Investigation of TRPM2 channel involvement in lipopolysaccharide and experimental alzheimer's disease-induced inflammation, oxidative stress, and apoptosis in microglia cells by using molecular techniques

Investigation of TRPM2 Channel Involvement in Lipopolysaccharide and Experimental Alzheimer's Disease-Induced Inflammation, Oxidative Stress, and Apoptosis in Microglia Cells by Using Molecular Techniques Alzheimer's disease (AD) is characterized by a decline in cognitive function and a decrease in daily activities as a result of the deterioration of neurons and inter-neuronal synaptic structure. BV2 cells are a useful microglial model in terms of morphological, phenotypic, and functional properties. Lipopolysaccharide (LPS) is widely used in experimental models of neuroinflammation and amyloidosis. The TRPM2 channel is a non-selective Ca+2 permeable channel. TRPM2 stimulation modulates oxidative stress and inflammation through interaction with microglial cells, while glutathione (GSH) treatment reduces oxidative stress and inflammation through TRPM2 inhibition. However, the effect of TRPM2 channel inhibitors and GSH treatment on oxidative stress, inflammation, and apoptotic values in BV2 microglia cells activated with LPS and amyloid beta (A) has not been investigated yet. In our study, we aimed to investigate the effect of TRPM2 channel inhibition on inflammation, oxidative stress, and apoptosis in BV2 microglia cells activated with LPS and Ab by molecular techniques. BV2 cells were divided into five groups as control, LPS, A, A+ LPS, and A + LPS + GSH. Laser confocal microscope, ELISA, spectrofluorometer, and spectrophotometer analyses were performed on the obtained cells. Increased levels of inflammation biomarkers (TNF-, IL-1 ve IL-6), intracellular Ca+2 level, cytosolic reactive oxygen species, mitochondrial membrane depolarization, cell death, apoptosis and caspase (caspase -3, -8, and -9) were observed in microglia stimulated with LPS and Aβ. These values increased more when LPS and Aβ were used together. However, these apoptotic, inflammatory, and oxidant values decreased in cells treated with GSH and TRPM2 channel blockers. In conclusion, TRPM2 channel activity was demonstrated on Aβ and LPS-induced Ca+2 entry, oxidative stress, inflammation, and apoptosis parameters in microglia cells. TRPM2 channel inhibition by GSH treatment seems to be a potential source for the prevention of Aβ and LPS-induced oxidative stress, apoptosis, and inflammation. Keywords: Alzheimer's disease, Apoptosis, BV2 Microglia, Inflammation, TRPM2, Lipopolysaccharide, Oxidative stress,