穿心莲内酯通过靶向动力相关蛋白 1 介导的线粒体分裂缓解 MPTP-PD 小鼠的帕金森病。
Andrographolide alleviates Parkinsonism in MPTP-PD mice via targeting mitochondrial fission mediated by dynamin-related protein 1.
机构信息
School of Pharmacy, Jiangsu University, Zhenjiang, China.
Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases, College of Pharmaceutical Science, Soochow University, Suzhou, China.
出版信息
Br J Pharmacol. 2019 Dec;176(23):4574-4591. doi: 10.1111/bph.14823. Epub 2019 Nov 14.
BACKGROUND AND PURPOSE
Accumulating evidence indicates that mitochondrial dynamics play an important role in the progressive deterioration of dopaminergic neurons. Andrographolide has been found to exert neuroprotective effects in several models of neurological diseases. However, the mechanism of how andrographolide protects neurons in Parkinson's disease (PD) remains not fully understood.
EXPERIMENTAL APPROACH
Behavioural experiments were performed to examine the effect of andrographolide in 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-PD mice. Mitochondrial mass and morphology were visualized using transmission electron microscopy (TEM). SH-SY5Y cells and primary mouse neurons were exposed to rotenone to mimic PD in vitro. Western blotting, co-immunoprecipitation and immunofluorescence were performed. The target protein of andrographolide was identified by biotin-andrographolide pulldown assay as well as drug affinity responsive target stability (DARTS), cellular thermal shift (CETSA), and surface plasmon resonance (SPR) assays.
KEY RESULTS
Andrographolide administration improved behavioural deficits and attenuated loss of dopaminergic neurons in MPTP-exposed mice and reduced cell death induced by rotenone in vitro. An increased mitochondrial mass, and decreased surface area were found in the striatum from MPTP-PD mice, as well as in rotenone-treated primary neurons and SH-SY5Y cells, while andrographolide treatment preserved mitochondrial mass and morphology. Dynamin-related protein 1 (DRP1) was identified as a target protein of andrographolide. Andrographolide bound to DRP1 and inhibited its GTPase activity, thereby preventing excessive mitochondria fission and neuronal damage in PD.
CONCLUSIONS AND IMPLICATIONS
Our findings suggest that andrographolide may protect neurons against rotenone- or MPTP-induced damage in vitro and in vivo through inhibiting mitochondrial fission.
背景与目的
越来越多的证据表明，线粒体动力学在多巴胺能神经元的进行性恶化中起着重要作用。穿心莲内酯已被发现对几种神经疾病模型具有神经保护作用。然而，穿心莲内酯如何保护帕金森病（PD）神经元的机制仍不完全清楚。
实验方法
通过行为学实验研究了穿心莲内酯在 1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）诱导的 PD 小鼠中的作用。使用透射电子显微镜（TEM）观察线粒体质量和形态。用鱼藤酮处理 SH-SY5Y 细胞和原代小鼠神经元，在体外模拟 PD。进行 Western blot、共免疫沉淀和免疫荧光实验。通过生物素穿心莲内酯下拉实验、药物亲和反应靶标稳定性（DARTS）、细胞热转移（CETSA）和表面等离子体共振（SPR）实验鉴定穿心莲内酯的靶蛋白。
主要结果
穿心莲内酯给药可改善 MPTP 暴露小鼠的行为缺陷和减少多巴胺能神经元的丢失，并减少体外鱼藤酮诱导的细胞死亡。MPTP-PD 小鼠纹状体以及鱼藤酮处理的原代神经元和 SH-SY5Y 细胞中发现线粒体质量增加，表面积减少，而穿心莲内酯处理可维持线粒体质量和形态。动力相关蛋白 1（DRP1）被鉴定为穿心莲内酯的靶蛋白。穿心莲内酯与 DRP1 结合并抑制其 GTP 酶活性，从而防止 PD 中过度的线粒体裂变和神经元损伤。
结论和意义
我们的研究结果表明，穿心莲内酯可能通过抑制线粒体裂变来保护神经元免受鱼藤酮或 MPTP 诱导的损伤，无论是在体外还是体内。
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