天麻通过调控线粒体形态来减轻突变型亨廷顿蛋白聚集的新机制。
Morphological control of mitochondria as the novel mechanism of Gastrodia elata in attenuating mutant huntingtin-induced protein aggregations.
机构信息
National Research Institute of Chinese Medicine, Ministry of Health and Welfare, Taipei, Taiwan, ROC; Program for Neural Regenerative Medicine, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan, ROC.
Brain Research Center, National Yang-Ming University, Taipei, Taiwan, ROC; Department of Life Sciences and Institute of Genome Sciences, National Yang-Ming University, Taipei, Taiwan, ROC; Biophotonics Interdisciplinary Research Center, National Yang-Ming University, Taipei, Taiwan, ROC.
出版信息
Phytomedicine. 2019 Jun;59:152756. doi: 10.1016/j.phymed.2018.11.016. Epub 2018 Nov 14.
BACKGROUND
According to Compendium of Materia Medica, Gastrodia elata (GE) Blume as a top grade and frequently prescribed herbal medicine has been used in treating dizziness, headaches, and epilepsy, indicating a neuroprotective effect. Because GE is capable of suppressing a hyperactive liver and thus calming endogenous wind, and because Huntington's disease (HD) can be classified as a phenomenon of disturbed liver wind, it is suggested that GE might be beneficial in treating HD. However, although current studies support GE for the prevention of diverse neurodegenerations such as HD, its detailed mechanisms remain elusive.
PURPOSE
To investigate the molecular mechanism of GE in preventing HD by focusing on mitochondrial morphology, which is highly associated with HD etiology and thus proposed as a therapeutic target of neurodegenerations.
STUDY DESIGN/METHODS: The overexpression of the mutant huntingtin (mHTT) gene in rat pheochromocytoma (PC12) cells was used as an in vitro cell model of HD. A filter retardation assay was applied to measure protein aggregations during HTT expression. Cotransfection with mitochondrial fusion and fission genes was used to test their relationships with HTT aggregates by monitoring with a confocal laser scanning microscope and filter retardation assay. Western blot analysis was used to estimate protein expression under different drug treatments or cotransfections with other related genes.
RESULTS
The overexpression of mutant but not normal HTT genes significantly resulted in protein aggregations in PC12 cells. GE dose-dependently attenuated mHTT-induced protein aggregations and free radical formations. GE significantly reversed mHTT-induced mitochondrial fragmentation and dysregulation of mitochondrial fusion and fission molecules. The overexpression of mitochondrial fusion genes attenuated mHTT-induced protein aggregations. Further, Mdivi-1, a DRP1 fission molecule inhibitor, significantly reversed mHTT-induced protein aggregations and mitochondrial fragmentation.
CONCLUSION
GE attenuated mHTT aggregations through the control of mitochondrial fusion and the fission pathway.
背景
根据《本草纲目》记载，天麻（GE）作为一种高档、常用的草药，已被用于治疗头晕、头痛和癫痫等疾病，具有神经保护作用。由于 GE 能够抑制肝脏亢进，从而平息内风，而亨廷顿病（HD）可归类为肝风失调现象，因此推测 GE 可能对治疗 HD 有益。然而，尽管目前的研究支持 GE 预防多种神经退行性疾病，如 HD，但其详细机制仍不清楚。
目的
通过关注与 HD 病因高度相关的线粒体形态，研究 GE 预防 HD 的分子机制，线粒体形态被提出作为神经退行性疾病的治疗靶点。
研究设计/方法：在体外细胞模型中，使用大鼠嗜铬细胞瘤（PC12）细胞中转染突变型 huntingtin（mHTT）基因来模拟 HD。采用滤膜阻滞试验来测量 HTT 表达过程中的蛋白聚集。通过共转染线粒体融合和分裂基因，使用共聚焦激光扫描显微镜和滤膜阻滞试验监测它们与 HTT 聚集物的关系。Western blot 分析用于评估不同药物处理或与其他相关基因共转染后的蛋白表达。
结果
突变型而非正常型 HTT 基因的过表达显著导致 PC12 细胞中的蛋白聚集。GE 呈剂量依赖性地减弱 mHTT 诱导的蛋白聚集和自由基形成。GE 显著逆转 mHTT 诱导的线粒体碎片化和线粒体融合与分裂分子的失调。线粒体融合基因的过表达减弱了 mHTT 诱导的蛋白聚集。此外，DRP1 分裂分子抑制剂 Mdivi-1 显著逆转 mHTT 诱导的蛋白聚集和线粒体碎片化。
结论
GE 通过控制线粒体融合和分裂途径来减弱 mHTT 聚集。
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