Effect Of Anshen Yizhi Method On Cognitive And Rhythmic Genes In AD Model Mice And Mechanism Analysis

Objective:The common etiology,pathogenesis and correlation between AD and senile insomnia were discussed in TCM theory,and the rationality of the method of calming the mind and promoting the mind was further explored in combination with the time medicine theory of Neijing.To verify its effectiveness,animal experiments were conducted to observe the effects of selective administration of tranquilizing mind and promoting intelligence on cognitive and rhythmic genes in AD mice.In order to further elucidate the relationship between AD and insomnia and the mechanism of Anshen Yizhi method,GO enrichment analysis,network pharmacology,molecular docking and kinetic simulation were used to study the related genes,target of Suanzaoren Decoction,gamma-aminobutyric acid receptor and acetylcholinesterase.Method:1.Consult the literature and theory of AD and senile insomnia,and discuss the common etiology and pathogenesis of AD and senile insomnia.The research on meridian flow and Yingwei theory in TCM chronomedicine was sorted out,and the rationality of the method of tranquilizing mind and promoting intelligence was discussed in combination with the relevant literature on timing administration.2.Ninety mice were divided into blank group,model group,tranquilizing mind group,mental retardation group,mental retardation group and mental retardation group.Compound AD mice were made by intraperitoneal injection of D-galactose and intraventricular injection of Abeta 1-42.The mice in the administration group were administered at 7:00 and 19:00 for 3 weeks.Anshen Decoction was given by intragastric administration as Suanzaoren Decoction and Yizhi Decoction as Yizhi Wendan Granule.Morris water maze was used to test the spatial learning and memory ability of mice in each group.The expression levels of Per1,Bmall,Clock circadianrhythm genes and IDE genes in hypothalamus were detected by fluorescence quantitative Pcr.The level of IDE protein in hippocampus was detected by ELISA.3.Search the genes related to AD and insomnia in Gene Cards database,screen out the duplicate genes between them,and establish the database of genes related to AD and insomnia.Cluego was used to analyze the GO biological process enrichment and KEGG signaling pathway enrichment of related genes.The signaling pathways that have important effects on both diseases were obtained,and the correlation between them was elaborated at the level of gene and signaling pathway.4.Go bioaccumulation analysis and KEGG signaling pathway enrichment analysis were carried out for the target of AD and insomnia related genes in the active ingredients of Suanzaoren Decoction,so as to explore the interaction mechanism of Suanzaoren Decoction in improving AD and insomnia.5.In order to further elucidate the mechanism of Suanzaoren Decoction in improving AD and insomnia,two target proteins,GABA receptor and Ache E,were selected to perform semi-flexible docking and kinetic simulation of the components acting on the two targets by using molecular docking and molecular dynamics simulation methods,in order to explore the interaction structure and stability between small effective components and target proteins at the micro level.Result:1.Theoretical studies show that the common pathogenesis of AD and senile insomnia is mainly organ deficiency,in which kidney deficiency is the main pathogenesis.According to the meridian flow theory and Yingwei theory,the method of soothing the mind and improving the intelligence is put forward to treat AD at the right time,which has a certain theoretical basis.2.Morris water maze showed that compared with the blank group,the learning and memory abilities of AD model group decreased(P < 0.05);compared with AD model group,the learning and memory abilities of mice in the administration group were improved in varying degrees(P < 0.05,P < 0.01),especially in the morning and evening group(P < 0.01).RT-PCR results showed that compared with the blank group,the expression of Per1,Bmall and Clock in the model group was down-regulated,while the expression of IDE was up-regulated.Compared with the model group,theexpressions of Per1,Bmall,Clock and IDE in mice in the administration group were all up-regulated,and the most obvious up-regulated expression was found in the morning,mind and evening mental retardation group.The results of IDE ELISA in hippocampus tissue were consistent with the trend of PCR.Compared with the blank group,the content of IDE in hippocampus tissue of other groups increased significantly(P < 0.05),and the content of IDE was the highest in the group of Morning Good Spirit and Evening Yizhi(P < 0.01).3.Enrichment analysis of genes associated with AD and insomnia showed that the biological processes of AD and insomnia were mainly related to neuropeptide signaling pathway,nerve impulse transmission,regulation of postsynaptic membrane potential,apoptosis of myeloid cells,inflammatory response,etc.The connections in KEGG signaling pathway were mainly concentrated on the interaction of neuroactive ligand-receptor,serotoninergic synapse,insulin secretion,and so on.Cytokine-cytokine receptor interaction,JAK-STAT signaling pathway.4.Suanzaoren Decoction's enrichment analysis of AD and insomnia-related genes showed that its mechanism mainly focused on steroid decomposition process,dopamine transport,neurotransmitter biosynthesis process,coagulation regulation,fibrinolysis regulation.The connection of KEGG signaling pathway mainly focused on complement and coagulation cascade signaling pathway,serotoninergic synapse,steroids.Hormone biosynthesis and other signaling pathways.5.GABA receptor docking results showed that the optimal ligand was MOL005018,and Ach E docking results showed that the optimal ligand was MOL004966.Molecular dynamics simulation results showed that both MOL005018 and MOL004966 could bind stably to the corresponding receptors.Conclusion:1.The common etiology and pathogenesis of AD and senile insomnia are kidney deficiency and phlegm turbidity,of which kidney deficiency is the main cause.2.Morning soothing and evening soothing method is the best way to improve the cognitive ability of AD mice.3.AD is closely related to insomnia in a variety of biological processes and KEGG signaling pathways.4.The mechanism of Suanzaoren Decoction on AD and insomnia is various,mainly concentrated in steroid decomposition,dopamine transport,neurotransmitter biosynthesis,coagulation regulation,fibrinolysis regulation,complement and coagulation cascade signaling pathway,serotoninergic synapse,steroid hormone biosynthesis and other signaling pathways.5.There are many active ingredients of Suanzaoren Decoction on GABA receptor and Ach E,mainly small molecules MOL005018 and MOL004966.