Institute of Microbiology and Immunology, Faculty of Medicine, University of Belgrade, Dr. Subotica 1, 11000 Belgrade, Serbia.
Institute for Biological Research "Sinisa Stankovic"-National Institute of Republic of Serbia, University of Belgrade, Bulevar Despota Stefana 142, 11000 Belgrade, Serbia.
Int J Mol Sci. 2024 Oct 3;25(19):10659. doi: 10.3390/ijms251910659.
The effects of trehalose, an autophagy-inducing disaccharide with neuroprotective properties, on the neurotoxicity of parkinsonian mimetics 6-hydroxydopamine (6-OHDA) and 1-methyl-4-phenylpiridinium (MPP) are poorly understood. In our study, trehalose suppressed 6-OHDA-induced caspase-3/PARP1 cleavage (detected by immunoblotting), apoptotic DNA fragmentation/phosphatidylserine externalization, oxidative stress, mitochondrial depolarization (flow cytometry), and mitochondrial damage (electron microscopy) in SH-SY5Y neuroblastoma cells. The protection was not mediated by autophagy, autophagic receptor p62, or antioxidant enzymes superoxide dismutase and catalase. Trehalose suppressed 6-OHDA-induced activation of c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (MAPK), and AMP-activated protein kinase (AMPK), as revealed by immunoblotting. Pharmacological/genetic inhibition of JNK, p38 MAPK, or AMPK mimicked the trehalose-mediated cytoprotection. Trehalose did not affect the extracellular signal-regulated kinase (ERK) and mechanistic target of rapamycin complex 1 (mTORC1)/4EBP1 pathways, while it reduced the prosurvival mTORC2/AKT signaling. Finally, trehalose enhanced oxidative stress, mitochondrial damage, and apoptosis without decreasing JNK, p38 MAPK, AMPK, or AKT activation in SH-SY5Y cells exposed to MPP. In conclusion, trehalose protects SH-SY5Y cells from 6-OHDA-induced oxidative stress, mitochondrial damage, and apoptosis through autophagy/p62-independent inhibition of JNK, p38 MAPK, and AMPK. The opposite effects of trehalose on the neurotoxicity of 6-OHDA and MPP+ suggest caution in its potential development as a neuroprotective agent.
海藻糖是一种具有神经保护作用的自噬诱导二糖，其对帕金森病模拟物 6-羟多巴胺（6-OHDA）和 1-甲基-4-苯基吡啶（MPP+）神经毒性的影响知之甚少。在我们的研究中，海藻糖抑制了 SH-SY5Y 神经母细胞瘤细胞中 6-OHDA 诱导的半胱天冬酶-3/PARP1 裂解（通过免疫印迹检测）、凋亡性 DNA 片段化/磷脂酰丝氨酸外翻、氧化应激、线粒体去极化（流式细胞术）和线粒体损伤（电子显微镜）。这种保护不是通过自噬、自噬受体 p62 或抗氧化酶超氧化物歧化酶和过氧化氢酶介导的。海藻糖抑制了 6-OHDA 诱导的 c-Jun N-末端激酶（JNK）、p38 丝裂原激活蛋白激酶（p38 MAPK）和 AMP 激活蛋白激酶（AMPK）的激活，这是通过免疫印迹显示的。JNK、p38 MAPK 或 AMPK 的药理学/遗传学抑制模拟了海藻糖介导的细胞保护作用。海藻糖不影响细胞外信号调节激酶（ERK）和雷帕霉素机制靶蛋白复合物 1（mTORC1）/4EBP1 通路，而减少了促生存的 mTORC2/AKT 信号。最后，海藻糖增强了氧化应激、线粒体损伤和细胞凋亡，而不会降低暴露于 MPP+的 SH-SY5Y 细胞中 JNK、p38 MAPK、AMPK 或 AKT 的激活。总之，海藻糖通过非依赖自噬/p62 的 JNK、p38 MAPK 和 AMPK 抑制来保护 SH-SY5Y 细胞免受 6-OHDA 诱导的氧化应激、线粒体损伤和细胞凋亡。海藻糖对 6-OHDA 和 MPP+神经毒性的相反作用表明，在将其开发为神经保护剂时需要谨慎。
