Main active components and mechanism of Sanmiao Pills in treating rheumatoid arthritis
UPLC-Q-TOF-MS was employed to analyze the chemical components of Sanmiao Pills(SMP)and network pharmacology and animal experiments were employed to investigate the mechanism of SMP in treating rheumatoid arthritis(RA).DisGeNET and SwissTargetPrediction were used to identify the overlapping targets between RA and SMP.Subsequently,a protein-protein interaction(PPI)network was built based on the key targets.The identified targets were further subjected to Gene Ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway annotation in MATESCAPE.Cytoscape 3.8 was used to build the"component-target-pathway"network,on the basis of which the main active components,key targets,and pathways involved in the treatment of RA with SMP were predicted.Additionally,an HPLC-DAD method was established to simultaneously determine the content of berberine,atractylolide Ⅰ,atractylolide Ⅱ,ginsenoside Ro,and notoginsenoside R1 in SMP.Furthermore,a rat model of collagen-induced arthritis(CIA)was established and the behavior test was carried out.Hematoxylin-eosin(HE)staining was conducted to reveal the pathological changes in the synovial tissue.Western blot was employed to determine the expression levels of proteins in the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)pathway.A total of 126 chemical components were identified in SMP,and 332 targets were shared by SMP and RA.The PPI network predicted the key targets including AKT1 and TNF,which were mainly involved in 20 pathways including the PI3K/AKT pathway.Five components showed good linear relationships within the test concentration ranges,with RSD≤2%.The animal experiments revealed that SMP ameliorated synovial hyperplasia and down-regulated the expression levels of PI3K and p-AKT in CIA rats.The findings suggest that SMP may alleviate RA by modulating the PI3K/AKT pathway.
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