To explore how MTX induces autophagy in RA-FLS and OA-FLS treated with MTX at times ranging from 12 h to 96 h at a concentration of 0.1 μM, two important autophagy-related signaling pathways were investigated: the Akt/mTOR signaling pathway and the HMGB1/Beclin-1 pathway. This evidence concerns the gene HMGB1 and rheumatoid arthritis.