Circ-gabra5 exacerbates cognitive impairment in Alzheimer’s disease mice by regulating AKT through miR-455-3p- Journal of Army Medical University Vol. 48, Issue 2, Pages: 128-137(2026)
- 作者机构：
1.蚌埠医科大学第一附属医院神经内科，安徽蚌埠
2.陆军特色医学中心神经内科2.，重庆
- 作者简介：
LU Yi， E-mail： luyi4532@126.com
- 基金信息：the General Program of National Natural Science Foundation of China(81771177)
DOI：10.16016/j.2097-0927.202509106
CLC： R363.21;R394.3;R745.7Received：30 September 2025，
Revised：2025-12-31，
Accepted：04 January 2026，
Published：30 January 2026
- 稿件说明：
移动端阅览
王东林,冯祥谦,卢静,等.circ-gabra5通过miR-455-3p调节AKT蛋白加重阿尔茨海默病的认知损害[J].陆军军医大学学报,2026,48(02):128-137.
WANG D L,FENG X Q,LU J,et al.Circ-gabra5 exacerbates cognitive impairment in Alzheimer’s disease mice by regulating AKT through miR-455-3p[J].J Army Med Univ,2026,48(02):128-137.
王东林,冯祥谦,卢静,等.circ-gabra5通过miR-455-3p调节AKT蛋白加重阿尔茨海默病的认知损害[J].陆军军医大学学报,2026,48(02):128-137. DOI：10.16016/j.2097-0927.202509106.
WANG D L,FENG X Q,LU J,et al.Circ-gabra5 exacerbates cognitive impairment in Alzheimer’s disease mice by regulating AKT through miR-455-3p[J].J Army Med Univ,2026,48(02):128-137. DOI：10.16016/j.2097-0927.202509106.
摘要
目的
2
本研究探讨
circ-gabra5
在阿尔茨海默病（Alzheimer's disease， AD）小鼠的发病机制中的作用。
方法
2
选用6月龄APP/PS1双转基因 AD小鼠为实验组，C57BL/6野生型（WT）小鼠为对照组，根据随机数字表法分为2组（
n
=5）：双转基因 AD小鼠（AD组）和敲除
circ-gabra5
基因的双转基因AD小鼠（AD+敲除
circ-gabra5
组）。提取小鼠脑总RNA后，检测circRNA的差异表达，RT-qPCR验证差异前6的circRNA基因，并进行基因本体论（gene ontology， GO）和京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路分析。采用海马立体注射慢病毒敲除
circ
-
gabra5
基因，通过Morris水迷宫检测小鼠的学习记忆功能，免疫组化观察小鼠脑组织的Aβ斑块数量。透射电镜观察小鼠神经元的线粒体损伤以及神经突触数量。使用双荧光素酶报告实验和RNA结合蛋白免疫沉淀法（RIP测定）验证
circ-gabra5
和miR-455-3P之间的相互作用。Western Blot检测AKT蛋白表达水平。
结果
2
AD小鼠和WT小鼠基因检测筛选出有差异性的273个基因，其中表达上调的基因有163个，表达下调的有110个，
circ-gabra5
相差倍数最大，为1.39（
P
<
0.05）。GO分析及KEGG分析提示，AD小鼠与WT小鼠之间的功能表达差异富集在甲状腺激素、PI3K/AKT、RNA代谢等信号通路。AD小鼠敲除
circ-gabra5
基因后通过Morris水迷宫检测显示认知障碍改善，免疫组化提示β-淀粉样蛋白（amyloid-β， Aβ）斑块数量减少，透射电镜检测显示AD小鼠神经元线粒体损伤减少，脑皮质神经突触数量增加（
P
<
0.05）。通过starBase数据库预测，
circ-gabra5
和miR-455-3P具有互补序列，双荧光素酶报告实验和RIP测定证实
circ-gabra5
和miR-455-3p之间存在相互作用。数据库微生信预测miR-455-3p和AKT蛋白具有互补序列。qPCR方法显示，在AD鼠原代培养神经元敲低
circ-gabra5
基因后miR-455-3p表达下降，并在后续Western Blot检测显示AKT蛋白表达降低（
P
<
0.01）。
结论
2
circ-gabra5
通过miR-455-3P调节AKT蛋白加重AD小鼠认知损害，增加脑组织Aβ沉积，加重神经元线粒体损伤，减少神经突触数量。
Abstract
Objective
2
TTo explore the role of circ-gabra5 in the pathogenesis of Alzheimer’s disease (AD) in mice.
Methods
2
APP/PS1 double transgenic AD mice (6 months old) were selected as the experimental group
and C57BL/6 wild-type (WT) mice were enrolled as the control group. The mice were divided into an AD group and an AD mice with circ-gabra5 knockout (AD+circ-gabra5 knockout) group
with 5 mice in each group. After total RNA was extracted from mouse brain
differential expression of circRNA was detected
and real-time quantitative PCR (RT-qPCR) was used to validate the top 6 circRNA genes with differences. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed. Using hippocampal stereo injection of lentivirus to knock out the circ-gabra5 gene
the learning and memory function of mice was detected with Morris water maze test
and the number of Aβ plaques in mouse brain tissue was observed through immunohistochemistry. Transmission electron microscopy was utilized to observe the mitochondrial damage and synaptic count in mouse neurons. Dual luciferase reporter assay and RNA binding protein immunoprecipitation (RIP) assay were employed to verify the interaction between circ-gabra5 and miR-455-3p. Western blotting was apllied to detect the expression level of AKT protein.
Results
2
There were 273 differentially expressed genes screened between AD mice and WT mice
including 163 up-regulated and 110 down-regulated genes. Circ-gabra5 had the largest fold difference of 1.39 (
P
<
0.05). GO and KEGG analyses suggested that the functional expression differences between AD mice and WT mice were enriched in thyroid hormone
PI3K/AKT
RNA metabolism and other signaling pathways. Knocking out of circ-gabra5 gene in AD mice resulted in obvious improvement in cognitive impairment in Morris water maze test
decreased Aβ plaques by immunohistochemistry
and attenuated neuronal mitochondrial damage and more cortical synapses in AD mice by transmission electron microscopy (P
<
0.05). According to the starBase database prediction
circ-gabra5 and miR-455-3p have complementary sequences. Dual luciferase reporter assay and RIP assay confirmed the interaction between circ-gabra5 and miR-455-3p. Database bioinformatics analysis predicted that miR-455-3p and AKT have complementary sequences. qPCR indicated that knocking down the circ-gabra5 gene in primary cultured neurons of AD mice resulted in a decrease in m
iR-455-3p expression
and subsequent Western blot analysis showed a decrease in AKT protein level (P
<
0.01).
Conclusion
2
Circ-gabra
5 exacerbates cognitive impairment in AD mice by regulating AKT protein through miR-455-3p
increases Aβ deposition in brain tissue
aggravates neuronal mitochondrial damage
and reduces the number of neural synapses.
关键词
Keywords
references
王刚 ， 齐金蕾 ， 刘馨雅 ， 等 . 中国阿尔茨海默病报告2024 ［J］. 诊断学理论与实践 ， 2024 ， 23 （ 3 ）： 219 - 256 . DOI： 10.16150/j.1671-2870.2024.03.001 http://dx.doi.org/10.16150/j.1671-2870.2024.03.001 .
WANG G ， QI J L ， LIU X Y ， et al . China Alzheimer disease report 2024 ［J］. J Diagn Concepts Pract ， 2024 ， 23 （ 3 ）： 219 - 256 . DO I： 10.16150/j.1671-2870.2024.03.001 http://dx.doi.org/10.16150/j.1671-2870.2024.03.001 .
LIU E J ， ZHANG Y ， WANG J Z . Updates in Alzheimer’ s disease： from basic research to diagnosis and therapies ［J］. Transl Neurodegener ， 2024 ， 13 （ 1 ）： 45 . DOI： 10.1186/s40035-024-00432-x http://dx.doi.org/10.1186/s40035-024-00432-x .
SINGH A ， TIWARI S ， SINGH S . Pirh2 modulates the mitochondrial function and cytochrome c-mediated neuronal death during Alzheimer’s disease ［J］. Cell Death Dis . 2024 ， 15 （ 5 ）： 331 . doi： 10.1038/s41419-024-06662-1 http://dx.doi.org/10.1038/s41419-024-06662-1 .
SOUTSCHEK M ， SCHRATT G . Non-coding RNA in the wiring and remodeling of neural circuits ［J］. Neuron ， 2023 ， 111 （ 14 ）： 2140 - 2154 . DOI： 10.1016/j.neuron.2023.04.031 http://dx.doi.org/10.1016/j.neuron.2023.04.031 .
HUANG A Q ， ZHENG H X ， WU Z Y ， et al . Circular RNA-protein interactions： functions， mechanisms， and identification ［J］. Theranostics ， 2020 ， 10 （ 8 ）： 3503 - 3517 . DOI： 10.7150/thno.42174 http://dx.doi.org/10.7150/thno.42174 .
KIM E ， KIM Y K ， LEE S V . Emerging functions of circular RNA in aging ［J］. Trends Genet ， 2021 ， 37 （ 9 ）： 819 - 829 . DOI： 10.1016/j.tig.2021.04.014 http://dx.doi.org/10.1016/j.tig.2021.04.014 .
WU D P ， ZHAO Y D ， YAN Q Q ， et al . Circular RNAs： emerging players in brain aging and neurodegenerative diseases ［J］. J Pathol ， 2023 ， 259 （ 1 ）： 1 - 9 . DOI： 10.1002/path.6021 http://dx.doi.org/10.1002/path.6021 .
LONG H Z ， CHENG Y ， ZHOU Z W ， et al . PI3K/AKT signal pathway： a target of natural products in the prevention and treatment of Alzheimer’ s disease and Parkinson’ s disease ［J］. Front Pharmacol ， 2021 ， 12 ： 648636 . DOI： 10.3389/fphar.2021.648636 http://dx.doi.org/10.3389/fphar.2021.648636 .
STEKIC A ， ZELJKOVIC M ， ZARIC KONTIC M ， et al . Intermittent Theta Burst stimulation ameliorates cognitive deficit and attenuates neuroinflammation via PI3K/Akt/mTOR signaling pathway in Alzheimer’s-like disease model ［J］. Front Aging Neurosci ， 2022 ， 14 ： 889983 . DOI： 10.3389/fnagi.2022.889983 http://dx.doi.org/10.3389/fnagi.2022.889983 .
RAZANI E ， POURBAGHERI-SIGAROODI A ， SAFAROGHLI-AZAR A ， et al . The PI3K/Akt signaling axis in Alzheimer’s disease： a valuable target to stimulate or suppress ［J］. Cell Stress Chaperones ， 2021 ， 26 （ 6 ）： 871 - 887 . DOI： 10.1007/s12192-021-01231-3 http://dx.doi.org/10.1007/s12192-021-01231-3 .
DOI Y ， MIZUNO T ， MAKI Y ， et al . Microglia activated with the toll-like receptor 9 ligand CpG attenuate oligomeric amyloid｛beta｝neurotoxicity in in vitro and in vivo models of Alzheimer’ s disease ［J］. Am J Pathol ， 2009 ， 175 （ 5 ）： 2121 - 2132 . DOI： 10.2353/ajpath.2009.090418 http://dx.doi.org/10.2353/ajpath.2009.090418 .
WANG P ， ANDERSON D E ， YE Y H . PI3K-AKT activation resculpts integrin signaling to drive filamentous tau-induced proinflammatory astrogliosis ［J］. Res Sq ， 2023 ： rs.3.rs-rs.3253118. DOI： 10.21203/rs.3.rs-3253118/v1 http://dx.doi.org/10.21203/rs.3.rs-3253118/v1 .
LUKIW W J . Circular RNA （circRNA） in Alzheimer’ s disease （AD） ［J］. Front Genet ， 2013 ， 4 ： 307 . DOI： 10.3389/fgene.2013.00307 http://dx.doi.org/10.3389/fgene.2013.00307 .
CHEN H H ， ETELEEB A ， WANG C Y ， et al . Circular RNA detection identifies circPSEN1 alterations in brain specific to autosomal dominant Alzheimer’ s disease ［J］. Acta Neuropathol Commun ， 2022 ， 10 （ 1 ）： 29 . DOI： 10.1186/s40478-022-01328-5 http://dx.doi.org/10.1186/s40478-022-01328-5 .
SA M ， YOO E S ， KOH W ， et al . Hypothalamic GABRA5-positive neurons control obesity via astrocytic GABA ［J］. Nat Metab ， 2023 ， 5 （ 9 ）： 1506 - 1525 .
BUTLER K M ， MOODY O A ， SCHULER E ， et al . De novo variants in GABRA2 and GABRA5 alter receptor function and contribute to early-onset epilepsy ［J］. Brain ， 2018 ， 141 （ 8 ）： 2392 - 2405 . DOI： 10.1093/brain/awy171 http://dx.doi.org/10.1093/brain/awy171 .
WANG Z ， DONG J P ， ZHANG M X ， et al . Sevoflurane-induced overexpression of extrasynaptic α5-GABA（a）R via the RhoA/ROCK2 pathway impairs cognitive function in aged mice ［J］. Aging Cell ， 2024 ， 23 （ 9 ）： e14209 . DOI： 10.1111/acel.14209 http://dx.doi.org/10.1111/acel.14209 .
YANG H ， WANG H ， SHANG H ， et al . Circular RNA circ_0000950 promotes neuron apoptosis， suppresses neurite outgrowth and elevates inflammatory cytokines levels via directly sponging miR-103 in Alzheimer’ s disease ［J］. Cell Cycle ， 2019 ， 18 （ 18 ）： 2197 - 2214 . DOI： 10.1080/15384101.2019.1629773 http://dx.doi.org/10.1080/15384101.2019.1629773 .
MENG T ， CHEN Y L ， WANG P ， et al . Circ-HUWE1 knockdown alleviates amyloid-β-induced neuronal injury in SK-N-SH cells via miR-433-3p release-mediated FGF7 downregulation ［J］. Neurotox Res ， 2022 ， 40 （ 4 ）： 913 - 924 . DOI： 10.1007/s12640-022-00523-5 http://dx.doi.org/10.1007/s12640-022-00523-5 .
LI Y ， WANG H L ， CHEN L ， et al . Circ_0003611 regulates apoptosis and oxidative stress injury of Alzheimer’s disease via miR-383-5p/KIF1B axis ［J］. Metab Brain Dis ， 2022 ， 37 （ 8 ）： 2915 - 2924 . DOI： 10.1007/s11011-022-01051-z http://dx.doi.org/10.1007/s11011-022-01051-z .
FENG J H ， ZHAO J ， KUANG Y ， et al . Hsa_Circ_0105596/FTO inhibits progression of Parkinson’s disease by sponging miR-187-3p and regulating eEF2 ［J］. Heliyon ， 2024 ， 10 （ 23 ）： e39830 . DOI： 10.1016/j.heliyon.2024.e39830 http://dx.doi.org/10.1016/j.heliyon.2024.e39830 .
LEI J ， LIAO J J ， LIU Y Y ， et al . circ_0005654 as a sponge of the miR-588 dual pathway to promote α-synuclein expression ［J］. FASEB J ， 2025 ， 39 （ 11 ）： e70684 . DOI： 10.1096/fj.202500556R http://dx.doi.org/10.1096/fj.202500556R .
ISLAM M A ， KSHIRSAGAR S ， ALVIR R V ， et al . Overexpressed microRNA-455-3p enhances cognitive function in late-onset Alzheimer’ s disease ［J］. Brain Res ， 2025 ， 1865 ： 149853 . DOI： 10.1016/j.brainres.2025.149853 http://dx.doi.org/10.1016/j.brainres.2025.149853 .
WANG Y P ， LIN Y ， WANG L H ， et al . TREM2 ameliorates neuroinflammatory response and cognitive impairment via PI3K/AKT/FoxO3a signaling pathway in Alzheimer’ s disease mice ［J］. Aging ， 2020 ， 12 （ 20 ）： 20862 - 20879 . DOI： 10.18632/aging.104104 http://dx.doi.org/10.18632/aging.104104 .
SUN Z Q ， SUN L ， TU L X . GABAB receptor-mediated PI3K/Akt signaling pathway alleviates oxidative stress and neuronal cell injury in a rat model of Alzheimer’ s disease ［J］. J Alzheimers Dis ， 2020 ， 76 （ 4 ）： 1513 - 1526 . DOI： 10.3233/JAD-191032 http://dx.doi.org/10.3233/JAD-191032 .
WEN X Z ， LI H Y ， SUN H ， et al . MiR-455-3p reduces apoptosis and alleviates degeneration of chondrocyte through regulating PI3K/AKT pathway ［J］. Life Sci ， 2020 ， 253 ： 117718 . DOI： 10.1016/j.lfs.2020.117718 http://dx.doi.org/10.1016/j.lfs.2020.117718 .
YI J H ， BAEK S J ， HEO S ， et al . Direct pharmacological Akt activation rescues Alzheimer’ s disease like memory impairments and aberrant synaptic plasticity ［J］. Neuropharmacology ， 2018 ， 128 ： 282 - 292 . DOI： 10.1016/j.neuropharm.2017.10.028 http://dx.doi.org/10.1016/j.neuropharm.2017.10.028 .
SHARMA C ， KIM S ， NAM Y ， et al . Mitochondrial dysfunction as a driver of cognitive impairment in Alzheimer’ s disease ［J］. Int J Mol Sci ， 2021 ， 22 （ 9 ）： 4850 . DOI： 10.3390/ijms22094850 http://dx.doi.org/10.3390/ijms22094850 .
GOWDA P ， REDDY P H ， KUMAR S . Deregulated mitochondrial microRNAs in Alzheimer’ s disease： Focus on synapse and mitochondria ［J］. Ageing Res Rev ， 2022 ， 73 ： 101529 . DOI： 10.1016/j.arr.2021.101529 http://dx.doi.org/10.1016/j.arr.2021.101529 .
ASHLEIGH T ， SWERDLOW R H ， BEAL M F . The role of mitochondrial dysfunction in Alzheimer’s disease pathogenesis ［J］. Alzheimers Dement ， 2023 ， 19 （ 1 ）： 333 - 342 . DOI： 10.1002/alz.12683 http://dx.doi.org/10.1002/alz.12683 .
0
Views
9
下载量
0
CSCD
- L-PDFDownload
- WORDDownload
- Meta-XMLDownload
- BibTeXDownload
- EndnoteDownload
- RefMan Download
- NoteExpressDownload
- NoteFirstDownload
Publicity Resources
Related Articles
Related Author
Related Institution
- Technical support is provided by Beijing Founder electronics co., LTD 京ICP备09064830号-19
京公网安备11010802024621 - It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
- Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.
