The Second Clinical College of Guangzhou University of Chinese Medicine, No. 232, East Outer Ring Road, University Town, Panyu District, Guangzhou, 510006, China.
Department of Neurology, Guangdong Provincial Hospital of Chinese Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, No. 111, Dade Lu, Yuexiu District, Guangzhou, 510120, China.
J Ethnopharmacol. 2025 Jan 10;336:118741. doi: 10.1016/j.jep.2024.118741. Epub 2024 Aug 27.
Amyotrophic lateral sclerosis (ALS) is a fetal neuromuscular disorder characterized by the gradual deterioration of motor neurons. Semen Strychni pulveratum (SSP), a processed version of Semen Strychni (SS) powder, is widely used to treat ALS in China. Vomicine is one of the most primary components of SS. However, their pharmacological effects and mechanisms for ALS remain elusive.
This study aimed to evaluate the neuroprotective and anti-neuroinflammatory effects of SSP and vomicine, as well as to explore their protective roles in ALS and the underlying mechanisms.
In vivo, 8-week-old hSOD1-WT mice and hSOD1-G93A mice were orally administered different concentrations of SSP (SSP-L = 5.46 mg/ml, SSP-M = 10.92 mg/ml or SSP-H = 16.38 mg/ml) once every other day for 8 weeks. A series of experiments, including body weight measurement, footprint tests, Hematoxylin & Eosin staining, and Nissl staining, were performed to evaluate the preventive effect of SSP. Immunofluorescence staining, western blotting, and RT-qPCR were subsequently performed to evaluate activation of the cGAS-STING-TBK1 pathway in the spinal cord. In vitro, hSOD1 NSC-34 cells were treated with vomicine to further explore the pharmacological mechanism of vomicine in the treatment of ALS via the cGAS-STING-TBK1 pathway.
SSP improved motor function, body weight loss, gastrocnemius muscle atrophy, and motor neuron loss in the spine and cortex of hSOD1-G93A mice. Furthermore, the cGAS-STING-TBK1 pathway was activated in the spinal cord of hSOD1-G93A mice, with activation predominantly observed in neurons and microglia. However, the levels of cGAS, STING, and pTBK1 proteins and cGAS, IRF3, IL-6, and IL-1β mRNA were reversed following intervention with SSP. Vomicine not only downregulated the levels of cGAS, TBK1, IL-6 and IFN-β mRNA, but also the levels of cGAS and STING protein in hSOD1 NSC-34 cells.
This study demonstrated that SSP and vomicine exert neuroprotective and anti-neuroinflammatory effects in the treatment of ALS. SSP and vomicine may reduce neuroinflammation by regulating the cGAS-STING-TBK1 pathway, and could thereby play a role in ALS treatment.
肌萎缩侧索硬化症（ALS）是一种胎儿神经肌肉疾病，其特征是运动神经元逐渐恶化。经过炮制的马钱子粉（SSP）是一种广泛用于治疗中国 ALS 的药物。马钱子碱是 SS 的主要成分之一。然而，它们在 ALS 中的药理作用和机制仍不清楚。
本研究旨在评估 SSP 和马钱子碱的神经保护和抗炎作用，并探讨其在 ALS 中的保护作用及其潜在机制。
在体内实验中，使用 hSOD1-WT 小鼠和 hSOD1-G93A 小鼠进行实验，将不同浓度的 SSP（SSP-L=5.46mg/ml、SSP-M=10.92mg/ml 或 SSP-H=16.38mg/ml）每隔一天口服给药一次，共 8 周。进行一系列实验，包括体重测量、足迹测试、苏木精和伊红染色和尼氏染色，以评估 SSP 的预防作用。随后进行免疫荧光染色、Western blot 和 RT-qPCR，以评估脊髓中 cGAS-STING-TBK1 通路的激活情况。在体外，使用马钱子碱处理 hSOD1 NSC-34 细胞，通过 cGAS-STING-TBK1 通路进一步探讨马钱子碱治疗 ALS 的药理机制。
SSP 改善了 hSOD1-G93A 小鼠的运动功能、体重减轻、腓肠肌萎缩和脊髓和皮质中的运动神经元丢失。此外，cGAS-STING-TBK1 通路在 hSOD1-G93A 小鼠的脊髓中被激活，激活主要发生在神经元和小胶质细胞中。然而，SSP 干预后，cGAS、STING 和 pTBK1 蛋白以及 cGAS、IRF3、IL-6 和 IL-1βmRNA 的水平均被逆转。马钱子碱不仅下调了 hSOD1 NSC-34 细胞中 cGAS、TBK1、IL-6 和 IFN-βmRNA 的水平，还下调了 cGAS 和 STING 蛋白的水平。
本研究表明，SSP 和马钱子碱在治疗 ALS 中具有神经保护和抗炎作用。SSP 和马钱子碱可能通过调节 cGAS-STING-TBK1 通路来减轻神经炎症，从而在 ALS 治疗中发挥作用。
