中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (30): 6466-6473.doi: 10.12307/2025.917

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

丁苯酞对帕金森病细胞模型的保护作用及机制

张 鑫 1 ，郭宝娟 2 ，徐慧鑫 1 ，沈玉珍 1 ，杨晓帆 3 ，杨旭芳 1 ，陈 培 1
  

  1. 1牡丹江医科大学基础医学院病理生理学教研室，黑龙江省牡丹江市  157011；2包头市蒙医中医医院病理科，内蒙古自治区包头市  014000；3牡丹江医科大学附属红旗医院康复医学科，黑龙江省牡丹江市  157011
  • 收稿日期:2024-09-10 接受日期:2024-11-26 出版日期:2025-10-28 发布日期:2025-03-28
  • 通讯作者: 陈培，女，副教授，硕士生导师，牡丹江医科大学基础医学院病理生理学教研室，黑龙江省牡丹江市 157011
  • 作者简介:张鑫，男，1999年生，山东省泰安市人，汉族，硕士，主要从事神经退行性疾病的发病机制及治疗研究。 共同第一作者：郭宝娟，女，1997年生，内蒙古自治区包头市人，汉族，硕士，主要从事神经退行性疾病的发病机制及治疗研究。
  • 基金资助:
    黑龙江省省属高校基本科研业务费科研项目(2023-KYYWF-0936)，项目负责人：陈培

Protective effects and mechanisms of 3-N-butylphthalide in Parkinson’s disease cell models

Zhang Xin1, Guo Baojuan2, Xu Huixin1, Shen Yuzhen1, Yang Xiaofan3, Yang Xufang1, Chen Pei1   

  1. 1Department of Pathophysiology, School of Basic Medicine, Mudanjiang Medical University, Mudanjiang 157011, Heilongjiang Province, China; 2Department of Pathology, Baotou Mongolian Medicine and Traditional Chinese Medicine Hospital, Baotou 014000, Inner Mongolia Autonomous Region, China; 3Department of Rehabilitation Medicine, Hongqi Hospital Affiliated to Mudanjiang Medical University, Mudanjiang 157011, Heilongjiang Province, China
  • Received:2024-09-10 Accepted:2024-11-26 Online:2025-10-28 Published:2025-03-28
  • Contact: Chen Pei, Associate professor, Master’s supervisor, Department of Pathophysiology, School of Basic Medicine, Mudanjiang Medical University, Mudanjiang 157011, Heilongjiang Province, China
  • About author:Zhang Xin, MS, Department of Pathophysiology, School of Basic Medicine, Mudanjiang Medical University, Mudanjiang 157011, Heilongjiang Province, China Guo Baojuan, MS, Department of Pathology, Baotou Mongolian Medicine and Traditional Chinese Medicine Hospital, Baotou 014000, Inner Mongolia Autonomous Region, China Zhang Xin and Guo Baojuan contributed equally to this article.
  • Supported by:
    Basic Scientific Research Business Expense Project of Provincial Colleges and Universities in Heilongjiang, No. 2023-KYYWF-0936 (to CP)

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摘要：


文题释义：
帕金森病：也常被称为“震颤麻痹”，是一种神经系统退行性疾病，主要病因是由于黑质多巴胺能神经元的退化和死亡，可能与遗传、环境因素及神经系统老化等多种因素有关。
丁苯酞：是芹菜籽中的一种化学成分，具有重建微循环、保护线粒体功能、抑制氧化应激和神经元凋亡等多种药理作用。研究表明，丁苯酞能够通过抗氧化、抗炎和改善脑微循环等机制保护多巴胺能神经元，但丁苯酞在改善帕金森病中的具体机制尚不清楚。

背景：丁苯酞具有抗氧化和抗炎作用，有研究证实其在帕金森病中具有保护作用，但具体机制尚不清楚。
目的：通过网络药理学、分子对接及细胞实验探讨丁苯酞的保护机制。
方法：①网络药理学、分子对接实验：利用数据库筛选丁苯酞的作用靶点及帕金森病靶点，取交集后构建交集靶点蛋白互作网络并筛选核心靶点，对核心靶点进行GO和KEGG通路富集分析，将目标靶点蛋白与丁苯酞进行分子对接验证。②细胞验证：将第6代PC12细胞分6组培养：对照组使用常规培养基培养，模型组采用N-甲基-4-苯基吡啶碘化物诱导帕金森病模型，ML385抑制剂组在诱导帕金森病模型的基础上加入核因子E2相关因子2抑制剂ML385，丁苯酞治疗组在诱导帕金森病模型的基础上加入丁苯酞，丁苯酞联合ML385处理组诱导帕金森病模型的基础上加入丁苯酞与ML385，丁苯酞组使用单独含丁酚酞的常规培养基培养，检测细胞增殖、细胞内还原型谷胱甘肽和丙二醛水平以及蛋白激酶B/糖原合酶激酶3β/核因子E2相关因子2(AKT/GSK-3β/Nrf2)信号通路的蛋白表达。
结果与结论：①筛选出药物与疾病靶点的交集靶点共52个，核心靶点包括基质金属蛋白酶9、GSK-3β等，涉及磷脂酰肌醇3-激酶/AKT、氧化应激相关信号通路；丁苯酞与GSK-3β的分子对接结合能为-18.27 kJ/mol，表明丁苯酞与GSK-3β具有一定的结合能力。②与模型组相比，丁苯酞治疗组PC12细胞活性与还原型谷胱甘肽水平升高(P < 0.05)，丙二醛水平降低(P < 0.05)，p-AKT、p-GSK-3β、Nu-Nrf2和T-Nrf2蛋白表达升高(P < 0.05)；与丁苯酞组相比，丁苯酞联合ML385处理组PC12细胞活性与还原型谷胱甘肽水平降低(P < 0.05)，丙二醛水平升高(P < 0.05)，Nu-Nrf2和T-Nrf2蛋白表达降低(P < 0.05)。③结果表明，丁苯酞可通过AKT/GSK-3β/Nrf2信号通路抑制氧化应激、改善细胞活性，对N-甲基-4-苯基吡啶碘化物诱导的帕金森细胞模型具有保护作用。
https://orcid.org/0009-0006-8605-3040(张鑫)；https://orcid.org/0000-0002-1084-7366(郭宝娟)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 网络药理学, 分子对接, 帕金森病, 丁苯酞, 氧化应激, 信号通路, 工程化组织构建

Abstract: BACKGROUND: D1-3-n-butylphthalide has antioxidant and anti-inflammatory effects and has been explored to have protective role in Parkinson’s disease, but the underlying mechanisms are unknown.
OBJECTIVE: To investigate the protective effect of D1-3-n-butylphthalide by the approach of network pharmacology, molecular docking, and cellular experimental validation.
METHODS: (1) Network pharmacology and molecular docking: The database was used to screen the targets of D1-3-n-butylphthalide and Parkinson’s disease. The intersection was taken from the construction of the target protein interaction network, and then screen the core targets. The GO and KEGG pathway enrichment was used to further analyze the core targets. The interaction between the target proteins and D1-3-n-butylphthalide was verified by molecular docking. (2) Cell validation: The passage 6 PC12 cells were divided into six groups for culture. The control group was cultured with conventional culture medium. The model group was cultured with N-methyl-4-phenylpyridinium iodide to induce Parkinson’s disease model. The ML385 inhibitor group was added with nuclear factor E2-related factor 2 inhibitor ML385 on the basis of inducing Parkinson’s disease model. The D1-3-n-butylphthalide treatment group was added with butylphthalide on the basis of inducing Parkinson’s disease model. The D1-3-n-butylphthalide combined with ML385 treatment group was added with D1-3-n-butylphthalide and ML385 on the basis of inducing Parkinson’s disease model. The D1-3-n-butylphthalide group was cultured with conventional culture medium containing butylphthalide alone. Cell proliferation, intracellular reduced glutathione and malondialdehyde levels, and protein expression of protein kinase B/glycogen synthase kinase 3β/nuclear factor E2-related factor 2 (AKT/GSK-3β/Nrf2) signaling pathway were detected.
RESULTS AND CONCLUSION: (1) A total of 52 targets were screened for the intersection of drugs and disease targets, and the core targets including the matrix metalloproteinase 9 and GSK-3β were involved the phosphatidylinositol 3-kinase (PI3K)/AKT and oxidative stress-related signaling pathways. The molecular docking binding energy of D1-3-n-butylphthalide and GSK-3β was -18.27 kJ/mol, which indicated that D1-3-n-butylphthalide had a good binding ability with GSK-3β. (2) Compared with the model group, the PC12 cell activity and reduced glutathione level in the D1-3-n-butylphthalide treatment group were increased (P < 0.05), the malondialdehyde level was decreased (P < 0.05), and the expression of p-AKT, p-GSK-3β, Nu-Nrf2, and T-Nrf2 proteins was increased (P < 0.05). Compared with the D1-3-n-butylphthalide group, the PC12 cell activity and reduced glutathione level in the D1-3-n-butylphthalide combined with ML385 treatment group were decreased (P < 0.05), the malondialdehyde level was increased (P < 0.05), and the expression of Nu-Nrf2 and T-Nrf2 proteins was decreased (P < 0.05). (3) These results demonstrate that D1-3-n-butylphthalide can inhibit oxidative stress and improve cell activity through the AKT/GSK-3β/Nrf2 signaling pathway, and has a protective effect on the Parkinson’s cell model induced by N-methyl-4-phenylpyridinium iodide.

Key words: network pharmacology, molecular docking, Parkinson’s disease, D1-3-n-butylphthalide, oxidative stress, signaling pathway, engineered tissue construction

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