Qi Zhengyu, Chu Xinge, Li Sha, Bai Qiaoyun, Ding Ningpo, Yan Guanghai, Yu Hailing, Cui Chunai
Department of Anatomy, Histology and Embryology, Yanbian University Medical College, Yanji, 133002, PR China; Jilin Key Laboratory for Immune and Targeting Research on Common Allergic Diseases, Yanbian University, Yanji, 133002, PR China.
Jilin Key Laboratory for Immune and Targeting Research on Common Allergic Diseases, Yanbian University, Yanji, 133002, PR China; Department of Pharmacology, School of Medicine, Ningbo University, Ningbo, 315211, PR China.
J Ethnopharmacol. 2026 Jan 30;355(Pt A):120603. doi: 10.1016/j.jep.2025.120603. Epub 2025 Sep 12.
Myristica fragrans Houtt. has a long history of use in traditional medicine as a nervine tonic for enhancing cognitive function, relieving anxiety, and managing neurological symptoms associated with aging. Given its established ethnopharmacological profile, we postulated that its principal active lignan, Macelignan, could exert potent neuroprotective effects in the context of neurodegenerative diseases like vascular dementia (VaD).
This study was designed to investigate the neuroprotective properties of Macelignan in a preclinical model of vascular dementia (VaD) and to elucidate the underlying molecular mechanisms, with a particular focus on its modulation of the mTOR signaling pathway and mitochondrial homeostasis.
In this study, in vivo and in vitro models of ischemia-hypoxia were established in Wistar rats by bilateral common carotid artery occlusion (BCCAo) and in HT22 neuronal cells with cobalt chloride (CoCl) treatment, respectively. The resulting cognitive, pathological, and cell survival damages were comprehensively assessed using a battery of behavioral tests, histological staining (H&E and Nissl), and the CCK-8 assay. To elucidate the underlying mechanisms, we first predicted and validated the direct interaction between the drug and its core target protein using transcriptome sequencing (RNA-seq) combined with molecular docking and dynamics simulations. Subsequently, changes in key signaling pathways, including mTOR, mitochondrial dynamics, autophagy, and apoptosis, were systematically investigated utilizing Seahorse metabolic flux analysis, transmission electron microscopy (TEM), various fluorescent probes (JC-1, MitoSOX, ROS, Ca), Western blotting, and immunofluorescence (IF). All data were statistically analyzed using GraphPad Prism 10.1.2.
In vivo, we demonstrate that Macelignan ameliorates neuronal damage and cognitive deficits in a model of vascular dementia by directly targeting and activating the mTOR signaling pathway. At the cellular level, this mTOR activation orchestrates a multifaceted protective response, which includes restoring mitochondrial function and homeostasis, enhancing antioxidant defenses, suppressing the stress-induced expression of mitophagy-related proteins Beclin-1 and Parkin, and potently inhibiting apoptosis. Critically, these neuroprotective effects of Mace were completely abrogated by the mTORC1-specific inhibitor rapamycin, definitively establishing that its therapeutic efficacy is dependent on mTOR activation.
Macelignan targets and activates mTOR to restore mitochondrial homeostasis, thereby ameliorating vascular dementia.
肉豆蔻在传统医学中有着悠久的使用历史，作为一种神经滋补剂，可增强认知功能、缓解焦虑并管理与衰老相关的神经症状。鉴于其已确立的民族药理学特征，我们推测其主要活性木脂素——马塞林，在血管性痴呆（VaD）等神经退行性疾病的背景下可能发挥强大的神经保护作用。
本研究旨在调查马塞林在血管性痴呆（VaD）临床前模型中的神经保护特性，并阐明其潜在的分子机制，特别关注其对mTOR信号通路和线粒体稳态的调节作用。
在本研究中，分别通过双侧颈总动脉闭塞（BCCAo）在Wistar大鼠中建立体内缺血缺氧模型，以及用氯化钴（CoCl）处理HT22神经元细胞建立体外缺血缺氧模型。使用一系列行为测试、组织学染色（苏木精-伊红染色和尼氏染色）和CCK-8测定法全面评估由此产生的认知、病理和细胞存活损伤。为了阐明潜在机制，我们首先使用转录组测序（RNA-seq）结合分子对接和动力学模拟预测并验证了药物与其核心靶蛋白之间的直接相互作用。随后，利用海马代谢通量分析、透射电子显微镜（TEM）、各种荧光探针（JC-1、MitoSOX、ROS、Ca）、蛋白质免疫印迹法和免疫荧光法（IF）系统地研究了关键信号通路的变化，包括mTOR、线粒体动力学、自噬和凋亡。所有数据均使用GraphPad Prism 10.1.2进行统计分析。
在体内，我们证明马塞林通过直接靶向并激活mTOR信号通路，改善血管性痴呆模型中的神经元损伤和认知缺陷。在细胞水平上，这种mTOR激活协调了多方面的保护反应，包括恢复线粒体功能和稳态、增强抗氧化防御、抑制应激诱导的与线粒体自噬相关蛋白Beclin-1和Parkin的表达，并有效抑制细胞凋亡。至关重要的是，mTORC1特异性抑制剂雷帕霉素完全消除了马塞林的这些神经保护作用，明确证实其治疗效果依赖于mTOR激活。
马塞林靶向并激活mTOR以恢复线粒体稳态，从而改善血管性痴呆。
