@article{ZHANG2024, 
author = {Xiaoyuan ZHANG and Ziteng MA and Yunfang JIA and Guiqiong HE},
title = {Knockdown of equilibrative nucleotide transporter 1 protects against Alzheimer's disease by reducing inflammatory response},
year = {2024},
journal = {Journal of Army Medical University},
volume = {46},
number = {23},
pages = {2588-2598},
keywords = {equilibrative nucleotide transporter 1, Alzheimer's disease, inflammatory factor, cell viability},
url = {https://www.sciopen.com/article/10.16016/j.2097-0927.202403052},
doi = {10.16016/j.2097-0927.202403052},
abstract = {ObjectiveTo study the role and mechanisms of equilibrative nucleotide transporter 1 (ENT1) on Alzheimer's disease (AD) by constructing ENT1 overexpression and knockdown plasmids.MethodsMolecular cloning was used to construct the ENT1 overexpression (pAAV-ENT1-mCherry) and knockdown (pAAV-ENT1shRNA-ZsGreen) plasmids. The overexpression plasmids and the knockdown plasmids were transfected into N2A cells (mouse Neuro A2 cells) and N2A-APP cells (N2A cells stably expressing human APP695). The expression of ENT1 and inflammatory factors at mRNA and protein levels were detected by real-time qPCR and Western blotting, respectively, and the change in cell viability were measured with CCK-8 assay.ResultsSequencing and real-time qPCR indicated that ENT1 overexpression and knockdown plasmids were successfully constructed. CCK-8 assay showed that ENT1 overexpression significantly reduced the cell survival rate within 24 h (P < 0.05), while its knockdown increased the cell survival rate (P < 0.01). Real-time qPCR displayed that overexpression of ENT1 enhanced the expression levels of inflammatory factors, such as IL-1β, TNF-α, C1q-a and C1q-b in N2A cells (P < 0.05), while ENT1 knockdown reversed the above changes in inflammatory factors in N2A-APP cells (P < 0.05).ConclusionKnockdown of ENT1 attenuates pathological changes in AD by reducing the inflammatory response. ENT1 may be a potential target in the pathological mechanism of AD.}
}