Knockdown of equilibrative nucleotide transporter 1 protects against Alzheimer's disease by reducing inflammatory response
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-EN...
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Editorial Office of Journal of Army Medical University
2024-12-01
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| Series: | 陆军军医大学学报 |
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| Online Access: | https://aammt.tmmu.edu.cn/html/202403052.htm |
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| _version_ | 1846121545286549504 |
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| author | ZHANG Xiaoyuan MA Ziteng JIA Yunfang |
| author_facet | ZHANG Xiaoyuan MA Ziteng JIA Yunfang |
| author_sort | ZHANG Xiaoyuan |
| collection | DOAJ |
| description | 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.
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| format | Article |
| id | doaj-art-fb6a64c6b4cc4318924f6a95da74959f |
| institution | Kabale University |
| issn | 2097-0927 |
| language | zho |
| publishDate | 2024-12-01 |
| publisher | Editorial Office of Journal of Army Medical University |
| record_format | Article |
| series | 陆军军医大学学报 |
| spelling | doaj-art-fb6a64c6b4cc4318924f6a95da74959f2024-12-16T01:15:14ZzhoEditorial Office of Journal of Army Medical University陆军军医大学学报2097-09272024-12-0146232588259810.16016/j.2097-0927.202403052Knockdown of equilibrative nucleotide transporter 1 protects against Alzheimer's disease by reducing inflammatory responseZHANG Xiaoyuan0MA Ziteng1JIA Yunfang2Neuroscience Center, College of Basic Medical Sciences, Chongqing Medical University, Chongqing, 400016, ChinaNeuroscience Center, College of Basic Medical Sciences, Chongqing Medical University, Chongqing, 400016, ChinaNeuroscience Center, College of Basic Medical Sciences, Chongqing Medical University, Chongqing, 400016, China 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. https://aammt.tmmu.edu.cn/html/202403052.htmequilibrative nucleotide transporter 1alzheimer's diseaseinflammatory factorcell viability |
| spellingShingle | ZHANG Xiaoyuan MA Ziteng JIA Yunfang Knockdown of equilibrative nucleotide transporter 1 protects against Alzheimer's disease by reducing inflammatory response 陆军军医大学学报 equilibrative nucleotide transporter 1 alzheimer's disease inflammatory factor cell viability |
| title | Knockdown of equilibrative nucleotide transporter 1 protects against Alzheimer's disease by reducing inflammatory response |
| title_full | Knockdown of equilibrative nucleotide transporter 1 protects against Alzheimer's disease by reducing inflammatory response |
| title_fullStr | Knockdown of equilibrative nucleotide transporter 1 protects against Alzheimer's disease by reducing inflammatory response |
| title_full_unstemmed | Knockdown of equilibrative nucleotide transporter 1 protects against Alzheimer's disease by reducing inflammatory response |
| title_short | Knockdown of equilibrative nucleotide transporter 1 protects against Alzheimer's disease by reducing inflammatory response |
| title_sort | knockdown of equilibrative nucleotide transporter 1 protects against alzheimer s disease by reducing inflammatory response |
| topic | equilibrative nucleotide transporter 1 alzheimer's disease inflammatory factor cell viability |
| url | https://aammt.tmmu.edu.cn/html/202403052.htm |
| work_keys_str_mv | AT zhangxiaoyuan knockdownofequilibrativenucleotidetransporter1protectsagainstalzheimersdiseasebyreducinginflammatoryresponse AT maziteng knockdownofequilibrativenucleotidetransporter1protectsagainstalzheimersdiseasebyreducinginflammatoryresponse AT jiayunfang knockdownofequilibrativenucleotidetransporter1protectsagainstalzheimersdiseasebyreducinginflammatoryresponse |