Daurisoline 通过激活 PI3K/Akt/mTOR 信号通路减弱 H2O2 诱导的软骨细胞自噬,Journal of Orthopaedic Surgery and Research

当前位置： X-MOL 学术 › J. Orthop. Surg. Res. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Daurisoline 通过激活 PI3K/Akt/mTOR 信号通路减弱 H2O2 诱导的软骨细胞自噬
Journal of Orthopaedic Surgery and Research ( IF 2.8 ) Pub Date : 2023-03-27 , DOI: 10.1186/s13018-023-03717-5
Yang Zhang ₁ , Wenguang Liu ₁ , Zhonghao Liu ₁ , Yi Liu ₁

Affiliation

骨关节炎（OA）是一种以软骨退变和关节内炎症为特征的慢性退行性关节病。Daurisoline (DAS) 是一种从北豆根中分离出来的异喹啉生物碱，其抗肿瘤和抗炎药理作用已得到证实，但 DAS 对 OA 的影响却鲜有研究。在本研究中，我们旨在探讨 DAS 在 OA 中的潜在作用及其部分机制。通过 Cell Counting Kit-8 测定法检测 H2O2 和 DAS 对软骨细胞的细胞毒性。番红 O 染色用于检测软骨细胞表型变化。通过流式细胞术和通过蛋白质印迹对细胞凋亡相关因子 Bax、Bcl-2 和裂解的 caspase-3 的蛋白质水平进行定量分析来测量细胞凋亡。蛋白质印迹和免疫荧光用于评估自噬相关蛋白 LC3、Beclin-1 和 p62 的表达。此外，通过蛋白质印迹测量了关键信号通路目标和基质降解指标。我们的结果表明，H2O2 以剂量依赖的方式诱导人软骨细胞凋亡和激活自噬。DAS 处理剂量依赖性地逆转凋亡相关蛋白（Bax、Bcl-2 和裂解的半胱天冬酶 3）的表达和 H2O2 诱导的细胞凋亡率。蛋白质印迹和免疫荧光分析表明，DAS 降低了 H2O2 诱导的自噬标记物 Beclin-1 和 LC3 II/LC3 I 比率的上调，并上调了 p62 蛋白水平。从机械上讲，DAS 通过激活经典的 PI3K/AKT/mTOR 信号通路抑制自噬，并保护软骨细胞免于凋亡。此外，DAS 减轻了 H2O2 诱导的 II 型胶原降解和基质金属蛋白酶 3 (MMP3) 和 MMP13 的高表达。我们的研究表明，DAS 通过激活 PI3K/AKT/mTOR 信号通路减轻 H2O2 引起的软骨细胞自噬，并保护软骨细胞免于凋亡和基质降解。总之，这些发现表明 DAS 可作为一种有前途的 OA 治疗策略。我们的研究表明，DAS 通过激活 PI3K/AKT/mTOR 信号通路减轻 H2O2 引起的软骨细胞自噬，并保护软骨细胞免于凋亡和基质降解。总之，这些发现表明 DAS 可作为一种有前途的 OA 治疗策略。我们的研究表明，DAS 通过激活 PI3K/AKT/mTOR 信号通路减轻 H2O2 引起的软骨细胞自噬，并保护软骨细胞免于凋亡和基质降解。总之，这些发现表明 DAS 可作为一种有前途的 OA 治疗策略。

"点击查看英文标题和摘要"

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

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. 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. 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. 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.

更新日期：2023-03-27

点击分享查看原文

点击收藏

公开下载

阅读更多本刊新发论文本刊介绍/投稿指南