Knockdown of equilibrative nucleotide transporter 1 protects against Alzheimer's disease by reducing inflammatory response
10.16016/j.2097-0927.202403052
- VernacularTitle:靶向敲低腺苷转运体1通过降低炎症反应对阿尔茨海默病的保护作用研究
- Author:
Xiaoyuan ZHANG
1
;
Ziteng MA
;
Yunfang JIA
;
Guiqiong HE
Author Information
1. 400016 重庆,重庆医科大学:神经科学研究中心
- Keywords:
equilibrative nucleotide transporter 1;
Alzheimer's disease;
inflammatory factor;
cell viability
- From:
Journal of Army Medical University
2024;46(23):2588-2598
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the role and mechanisms of equilibrative nucleotide transporter 1(ENT1)on Alzheimer's disease(AD)by constructing ENT1 overexpression and knockdown plasmids.Methods Molecular 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.Results Sequencing 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).Conclusion Knockdown of ENT1 attenuates pathological changes in AD by reducing the inflammatory response.ENT1 may be a potential target in the pathological mechanism of AD.
