基于网络药理学与细胞实验探讨加味黄芪桂枝五物汤治疗类风湿关节炎的作用机制OA

Objective:To explore the action mechanism of Supplemented Astragalus and Cinnamon Twig Five Substances Decoction in the treatment of rheumatoid arthritis(RA)based on network pharmacology and cell experiments.Methods:The active components and tar-gets of Supplemented Astragalus and Cinnamon Twig Five Substances Decoction were retrieved and screened through Traditional Chi-nese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and BATMAN-TCM databases.The targets of RA were retrieved from GeneCards and OMIM databases,and the targets of Supplemented Astragalus and Cinnamon Twig Five Substances Decoc-tion and RA were taken as the intersection.The gene ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis of the intersection genes were performed through Bioconductor platform and R software(P<0.01).Twenty SD rats were ran-domly divided into the group of Supplemented Astragalus and Cinnamon Twig Five Substances Decoction(1.8 g·kg-1)and the nor-mal group,with 10 rats in each group.After 7 days of gavage,the drug-containing serum with volume fractions of 5%,10%and 20%were prepared respectively.The proliferation of MH7A cells was detected by CCK-8 method;the apoptosis of cells was detected by An-nexin V-PI double staining method;the protein expression levels of phosphatidylinositol-3-hydroxykinase(PI3K)and phosphorylated protein kinase B(p-AKT)were detected by Western Blot.ELISA was used to detect the contents of interleukin-17(IL-17),interleu-kin-6(IL-6),and tumor necrosis factor alpha(TNF-α).Results:There were 141 active components and 303 targets in Supplemented Astragalus and Cinnamon Twig Five Substances Decoction.There were 4 937 targets in RA,and 127 targets were shared by Supplemen-ted Astragalus and Cinnamon Twig Five Substances Decoction and RA.GO analysis mainly involved the response to lipopolysaccharide,negative feedback regulation of cell proliferation,response to cytokines,regulation of cell activation,and positive regulation of apoptosis.KEGG pathway mainly included PI3K/AKT signaling pathway,TNF signaling pathway,and calcium signaling pathway.At the 12th,24th,and 48th hour after cell culture,compared with the control group,the cell proliferation inhibition rate of Supplemented Astragalus and Cinnamon Twig Five Substances Decoction high-dose group,medium-dose group,and low-dose group was increased respectively(P0.05).At 24th hour after cell culture,compared with the control group,the apoptosis rate of Supplemented Astragalus and Cinnamon Twig Five Substances Decoction low-dose group,medium-dose group,and high-dose group was increased respectively(P<0.05).Com-pared with the control group,the expression levels of PI3K and p-AKT in the model group were increased(P<0.05).Compared with the model group,the expression levels of PI3K and p-AKT in the Supplemented Astragalus and Cinnamon Twig Five Substances Decoc-tion low-dose group,medium-dose group,and high-dose group were decreased respectively(P<0.05).Compared with the control group,the contents of IL-17,TNF-α,and IL-6 in the model group were increased(P<0.05).Compared with the model group,the con-tents of IL-17,TNF-α and IL-6 in the Supplemented Astragalus and Cinnamon Twig Five Substances Decoction low-dose,medium-dose and high-dose groups were decreased(P<0.05).Conclusion:Supplemented Astragalus and Cinnamon Twig Five Substances Decoction may inhibit cell proliferation,promote cell apoptosis,and reduce inflammatory factor levels by regulating PI3K/AKT signaling pathway and other related pathways,thereby exerting anti-RA effects.