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Pro-inflammatory and pro-apoptotic effects of the non-protein amino acid L-Azetidine-2-carboxylic acid in BV2 microglial cells

Citation

Piper, JA and Jansen, MI and Broome, ST and Rodgers, KJ and Musumeci, G and Castorina, A, Pro-inflammatory and pro-apoptotic effects of the non-protein amino acid L-Azetidine-2-carboxylic acid in BV2 microglial cells, Current Issues in Molecular Biology, 44, (10) pp. 4500-4516. ISSN 1467-3045 (2022) [Refereed Article]


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© 2022. The Authors. Licensee MDPI, Basel, Switzerland. This article is licensed under a Creative Commons Attribution 4.0 International (CC BY 4.0) License (https://creativecommons.org/licenses/by/4.0/), which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

DOI: doi:10.3390/cimb44100308

Abstract

L-Azetidine-2-carboxylic acid (AZE) is a toxic non-protein coding amino acid (npAA) that is highly abundant in sugar and table beets. Due to its structural similarity with the amino acid L-proline, AZE can evade the editing process during protein assembly in eukaryotic cells and be misincorporated into L-proline-rich proteins, potentially causing protein misfolding and other detrimental effects to cells. In this study, we sought to determine if AZE treatment triggered pro-inflammatory and pro-apoptotic responses in BV2 microglial cells. BV2 microglial cells exposed to AZE at increasing concentrations (0–2000 µM) at 0, 3, 6, 12 and 24 h were assayed for cell viability (MTT) and nitric oxide release (Griess assay). Annexin V-FITC/propidium iodide (PI) staining was used to assess apoptosis. Real-time qPCR, Western blot and immunocytochemistry were used to interrogate relevant pro- and anti-inflammatory and other molecular targets of cell survival response. AZE (at concentrations > 1000 µM) significantly reduced cell viability, increased BAX/Bcl2 ratio and caused cell death. Results were mirrored by a robust increase in nitric oxide release, percentage of activated/polarised cells and expression of pro-inflammatory markers (IL-1β, IL-6, NOS2, CD68 and MHC-2a). Additionally, we found that AZE induced the expression of the extracellular matrix degrading enzyme matrix metalloproteinase 9 (MMP-9) and brain derived neurotrophic factor (BDNF), two critical regulators of microglial motility and structural plasticity. Collectively, these data indicate that AZE-induced toxicity is associated with increased pro-inflammatory activity and reduced survival in BV2 microglia. This evidence may prompt for an increased monitoring of AZE consumption by humans.

Item Details

Item Type:Refereed Article
Keywords:azetidine, proinflammatory, neuropathology, neuroscience,
Research Division:Biomedical and Clinical Sciences
Research Group:Neurosciences
Research Field:Central nervous system
Objective Division:Health
Objective Group:Clinical health
Objective Field:Diagnosis of human diseases and conditions
UTAS Author:Piper, JA (Mr Jordan Piper)
ID Code:153963
Year Published:2022
Deposited By:Health Sciences
Deposited On:2022-10-18
Last Modified:2022-12-11
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