Affiliation:
1. State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Key Laboratory for Zoonosis
Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine Jilin University, Center of
Infectious Diseases and Pathogen Biology, Department of Infectious Diseases, First Hospital of Jilin University,
Changchun, China
2. Shenzhen Liyunde Biotechnology Co., Ltd., Shenzhen 518057, China
3. Key Lab for New Drug Research of TCM, Research Institute of Tsinghua University in Shenzhen, Shenzhen 518057, China
4. Beijing Center for Animal Disease Control and Prevention Beijing Beijing China
Abstract
Background:
To date, disease-modifying antirheumatic drugs (DMARDs) are widely
used as the primary first-line treatment option for patients with rheumatoid arthritis (RA), and
the curative effect of methotrexate (MTX) and leflunomide (LEF; MTX + LEF) is greater than
that of single-agent MTX therapy, but the synergistic mechanism of MTX + LEF is unclear.
Methods:
First, we explored the mechanism of action of MTX + LEF in RA through network
pharmacology and molecular docking. Venn diagram analysis revealed 97 overlapping gene
targets of MTX + LEF-RA and STRING, along with Cytoscape plug-in MOCDE and cytoHubba;
and GO enrichment analysis revealed that the functions of 97 synergistic targets were related
to 123 molecular functions (MF), 63 cell components (CC), and 1,068 biological processes
(BP). The Cytoscape plug-in ClueGO demonstrated that these targets were enriched in KEGG
pathways of 52 terms, whereas 9 pivotal genes were mainly involved in the signaling pathways
of estrogen, Ras, Rap1, PI3K-Akt, relaxin, TNF, AMPK, FoxO, prolactin, IL-17, and adherens
junction. Finally, CETSA and DARTS validated the direct binding of MTX or LEF to the selected
target proteins EGFR, PPARG, MMP9, and SRC in RAW264.7 cells.
Results:
We identified 292 MTX targets and 247 LEF targets from 7 databases. Furthermore,
2,814 potential targets of RA were identified by merging 1,925 targets from 7 databases and 999
differentially expressed genes (DEGs) between normal controls and patients with RA extracted
from 5 GEO databases. Nine pivotal genes, ESR1, ALB, CASP3, EGFR, HSP90AA1, SRC,
MMP9, PPARG, and IGF1, were identified. Molecular docking verified that both MTX and
LEF strongly bind to most of the 9 pivotal proteins except ESR1 and IGF1.
Conclusion:
These results contribute to our understanding of the enhancement mechanism of
MTX combined with LEF and provide a targeted basis for the clinical treatment of RA.
Publisher
Bentham Science Publishers Ltd.