Abstract：
BACKGROUND: Rheum tanguticum root, cataloged as \"Daehwang\" in the Korean Pharmacopeia, is rich in various anthraquinones known for their anti-inflammatory and antioxidant properties. Formulations containing Daehwang are traditionally employed for treating neurological conditions. This study aimed to substantiate the antiepileptic and neuroprotective efficacy of R. tanguticum root extract (RTE) against trimethyltin (TMT)-induced epileptic seizures and hippocampal neurodegeneration.
METHODS: The constituents of RTE were identified by ultra-performance liquid chromatography (UPLC). Experimental animals were grouped into the following five categories: control, TMT, and three TMT+RTE groups with dosages of 10, 30, and 100 mg/kg. Seizure severity was assessed daily for comparison between the groups. Brain tissue samples were examined to determine the extent of neurodegeneration and neuroinflammation using histological and molecular biology techniques. Network pharmacology analysis involved extracting herbal targets for Daehwang and disease targets for epilepsy from multiple databases. A protein-protein interaction network was built using the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database, and pivotal targets were determined by topological analysis. Enrichment analysis was performed using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) tool to elucidate the underlying mechanisms.
RESULTS: The RTE formulation was found to contain sennoside A, sennoside B, chrysophanol, emodin, physcion, (+)-catechin, and quercetin-3-O-glucuronoid. RTE effectively inhibited TMT-induced seizures at 10, 30, and 100 mg/kg dosages and attenuated hippocampal neuronal decay and neuroinflammation at 30 and 100 mg/kg dosages. Furthermore, RTE significantly reduced mRNA levels of tumor necrosis factor (TNF-α), glial fibrillary acidic protein (GFAP), and c-fos in hippocampal tissues. Network analysis revealed TNF, Interleukin-1 beta (IL-1β), Interleukin-6 (IL-6), Protein c-fos (FOS), RAC-alpha serine/threonine-protein kinase (AKT1), and Mammalian target of rapamycin (mTOR) as the core targets. Enrichment analysis demonstrated significant involvement of R. tanguticum components in neurodegeneration (p = 4.35 × 10-5) and TNF signaling pathway (p = 9.94 × 10-5).
CONCLUSIONS: The in vivo and in silico analyses performed in this study suggests that RTE can potentially modulate TMT-induced epileptic seizures and neurodegeneration. Therefore, R. tanguticum root is a promising herbal treatment option for antiepileptic and neuroprotective applications.
METHODS: The constituents of RTE were identified by ultra-performance liquid chromatography (UPLC). Experimental animals were grouped into the following five categories: control, TMT, and three TMT+RTE groups with dosages of 10, 30, and 100 mg/kg. Seizure severity was assessed daily for comparison between the groups. Brain tissue samples were examined to determine the extent of neurodegeneration and neuroinflammation using histological and molecular biology techniques. Network pharmacology analysis involved extracting herbal targets for Daehwang and disease targets for epilepsy from multiple databases. A protein-protein interaction network was built using the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database, and pivotal targets were determined by topological analysis. Enrichment analysis was performed using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) tool to elucidate the underlying mechanisms.
RESULTS: The RTE formulation was found to contain sennoside A, sennoside B, chrysophanol, emodin, physcion, (+)-catechin, and quercetin-3-O-glucuronoid. RTE effectively inhibited TMT-induced seizures at 10, 30, and 100 mg/kg dosages and attenuated hippocampal neuronal decay and neuroinflammation at 30 and 100 mg/kg dosages. Furthermore, RTE significantly reduced mRNA levels of tumor necrosis factor (TNF-α), glial fibrillary acidic protein (GFAP), and c-fos in hippocampal tissues. Network analysis revealed TNF, Interleukin-1 beta (IL-1β), Interleukin-6 (IL-6), Protein c-fos (FOS), RAC-alpha serine/threonine-protein kinase (AKT1), and Mammalian target of rapamycin (mTOR) as the core targets. Enrichment analysis demonstrated significant involvement of R. tanguticum components in neurodegeneration (p = 4.35 × 10-5) and TNF signaling pathway (p = 9.94 × 10-5).
CONCLUSIONS: The in vivo and in silico analyses performed in this study suggests that RTE can potentially modulate TMT-induced epileptic seizures and neurodegeneration. Therefore, R. tanguticum root is a promising herbal treatment option for antiepileptic and neuroprotective applications.
摘要：
背景：根根大黄，在韩国药典中被列为“Daehwang”，富含各种蒽醌，以其抗炎和抗氧化特性而闻名。含有Daehwang的制剂传统上用于治疗神经病症。本研究旨在证实丹参根提取物（RTE）对三甲基锡（TMT）诱导的癫痫发作和海马神经变性的抗癫痫和神经保护功效。
方法：采用超高效液相色谱法（UPLC）对RTE的成分进行鉴定。实验动物分为以下五类：对照，TMT,和三个TMT+RTE组,剂量为10、30和100mg/kg。每天评估癫痫发作的严重程度，以进行组间比较。使用组织学和分子生物学技术检查脑组织样品以确定神经变性和神经炎症的程度。网络药理学分析涉及从多个数据库中提取大王的草药靶标和癫痫的疾病靶标。使用用于检索相互作用基因/蛋白质（STRING）数据库的搜索工具建立了蛋白质-蛋白质相互作用网络，并通过拓扑分析确定关键目标。使用注释数据库进行富集分析，可视化,和集成发现(DAVID)工具来阐明底层机制。
结果：发现RTE制剂中含有皂甙A，森诺赛德B,大黄酚,大黄素,physcion,(+)-儿茶素,和槲皮素-3-O-葡糖醛酸。RTE在10、30和100mg/kg剂量下有效抑制TMT诱导的癫痫发作，并在30和100mg/kg剂量下减轻海马神经元衰变和神经炎症。此外，RTE显著降低肿瘤坏死因子（TNF-α）的mRNA水平，胶质纤维酸性蛋白（GFAP），和海马组织中的c-fos。网络分析显示TNF，白细胞介素-1β（IL-1β），白细胞介素-6(IL-6),蛋白质c-fos（FOS），RAC-α丝氨酸/苏氨酸蛋白激酶（AKT1），以哺乳动物雷帕霉素靶蛋白(mTOR)为核心靶点。富集分析显示，唐古汀菌成分显著参与神经变性（p=4.35×10-5）和TNF信号通路（p=9.94×10-5）。
结论：本研究中进行的体内和计算机模拟分析表明，RTE可以潜在地调节TMT诱导的癫痫发作和神经变性。因此,根根是一种有前途的草药治疗选择，用于抗癫痫和神经保护应用。
方法：采用超高效液相色谱法（UPLC）对RTE的成分进行鉴定。实验动物分为以下五类：对照，TMT,和三个TMT+RTE组,剂量为10、30和100mg/kg。每天评估癫痫发作的严重程度，以进行组间比较。使用组织学和分子生物学技术检查脑组织样品以确定神经变性和神经炎症的程度。网络药理学分析涉及从多个数据库中提取大王的草药靶标和癫痫的疾病靶标。使用用于检索相互作用基因/蛋白质（STRING）数据库的搜索工具建立了蛋白质-蛋白质相互作用网络，并通过拓扑分析确定关键目标。使用注释数据库进行富集分析，可视化,和集成发现(DAVID)工具来阐明底层机制。
结果：发现RTE制剂中含有皂甙A，森诺赛德B,大黄酚,大黄素,physcion,(+)-儿茶素,和槲皮素-3-O-葡糖醛酸。RTE在10、30和100mg/kg剂量下有效抑制TMT诱导的癫痫发作，并在30和100mg/kg剂量下减轻海马神经元衰变和神经炎症。此外，RTE显著降低肿瘤坏死因子（TNF-α）的mRNA水平，胶质纤维酸性蛋白（GFAP），和海马组织中的c-fos。网络分析显示TNF，白细胞介素-1β（IL-1β），白细胞介素-6(IL-6),蛋白质c-fos（FOS），RAC-α丝氨酸/苏氨酸蛋白激酶（AKT1），以哺乳动物雷帕霉素靶蛋白(mTOR)为核心靶点。富集分析显示，唐古汀菌成分显著参与神经变性（p=4.35×10-5）和TNF信号通路（p=9.94×10-5）。
结论：本研究中进行的体内和计算机模拟分析表明，RTE可以潜在地调节TMT诱导的癫痫发作和神经变性。因此,根根是一种有前途的草药治疗选择，用于抗癫痫和神经保护应用。
