Molecular Endocrinology Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology Roorkee, Roorkee, Uttarakhand, 247667, India; Biomaterials and Multiscale Mechanics Laboratory, Department of Metallurgical and Materials Engineering, Indian Institute of Technology Roorkee, Roorkee, Uttarakhand, 247667, India; Centre of Nanotechnology, Indian Institute of Technology Roorkee, Roorkee, Uttarakhand, 247667, India.
Molecular Endocrinology Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology Roorkee, Roorkee, Uttarakhand, 247667, India.
J Ethnopharmacol. 2024 Jun 28;328:117899. doi: 10.1016/j.jep.2024.117899. Epub 2024 Feb 9.
This study has important ethnopharmacological implications since it systematically investigated the therapeutic potential of Bacopa monnieri(L.) Wettst. (Brahmi) in treating neurological disorders characterized by oxidative stress-a growing issue in the aging population. Bacopa monnieri, which is strongly rooted in Ayurveda, has long been recognized for its neuroprotective and cognitive advantages. The study goes beyond conventional wisdom by delving into the molecular complexities of Bacopa monnieri, particularly its active ingredient, Bacoside-A, in countering oxidative stress. The study adds to the ethnopharmacological foundation for using this herbal remedy in the context of neurodegenerative disorders by unravelling the scientific underpinnings of Bacopa monnieri's effectiveness, particularly at the molecular level, against brain damage and related conditions influenced by oxidative stress. This dual approach, which bridges traditional wisdom and modern investigation, highlights Bacopa monnieri's potential as a helpful natural remedy for oxidative stress-related neurological diseases.
The aim of this study is to investigate the detailed molecular mechanism of action (in vitro, in silico and in vivo) of Bacopa monnieri (L.) Wettst. methanolic extract and its active compound, Bacoside-A, against oxidative stress in neurodegenerative disorders.
ROS generation activity, mitochondrial membrane potential, calcium deposition and apoptosis were studied through DCFDA, Rhodamine-123, FURA-2 AM and AO/EtBr staining respectively. In silico study to check the effect of Bacoside-A on the Nrf-2 and Keap1 axis was performed through molecular docking study and validated experimentally through immunofluorescence co-localization study. In vivo antioxidant activity of Bacopa monnieri extract was assessed by screening the oxidative stress markers and stress-inducing hormone levels as well as through histopathological analysis of tissues.
The key outcome of this study is that the methanolic extract of Bacopa monnieri (BME) and its active component, Bacoside-A, protect against oxidative stress in neurodegenerative diseases. At 100 and 20 μg/ml, BME and Bacoside-A respectively quenched ROS, preserved mitochondrial membrane potential, decreased calcium deposition, and inhibited HT-22 mouse hippocampus cell death. BME and Bacoside-A regulated the Keap1 and Nrf-2 axis and their downstream antioxidant enzyme-specific genes to modify cellular antioxidant machinery. In vivo experiments utilizing rats subjected to restrained stress indicated that pre-treatment with BME (50 mg/kg) downregulated oxidative stress markers and stress-inducing hormones, and histological staining demonstrated that BME protected the neuronal cells of the Cornu Ammonis (CA1) area in the hippocampus.
Overall, the study suggests that Bacopa monnieri(L.) Wettst. has significant potential as a natural remedy for neurodegenerative disorders, and its active compounds could be developed as new drugs for the prevention and treatment of oxidative stress-related diseases.
本研究具有重要的民族药理学意义，因为它系统地研究了假马齿苋（Bacopa monnieri（L.） Wettst.）（ Brahmi）在治疗以氧化应激为特征的神经病变中的治疗潜力-这是老龄化人口中的一个日益严重的问题。假马齿苋在阿育吠陀中根深蒂固，长期以来一直因其神经保护和认知优势而闻名。该研究通过深入研究假马齿苋的分子复杂性，特别是其活性成分 Bacoside-A 来对抗氧化应激，超越了传统智慧。该研究通过揭示假马齿苋在对抗与氧化应激相关的脑损伤和相关疾病中的有效性的科学基础，特别是在分子水平上，为使用这种草药疗法作为神经退行性疾病的基础提供了民族药理学基础。这种融合传统智慧和现代研究的双重方法突显了假马齿苋作为治疗与氧化应激相关的神经疾病的有益天然疗法的潜力。
本研究的目的是研究假马齿苋（L.） Wettst.甲醇提取物及其活性化合物 Bacoside-A 对神经退行性疾病中氧化应激的详细分子作用机制（体外，体内和体内）。
通过 DCFDA、Rhodamine-123、FURA-2 AM 和 AO/EtBr 染色分别研究 ROS 生成活性，线粒体膜电位，钙沉积和细胞凋亡。通过分子对接研究和通过免疫荧光共定位研究进行实验验证，研究了 Bacoside-A 对 Nrf-2 和 Keap1 轴的影响。通过筛选氧化应激标志物和应激诱导激素水平以及通过组织学分析，评估了假马齿苋提取物的体内抗氧化活性。
这项研究的关键结果是，假马齿苋的甲醇提取物（BME）及其活性成分 Bacoside-A 可预防神经退行性疾病中的氧化应激。在 100 和 20 μg/ml 时，BME 和 Bacoside-A 分别猝灭了 ROS，维持了线粒体膜电位，减少了钙沉积，并抑制了 HT-22 小鼠海马细胞死亡。BME 和 Bacoside-A 调节了 Keap1 和 Nrf-2 轴及其下游抗氧化酶特异性基因，以改变细胞抗氧化机制。使用受约束应激的大鼠进行的体内实验表明，BME（50mg/kg）预处理可下调氧化应激标志物和应激诱导激素，组织学染色表明 BME 可保护海马 Cornu Ammonis（CA1）区的神经元细胞。
总的来说，该研究表明假马齿苋（L.） Wettst.具有作为神经退行性疾病天然疗法的重要潜力，其活性化合物可开发为预防和治疗与氧化应激相关疾病的新药。
