Long-term protective effects of AAV9-mesencephalic astrocyte-derived neurotrophic factor gene transfer in parkinsonian rats
Long-term protective effects of AAV9-mesencephalic astrocyte-derived neurotrophic factor gene transfer in parkinsonian rats
复制标题
AAV 9-中脑星形胶质细胞源性神经营养因子基因转移对帕金森病大鼠的长期保护作用
DOI：
10.1016/j.expneurol.2017.01.008
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发表时间：
2017-05-01
影响因子：
5.3
通讯作者：
Xu, Qun-Yuan
中科院分区：
文献类型：
--
作者：
Hao, Fei;Yang, Chun;Xu, Qun-Yuan
纹状体内注射中脑星形胶质细胞源性神经营养因子（MANF）蛋白已被证明在6-羟基多巴胺（6-OHDA）损伤的帕金森病大鼠模型中提供神经保护和神经恢复作用。在这里，我们使用腺相关病毒血清型9（AAV 9）载体，将人MANF（hMANF）基因导入6-OHDA损伤后10天的大鼠纹状体，以研究hMANF对黑质多巴胺能神经元的长期影响和MANF神经保护的机制。纹状体内注射AAV 9-hMANF载体导致hMANF基因在注射的纹状体中稳健且广泛的表达长达24周。在同侧黑质中也检测到hMANF蛋白水平升高。hMANF基因转移促进了黑质多巴胺能神经元的存活，纹状体多巴胺能纤维的再生和纹状体多巴胺水平的上调，导致病毒注射后旋转行为的长期改善长达16周。通过使用SH-SY 5 Y细胞，我们发现细胞内和细胞外应用MANF通过抑制内质网应激和激活PI 3 K/Akt/mTOR通路来保护细胞免受6-OHDA诱导的毒性。我们的研究结果表明，AAV 9介导的hMANF基因递送到纹状体对帕金森病大鼠黑质纹状体多巴胺能系统产生长期的神经保护和神经再生作用，并提供了对MANF神经保护机制的见解。(C)2017爱思唯尔公司All rights reserved.
Intrastriatal injection of mesencephalic astrocyte-derived neurotrophic factor (MANF) protein has been shown to provide neuroprotective and neurorestorative effects in a 6-hydroxydopamine (6-OHDA) - lesioned rat model of Parkinson's disease. Here, we used an adeno-associated virus serotype 9 (AAV9) vector to deliver the human MANF (hMANF) gene into the rat striatum 10 days after a 6-OHDA lesion to examine long-term effects of hMANF on nigral dopaminergic neurons and mechanisms underlying MANF neuroprotection. Intrastriatal injection of AAV9-hMANF vectors led to a robust and widespread expression of the hMANF gene in the injected striatum up to 24 weeks. Increased levels of hMANF protein were also detected in the ipsilateral substantia nigra. The hMANF gene transfer promoted the survival of nigral dopaminergic neurons, regeneration of striatal dopaminergic fibers and an upregulation of striatal dopamine levels, resulting in a long-term improvement of rotational behavior up to 16 weeks after viral injections. By using SH-SY5Y cells, we found that intra- and extracellular application of MANF protected cells against 6-OHDA-induced toxicity via inhibiting the endoplasmic reticulum stress and activating the PI3K/Akt/mTOR pathway. Our results suggest that AAV9-mediated hMANFgene delivery into the striatum exerts long-term neuroprotective and neuroregenerative effects on the nigrostriatal dopaminergic system in parkinsonian rats, and provide insights into mechanisms responsible for MANF neuroprotection. (C) 2017 Elsevier Inc. All rights reserved.