Dapagliflozin targets the crosstalk between apoptosis, autophagy, and Hedgehog signaling pathways through AMPK activation in the adjuvant-induced arthritic rat model

Abstract

Rheumatoid arthritis is a long-term autoimmune disorder, causes joint capsule, cartilage, and bone damage. Dapagliflozin, a novel antidiabetic drug, demonstrated promising effects against different disorders. Herein, we aimed to detect the dose-dependent antiarthritic impact of dapagliflozin alone and in combination with methotrexate standard treatment. Complete Freund’s adjuvant-induced arthritic rats were treated with three doses of dapagliflozin (1, 5, or 10 mg/kg/day, p.o.) for 3 weeks, in which 10 mg dose showed eminent anti-arthritic effects according to gait score, paw diameter, arthritic index (AI), morphological and histological results. To reveal dapagliflozin mechanism, locomotor, biochemical, and histological measures were assessed in dapagliflozin (10 mg/kg/day) and/or methotrexate (0.75 mg/kg/week, i.p.)-treated arthritic rats. Radiography and histology confirmed the prominent anti-arthritic effect of dapagliflozin via reduced RF, MMP-1, and MMP-3, and improved gait score, ankle diameter, and AI. Anti-inflammatory impact was confirmed by the downregulation of TNF-α, IL-1β, IL-6, and NF-κb p65 expression. Upregulation of autophagy was detected through; Beclin-1, ULK-1, and ATG-7, in dapagliflozin treated arthritic rats. Furtherly, dapagliflozin stimulated apoptotic activity, by boosting articular levels of CASP-3, CASP-9, cartilage gene expression of p53, and Bax/Bcl2 ratio. Interestingly, dapagliflozin upregulates p-AMPK/t-AMPK articular activity. Additionally, dapagliflozin inhibited the Hedgehog signaling pathway, through the downregulation of cartilage Shh, ptch1, Smo, and Gli-1 expression. Dapagliflozin/methotrexate combination therapy exhibited greater anti-arthritic benefits compared to methotrexate alone. These data highlight dapagliflozin as an anti-rheumatic drug, either alone or with methotrexate.