Treatment and mechanism of Yigansan on Alzheimer's disease
Objective To explore the therapeutic effect of Yigansan on Alzheimer's disease(AD),to predict its mechanism through network pharmacology,and to carry out experimental verification.Methods The effects of Yigansan(400 and 800 mg·kg-1)on the learning and memory function of AD mice were investigated in a D-galactose-induced AD mouse model,and the mRNA expressions of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-2(IL-2)and interleukin-6(IL-6)in the brain tissues of AD mice were analyzed by real-time quantitative PCR(qRT-PCR).Through the TCM Systematic Pharmacology Data Analysis Platform(TCMSP),the active ingredients of the herbal medicines Bupleuri Radix,Glycyrrhizae Radix et Rhizoma,Chuanxiong Rhizoma,Angelicae Sinensis Radix,Atractylodis Macrocephalae Rhizoma,Poria and Uncariae Ramulus cum Uncis were screened,and the AD targets were obtained by GeneCards and other databases,and the core targets were enriched by gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways.The biological processes(BP),molecular function(MF)and cellular composition(CC)and KEGG signaling pathwayinvolved in the core targets were obtained,respectively.Mouse neuroblastoma Neuro-2a cells and RAW264.7 macrophages were co-cultured with H2O2 and hepatosuppress san(3 and 6 μg·mL-1),respectively,to observe the effects of Yigansan on the proliferation and apoptosis of Neuro-2a cells in vitro,and the activity of Caspase-3 and Caspase-8,reactive oxygen species(ROS),glutathione(GSH),malondialdehyde(MDA),effects on superoxide dismutase(SOD)and mitochondrial membrane potential(MMP).The effects of Yigansan on the expression of TNF-α,IL-1β,IL-2 and IL-6 in RAW264.7 cells were observed,and the network pharmacology results were verified.Results A total of 159 active ingredients were screened for the treatment of AD by network pharmacology technology,and the top 10 compounds were quercetin,kaempferol,isorhamnetin,β-sitosterol,stigmasterol,7-methoxyisoflavone,mangopertin,naringenin,meditalpin,and licochalcone A,and 227 core targets were screened,and the top 10 core targets were serine/threonine kinase 1(AKT1),tumor necrosis factor(TNF),interleukin 6(IL6),tumor protein P53(TP53),interleukin 1β(IL1B),estrogen receptor 1(ESR),proto-oncogene(JUN),prostaglandin-endoperoxide synthase 2(PTGS),caspase 3(CASP3),signal transducer and activator of transcription 3(STAT3).Bioinformatics analysis showed that the treatment of AD was related to glycosylation end products/glycation end product receptors and lipid and atherosclerosis signaling pathways.The results of animal experiments showed that Yigansan could improve the impairment of learning and memory function in AD model mice,and down-regulate the mRNA expression of IL-1β,IL-2,IL-6 and TNF-α in the brain tissues of AD mice.The results of cellular experiments showed that Yigansan significantly alleviated the proliferation inhibitory and apoptosis-inducing effects of H2O2 on Neuro-2a cells(P＜0.05),decreased the activities of Caspase-3 and Caspase-8(P＜0.05),lowered the levels of cellular MDA and ROS(P＜0.05),and elevated the levels of MMP as well as GSH.Yigansan significantly suppressed the H2O2-induced elevation of TNF-α and IL-1β,IL-2 and IL-6 in RAW264.7 cells(P＜0.05).Conclusion Yigansan exerts therapeutic effects on AD by alleviating learning memory impairment and oxidative damage.
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