Study On The Anti-RA Mechanism Of FuFangDuZhengPian Based On Network Pharmacology Of Blood Components And 16SrDNA

Objective: This study usesd network pharmacology of blood components and 16 S r DNA analysis methods to comprehensively explore the therapeutic efficacy,molecular mechanisms,and regulatory effects on intestinal microbiota of Fufang Duzheng Pian(referred to as FFDZP)in the treatment of rheumatoid arthritis(RA).Methods:(1)A collagen-induced arthritis model was constructed in rats,and they were given FFDZP continuously for three weeks.The pathological changes in the tissues of each group of rats were observed,and indicators such as body weight,toe thickness,and the content of TNF-α and IL-1β in the serum were measured.(2)UPLC/ MS-MS technology was used to analyze the blood components of FFDZP,and network pharmacology analysis method was used to predict the targets of these components.Protein interaction network(PPI)and ’ disease-target-component ’network were constructed to analyze the main components and targets.GO and KEGG functions of common targets were analyzed,and molecular docking simulation was carried out.(3)16Sr DNA was used to explore the differences of intestinal microorganisms in each treatment group.Firstly,the operational taxonomic units(OTUs)were classified and counted.On this basis,their α diversity and βdiversity were analyzed,and the differential microorganisms were compared.Results:(1)Compared with the normal group,the body weight of rats after modeling decreased,but the body weight of rats increased after treatment with FFDZP and MTX groups;compared with the model group,the toe thickness and serum concentrations of TNF-α and IL-1β in the FFDZP and MTX groups decreased,and the pathological manifestations were alleviated.(2)A total of 105 metabolites were identified,and 18 blood components were obtained through cometabolite analysis.The main components of FFDZP were 3,4’-dihydroxyflavone,7,4-dihydroxyflavone,α-acetylcinnamic acid,dehydroabietic acid,and 3,4-O-dicaffeoylquinic acid methylester.VEGFA,TP53,EGFR,and AKT1 were the main therapeutic targets of FFDZP for treating rheumatoid arthritis.GO and KEGG analyses identified 456 BPs,54 CCs,117 MFs,and 128 KEGG pathways.Molecular docking showed that there was good binding efficiency between the core targets and the main targets(3)Intestinal microbial sequencing showed that more than 97% of the species were clustered in4,796 OTUs.There was no significant difference in species diversity among the three groups,but the population structure changed.Relative abundance analysis showed that the intestinal microbial imbalance in CIA rats was characterized by a significant increase in Proteobacteria and Bacteroidetes and a decrease in Firmicutes.FFDZP could significantly reduce the abundance of Proteobacteria and Pseudomonas and increase the relative abundance of Prevotella and Alloprevotella.Conclusion:(1)FFDZP can improve joint inflammation in CIA rats,reduce the levels of TNF-α and IL-1β in serum,and reduce synovial hyperplasia.(2)FFDZP may treat RA by inhibiting abnormal activation of the PI3K-Akt signaling pathway,suppressing abnormal activation of the MAPK signaling pathway,alleviating local hypoxia in the joint cavity,and regulating the normal expression of TP53.(3)FFDZP can regulate the intestinal microecological balance,inhibit the abundance of harmful bacteria such as Proteobacteria,and increase the abundance of beneficial bacteria such as Prevotella and Alloprevotella.