Attenuation of Spatial Memory in 5xFAD Mice by Halting Cholinesterases, Oxidative Stress and Neuroinflammation Using a Cyclopentanone Derivative

Abstract

Alzheimer’s disease (AD) is an irreversible and chronic neurological disorder that gradually destroys memory and thinking skills. The research study was designed to investigate the underlying molecular signaling involved in the neuroprotective effects of cyclopentanone derivative i.e., 2-(hydroxyl-(3-nitrophenyl)methyl)cyclopentanone (3NCP) as a therapeutic agent for AD. In this study, In vivo studies were carried out on a well-known 5xFAD mice model using different behavioural test models such as open field, rotarod, Morris water maze (MWM), and Y-maze tests. Furthermore, in vitro cholinesterase inhibition activity assays were carried out. The frontal cortex (FC) and hippocampus (HC) homogenates were tested for the levels/activities of cholinesterases, glutathione (GSH), glutathione S-transferase (GST), and catalase. Furthermore, the hippocampal expression of inflammatory cytokines was observed via RT-PCR and western blot. The results of in vivo studies show an enhancement in the learning behavior. The 3NCP treatment reduced latency time in MWM and Y-maze tests, also increase spontaneous alternation indicate significant effect of 3NCP on memory. Furthermore, open field and rotarod studies revealed that 3NCP does not cause motor coordination deficit. The results of the in vitro studies revealed that the IC50 values of the 3NCP against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) were 16.17 and 20.51 µg/mL, respectively. This decline in AChE and BChE was further supported by ex vivo studies. Further, the 3NCP mitigates the GSH level, GST, and catalase activities in HC and FC. The mRNA and protein expression of inflammatory cytokines (IL-1β, IL-6, TNF-α) markedly declined in RT-PCR and western blotting. The results of the current study conclusively demonstrate that 3NCP reduces oxidative stress and mitigates neuroinflammation in 5xFAD mice, implying that 3NCP may be a potential therapeutic candidate for AD treatment in the future.

Alleviation of Memory Deficit by Bergenin via the Regulation of Reelin and Nrf-2/NF-κB Pathway in Transgenic Mouse Model

“…The effect of the Bergenin on the spatial learning and memory of 5xFAD Tg mice was evaluated through the Morris water maze apparatus (MWM, diameter: 110 cm, height: 40 cm) containing opaque water (depth: 20 cm) maintained at 22–23 °C, following previously reported method [ 61 ]. The maze consisted of four quadrants with a hidden platform (diameter: 7 cm, 1 cm below the water surface) in one of the four quadrants.…”

Cyclopentanone Derivative Attenuates Memory Impairment by Inhibiting Amyloid Plaques Formation in the 5xFAD Mice

“…Indeed, the real promise of metabolic/ketogenic approaches is that by altering cellular energetics, oxidative stress, and inflammation, ketogenic (and possible 3NCP) have the potential to do more than provide symptomatic relief but also have the potential to alter disease trajectory. The cyclopentanone derivatives including 3NCP that were tested in our lab exhibited anti-cholinesterase, anti-oxidant, and anti-neuroinflammatory potential [23,24]. The 3NCP has a nitrophenyl group, and the literature studies revealed that compounds containing nitrophenyl group have the potential to inhibit AChE, BChE, MAO-B, and relieve oxidative stress [25].…”

“…Methodology of genotyping as specified by Jackson Lab U.S. for strain transgenic mice was followed with slight modifications [23]. DNA extraction was carried out by using a DNA extraction kit following the manufacturer's protocol.…”

“…The primers used in this study were BACE-1 Forward: 5 -ACCTGGTGAGCATCCCTCAT-3 Reverse: 5 -CCTCCCAGTTGG-AACCATTG-3 , and GAPDH Forward: 5 -TGCACCACCAACTGCTTAGC-3 , Reverse: 5 -GGCATGGACTGTGGTCATGAG-3 . The amplification products were separated via 1.5% agarose gel and visualized through a UV transilluminator [23,72].…”

Attenuation of nociceptive and paclitaxel-induced neuropathic pain by targeting inflammatory, CGRP and substance P signaling using 3-Hydroxyflavone

“…Agarose gel (1.5%) was used for amplified product separation. The mRNA expression was calculated in the arbitrary unit using ImageJ software (Ullah et al, 2020).…”