State Key Laboratory of Southwestern Chinese Medicine Resources, School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, 611137, China.
School of Intelligent Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, 611137, China.
J Ethnopharmacol. 2024 May 23;326:117937. doi: 10.1016/j.jep.2024.117937. Epub 2024 Feb 27.
Moschus, first described in the Shennong's Classic of the Materia medicine, is a scarce and precious animal medicine. Modern pharmacological researches have suggested that Moschus has neuroprotective actions, and its mechanism is related to anti-inflammatory, antioxidant, and anti-apoptosis effects. Ferroptosis is one of the major pathologies of Alzheimer's disease (AD) and is widely implicated in the pathogenesis and progression of AD. Although previous studies have suggested that Moschus possesses neuroprotective effect, whether Moschus could mitigate neuronal damages by inhibiting the onset of ferroptosis is unknown in model cells of AD.
The aim of study was to explore the water extract of Moschus (WEM) on ferroptosis caused by erastin and the potential mechanism.
Erastin was used to stimulate HT22 cells to form ferroptosis model to evaluate the anti-ferroptosis effect of WEM by cell counting kit-8 and lactic dehydrogenase (LDH) tests. The malondialdehyde (MDA) and glutathione (GSH) kits are used for detection of MDA and GSH levels, and 2',7'-dichlorofluorescein diacetate and C11 BODIPY 581/591 fluorescence probe are used for evaluation of reactive oxygen species (ROS) and lipid peroxide (LOOH) levels. And Western blot was used to test nuclear factor erythroid 2-related factor 2 (Nrf2), Kelch-like ECH-associated protein 1 (Keap1), heme oxygenase-1 (HO-1), and ferroptosis associated proteins including glutathione peroxidase 4 (GPX4), cystine/glutamate antiporter subunit (SLC7A11), ferritin heavy chain 1 (FTH1), ferroportin1 (FPN1), transferrin receptor (TFRC). In addition, the Nrf2 inhibitor ML385 was applied to verify whether WEM prevents erastin-induced ferroptosis by activating the Keap1/Nrf2 pathway.
After WEM treatment, erastin-induced HT22 cell survival was significantly elevated, the accumulation of intracellular MDA, ROS, and LOOH were significantly reduced, the level of GSH and expressions of ferroptosis inhibitors GPX4 and SLC7A11 were significantly increased, and iron metabolism-related proteins TFRC, FPN1, and FTH1 were regulated. These effects of WEM are implemented by activating the Keap1/Nrf2 pathway.
This study demonstrated that WEM could perform neuroprotective effects by alleviating ferroptosis, verified that WEM treatment of AD can be mediated by the Keap1/Nrf2 pathway, and provided theoretical support for the application of WEM in the treatment of AD.
麝香，最早在《神农本草经》中被描述，是一种稀缺而珍贵的动物药材。现代药理学研究表明，麝香具有神经保护作用，其机制与抗炎、抗氧化和抗细胞凋亡作用有关。铁死亡是阿尔茨海默病（AD）的主要病理学之一，广泛涉及 AD 的发病机制和进展。尽管先前的研究表明麝香具有神经保护作用，但麝香是否能通过抑制铁死亡的发生来减轻 AD 模型细胞中的神经元损伤尚不清楚。
本研究旨在探讨麝香水提物（WEM）对依拉司琼诱导的铁死亡的影响及其潜在机制。
用依拉司琼刺激 HT22 细胞形成铁死亡模型，通过细胞计数试剂盒-8 和乳酸脱氢酶（LDH）试验评价 WEM 的抗铁死亡作用。采用丙二醛（MDA）和谷胱甘肽（GSH）试剂盒检测 MDA 和 GSH 水平，采用 2'，7'-二氯荧光素二乙酸酯和 C11 BODIPY 581/591 荧光探针评价活性氧（ROS）和脂质过氧化物（LOOH）水平。Western blot 法检测核因子红细胞 2 相关因子 2（Nrf2）、Kelch 样 ECH 相关蛋白 1（Keap1）、血红素加氧酶-1（HO-1）和铁死亡相关蛋白，包括谷胱甘肽过氧化物酶 4（GPX4）、胱氨酸/谷氨酸反向转运蛋白亚基（SLC7A11）、铁蛋白重链 1（FTH1）、铁蛋白 1（FPN1）、转铁蛋白受体（TFRC）。此外，应用 Nrf2 抑制剂 ML385 验证 WEM 是否通过激活 Keap1/Nrf2 通路来预防依拉司琼诱导的铁死亡。
WEM 处理后，依拉司琼诱导的 HT22 细胞存活率明显升高，细胞内 MDA、ROS 和 LOOH 的积累明显减少，GSH 水平和铁死亡抑制剂 GPX4 和 SLC7A11 的表达明显升高，调节铁代谢相关蛋白 TFRC、FPN1 和 FTH1。WEM 的这些作用是通过激活 Keap1/Nrf2 通路来实现的。
本研究表明，WEM 通过减轻铁死亡发挥神经保护作用，验证了 WEM 治疗 AD 可以通过 Keap1/Nrf2 通路介导，并为 WEM 在 AD 治疗中的应用提供了理论支持。
