Abstract：
BACKGROUND: Parkinson\'s disease (PD), a neurodegenerative disorder, is characterized by motor symptoms due to the progressive loss of dopaminergic neurons in the substantia nigra (SN) and striatum (STR), alongside neuroinflammation. Asiaticoside (AS), a primary active component with anti-inflammatory and neuroprotective properties, is derived from Centella asiatica. However, the precise mechanisms through which AS influences PD associated with inflammation are not yet fully understood.
OBJECTIVE: This study aimed to explore the protective mechanism of AS in PD.
METHODS: Targets associated with AS and PD were identified from the Swiss Target Prediction, Similarity Ensemble Approach, PharmMapper, and GeneCards database. A protein-protein interaction (PPI) network was constructed to identify potential therapeutic targets. Concurrently, GO and KEGG analyses were performed to predict potential signaling pathways. To validate these mechanisms, the effects of AS on 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD in mice were investigated. Furthermore, neuroinflammation and the activation of the NLRP3 inflammasome were assessed to confirm the anti-inflammatory properties of AS. In vitro experiments in BV2 cells were then performed to investigate the mechanisms of AS in PD. Moreover, CETSA, molecular docking, and molecular dynamics simulations (MDs) were performed for further validation.
RESULTS: Network pharmacology analysis identified 17 potential targets affected by AS in PD. GO and KEGG analyses suggested the biological roles of these targets, demonstrating that AS interacts with 149 pathways in PD. Notably, the NOD-like receptor signaling pathway was identified as a key pathway mediating AS\'s effect on PD. In vivo studies demonstrated that AS alleviated motor dysfunction and reduced the loss of dopaminergic neurons in MPTP-induced PD mice. In vitro experiments demonstrated that AS substantially decreased IL-1β release in BV2 cells, attributing this to the modulation of the NLRP3 signaling pathway. CETSA and molecular docking studies indicated that AS forms a stable complex with NLRP3. MDs suggested that ARG578 played an important role in the formation of the complex.
CONCLUSIONS: In this study, we first predicted that the potential target and pathway of AS\'s effect on PD could be NLRP3 protein and NOD-like receptor signaling pathway by network pharmacology analysis. Further, we demonstrated that AS could alleviate symptoms of PD induced by MPTP through its interaction with the NLRP3 protein for the first time by in vivo and in vitro experiments. By binding to NLRP3, AS effectively inhibits the assembly and activation of the inflammasome. These findings suggest that AS is a promising inhibitor for PD driven by NLRP3 overactivation.
OBJECTIVE: This study aimed to explore the protective mechanism of AS in PD.
METHODS: Targets associated with AS and PD were identified from the Swiss Target Prediction, Similarity Ensemble Approach, PharmMapper, and GeneCards database. A protein-protein interaction (PPI) network was constructed to identify potential therapeutic targets. Concurrently, GO and KEGG analyses were performed to predict potential signaling pathways. To validate these mechanisms, the effects of AS on 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD in mice were investigated. Furthermore, neuroinflammation and the activation of the NLRP3 inflammasome were assessed to confirm the anti-inflammatory properties of AS. In vitro experiments in BV2 cells were then performed to investigate the mechanisms of AS in PD. Moreover, CETSA, molecular docking, and molecular dynamics simulations (MDs) were performed for further validation.
RESULTS: Network pharmacology analysis identified 17 potential targets affected by AS in PD. GO and KEGG analyses suggested the biological roles of these targets, demonstrating that AS interacts with 149 pathways in PD. Notably, the NOD-like receptor signaling pathway was identified as a key pathway mediating AS\'s effect on PD. In vivo studies demonstrated that AS alleviated motor dysfunction and reduced the loss of dopaminergic neurons in MPTP-induced PD mice. In vitro experiments demonstrated that AS substantially decreased IL-1β release in BV2 cells, attributing this to the modulation of the NLRP3 signaling pathway. CETSA and molecular docking studies indicated that AS forms a stable complex with NLRP3. MDs suggested that ARG578 played an important role in the formation of the complex.
CONCLUSIONS: In this study, we first predicted that the potential target and pathway of AS\'s effect on PD could be NLRP3 protein and NOD-like receptor signaling pathway by network pharmacology analysis. Further, we demonstrated that AS could alleviate symptoms of PD induced by MPTP through its interaction with the NLRP3 protein for the first time by in vivo and in vitro experiments. By binding to NLRP3, AS effectively inhibits the assembly and activation of the inflammasome. These findings suggest that AS is a promising inhibitor for PD driven by NLRP3 overactivation.
摘要：
背景：帕金森病（PD），神经退行性疾病,由于黑质（SN）和纹状体（STR）中多巴胺能神经元的进行性丧失，其特征是运动症状，除了神经炎症。Asiaticoside（AS），具有抗炎和神经保护特性的主要活性成分，来自积雪草。然而,AS影响与炎症相关的PD的确切机制尚不完全清楚.
目的：本研究旨在探讨AS在PD中的保护机制。
方法：从瑞士目标预测中确定了与AS和PD相关的目标，相似性集成方法，PharmMapper,和GeneCards数据库。构建蛋白质-蛋白质相互作用（PPI）网络以识别潜在的治疗靶标。同时,进行GO和KEGG分析以预测潜在的信号通路。为了验证这些机制，研究了AS对1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）诱导的小鼠PD的影响。此外，评估了神经炎症和NLRP3炎症小体的激活,以确认AS的抗炎特性.然后在BV2细胞中进行体外实验以研究AS在PD中的机制。此外，CETSA,分子对接,和分子动力学模拟（MD）进行了进一步验证。
结果：网络药理学分析确定了PD中受AS影响的17个潜在目标。GO和KEGG分析提出了这些靶标的生物学作用，证明AS与PD中的149条通路相互作用。值得注意的是,NOD样受体信号通路被确定为介导AS对PD作用的关键通路。体内研究表明，AS减轻了MPTP诱导的PD小鼠的运动功能障碍并减少了多巴胺能神经元的丢失。体外实验表明，AS显著降低了BV2细胞中IL-1β的释放，将此归因于NLRP3信号通路的调节。CETSA和分子对接研究表明AS与NLRP3形成稳定的复合物。MDs提示ARG578在复合物的形成中起重要作用。
结论：在这项研究中，我们首先通过网络药理学分析预测了AS对PD的潜在作用靶点和途径可能是NLRP3蛋白和NOD样受体信号通路。Further,我们首次通过体内和体外实验证明，AS可以通过与NLRP3蛋白的相互作用来缓解MPTP诱导的PD症状。通过与NLRP3结合,AS有效地抑制炎性体的组装和活化。这些发现表明AS是由NLRP3过度激活驱动的PD的有希望的抑制剂。
目的：本研究旨在探讨AS在PD中的保护机制。
方法：从瑞士目标预测中确定了与AS和PD相关的目标，相似性集成方法，PharmMapper,和GeneCards数据库。构建蛋白质-蛋白质相互作用（PPI）网络以识别潜在的治疗靶标。同时,进行GO和KEGG分析以预测潜在的信号通路。为了验证这些机制，研究了AS对1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）诱导的小鼠PD的影响。此外，评估了神经炎症和NLRP3炎症小体的激活,以确认AS的抗炎特性.然后在BV2细胞中进行体外实验以研究AS在PD中的机制。此外，CETSA,分子对接,和分子动力学模拟（MD）进行了进一步验证。
结果：网络药理学分析确定了PD中受AS影响的17个潜在目标。GO和KEGG分析提出了这些靶标的生物学作用，证明AS与PD中的149条通路相互作用。值得注意的是,NOD样受体信号通路被确定为介导AS对PD作用的关键通路。体内研究表明，AS减轻了MPTP诱导的PD小鼠的运动功能障碍并减少了多巴胺能神经元的丢失。体外实验表明，AS显著降低了BV2细胞中IL-1β的释放，将此归因于NLRP3信号通路的调节。CETSA和分子对接研究表明AS与NLRP3形成稳定的复合物。MDs提示ARG578在复合物的形成中起重要作用。
结论：在这项研究中，我们首先通过网络药理学分析预测了AS对PD的潜在作用靶点和途径可能是NLRP3蛋白和NOD样受体信号通路。Further,我们首次通过体内和体外实验证明，AS可以通过与NLRP3蛋白的相互作用来缓解MPTP诱导的PD症状。通过与NLRP3结合,AS有效地抑制炎性体的组装和活化。这些发现表明AS是由NLRP3过度激活驱动的PD的有希望的抑制剂。
