Gnetupendin A protects against ischemic stroke through activating the PI3K/AKT/mTOR-dependent autophagy pathway.

IF 6.7 1区医学 Q1 CHEMISTRY, MEDICINAL

Phytomedicine Pub Date : 2024-11-08 DOI:10.1016/j.phymed.2024.156233

Danyang Mu, Jingyu Liu, Yan Mi, Dequan Wang, Libin Xu, Yuxin Yang, Yueyang Liu, Dong Liang, Yue Hou

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引用次数: 0

Abstract

Background: Autophagy has been recently emerged as a prominent factor in the pathogenesis of ischemic stroke (IS) and is increasingly being considered as a potential therapeutic target for IS. Gnetum parvifolium has been identified as a potential therapeutic agent for inflammatory diseases such as rheumatism and traumatic injuries. However, the pharmacological effects of Gnetupindin A (GA), a stilbene compound isolated from Gnetum parvifolium, have not been fully elucidated until now.

Objective: Here we identified the therapeutic potential of GA for IS, deeply exploring the possible mechanisms related to its regulation of autophagy.

Methods: The mouse model of middle cerebral artery occlusion-reperfusion (MCAO/R) and the oxygen-glucose deprivation reperfusion (OGD/R)-exposed cells served as models to study the protection of GA against IS. The adeno-associated virus (AAV) encoding shAtg5, in conjunction with autophagy inhibitor 3-Methyladenine (3-MA) were utilized to explore the role of GA in regulating autophagy following IS. Molecular docking, CETSA, and DARTS were used to identify the specific therapeutic target of GA. PI3K inhibitor LY294002 was employed to test the participation of PI3K in GA-mediated autophagy and neuroprotective effects following IS.

Results: Our findings revealed that treatment with GA significantly alleviated the brain infract volume, edema, improved neurological deficits and attenuated apoptosis. Mechanistically, we found that GA promoted autophagic flow both in vivo and in vitro after IS. Notably, neural-targeted knockdown of Atg5 abolished the neuroprotective effects mediated by GA. Inhibition of autophagy using 3-MA blocked the attenuation on apoptosis induced by GA. Moreover, molecular docking, CETSA, and DARTS analysis demonstrated that GA specifically targeted PI3K and further inhibited the activation of PI3K/AKT/mTOR signaling pathway. LY294002, which inhibits PI3K, reversed GA-induced autophagy and neuroprotective effects on OGD/R-treated cells.

Conclusion: We demonstrated, for the first time, that GA protects against IS through promoting the PI3K/AKT/mTOR-dependent autophagy pathway. Our findings provide a novel mechanistic insight into the anti-IS effect of GA in regulating autophagy.

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Gnetupendin A可通过激活PI3K/AKT/mTOR依赖性自噬途径预防缺血性中风。

背景：自噬最近被认为是缺血性中风（IS）发病机制中的一个重要因素，并逐渐被认为是IS的潜在治疗靶点。Gnetum parvifolium 已被确定为治疗风湿和创伤等炎症性疾病的潜在药物。目的：在此，我们确定了 Gnetupindin A（GA）对 IS 的治疗潜力，并深入探讨了其调控自噬的可能机制：方法：以大脑中动脉闭塞再灌注（MCAO/R）小鼠模型和氧-葡萄糖剥夺再灌注（OGD/R）暴露细胞为模型，研究GA对IS的保护作用。研究人员利用编码 shAtg5 的腺相关病毒（AAV）和自噬抑制剂 3-甲基腺嘌呤（3-MA）来探讨 GA 在 IS 发生后调节自噬的作用。分子对接、CETSA 和 DARTS 被用来确定 GA 的特定治疗靶点。采用PI3K抑制剂LY294002检测PI3K在GA介导的IS后自噬和神经保护作用中的参与情况：结果：我们的研究结果表明，GA能明显减轻脑梗死的体积和水肿，改善神经功能缺损，减少细胞凋亡。从机理上讲，我们发现 GA 在 IS 后促进了体内和体外的自噬流。值得注意的是，神经靶向敲除 Atg5 会取消 GA 介导的神经保护效应。使用 3-MA 抑制自噬可以阻止 GA 诱导的细胞凋亡。此外，分子对接、CETSA 和 DARTS 分析表明，GA 能特异性地靶向 PI3K，并进一步抑制 PI3K/AKT/mTOR 信号通路的激活。抑制 PI3K 的 LY294002 逆转了 GA 诱导的自噬和对 OGD/R 处理细胞的神经保护作用：结论：我们首次证明了 GA 可通过促进 PI3K/AKT/mTOR 依赖性自噬途径保护细胞免受 IS 的伤害。我们的研究结果为了解 GA 在调节自噬过程中的抗 IS 作用提供了一种新的机制。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Phytomedicine 医学-药学

CiteScore

10.30

自引率

5.10%

发文量

670

审稿时长

91 days

期刊介绍： Phytomedicine is a therapy-oriented journal that publishes innovative studies on the efficacy, safety, quality, and mechanisms of action of specified plant extracts, phytopharmaceuticals, and their isolated constituents. This includes clinical, pharmacological, pharmacokinetic, and toxicological studies of herbal medicinal products, preparations, and purified compounds with defined and consistent quality, ensuring reproducible pharmacological activity. Founded in 1994, Phytomedicine aims to focus and stimulate research in this field and establish internationally accepted scientific standards for pharmacological studies, proof of clinical efficacy, and safety of phytomedicines.