[TFEB activator 1 enhances autophagic degradation of oligomeric amyloid-β in microglia].

Q3 Medicine
生理学报 Pub Date : 2024-06-25
Yu-Qi Xie, Li Zhu, Xue-Ting Wang
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引用次数: 0

Abstract

The purpose of the study was to investigate the mechanism of TFEB activator 1 (TA1) improving the autophagic degradation of oligomeric amyloid-β (oAβ) in microglia, and to explore the therapeutic effect of TA1 on an in vitro model of microglia in Alzheimer's disease (AD). Primary microglia were exposed to 1 μmol/L oAβ for 0, 3, 12, and 24 h respectively to construct the in vitro model of microglia in AD. In order to explore the therapeutic effect of TA1, primary microglia were co-treated with 1 μmol/L oAβ and 1 μmol/L TA1 for 12 h. To determine the autophagy flux, the above cells were further treated with 100 nmol/L Bafilomycin A1 for 1 h before fixation. Fluorescent probes were used to detect the endocytosis or degradation of oAβ1-42 by microglia. The autophagic flux was determined by infection of lentivirus mCherry-EGFP-LC3. The nuclear TFEB intensity, the autophagosomes number, and the colocalization ratio of oAβ1-42 with lysosome-associated membrane protein 1 (LAMP1) or microtubule-associated protein light chain 3 (LC3), were detected by immunofluorescence assay. Expressions of autophagy-related-genes, including Lamp1, Atg5, and Map1lc3b, were detected by qRT-PCR. Results showed that prolonged oAβ exposure inhibited the endocytosis and degradation of oAβ by microglia. Meanwhile, the number of autophagosomes and autophagy flux in microglia decreased after 12 h of oAβ treatment. We further found that the nuclear expression of autophagy regulator TFEB decreased after 12 h of oAβ exposure, resulting in the decrease of autophagy genes, thus leading to the damage of autophagic degradation of oAβ. Therefore, long-term oAβ exposure was considered to construct the in vitro model of microglia in AD. After TA1 treatment, the nuclear expression of TFEB in cells was obviously upregulated. TA1 treatment upregulated the expressions of autophagy-related genes, leading to the recovery of autophagy flux. TA1 also recovered the endocytosis and degradation of oAβ by microglia. In conclusion, TA1 could improve oAβ clearance by microglia in AD by upregulating microglial TFEB-mediated autophagy, suggesting TA1 as a potential therapeutic drug for AD.

[TFEB激活剂1能增强小胶质细胞中低聚淀粉样蛋白-β的自噬降解能力】。]
该研究旨在探讨TFEB激活剂1(TA1)改善小胶质细胞自噬降解低聚淀粉样蛋白-β(oAβ)的机制,并探索TA1对阿尔茨海默病(AD)小胶质细胞体外模型的治疗效果。将原代小胶质细胞分别暴露于1 μmol/L oAβ中0、3、12和24小时,构建AD小胶质细胞体外模型。为了探究TA1的治疗效果,原代小胶质细胞用1 μmol/L oAβ和1 μmol/L TA1共同处理12小时。为了测定自噬通量,上述细胞在固定前再用100 nmol/L Bafilomycin A1处理1小时。荧光探针用于检测小胶质细胞对 oAβ1-42 的内吞或降解。自噬通量通过感染慢病毒 mCherry-EGFP-LC3 进行检测。免疫荧光检测了核TFEB强度、自噬体数量以及oAβ1-42与溶酶体相关膜蛋白1(LAMP1)或微管相关蛋白轻链3(LC3)的共定位比率。通过 qRT-PCR 检测自噬相关基因的表达,包括 Lamp1、Atg5 和 Map1lc3b。结果表明,长时间暴露于 oAβ 会抑制小胶质细胞对 oAβ 的内吞和降解。同时,oAβ处理12小时后,小胶质细胞中自噬体的数量和自噬通量减少。我们进一步发现,自噬调节因子 TFEB 的核表达在 oAβ 暴露 12 h 后下降,导致自噬基因的减少,从而导致 oAβ 的自噬降解受损。因此,长期暴露于oAβ被认为是构建AD小胶质细胞体外模型的考虑因素。TA1处理后,细胞核中TFEB的表达明显上调。TA1处理后,自噬相关基因表达上调,自噬通量恢复。TA1 还能恢复小胶质细胞对 oAβ 的内吞和降解。总之,TA1可通过上调小胶质细胞TFEB介导的自噬,改善小胶质细胞对AD中oAβ的清除,提示TA1是一种潜在的AD治疗药物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
生理学报
生理学报 Medicine-Medicine (all)
CiteScore
1.20
自引率
0.00%
发文量
4820
期刊介绍: Acta Physiologica Sinica (APS) is sponsored by the Chinese Association for Physiological Sciences and Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences (CAS), and is published bimonthly by the Science Press, China. APS publishes original research articles in the field of physiology as well as research contributions from other biomedical disciplines and proceedings of conferences and symposia of physiological sciences. Besides “Original Research Articles”, the journal also provides columns as “Brief Review”, “Rapid Communication”, “Experimental Technique”, and “Letter to the Editor”. Articles are published in either Chinese or English according to authors’ submission.
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