Background: Osteo joint disease (OA) can be a chronic degenerative osteo-joint disease characterised by cartilage degeneration and intra-articular inflammation. Daurisoline (DAS) is certainly an isoquinoline alkaloid isolated from Rhizoma Menispermi, whose antitumor and anti-inflammatory medicinal effects are actually proven, nevertheless the outcomes of DAS on OA have rarely been researched. In this particular study, we aimed to research the potential role of DAS in OA which is partial mechanism.
Techniques and materials: The cytotoxicity of H2O2 and DAS toward chondrocytes was detected with the Cell Counting Package-8 assay. Safranin O staining was applied to recognize chondrocyte phenotype changes. Cell apoptosis was measured by flow cytometry and quantitative research in to the protein amount apoptosis-related factors Bax, Bcl-2 and cleaved caspase-3 by western blot. Western blotting and immunofluorescence were chosen to judge the expression in the autophagy-related proteins LC3, Beclin-1 and p62. Furthermore, key signal path targets and matrix-degrading indicators were measured by western blot.
Results: Our results revealed that H2O2 caused human chondrocyte apoptosis and activated autophagy in the dose-dependent manner. DAS treatment dose-dependently reversed the expression of apoptosis-related proteins (Bax, Bcl-2 and cleaved caspase3) as well as the apoptosis rate brought on by H2O2. Western blot and immunofluorescence analyses shown that DAS decreased the H2O2-caused upregulation in the autophagy marker Beclin-1 as well as the LC3 II/LC3 I ratio and upregulated the p62 protein level. Mechanistically, DAS inhibited autophagy using the activation in the classical PI3K/AKT/mTOR signaling path and guarded chondrocytes from apoptosis. Furthermore, DAS alleviated the H2O2-caused degradation of type II bovine bovine collagen as well as the high expression of matrix metalloproteinase 3 (MMP3) and MMP13.
Conclusion: Our research proven that DAS alleviated chondrocyte autophagy introduced on by H2O2 through activation in the PI3K/AKT/mTOR signaling path and guarded chondrocytes from apoptosis and matrix degradation. To summarize, these items of information declare that DAS is definitely an encouraging therapeutic way of OA.Daurisoline