α-突触核蛋白通过 STAT1 依赖性抑制 Ulk1 转录来破坏小胶质细胞自噬。

IF 9.3 1区 医学 Q1 IMMUNOLOGY
Chong-Shuang Pei, Xiao-Ou Hou, Zhen-Yuan Ma, Hai-Yue Tu, Hai-Chun Qian, Yang Li, Kai Li, Chun-Feng Liu, Liang Ouyang, Jun-Yi Liu, Li-Fang Hu
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引用次数: 0

摘要

背景:神经胶质细胞的自噬功能障碍与帕金森病(PD)的发病机制有关。之前的研究报告称,α-突触核蛋白(α-Syn)会破坏培养小胶质细胞的自噬功能。然而,人们对小胶质细胞自噬失调的机制知之甚少:方法:通过 AAV 介导将 α-Syn 运送到小鼠黑质和纹状体注射 α-Syn 预成纤维(PFF),产生了两种基于 α-Syn 的帕金森病模型。通过定量 PCR、Western 印迹和免疫染色法测定了小胶质细胞 UNC-51 样激酶 1(Ulk1)和其他自噬相关基因在体外、PD 小鼠体内以及 PD 患者和健康对照组外周血单核细胞中的水平。通过荧光素酶报告实验和其他生化研究确定了信号转导和激活转录1(STAT1)对Ulk1转录的调控作用,并通过敲除或过表达Stat1进行了验证。α-Syn对神经胶质STAT1活化的影响通过免疫组化和免疫印迹进行了评估。通过免疫染色和免疫印迹法评估了小胶质细胞Stat1条件性敲除(cKO)或ULK1激活剂BL-918治疗后,PD小鼠和对照组小鼠黑质中的小胶质细胞状态、促炎症分子表达和多巴胺能神经元丢失的变化。运动行为通过开阔地测试、旋转木马测试和平衡木穿越进行测定:结果:在体外和体内的帕金森病小鼠模型中,控制自噬启动的激酶--小胶质细胞ULK1的转录均有所下降。STAT1在抑制Ulk1转录方面起着关键作用。具体来说,Stat1过表达会下调Ulk1的转录,而敲除Stat1会增加ULK1的表达,同时增加LC3II和减少SQSTM1/p62蛋白。Stat1的消减减轻了α-Syn PFF引起的小胶质细胞ULK1表达的减少和自噬的破坏。重要的是,ULK1激活剂BL-918和小胶质细胞Stat1 cKO减轻了PD模型的神经炎症、多巴胺能神经元损伤和运动缺陷:这些发现揭示了α-Syn损害小胶质细胞自噬的新机制,并表明靶向STAT1或ULK1可能是一种治疗帕金森病的策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
α-Synuclein disrupts microglial autophagy through STAT1-dependent suppression of Ulk1 transcription.

Background: Autophagy dysfunction in glial cells is implicated in the pathogenesis of Parkinson's disease (PD). The previous study reported that α-synuclein (α-Syn) disrupted autophagy in cultured microglia. However, the mechanism of microglial autophagy dysregulation is poorly understood.

Methods: Two α-Syn-based PD models were generated via AAV-mediated α-Syn delivery into the mouse substantia nigra and striatal α-Syn preformed fibril (PFF) injection. The levels of microglial UNC-51-like kinase 1 (Ulk1) and other autophagy-related genes in vitro and in PD mice, as well as in the peripheral blood mononuclear cells of PD patients and healthy controls, were determined via quantitative PCR, western blotting and immunostaining. The regulatory effect of signal transducer and activator of transcription 1 (STAT1) on Ulk1 transcription was determined via a luciferase reporter assay and other biochemical studies and was verified through Stat1 knockdown or overexpression. The effect of α-Syn on glial STAT1 activation was assessed by immunohistochemistry and western blotting. Changes in microglial status, proinflammatory molecule expression and dopaminergic neuron loss in the nigrostriatum of PD and control mice following microglial Stat1 conditional knockout (cKO) or treatment with the ULK1 activator BL-918 were evaluated by immunostaining and western blotting. Motor behaviors were determined via open field tests, rotarod tests and balance beam crossing.

Results: The transcription of microglial ULK1, a kinase that controls autophagy initiation, decreased in both in vitro and in vivo PD mouse models. STAT1 plays a critical role in suppressing Ulk1 transcription. Specifically, Stat1 overexpression downregulated Ulk1 transcription, while Stat1 knockdown increased ULK1 expression, along with an increase in LC3II and a decrease in the SQSTM1/p62 protein. α-Syn PFF caused toll-like receptor 4-dependent activation of STAT1 in microglia. Ablation of Stat1 alleviated the decrease in microglial ULK1 expression and disruption of autophagy caused by α-Syn PFF. Importantly, the ULK1 activator BL-918 and microglial Stat1 cKO attenuated neuroinflammation, dopaminergic neuronal damage and motor defects in PD models.

Conclusions: These findings reveal a novel mechanism by which α-Syn impairs microglial autophagy and indicate that targeting STAT1 or ULK1 may be a therapeutic strategy for PD.

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来源期刊
Journal of Neuroinflammation
Journal of Neuroinflammation 医学-神经科学
CiteScore
15.90
自引率
3.20%
发文量
276
审稿时长
1 months
期刊介绍: The Journal of Neuroinflammation is a peer-reviewed, open access publication that emphasizes the interaction between the immune system, particularly the innate immune system, and the nervous system. It covers various aspects, including the involvement of CNS immune mediators like microglia and astrocytes, the cytokines and chemokines they produce, and the influence of peripheral neuro-immune interactions, T cells, monocytes, complement proteins, acute phase proteins, oxidative injury, and related molecular processes. Neuroinflammation is a rapidly expanding field that has significantly enhanced our knowledge of chronic neurological diseases. It attracts researchers from diverse disciplines such as pathology, biochemistry, molecular biology, genetics, clinical medicine, and epidemiology. Substantial contributions to this field have been made through studies involving populations, patients, postmortem tissues, animal models, and in vitro systems. The Journal of Neuroinflammation consolidates research that centers around common pathogenic processes. It serves as a platform for integrative reviews and commentaries in this field.
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