RESUMO
Objective::To explore the effect of Erjingwan on the biological basis of kidney yin deficiency Alzheimer' s disease(AD)rats induced by ovariectomy+ D-galactose combined with amyloid beta1-40 (Aβ1-40). Method::After ovariectomy, rats were randomly divided into five groups: model group, positive group, Erjingwan high, medium and low dose group, 11 rats in each group, and 11 rats in sham operation group. One week after operation, D-galactose was injected intraperitoneally for 7 weeks, and four weeks after operation, Aβ1-40 was injected unilaterally into hippocampus. The rats in model group and sham-operation group were given saline by intragastric administration 3 weeks after operation. The rats in high, middle and low dose groups of Erjingwan were given corresponding concentration (9.0, 4.5, 2.25 g·kg-1). The rats in positive control group were given Donepezil 1.0 mg·kg-1 with dosage of 10 mL·kg-1 once a day for 35 consecutive days. After 30 days of administration, the learning ability of the rats were examined using a Y-maze. The hippocampus tissues of rats were isolated. The morphology of hippocampus was observed by Nissl staining.The proteins were detected by Nanol-ESI liquid-mass spectrometry system and identified by protein Discovery software. Relative quantitative and qualitative analysis of differential proteins in hippocampus was performed by SIEVE software, and Gene Ontology of differential protein was performed by PANTHER Classification System database. String analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment were performed on the differential proteins. Result::Compared with model group, the correct rate of Y maze in the high and middle dose groups of Erjingwan was significantly raised(P<0.05), and the number of neurons in the hippocampal CA1 area was significantly increased(P<0.01).115 differential proteins (Ratio>1.5 or Ratio<0.5) such as Insulin-like growth factor 1 receptors(IGF-1R) were found in the high-dose group of the Erjingwan group as well as 94 differential proteins such as Synaptophysin expressed in the middle-dose group of the Erjingwan. And there are 87 differential proteins such as Acetyl-CoA acetyltransferase-cytosolic in the positive drug group. It showed that these proteins were mainly divided into tubule-related proteins, heat shock proteins, energy metabolism-related proteins and AD-related proteins with GO analysis. It was found that the above differential proteins involved 93 signaling pathways such as Dopaminergic synaps by KEGG analysis. Conclusion::Erjingwan can improve cognitive impairment and neuronal damage in rats with kidney yin deficiency, possibly by altering the expression of multiple pathway-associated proteins such as phosphatidylinositol 3-kinase/protein kinase B signaling pathway(PI3K/Akt), insulin signaling pathway, and adenylate-activated protein kinase (AMPK)signaling pathway, estrogen signaling pathway, and Dopaminergic synapse.
RESUMO
The present study is to explore the material basis and mechanism of Erzhi Wan the prevented Alzheimer's disease by using network pharmacology. The key target of Alzheimer's disease was docked with the Erzhi Wan compounds, and the drugs-target combined with target-signal pathway network model was established by Cytoscape 3.2.1 software. Thirty compounds have a strong interaction with key target of Alzheimer's disease and three key pathways related with Wnt, MAPK and PI3K-Akt-mTOR. There are 5 ingredients such as quercetin,geraniol,beta-sitosterol,nerol,eriodictyol that could be verified from literature.This result initially revealed the material basis for Erzhi Wan for Alzheimer's disease and the mechanism in terms of three signaling pathways. The network pharmacology method found that the active ingredients of Erzhi Wan for Alzheimer's disease may be quercetin,geraniol,beta-sitosterol,nerol,and eriodictyol, and the mechanism may be related to three signal pathways including Wnt, MAPK, and PI3K-Akt-mTOR.