Liu Yonglei, Mao Hongdie, Sha Zhengguang, Zhao Jishuai, Cai Hui, Xi Rong, Zhao Zhenzhu, Yin Xiaoling, Yang Lin, Liu Changyun
Department of Neurology, Fujian Medical University Union Hospital, Fuzhou, Fujian, China.
Department of Neurology, The First Affiliated Hospital of Dali University, Dali, Yunnan, China.
Front Pharmacol. 2025 Aug 21;16:1600435. doi: 10.3389/fphar.2025.1600435. eCollection 2025.
Ischemic stroke is a leading cause of mortality and disability worldwide, with limited therapeutic options and high rates of recurrence. Mitochondrial dysfunction plays a critical role in neuronal injury during ischemia-reperfusion, making mitochondrial autophagy a potential therapeutic target. Gypenoside XLIX, a major active metabolite of Gynostemma pentaphyllum, exhibits antioxidant and organ-protective properties, but its effects on neuronal mitochondrial damage in stroke remain unclear. This study aimed to explore the neuroprotective mechanisms of Gypenoside XLIX in ischemic stroke, focusing on the PI3K/AKT/FOXO1 signaling pathway.
Neuroprotective effects were evaluated in oxygen-glucose deprivation (OGD) neuronal cells and middle cerebral artery occlusion (MCAO) rat models. Cell viability, apoptosis, ROS production, mitochondrial membrane potential, and autophagic flux were assessed by CCK-8, flow cytometry, ELISA, TMRE staining, immunofluorescence, and Western blotting. Signaling pathway involvement was examined using PI3K inhibitor LY294002, AKT activator SC79, and FOXO1 knockdown.
Gypenoside XLIX significantly improved neuronal viability (p < 0.01), reduced apoptosis (p < 0.01), and decreased ROS levels (p < 0.001) in OGD cells. It enhanced p-PI3K and p-AKT expression while suppressing FOXO1 (p < 0.05), promoted Beclin-1, LC3, PINK1, and Parkin expression (p < 0.001), and reduced p62 (p < 0 .01). In MCAO rats, Gypenoside XLIX decreased infarct volume (p < 0.001), brain edema (p < 0.01), and TUNEL-positive cells (p < 0.001), while elevating mitochondrial membrane potential and antioxidant enzyme levels (SOD, GSH-Px, CAT; all p < 0.001).
Gypenoside XLIX alleviates ischemic stroke injury by activating the PI3K/AKT/FOXO1 pathway, enhancing mitochondrial autophagy, and reducing oxidative stress, supporting its potential as a novel neuroprotective agent in stroke management.
缺血性中风是全球范围内导致死亡和残疾的主要原因，治疗选择有限且复发率高。线粒体功能障碍在缺血再灌注期间的神经元损伤中起关键作用，使线粒体自噬成为一个潜在的治疗靶点。绞股蓝总苷XLIX是绞股蓝的一种主要活性代谢产物，具有抗氧化和器官保护特性，但其对中风中神经元线粒体损伤的影响尚不清楚。本研究旨在探讨绞股蓝总苷XLIX在缺血性中风中的神经保护机制，重点关注PI3K/AKT/FOXO1信号通路。
在氧糖剥夺（OGD）神经元细胞和大脑中动脉闭塞（MCAO）大鼠模型中评估神经保护作用。通过CCK-8、流式细胞术、ELISA、TMRE染色、免疫荧光和蛋白质印迹法评估细胞活力、凋亡、活性氧产生、线粒体膜电位和自噬通量。使用PI3K抑制剂LY294002、AKT激活剂SC79和FOXO1基因敲低来检测信号通路的参与情况。
绞股蓝总苷XLIX显著提高了OGD细胞中的神经元活力（p < 0.01），减少了凋亡（p < 0.01），并降低了活性氧水平（p < 0.001）。它增强了p-PI3K和p-AKT的表达，同时抑制了FOXO1（p < 0.05），促进了Beclin-1、LC3、PINK1和Parkin的表达（p < 0.001），并降低了p62（p < 0.01）。在MCAO大鼠中，绞股蓝总苷XLIX减小了梗死体积（p < 0.001）、脑水肿（p < 0.01）和TUNEL阳性细胞数量（p < 0.001），同时提高了线粒体膜电位和抗氧化酶水平（超氧化物歧化酶、谷胱甘肽过氧化物酶、过氧化氢酶；均p < 0.001）。
绞股蓝总苷XLIX通过激活PI3K/AKT/FOXO1通路减轻缺血性中风损伤，增强线粒体自噬并减少氧化应激，支持其作为中风治疗中新型神经保护剂的潜力。
