湖南中医药大学学报2025，Vol.45Issue(4)：638-646,9.DOI:10.3969/j.issn.1674-070X.2025.04.008
丹参多酚酸B通过NLRP3/Caspase-1/GSDMD通路抑制OGD诱导海马神经元焦亡的分子机制研究
Molecular mechanism of salvianolic acid B inhibiting OGD-induced pyroptosis of hippocampal neurons via NLRP3/Caspase-1/GSDMD pathway
摘要
Abstract
Objective To explore the potential molecular mechanisms of salvianolic acid B(Sal B)in treating vascular dementia(VD)through network pharmacology and experimental verification.Methods Network pharmacology was used to identify the potential targets of Sal B intervention in VD.A protein-protein interaction(PPI)network was constructed,followed by GO and KEGG pathway enrichment analyses to determine biologically relevant target pathways.Molecular docking was performed to evaluate the interaction between Sal B and its targets.The optimal concentration of Sal B was determined via CCK-8 assay,the cell morphology was observed under a microscope,and the release rate of lactate dehydrogenase(LDH)in the cell supernatant was measured using a microplate reader.The levels of inflammatory factors were measured by ELISA,the protein expression of NLRP3 was examined by immunofluorescence,and the protein expression levels of NOD-like receptor protein 3(NLRP3),apoptosis-associated speck-like protein containing a CARD(ASC),cysteinyl aspartate specificproteinase 1(Caspase-1),and GSDMD were checked by Western blot.Results A total of 37 targets for Sal B intervention in VD were obtained by bioinformatics analysis.Core target genes such as Caspase-1,NLRP3,and TNFR1 were screened by network pharmacology,and the pathways including the NOD-like receptors(NLRs)signaling pathway that may involve these target genes were identified through KEGG and GO analyses.In vitro experiments were performed using the classic NLRP3/Caspase-1/GSDMD pyroptosis pathway.The OGD-induced pyroptosis model of hippocampal neurons was established,and it was found that Sal B could significantly enhance the survival rate of HT22 cells after OGD(P＜0.05,P＜0.01),alleviate cell damage,reduce LDH release(P＜0.05 or P＜0.01),decrease the levels of IL-1β and IL-18 in OGD-damaged HT22 cells(P＜0.01),inhibit the activation of NLRP3 inflammasome,and reduce the protein expression levels of NLRP3,GSDMD-N,cleaved Caspase-1,and ASC(P＜0.05,P＜0.01).Conclusion Sal B may inhibit OGD-induced pyroptosis of hippocampal neurons through NLRP3/Caspase-1/GSDMD pathway,providing experimental evidence for preventing and treating VD with Sal B.关键词
血管性痴呆/丹参多酚酸B/氧糖剥夺/细胞焦亡/NLRP3/Caspase-1/GSDMD信号通路/网络药理学Key words
vascular dementia/salvianolic acid B/oxygen-glucose deprivation/pyroptosis/NLRP3/Caspase-1/GSDMD sig-naling pathway/network pharmacology分类
中医学引用本文复制引用
谭惠中,刘竹暄,唐洁,谢乐,张秀丽,朱新华,伍大华..丹参多酚酸B通过NLRP3/Caspase-1/GSDMD通路抑制OGD诱导海马神经元焦亡的分子机制研究[J].湖南中医药大学学报,2025,45(4):638-646,9.基金项目
国家自然科学基金项目(82374441) （82374441）
湖南省自然科学基金项目(2024JJ5317) （2024JJ5317）
湖南省卫生健康委卫生科研课题(W20243137) （W20243137）
湖南省研究生科研创新项目(CX20230809). （CX20230809）
