基于网络药理学和实验验证探讨涤痰汤治疗阿尔茨海默病作用机制OA

Objective To explore the mechanism of Ditan Decoction in the treatment of Alzheimer disease(AD)based on network pharmacology and animal experiments.Methods Active components and corresponding targets of Ditan Decoction were retrieved through the TCMSP databases;the targets of AD were searched through the OMIM and GeneCards databases,and protein-protein interaction network map was constructed through STRING online database.GO and KEGG enrichment analysis of key targets were carried out by using R language.AD model was established by injecting β-amyloid25-35 into bilateral hippocampus of rats.The rats were divided into sham-operation group,model group,donepezil group,Ditan Decoction low-and high-dosage groups,Ditan Decoction high-dosage+AMPK inhibitor group,and were intervened accordingly.The pathomorphological changes in the hippocampus were observed by HE staining.Transmission electron microscope was used to detect the ultrastructural observation of neurons and synapses in rat hippocampus.The content of inflammatory factors in hippocampus was detected by ELISA.The expressions of AMPK,p-AMPK,NLRP3,ASC,Caspase-1,GSDMD-N,GAP43,SYN,PSD95,BDNF proteins in the hippocampus were detected by Western blot.The mRNA expression of AMPK,NLRP3,ASC,Caspase-1,GSDMD-N,GAP43,SYN,PSD95 and BDNF were detected by RT-qPCR.Results A total of 129 active components of Ditan Decoction and 102 main targets for treating AD were screened,with the core targets including NLRP3,CASP1,GAP43,SYN1,GSDMD,IL1B,BDNF,TNF,CREB1 and APP.KEGG pathway enrichment analysis suggested that AMPK signaling pathway,NOD like receptor signaling pathway,cholinerqic synapse,AD and gluta-matergic synapse may play a core regulatory role.Animal experiments showed that Ditan Decoction could improve the learning and spatial memory abilities of model rats significantly(P<0.05,P<0.01),alleviate pathological morphology damage in hippocampal tissue,and improve the ultrastructure of neurons and synapses in hippocampus;decrease the contents of IL-1β,IL-18 and TNF-α in hippocampus(P<0.05,P<0.01),down-regulate the expressions of ASC,Caspase-1,GSDMD-N proteins and mRNA(P<0.05,P<0.01),and up-regulate the expressions of GAP43,SYN,PSD95,BDNF proteins and mRNA(P<0.05,P<0.01).Ditan Decoction could increase the phosphorylated level of AMPK,thus down-regulate the expression of NLRP3 protein and mRNA(P<0.05,P<0.01).However,the co-treatment of AMPK inhibitor partially reversed the effects of Ditan Decoction on pyroptosis and synaptic plasticity.Conclusion Ditan Decoction can improve synaptic plasticity in AD via inhibiting AMPK/NLRP3-mediated pyroptosis.