Mechanism of kaempferol in the treatment of Alzheimer's disease based on network pharmacology and cell experiment
Objective To explore the mechanism of kaempferol in the treatment of Alzheimer's disease based on network pharmacology and cell experiments.Methods The targets related to kaempferol were obtained using the Traditional Chinese Medicine Systems Pharmacology(TCMSP).Differentially expressed genes for Alzheimer's disease(GSE5281)were obtained from the Gene Expression Omnibus(GEO)database.Gene ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis of both overlapping targets were performed using the DAVID database.The molecular docking was performed through the Autodock Vina software.SH-SY5Y cells were divided into control group,Aβ group and kaempferol group.The cells in control group were not given any treatment,the cells in Aβ group were treated with 10 μmol/L Aβ1-42,and the cells in kaempferol group were treated with 10 μmol/L Aβ1-42 and then 10 μmol/L kaempferol.Apoptosis was detected by Annexin V-PI double staining.The expression levels of calmodulin-1(CALM1),muscarinic acetylcholine receptor M1(CHRM1)and protein phosphatase 3 catalytic subunit alpha(PPP3CA)were detected by real-time quantitative PCR and Western blot.Results A total of 60 targets related to kaempferol,2 155 differentially expressed genes for Alzheimer's disease,and 6 potential targets for kaempferol in the treat-ment of Alzheimer's disease were obtained.GO functional enrichment analysis showed that the potential targets mainly involved in the positive regulation of phosphoprotein phosphatase activity,the regulation of postsynaptic membrane potential,the response to calcium ion and the G-protein coupled receptor signaling pathway.KEGG pathway enrichment analysis showed that the potential targets mainly involved calcium signaling pathway,Alzheimer's disease,and neurodegenerative diseases.Molecular docking results showed that the binding energies of kaempferol with CHRM1,CALM1 and PPP3CA were all<-5.0 kcal/mol.Apoptosis was increased in Aβ group compared to control group(P<0.05),whereas apoptosis was decreased in kaempferol group compared to Aβ group(P<0.05).The mRNA and protein expressions of CHRM1,CALM1 and PPP3CA were significantly upregulated in kaempferol group compared with Aβ group(P<0.05).Conclusion Kaempferol may play a therapeutic role for Alzheimer's disease by ameliorating apoptosis of SH-SY5Y cells through regulating key genes(CHRM1,CALM1 and PPP3CA)in the calcium signaling pathway.
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