Objective: The study aimed to explore the mechanism of total flavonoids of Rhizoma Drynariae (TFRD) in the treatment of rheumatoid arthritis (RA) based on network pharmacology and experimental validation. Methods: The active components of TFRD were identified from TCMSP and TCMID databases. Relevant targets of the active compounds of TFRD and RA-related targets were predicted by public databases online. A component-target (C-T) regulatory network was constructed by Cytoscape. The genes of TFRD regulating RA were imported into STRING database to construct a protein-protein interaction (PPI) network in order to predict the key targets. KEGG enrichment analysis was performed to predict the crucial mechanism of TFRD against RA. The active components of TFRD underwent molecular docking with the key proteins. Collagen-induced arthritis (CIA) model of rats and inflammatory factors-stimulated fibroblast-like synoviocytes were used in vivo and in vitro to validate the efficacy and predicted critical mechanisms of TFRD. Results: Network Pharmacology analysis revealed that TFRD had 14 active compounds, corresponding to 213 targets, and RA related to 2814 genes. There were 137 intersection genes between TFRD and RA. KEGG indicated that therapeutic effects of TFRD on RA involves T cell receptor signaling pathway, Th17 cell differentiation, IL-17 signaling pathway, TNF signaling pathway, MAPK signaling pathway and PI3K/AKT signaling pathway. In vivo experiments suggested TFRD can alleviate the inflammatory response, joint swelling and synovial abnormality of CIA rats. TFRD contributed to the decrease of Th17 cells and the down-regulated secretion of IL-17A and TNF-α of activated lymphocyte in CIA model. In vitro experiments confirmed TFRD can effectively inhibit the inflammatory response of fibroblast-like synoviocytes and suppress the abnormal activation of MAPK, PI3K/AKT and NFκB signaling pathways. Conclusion: The treatment of RA with TFRD is closely related to inhibiting Th17 differentiation and inflammatory response of synoviocytes.

Objective: Previous clinical studies have found that total flavonoids of Rhizoma Drynariae (TFRD) have a good therapeutic effect on osteoarthritis (OA), but its therapeutic mechanism needs further research. Methods: OA rat model was established by Hulth method and was intervened by TFRD. Pathological assessments were conducted to assess the protective effect of TFRD on cartilage. Serum metabolomics and network pharmacology were detected to predict the mechanism of TFRD treating OA. In further experiments, molecular biology experiment was carried out to confirm the predicted mechanisms in vivo and in vitro. Results: TFRD can effectively reduce chondrocyte apoptosis and cartilage degeneration in OA model rats. Serum metabolomics revealed that the intervention effect may be closely related to arachidonic acid metabolism pathway. Network pharmacologic prediction showed that COX-2 was the key target of TFRD in treating OA, and its mechanism might be related with NFκB, apoptosis, AMPK and arachidonic acid metabolism pathway. In vivo experiments indicated that TFRD can inhibit the abnormal expression of COX-2 mRNA in OA model rats. In the in vitro studies, the expression of COX-2 mRNA and protein increased, AMPK phosphorylation was inhibited, and NFκB signaling pathway was activated in IL-1β-induced chondrocytes, and these changes can be reversed by TFRD. After the activation of AMPK signaling pathway or the block-down of NFκB signaling pathway, the effect of TFRD on COX-2 mRNA expression was significantly weakened. Conclusion: TFRD can inhibit COX-2-mediated arachidonic acid metabolites, and its mechanism is closely related to AMPK/NFκB pathway, which may be a key mechanism in the treatment of OA.

Objective: The mechanism of Wendan Decoction (WDD) against Generalized Anxiety Disorder (GAD) was predicted by network pharmacology and validated by in vivo and in vitro experiments. Methods: The targets of WDD for the treatment of GAD were obtained by a search of online databases. Further, PPI network and KEGG enrichment were used to identify the key targets and pathways. Ultimately, these key targets and pathways were validated by in vivo experiments on GAD mice modeled by repeated restraint stress (RRS) and in vitro experiments on inflammatory factor stimulated BV-2 cells. Results: Through searching the databases, the 137 ingredients of WDD that correspond to 938 targets and 4794 targets related to GAD were identified. Among them, 569 overlapping targets were considered as the therapeutic targets of WDD for GAD. PPI analysis showed that the inflammation-related proteins IL-6, TNF, SRC and AKT1 were the key targets, and KEGG enrichment suggested that PI3K/AKT and MAPK signaling pathways were key pathways of WDD in the treatment of GAD. In vivo experiments, RRS mice exhibited abnormality in behavioristics in open field test (OFT) and elevated plus maze (EPM) and increases in serum corticosterone and the percentage of lymphocytes positive for IL-6 in peripheral blood. These abnormal changes can be reversed by WDD and the positive control drug paroxetine. In vitro experiments, WDD can inhibit IL-6 induced activation of PI3K/AKT and MAPK signaling pathways in BV2 cells, and suppress the ensuing release of inflammatory factors TNF-α, IL-1β and PGE₂, and showed a dose-dependent effect. Conclusion: WDD is able to resist GAD by relieving inflammatory response in peripheral and central system.

Background: The co-occurrence of rheumatoid arthritis and depression typically exacerbates pain and leads to a range of adverse consequences, becoming a research hotspot in recent years. This study conducted the systematic retrieval of relevant articles within the past five years and employed bibliometric methods for scientometric analysis. Methods: Setting the keywords “Rheumatoid arthritis”, “Depression” and “Depressive Disorder”, relevant literature published between 2019 and 2023 was retrieved from the Web of Science database. Subsequently, the core information from the literature was subjected to visual analysis via CiteSpace software and bibliometric techniques. Results: A total of 974 articles related to rheumatoid arthritis and depression were identified through the search strategy, and 877 articles were retained for further analysis after duplicates. The United States (n=173), England (n=82), China (n=69), Canada (n=68), and Germany (n=54) ranked top five countries by publication count. The King’s College London was the leading institution with the highest number of publications (n = 20). LANCET PSYCHIATRY was the most frequently cited journal (n = 72) despite having only one article. The top five authors with the largest number of publications include CHARLES N BERNSTEIN (n=14), RUTH ANN MARRIE (n=13), JOHN D FISK (n=12), CAROL A HITCHON (n=12) and SCOTT B PATTEN (n=12), and all these are based in Canada. The keywords with a centrality score exceeding 0.1 were depression, rheumatoid arthritis, symptom, quality of life, impact, fibromyalgia, disease activity, prevalence, inflammation, health, anxiety, pain, fatigue, disease, arthritis and disability. Conclusion: Related research between the co-occurrence of rheumatoid arthritis and depression was a persistent hotspot, but it still lacks of international collaboration and in-depth mechanistic exploration.

Rhizoma Drynariae has been widely used for the treatment of osteoarthritis (OA), but its potential targets and molecular mechanisms remain to be further explored. Targets of Rhizoma Drynariae and OA were predicted by relevant databases, and a protein-protein interaction (PPI) network was constructed to identify key targets. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was performed to obtain related pathways and then select significant pathways associated with OA. The OA chondrocyte model was established by inflammatory factor-induced SW1353 chondrocytes, and molecular docking was conducted to verify the above theoretical prediction. The results showed that a total of 86 Rhizoma Drynariae-OA interaction targets were identified, among which IL-6 and AKT1 were the key targets in the PPI network. Luteolin was the most critical component of Rhizoma Drynariae. KEGG results indicated that the effects of Rhizoma Drynariae on OA are associated with the PI3K/AKT, TNF, IL-17, apoptosis, and HIF-1 signaling pathway. The PI3K/AKT pathway can activate the downstream NF-κB pathway and further regulate the transcription and expression of downstream IL-6, IL-17, HIF-1α, Bax, and TNF, suggesting that the PI3K/AKT/NF-κB pathway is the critical pathway in the treatment of OA with Rhizoma Drynariae. Active components of Rhizoma Drynariae and key proteins of the PI3K/AKT/NF-κB signaling pathway were subjected to molecular docking, whose results showed that luteolin and IKK-α played a critical role. In vitro experiments indicated that both aqueous extracts of Rhizoma Drynariae (AERD) and luteolin inhibited the expression of IL-6 and HIF-1α and suppressed the activation of PI3K/AKT/NF-κB, IL-17, and TNF pathways. The measurement of mitochondrial membrane potential (Δψm) indicated that AERD and luteolin can decrease the LPS-induced early apoptotic cells. Luteolin had a more prominent inhibitory effect than AERD in the abovementioned in vitro experiments. In conclusion, the therapeutic mechanism of Rhizoma Drynariae against OA may be closely related to the inhibition of the PI3K/AKT/NF-κB pathway and downstream pathways, and luteolin plays a vital role in the treatment.

Objective: Total glycoside of peony (TGP) enhances methotrexate efficacy and attenuates its hepatotoxicity in rheumatoid arthritis, but the mechanisms remain unclear. This study investigates the mechanisms of TGP against methotrexate-induced liver injury through a network pharmacology-based approach. Methods: A liver injury model was established in CD-1 mice by intraperitoneal injection of 20 mg/kg methotrexate. TGP and the positive control drug silybin were used to intervene in the methotrexate-induced liver injury model in mice. Serum ALT and AST activities, liver index test and histopathology was detected to evaluate the effects of the treatment on methotrexate-induced liver injury. Additionally, network pharmacology and serum metabolomics were employed to predict the mechanisms of TGP in treating methotrexate-induced liver injury. Experimental validation was conducted by RT-PCR, ELISA and Western blot. Results: TGP effectively alleviated the liver index and pathological liver damage induced by methotrexate and reduced the liver injury markers, serum ALT and AST, showing effects comparable to those of the positive control drug silybin. Network pharmacology predicted that the key targets and key signaling pathways of TGP in treating methotrexate-induced liver injury are closely associated with inflammatory response. Furthermore, serum metabolomics and network pharmacology analysis indicated a close association between effects of TGP on methotrexate-induced liver injury and arachidonic acid pathway. Experimental validation results confirmed that the expression levels of IL-6, TNF and COX-2 in liver tissues were significantly elevated, with the activation of the PI3K/AKT, MAPK, and NFκB pathways. TGP intervention can reverse these changes to a certain extent. Conclusion: TGP treatment effectively mitigates methotrexate-induced liver injury, and its mechanism is closely associated with the inhibition of hepatic inflammatory responses.