NLRP3 mediates the neuroprotective effects of SVHRSP derived from scorpion venom in rotenone-induced experimental Parkinson's disease model.
NLRP3 mediates the neuroprotective effects of SVHRSP derived from scorpion venom in rotenone-induced experimental Parkinson's disease model.
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DOI：
10.1016/j.jep.2023.116497
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发表时间：
2023-04
影响因子：
5.4
通讯作者：
Yu Zhang;Sheng Li;Liyan Hou;Ming‐yuan Wu;Jianing Liu;Ruonan Wang;Qingshan Wang;Jie Zhao
中科院分区：
文献类型：
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作者：
Yu Zhang;Sheng Li;Liyan Hou;Ming‐yuan Wu;Jianing Liu;Ruonan Wang;Qingshan Wang;Jie Zhao
民族药理学相关性在传统中医中，蝎子用于治疗颤抖、抽搐和痴呆等症状的疾病。我们实验室采用专利技术从蝎毒中提取和纯化活性单一成分。然后利用质谱法确定该多肽的氨基酸序列，并进行人工合成，得到纯度为99.3%的多肽，命名为SVHRSP（蝎毒耐热肽）。 SVHRSP已被证明对帕金森病具有强大的神经保护功效。本研究目的探讨SVHRSP在PD小鼠模型中提供神经保护作用的分子机制和潜在的分子靶点，以及研究NLRP3在SVHRSP介导的神经保护中的作用。材料与方法PD小鼠模型由鱼藤酮和 使用步态测试、转棒测试、多巴胺能神经元数量和小胶质细胞的激活来测量 SVHRSP 对 PD 小鼠模型的神经保护作用。通过 RNA 测序和 GSEA 分析来发现 SVHRSP 调节的差异生物学途径。采用原代中脑神经元胶质培养物和 NLRP3−/− 小鼠，通过 qRT-PCR、蛋白质印迹、酶联免疫吸附测定 (ELISA) 和免疫染色验证 NLRP3 的作用。 结果 SVHRSP 提供的多巴胺能神经保护作用伴随着小胶质细胞介导的抑制 神经炎症途径。重要的是，小胶质细胞的消耗显着降低了 SVHRSP 对体外鱼藤酮诱导的多巴胺能神经毒性的神经保护功效。 SVHRSP 抑制鱼藤酮 PD 小鼠小胶质细胞 NOD 样受体通路、NLRP3 mRNA 表达和蛋白水平。 SVHRSP 还减少了鱼藤酮诱导的 caspse-1 激活和 IL-1β 成熟，表明 SVHRSP 减轻了 NLRP3 炎性体的激活。此外，MCC950灭活NLRP3炎性体或NLRP3基因缺失几乎消除了SVHRSP提供的抗炎、神经保护作用和对鱼藤酮反应的运动表现的改善。结论NLRP3在鱼藤酮诱导的实验性PD模型中介导了SVHRSP的神经保护作用，为鱼藤酮诱导的实验性PD模型中的NLRP3介导了SVHRSP的神经保护作用，为鱼藤酮诱导的实验性PD模型提供了额外的证据。 SVHRSP 对 PD 具有抗炎和神经保护作用。
Ethnopharmacological relevanceIn traditional Chinese medicine, scorpion is used to treat diseases with symptoms such as trembling, convulsion and dementia. Our laboratory employs patented technology to extract and purify the active single component from scorpion venom. We then utilize mass spectrometry to determine the amino acid sequence of the polypeptide and synthesize it artificially to acquire the polypeptide with a purity of 99.3%, named SVHRSP (Scorpion Venom Heat-Resistant Peptide). SVHRSP has been demonstrated to display potent neuroprotective efficacy in Parkinson’s disease.Aim of the studyTo explore the molecular mechanisms and potential molecular targets of SVHRSP-afforded neuroprotection in PD mouse models, as well as to investigate the role of NLRP3 in SVHRSP-mediated neuroprotection.Materials and methodsThe PD mouse model was induced by rotenone and the neuroprotective role of SVHRSP on the PD mouse model was measured using the gait test, rotarod test, the number of dopaminergic neurons, and the activation of microglia. RNA sequencing and GSEA analysis were performed to find the differentially biological pathways regulated by SVHRSP. Primary mid-brain neuron-glial cultures and NLRP3−/− mice were applied to verify the role of NLRP3 by using qRT-PCR, western blotting, enzyme-linked immunosorbent assay (ELISA) and immunostaining.ResultsSVHRSP-afforded dopaminergic neuroprotection was accompanied with inhibition of microglia-mediated neuroinflammatory pathways. Importantly, depletion of microglia markedly reduced the neuroprotective efficacy of SVHRSP against rotenone-induced dopaminergic neurotoxicity in vitro. SVHRSP inhibited microglial NOD-like receptor pathway, mRNA expression and protein level of NLRP3 in rotenone PD mice. SVHRSP also reduced rotenone-induced caspse-1 activation and IL-1β maturation, indicating that SVHRSP mitigated activation of NLRP3 inflammasome. Moreover, inactivation of NLRP3 inflammasome by MCC950 or genetic deletion of NLRP3 almost abolished SVHRSP-afforded anti-inflammatory, neuroprotective effects and improvement of motor performance in response to rotenone.ConclusionsNLRP3 mediated the neuroprotective effects of SVHRSP in rotenone-induced experimental PD model, providing additional evidence for the mechanisms of SVHRSP-afforded anti-inflammatory and neuroprotective effects in PD.