Neuroprotective Effects of VGLUT1 Inhibition in HT22 Cells Overexpressing VGLUT1 Under Oxygen Glucose Deprivation Conditions

• Published in: Neuromolecular medicine 2024-08, Vol.26 (1), p.35
• Main Authors: Pomierny, B., Krzyżanowska, W., Skórkowska, A., Budziszewska, B., Pera, J.
• Format: Article
• Language: English
• Subjects: Animals; Apoptosis-inducing factor; Biomedical and Life Sciences; Biomedicine; Brain injury; Brief Report; Cell culture; Cell Hypoxia; Cell Line; Cell Survival - drug effects; Cell viability; Dehydrogenases; Excitotoxicity; Glucose; Glucose - deficiency; Glucose - metabolism; Glutamic Acid - metabolism; Hippocampus; Hippocampus - cytology; Hippocampus - metabolism; Hypoxia; Internal Medicine; Ischemia; L-Lactate dehydrogenase; L-Lactate Dehydrogenase - metabolism; Laboratories; Membrane potential; Membrane Potential, Mitochondrial - drug effects; Mice; Neurodegenerative diseases; Neurology; Neurons; Neurons - drug effects; Neurons - metabolism; Neuroplasticity; Neuroprotection; Neuroprotective Agents - pharmacology; Neurosciences; Oxygen - metabolism; Plasmids; Poly(ADP-ribose) polymerase; Proteins; Synaptic plasticity; Synaptic vesicles; Vesicular Glutamate Transport Protein 1 - biosynthesis; Vesicular Glutamate Transport Protein 1 - genetics; Vesicular Glutamate Transport Protein 1 - metabolism; Vesicular Glutamate Transport Protein 2
• ISSN: 1535-1084; 1559-1174; 1559-1174
• DOI: 10.1007/s12017-024-08803-3

Glutamate (Glu) is a major excitatory neurotransmitter in the brain, essential for synaptic plasticity, neuronal activity, and memory formation. However, its dysregulation leads to excitotoxicity, implicated in neurodegenerative diseases and brain ischemia. Vesicular glutamate transporters (VGLUTs) regulate Glu loading into synaptic vesicles, crucial for maintaining optimal extracellular Glu levels. This study investigates the neuroprotective effects of VGLUT1 inhibition in HT22 cells overexpressing VGLUT1 under oxygen glucose deprivation (OGD) conditions. HT22 cells, a hippocampal neuron model, were transduced with lentiviral vectors to overexpress VGLUT1. Cells were subjected to OGD, with pre-incubation of Chicago Sky Blue 6B (CSB6B), an unspecific VGLUT inhibitor. Cell viability, lactate dehydrogenase (LDH) release, mitochondrial membrane potential, and hypoxia-related protein markers (PARP1, AIF, NLRP3) were assessed. Results indicated that VGLUT1 overexpression increased vulnerability to OGD, evidenced by higher LDH release and reduced cell viability. CSB6B treatment improved cell viability and reduced LDH release in OGD conditions, particularly at 0.1 μM and 1.0 μM concentrations. Moreover, CSB6B preserved mitochondrial membrane potential and decreased levels of PARP1, AIF, and NLRP3 proteins, suggesting neuroprotective effects through mitigating excitotoxicity. This study demonstrates that VGLUT1 inhibition could be a promising therapeutic strategy for ischemic brain injury, warranting further investigation into selective VGLUT1 inhibitors.