Daurisoline attenuates H2O2-induced chondrocyte autophagy by activating the PI3K/Akt/mTOR signaling pathway

Abstract

Background Osteoarthritis (OA) is a chronic degenerative joint disease characterized by cartilage degeneration and intra-articular inflammation. Daurisoline (DAS) is an isoquinoline alkaloid isolated from Rhizoma Menispermi, whose antitumor and anti-inflammatory pharmacological effects have been demonstrated, but the effects of DAS on OA have rarely been researched. In this study, we aimed to explore the potential role of DAS in OA and its partial mechanism. Materials and methods The cytotoxicity of H2O2 and DAS toward chondrocytes was detected by the Cell Counting Kit-8 assay. Safranin O staining was used to detect chondrocyte phenotype changes. Cell apoptosis was measured by both flow cytometry and quantitative analysis of the protein levels of the apoptosis-related factors Bax, Bcl-2 and cleaved caspase-3 by western blot. Western blotting and immunofluorescence were used to assess the expression of the autophagy-related proteins LC3, Beclin-1 and p62. In addition, key signal pathway targets and matrix-degrading indicators were measured by western blot. Results Our results indicated that H2O2 induced human chondrocyte apoptosis and activated autophagy in a dose-dependent manner. DAS treatment dose-dependently reversed the expression of apoptosis-related proteins (Bax, Bcl-2 and cleaved caspase3) and the apoptosis rate induced by H2O2. Western blot and immunofluorescence analyses showed that DAS decreased the H2O2-induced upregulation of the autophagy marker Beclin-1 and the LC3 II/LC3 I ratio and upregulated the p62 protein level. Mechanistically, DAS inhibited autophagy through the activation of the classical PI3K/AKT/mTOR signaling pathway and protected chondrocytes from apoptosis. In addition, DAS alleviated the H2O2-induced degradation of type II collagen and the high expression of matrix metalloproteinase 3 (MMP3) and MMP13. Conclusion Our research demonstrated that DAS alleviated chondrocyte autophagy caused by H2O2 through activation of the PI3K/AKT/mTOR signaling pathway and protected chondrocytes from apoptosis and matrix degradation. In conclusion, these findings suggest that DAS may serve as a promising therapeutic strategy for OA.

Identification of EGFR as an essential regulator in chondrocytes ferroptosis of osteoarthritis using bioinformatics, in vivo, and in vitro study

“…Significantly, these genes were mainly enriched in cell death related BPs, including the cellular response to oxidative stress, superoxide metabolic process, and positive regulation of programmed cell death. Meanwhile, these genes were primarily engaged in pathways of cartilage homeostasis and chondrocytes death, such as MAPK, PI3K-Akt, and mTOR signaling pathways [ [28] , [29] , [30] ]. Further ClueGO analysis revealed that ferroptosis was associated with the pathways of fat metabolism.…”

Natural products in osteoarthritis treatment: bridging basic research to clinical applications

“…Oroxin B reduced the expression of MMP-3 and MMP-13 by downregulating the production of p-PI3K and p-AKT in chondrocytes [75]. Daurisoline, an isoquinoline alkaloid, alleviated the HO-induced high expression of MMP-3 and MMP-13 and degradation of type II collagen [117]. Urolithin A downregulated the production of p-PI3K and p-AKT in chondrocytes and reduced the expression of ADAMTS-5 and MMP-13 [76].…”

The emerging role of lncRNAs in osteoarthritis development and potential therapy

“…Salvianolic acid B reduced IL-1β-mediated inflammatory cytokine production in chondrocytes in osteoarthritis and protected osteoarthritis progression ( Lou et al, 2017 ). Daurisoline activated the PI3K/Akt/mTOR axis and led to inhibition of H2O2-mediated autophagy in chondrocytes ( Zhang et al, 2023b ). Oroxin B suppressed the PI3K/Akt/mTOR pathway and induced autophagy and anti-inflammation, leading to inhibition of osteoarthritis progression ( Lu et al, 2022 ).…”