Abstract
Leptin has been widely studied and found to have a significant impact on the development of osteoarthritis (OA). However, there are conflicting findings regarding the impact of leptin on chondrocytes. The study aimed to examine the impact of leptin on human chondrocytes and rats with OA. In the in vitro experiment, cartilage tissue obtained from patients hospitalized for knee replacement due to OA was collected for primary culture of chondrocytes. The proliferation and apoptosis of chondrocytes were assessed using cell counting kit-8 and flow cytometry. Autophagy levels were evaluated through monodansylcadaverine staining, mRFP-GFP-LC3 fluorescence, and transmission electron microscopy. Additionally, the expression of autophagy-related genes and proteins was analyzed using qRT-PCR and western blotting. In the in vivo experiment, an OA rat model was established. Following treatment with leptin and leptin antagonists, the cartilage tissues were examined using histology analysis (hematoxylin-eosin and Safranin O/fast green staining) and immunohistochemical. Mankin's score was utilized to assess the severity of OA, while qRT-PCR and western blotting were employed to detect the expression of autophagy-related genes and proteins in the cartilage. The ability of leptin to protect chondrocytes is achieved through the inhibition of autophagy via phosphoinositide 3 kinase/protein kinase B/mammalian target of rapamycin signaling pathway.