Protective Effect Of Rehmannia Glutinosa-Cornus Officinalis Combination On Aβ25-35-induced Apoptosis Of N2a Cells Based On PI3K-AKT Signaling Axi

Purpose:There is no record of the name of Alzheimer’s disease(AD)in ancient Chinese medicine literature.According to clinical symptoms,AD belongs to the category of diseases such as stupid disease,good memory and depression syndrome.Chinese medicine believes that AD is located in the brain,but has a close relationship with kidney essence deficiency As the pathological basis of AD,the deficiency of medulla sea has been recognized by most scholars,and the clinical treatment based on kidney tonification has also achieved good efficacy Disease,AD)is one of the commonly used drug pairs of kidney-tonifying compound.At present,the pharmacodynamic substance basis and mechanism of the compatibility of Rehmannia glutinosa and Cornus officinalis in the treatment of AD are still uncertain.Therefore,this paper is based on network pharmacology.Material and method:1.Employ TCMSP(traditional Chinese medicine database and analysis platforms ystem pharmacology,https://tcmspw.com/tcmsp.php)and BATMAN database(molecular mechanism of traditional Chinese medicine bioinformatics analysis tools,http://bionet.nc psb.org/batman-tcm/)prediction prepared rehmannia root,cornel of effective components and the corresponding target effective active ingredients;Use TDD database(Therape utic target database,http://db.idrblab.net/ttd/),GAD database(http://geneticassociationdb.nih.gov/)and TCMSP screening AD related targets;The network model of "ripe rehma nnia officinalis-effective active component-AD disease target" was verifide by Cytoscape software.GO note analysis as well as KEGG pathway analysis were executed on "Ripe Rehmannia glutinosa-effective active component of Cornus officinalis-AD disease target" by DAVID database.Screen out the possible pathway of rehmannia glutinosa-Cornus officinalis in the treatment of AD.2.N2 a cells were treated with Aβ25-35(20 μmol/L)to construct AD cell model,and treated with low and high concentration rehmannia officinalis ethanol extract for 24 h.The cell survival rate was determined by CCK8 method,and the optimal induction concentration of Aβ25-35 was sought.Apoptosis was checked by Hoechst 33342 and Annexin V-FITC/PI staining.The activity as well as protein show of caspase-3 containing cysteine were checked by colorimetry as well as Western blot.The protein show of p-PI3 K,PI3K,p-AKT and AKT were checked by Western blot.Results:1.A total of 41 active ingredients were screened for the drug pairs of rehmannia glutinosa and Cornus officinalis,among which 4 were the final effective ingredients of rehmannia glutinosa and 20 were the final effective ingredients of Cornus officinalis.The active components of mature rehmannia-Corni officinalis not only regulate the 10 targets of human prostaglandin peroxidase synthase 2(PTGS2),muscarinic choline receptor M1(CHRM1)and muscarinic choline receptor M2(CHRM2),but also regulate h uman prostaglandin peroxidase synthase 2(PTGS2),muscarinic choline receptor M2(C HRM2).It also affects 44 biological processes(such as acetylcholine receptor activity,acetylcholinesterase activity,cholinesterase activity,G-protein-coupled serotonin receptor activity,enzyme binding as well as serotonin binding)as well as 6 metabolic pathways(calcium signaling pathway as well as cholinergic synaptic signaling pathway).Comb ined with literature reports and the correlation with AD,we selected PI3K-AKT signal ing pathway as rehmannia-Cornus officinalis to improve AD nerve cell injury for subsequent experimental study.2.Stack up the vacuum control subset,the apoptosis speed of N2 a cells treated with Aβ25-35 prominent augmented,the relative activity as well as protein show of Caspase-3reduced,another key fact to remember the ratio of p-PI3 K /PI3 K and p-AKT /AKT decreased(P<0.05).After 24 h treatment with low and high concentration of cooked Rehmannia officinal-Cornus ethanol extract,apoptosis rate of N2 a cells decreased,caspase-3 activity and Cleaved caspase-3 protein expression decreased,p-PI3K/PI3 K,p-AKT/AKT ratio augmented.Contrasted with model subset,the divergence was statistically prominent(P<0.05).Contrasted with the low-dose subset,the apoptosis rate of N2 a cells and the relative activity of Caspase-3 reduced in the high-dose subset,another Key Fact to Remember the p-AKT/AKT ratio increased(P<0.05).Conclusion:Through network pharmacology study,the signaling pathway of rehmannia-Cornus officinalis on anti-AD was predicted,and the PI3K-AKT signaling pathway was selected as the target pathway for cell experiment verification.Cell model experiments proved that the ethanol extract of rehmannia glutinatum had anti-apoptotic effect on Aβ25-35 induced AD cell injury model,another key fact to remember the framework might be correlative to the activation of PI3K-AKT signaling pathway to inhibit capases-3 activity and protein expression.