陕西医学杂志2024，Vol.53Issue(10)：1314-1319,1325,7.DOI:10.3969/j.issn.1000-7377.2024.10.004
CTCFL/RPS3A通过抑制内质网应激减少神经元细胞凋亡缓解阿尔兹海默症大鼠认知功能障碍实验研究
CTCFL/RPS3A alleviates cognitive dysfunction in Alzheimer's disease rats by inhibiting endoplasmic reticulum stress and reducing neuronal apoptosis
摘要
Abstract
Objective:To explore the role of ribosomal protein S3 A(RPS3A)in Alzheimer's disease(AD)and its underlying molecular mechanism.Methods:The AD rat model was established by injecting Aβ into the bilateral hippocampus of rats.AD rats were injected with adenoviral containing RPS3A overexpression vector(Ad-RPS3A).After feeding for 14 days,the learning and memory abilities and spatial exploration abilities of rats were evaluated by water maze test and Y maze test.Moreover,neuronal HT22 cells were transfected with RPS3A overexpression vector(RPS3A)or shRNA(sh-RPS3A),or transfected with CTCFL overexpression vector(CTCFL)or shRNA(sh-CTC-FL)for 48 hours,and then with treated 30 μmol/L Aβ for 24 hours.The binding of CTCFL to RPS3A promoter was predicted by using J ASP AR database and verified with Ch-IP analysis and the luciferase reporter gene analysis.RT-qPCR was used to detect the overexpression and silencing efficiency.Western blot was used to detect the protein lev-els of RPS3A and CTCFL,as well as expression levels of Endoplasmic-reticulum(ER)stress-related proteins and ap-optosis related proteins.Cell viability and apoptosis were detected with CCK-8 and flow cytometry,respectively.Results:Compared with the sham group,the mRNA and protein levels of RPS3A in the hippocampus of AD model rats were significantly downregulated.Moreover,the protein expression of RPS3A was significantly downregulated in Aβ-induced HT22 cells.Overexpression of RPS3A promoted HT22 cell proliferation and inhibited Aβ-induced cell apoptosis,decreased the expression of Cleaved caspase-3 and Bax proteins,promoted Bcl-2 protein expression,inhibi-ted the expression of ER stress-related proteins CRP78,CHOP,ATF6 and XBP1,and decreased the phosphorylation levels of IRE1 and PERK.Mechanistic studies revealed that CTCFL promotes RPS3A transcriptional activation by binding to RPS3A promoter.Overexpression of CTCFL promoted RPS3A protein expression,while silencing CTCFL has the opposite effect.The in vivo results showed that overexpression of RPS3A reduced the time to reach the plat-form,the distance to reach the platform,and the time to enter the SW area for the first time in AD rats,increased the rate of spontaneous alternation and the ratio of entering the new opposite arm,and improved Aβ induced cognitive dysfunction in AD rats.Conclusion:CTCFL mediated upregulation of RPS3A alleviates cognitive dysfunction in Alzheimer's disease mice by inhibiting ER stress to reduce neuronal apoptosis.关键词
阿尔兹海默症/认知功能障碍/神经元凋亡/RPS3A/CTCFL/内质网应激Key words
Alzheimer's disease/Cognitive dysfunction/Neuronal apoptosis/RPS3 A/CTCFL/Endoplasmic-reticulum stress分类
医药卫生引用本文复制引用
孟清琳,潘娜,刘养凤,苟英之,吴雅欣,赵锦华..CTCFL/RPS3A通过抑制内质网应激减少神经元细胞凋亡缓解阿尔兹海默症大鼠认知功能障碍实验研究[J].陕西医学杂志,2024,53(10):1314-1319,1325,7.基金项目
陕西省自然科学基础研究计划项目(2020JQ-928) （2020JQ-928）
