Protective effect of 3-(naphthalen-2-yl(propoxy)methyl)azetidine hydrochloride on hypoxia-induced toxicity by suppressing microglial activation in BV-2 cells

Abstract

We recently reported the anti-inflammatory effects of 3-(naphthalen-2-yl(propoxy)methyl)azetidine hydrochloride (KHG26792) on the ATP-induced activation of the NFAT and MAPK pathways through the P2X7 receptor in microglia. To further investigate the underlying mechanism of KHG26792, we studied its protective effects on hypoxia-induced toxicity in microglia. The administration of KHG26792 significantly reduced the hypoxia-induced expression and activity of caspase-3 in BV-2 microglial cells. KHG26792 also reduced hypoxia-induced inducible nitric oxide synthase protein expression, which correlated with reduced nitric oxide accumulation. In addition, KHG26792 attenuated hypoxia-induced protein nitration, reactive oxygen species production, and NADPH oxidase activity. These effects were accompanied by the suppression of hypoxia-induced protein expression of hypoxia-inducible factor 1-alpha and NADPH oxidase-2. Although the clinical relevance of our findings remains to be determined, these data results suggest that KHG26792 prevents hypoxia-induced toxicity by suppressing microglial activation.

Anti-inflammatory and anti-oxidative effects of 3-(naphthalen-2-yl(propoxy)methyl)azetidine hydrochloride on β-amyloid-induced microglial activation

“…Briefly, cells were plated at a density of 5 × 10 5 cells in 96-well plates, and the extracellular and intracellular amounts of LDH were determined with the LDH toxicity detection kit from the Roche (Mannheim, Germany) according to the manufacturer’s instructions. MTT assay was performed to determine cell viability by quantifying the color formation at 595 nm with an enzyme-linked immunosorbent assay (ELISA) microplate reader as described before ( 11 ).…”

“…TNF-α, IL-6, and IL-1β levels were measured using commercial ELISA kits (R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions, as previously described ( 3 , 11 ). The optical density was detected with a microplate reader.…”

“…ROS levels were measured using 2′,7′-dichlorofluorescin diacetate (DCF-DA), which is converted by ROS to fluorescent DCF ( 11 ). The concentrations of ROS were determined using the oxidation-sensitive probe DCF-DA according to the manufacturer’s protocol.…”

“…The biological functions of azetidine derivatives against serotonin, norepinephrine, and dopamine transporters have also been reported ( 8 , 9 ). Recently, we described the protective effects of KHG26792—a novel azetidine derivative—on the ATP-induced activation of the NFAT and MAPK pathways through the P2X7 receptor in BV-2 cells and on hypoxia-induced toxicity through the suppression of microglial activation in BV-2 cells ( 10 , 11 ). Nevertheless, the mechanism responsible for the action of KHG26792 remains unclear.…”

3-(Naphthalen-2-yl(propoxy)methyl)azetidine hydrochloride attenuates MPP＋-induced cytotoxicity by regulating oxidative stress and mitochondrial dysfunction in SH-SY5Y cells

“…In addition, a previous study also showed the memory-restorative effect of ezetimibe, a well-known azetidine derivative, in memory dysfunctions associated with Alzheimer’s disease dementia ( 10 ). Previously, we synthesized 3-(naphthalen-2-yl(propoxy)methyl)azetidine hydrochloride (KHG26792) and reported that KHG26792 significantly decreased hypoxia-induced toxicity by attenuating oxidative stress ( 11 , 12 ). However, the precise mechanisms for the action of KHG26792 have not been elucidated, particularly for MPP + -induced neurotoxicity.…”