Wang Yunliang, Sun Xiangyun, He Biao, Yu Shaowen
College of Physical Education, AnHui Normal University, Wuhu, 241003, P.R. China.
The Department of Geriatrics, the First Affiliated Hospital of Wannan Medical College (Yijishan Hospital), Jinghu District, No. 2 West Zheshan Road, Anhui Province, Wuhu, 241001, P.R. China.
Mol Neurobiol. 2025 Jun 6. doi: 10.1007/s12035-025-05073-3.
This study aimed to investigate the mechanism of ginsenoside Rg1 in Alzheimer's disease (AD) via miR-9-5p/SIRT1-mediated mitochondrial function. The cognitive function of AD mice was assessed by Morris water maze experiment. The histopathological changes in the CA1 region were observed by H&E staining. TUNEL staining combined with the neuronal marker NeuN was used to detect neuronal apoptosis in hippocampal tissues. Aβ induced HT-22 cells were used as AD in vitro models. MiR-9-5p expression was detected by qRT-PCR, and SIRT1 protein and autophagy-related proteins (LC3B II/I, Beclin-1) levels were measured by western blot. The binding of miR-9-5p with SIRT1 was predicted and validated. Ginsenoside Rg1 treatment in AD mice reduced miR-9-5p expression, increased SIRT1 level, attenuated mitochondrial dysfunction, and effectively improved AD symptoms in mice, while such effect can be either reversed by miR-9-5p agomir or SIRT1 inhibitor (EX527). In vitro, Aβ-induced HT-22 cell activity was reduced, cell death was significantly increased, and mitochondrial dysfunction was progressed, but treatment of HT-22 cells with Aβ and ginsenoside Rg1 attenuated mitochondrial dysfunction and improved Aβ-induced HT-22 cell damage. Ginsenoside Rg1 ameliorated Aβ-induced HT-22 cell damage by down-regulating miR-9-5p to regulate SIRT1-mediated mitochondrial dysfunction. miR-9-5p negatively regulates SIRT1. Inhibition of mitochondrial autophagy partially reversed the ameliorative effect of ginsenoside Rg1 on mitochondrial dysfunction and cellular damage in HT-22 cells. Ginsenoside Rg1 down-regulates miR-9-5p expression to modulate SIRT1-mediated mitochondrial dysfunction, hereby attenuating Aβ induced cell injury in HT-22 cells and alleviating AD in mice.
本研究旨在通过miR-9-5p/SIRT1介导的线粒体功能来探究人参皂苷Rg1在阿尔茨海默病（AD）中的作用机制。通过莫里斯水迷宫实验评估AD小鼠的认知功能。采用苏木精-伊红（H&E）染色观察CA1区的组织病理学变化。运用TUNEL染色结合神经元标志物NeuN来检测海马组织中的神经元凋亡。用Aβ诱导的HT-22细胞作为AD体外模型。通过qRT-PCR检测miR-9-5p表达，采用蛋白质免疫印迹法检测SIRT1蛋白及自噬相关蛋白（LC3B II/I、Beclin-1）水平。预测并验证miR-9-5p与SIRT1的结合。对AD小鼠进行人参皂苷Rg1治疗可降低miR-9-5p表达，提高SIRT1水平，减轻线粒体功能障碍，并有效改善小鼠的AD症状，而miR-9-5p激动剂或SIRT1抑制剂（EX527）可逆转这种作用。在体外，Aβ诱导的HT-22细胞活性降低，细胞死亡显著增加，线粒体功能障碍进展，但用Aβ和人参皂苷Rg1处理HT-22细胞可减轻线粒体功能障碍并改善Aβ诱导的HT-22细胞损伤。人参皂苷Rg1通过下调miR-9-5p来调节SIRT1介导的线粒体功能障碍，从而改善Aβ诱导的HT-22细胞损伤。miR-9-5p负向调节SIRT1。抑制线粒体自噬可部分逆转人参皂苷Rg1对HT-22细胞线粒体功能障碍和细胞损伤的改善作用。人参皂苷Rg1下调miR-9-5p表达以调节SIRT1介导的线粒体功能障碍，从而减轻Aβ诱导的HT-22细胞损伤并缓解小鼠的AD症状。
