<p>Osteoarthritis (OA) is characterized by the deterioration of the articular cartilage in the joints, which activates the synthesis of pro-inflammatory cytokines, activating chondrocytes to release matrix metalloproteins (MMPs), thus exacerbating cartilage degradation. Risk factors for OA include age, gender, genetics, obesity, joint injuries, physical inactivity, metabolic disorders, etc. However, treatment options are still limited, with total joint replacement being the primary option for late-stage OA. Excessive and prolonged ER stress triggers apoptosis, whereas mild ER stress causes the induction of autophagy, offering the protection against apoptosis. However, the intricate interplay between ER stress, autophagy, and apoptosis in OA remains incompletely understood. So, in this study, we tried to elucidate the synergistic chondroprotective effect of ER stress inhibition and autophagy activation in ameliorating osteoarthritis. Our RT-qPCR results showed the significant upregulation of GRP78, PERK, ATF4, and CHOP genes of the UPR pathway along with the decrease in the expression of BECLIN 1, LC3, and ATG5 genes of autophagy in an IL-1β induced in vitro model of osteoarthritis. In line with these results, PERK inhibitor GSK2606414 caused significant downregulation of the PERK arm of the UPR pathway and rapamycin resulting in induction of autophagy in this in vitro OA model. Moreover, the combined usage of both the PERK arm inhibitor, GSK2606414, and autophagy activator, rapamycin, in IL-1β induced OA model significantly reduced the gene expression of COL1A1, RUNX2, and MMP13, and increased COL2A1 expression, which suggests prevention of OA progression. In summary, the study suggests the synergistic chondroprotective effect of PERK arm inhibition and autophagy activation in a mTOR dependent manner, which can be a promising strategy to ameliorate osteoarthritis progression.</p>