Andrographolide alleviates Parkinsonism in MPTP-PD mice via targeting mitochondrial fission mediated by dynamin-related protein 1
Andrographolide alleviates Parkinsonism in MPTP-PD mice via targeting mitochondrial fission mediated by dynamin-related protein 1
复制标题
穿心莲内酯通过靶向动力相关蛋白 1 介导的线粒体裂变减轻 MPTP-PD 小鼠的帕金森病
DOI：
10.1111/bph.14823
复制
发表时间：
2019-11-14
影响因子：
7.3
通讯作者：
Gao, Jing
中科院分区：
文献类型：
--
作者：
Geng, Ji;Liu, Wen;Gao, Jing
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退化中起重要作用。穿心莲内酯已被发现在几种神经系统疾病模型中发挥神经保护作用。然而，穿心莲内酯在帕金森病（PD）中如何保护神经元的机制尚不完全清楚。实验方法通过行为学实验研究穿心莲内酯对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)-PD小鼠的影响。透射电镜观察线粒体质量和形态。将SH-SY5Y细胞和小鼠原代神经元暴露于鱼藤酮体外模拟PD。Western blotting、共免疫沉淀、免疫荧光检测。通过生物素-穿心莲内酯下拉实验、药物亲和力响应靶稳定性（DARTS）、细胞热移（CETSA）、表面等离子体共振（SPR）等方法鉴定穿心莲内酯的靶蛋白。在体外实验中，给药穿心莲内酯改善了mptp暴露小鼠的行为缺陷，减轻了多巴胺能神经元的损失，减少了鱼藤酮诱导的细胞死亡。MPTP-PD小鼠纹状体、鱼藤酮处理的原代神经元和SH-SY5Y细胞线粒体质量增加，表面积减少，而穿心莲内酯处理的线粒体质量和形态保持不变。动力蛋白相关蛋白1 （DRP1）被确定为穿心莲内酯的靶蛋白。穿心术内酯与DRP1结合，抑制其GTPase活性，从而防止PD中线粒体过度分裂和神经元损伤。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诱导的体外和体内损伤。
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.