6-Hydroxydopamine Induces Neurodegeneration in Terminally Differentiated SH-SY5Y Neuroblastoma Cells via Enrichment of the Nucleosomal Degradation Pathway: a Global Proteomics Approach

Abstract

The SH-SY5Y human neuroblastoma cells have been used for decades as a cell-based model of dopaminergic neurons to explore the underlying science of cellular and molecular mechanisms of neurodegeneration in Parkinson’s disease (PD). However, data revealing the protein expression changes in 6-OHDA induced cytotoxicity in differentiated SH-SY5Y cells remain void. Therefore, we investigated the differentially regulated proteins expressed in terminally differentiated SH-SY5Y cells (differ-SH-SY5Y neural cells) exposed to 6-hydroxydopamine (6-OHDA) using the LC–MS/MS technology and construed the data using the online bioinformatics databases such as PANTHER, STRING, and KEGG. Our studies demonstrated that the neuronal development in differ-SH-SY5Y neural cells was indicated by the overexpression of proteins responsible for neurite formations such as calnexin (CANX) and calreticulin (CALR) besides significant downregulation of ribosomal proteins. The enrichment of the KEGG ribosome pathway was detected with significant downregulation (p < 0.05) of all the 21 ribosomal proteins in differ-SH-SY5Y neural cells compared with undifferentiated cells. Whereas in the PD model, the pathological changes induced by 6-OHDA were indicated by the presence of unfolded and misfolded proteins, which triggered the response of 10 kDa heat shock proteins (HSP), namely HSPE1 and HSPA9. Moreover, the 6-OHDA-induced neurodegeneration in differ-SH-SY5Y neural cells also upregulated the voltage-dependent anion-selective channel protein 1 (VDAC1) protein and enriched the KEGG systemic lupus erythematosus (SLE) pathway that was regulated by 17 histone proteins (p < 0.05) in differ-SH-SY5Y neural cells. These results suggest that the nucleosomal degradation pathway may have regulated the 6-OHDA induced neurodegeneration in PD cell-based model, which is reflected by increased apoptosis and histone release in differ-SH-SY5Y neural cells.

The Potential Role of Voltage-Dependent Anion Channel in the Treatment of Parkinson’s Disease

“…It can be suggested that activation of autophagy contributes to the alleviation of PD by regulating VDAC1; however, there is no VDAC1-specific autophagy activator. Studies on PD patients, research by Chu et al, and our current study have shown that VDAC1 expression is downregulated during PD [60], but VDAC1 expression is not downregulated in vivo and in vitro models of PD [41,42,59,[78][79][80][81]. We speculate that VDAC1 may be a double-edged sword playing different roles in PD.…”

“…However, the details of this protective mechanism remain unmapped. In addition, there is an increase in the expression of VDAC1 in the PD cell model induced by rotenone [ 78 ], 1-methyl-4-phenylpyridinium (MPP + ) [ 79 ], and 6-hydroxydopamine (6-OHDA) [ 80 ]. A recent study has revealed that vitamin D has neuroprotective effect on PD model of rats induced by 6-OHDA by decreasing the upregulation of VDAC1 [ 81 ].…”

Comparative proteomics study of mitochondrial electron transport system modulation in SH-SY5Y cells following MPP+ versus 6-OHDA-induced neurodegeneration

“…These proteins were analyzed based on changes in the biological processes of the GO term classification cluster. According to GO biological process analysis, upregulated proteins in 6-OHDAtreated SH-SY5Y cells respond to external stress, and as previous studies have shown (Magalingam et al 2022), proteins that regulate ribosomal assembly were upregulated. These results were consistent with reactome gene set analysis (Fig.…”

“…However, the mechanism underlying 6-OHDA-and MPP+-induced injury remains unclear. Previous studies have only reported the effects of 6-OHDA and MPP+ on neuronal cells separately (Blum et al 2001;Perfeito et al 2013;Xie et al 2011;Komatsubara et al 2012;Choi et al 2014;Alberio et al 2014;Magalingam et al 2022), and none of them have directly compared the effects of 6-OHDA and MPP+ on neuronal cells. Thus, large-scale proteome investigation of the effects of 6-OHDA and MPP+ on neuronal cells will bring new insights into PD.…”

“…Proteomics research using liquid chromatographytandem mass spectrometry (LC-MS/MS) involves largescale protein expression analysis that provides biological insights into cellular structures and functions. Because of the advantage of analyzing large-scale protein expression, proteomics research for PD is being actively conducted (Xie et al 2011;Komatsubara et al 2012;Choi et al 2014;Alberio et al 2014;Magalingam et al 2022). Proteins within the cell play a crucial role in providing cellular structure, movement, and communication, and participate in metabolism, respiration, signal transduction, and reproduction activities.…”