Regulatory role of ITCH-TXNIP-NLRP3 signaling pathway in Alzheimer disease-like lesions in vivo and in vitro
10.3969/j.issn.1000-4718.2025.06.007
- VernacularTitle:ITCH-TXNIP-NLRP3信号通路在体内外阿尔茨海默病样病变中的调控作用
- Author:
Qiuyu XIE
1
;
Jianfeng MA
;
Qiying SHEN
;
Yongxiang HE
;
Xiaobing LI
;
Shuo YANG
;
Yuke XIANG
;
Yuan QIN
;
Wei WEI
;
Yinghua LIU
Author Information
1. 广州医科大学药学院药理学教研室,神经致病基因与离子通道病教育部重点实验室,广东省分子靶标与临床药理学重点实验室,广东 广州 511436
- Publication Type:Journal Article
- Keywords:
Alzheimer disease;
ITCH protein;
thioredoxin-interacting protein;
nucleotide-binding oligomer-ization domain-like receptor protein 3
- From:
Chinese Journal of Pathophysiology
2025;41(6):1109-1117
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To investigate the modulatory role of E3 ubiquitin-protein ligase ITCH in Alzheimer disease(AD)-like pathology through the thioredoxin-interacting protein(TXNIP)-nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)signaling pathway using both in vivo and in vitro experimental models.METHODS:(1)Ten 5×FAD(AD model)mice and 10 wild-type(WT)mice at 2-,4-and 6-month-old were randomly allocated into AD and WT groups.Amyloid β-protein(Aβ)plaque burden in the brain was detected by thioflavin-S and immunofluorescence staining,with the latter method additionally applied to assess TXNIP protein expression.The protein levels of ITCH and TXNIP were determined by Western blot,while their interaction was verified by co-immunoprecipitation.(2)Mouse mi-croglia BV2 cells stimulated by lipopolysaccharide(LPS)were used to construct neuroinflammation model,and were di-vided into control(CON)group and LPS+ATP treatment group.The BV2 cells stimulated by Aβ were used to construct AD inflammation model.According to the different treatment time,they were divided into CON,and 12,24 and 48 h treatment groups.Western blot was used to evaluate the expression of ITCH,TXNIP,and NLRP3 inflammasome compo-nents(NLRP3 and caspase-1)as well as the downstream IL-1β.Adenovirus-mediated ITCH overexpression(OE-ITCH)in Aβ-stimulated BV2 cells comprised three experimental groups:negative control group,Aβ oligomer stimulation group,and OE-ITCH group,with subsequent immunoblotting of inflammatory mediators.RESULTS:The deposition of Aβ plaques in the cortex and hippocampus of 5×FAD transgenic mice exhibited an age-dependent progression(P<0.01).Compared with WT mice,the levels of TXNIP protein increased synchronously,and the levels of ubiquitin ligase ITCH was significantly down-regulated(P<0.05).Co-immunoprecipitation confirmed the interaction between ITCH and TXNIP proteins in the brain of 2-and 4-month-old 5×FAD mice,which exhibited marked attenuation by 4 months of age.In BV2 microglial models,Aβ/LPS stimulation provoked significant ITCH suppression,concurrently up-regulating TXNIP,core NLRP3 inflammasome components(NLRP3 and caspase-1),and downstream IL-1β(P<0.05).Overexpression of ITCH significantly inhibited Aβ-induced activation of TXNIP and NLRP3 and therelated inflammatory factors in BV2 cells.CONCLUSION:The results of in vitro and in vivo experiments showed that ITCH protein exerts effects against AD-like pathology by inhibiting the expression of TXNIP-NLRP3 signaling pathway.
