receptor signaling pathway,etc.).Conclusion This study suggests that Qingre Huoxue formula can effectively alleviate arthritis symptoms and pathological damage in CIA model rats.Its mechanism may be related to inhibiting METTL3-mediated RNA methylation and thereby regulating inflammatory signaling pathways.
Keywords：Active RAQingre Huoxue FormulaMETTL3m6A methylation modificationInflammation
Publication Date：2026-03-28
Online Publishing Date：2026-03-31(First online date of this platform, not the publication date of the document)
Pages：5( 584-588 )