Exploring The Mechanism Of Tianma Gouteng Decoction In The Treatment Of Epilepsy Based On Network Pharmacology And Experimental Validation

Background Epilepsy is a group of neuropathies characterized by transient and recurrent seizures,triggered by the sudden and abnormal release of neurons in brain tissue,resulting in functional disorders of the brain.Epilepsy has become the third largest problem in the world in terms of the burden on neurological patients.The clinical treatment of epilepsy is primarily pharmacological.Treatment with Chinese and Western medicine alone is not as effective as a combination of Chinese and Western medicine.Traditional Chinese medicine has become a popular form of complementary and alternative medicine,which has been confirmed by several studies.The use of traditional Chinese medicine to prevent and treat epilepsy has a long history.Tianma Gouteng Decoction is a traditional Chinese medicine with a wide range of uses for acute hypertension,Parkinson’s disease,Alzheimer’s disease,epilepsy and other neurological disorders.According to the theory of "disease-gene-target-drug" all-round and multi-angle influence network,the intervention and influence of the medication process on the pathogenic system is systematically and completely examined through network pharmacology,and the complexity of the relationship between biological system,medication and disease is elaborated by applying the method of mechanism of action of traditional Chinese medicine,and the molecular mechanism between medication and pathogenic system is explored,which brings to the diagnosis of epilepsy.This study was conducted to validate the main core targets and pathways of TCM for the treatment of epilepsy based on network pharmacology and experimental validation,in the hope of providing new approaches for the treatment of epilepsy.AimsThe mechanism of Tianma Gouteng Decoction in the treatment of epilepsy is still unclear.The aim of this study is to confirm the key therapeutic targets and pathways of TCM in epilepsy therapy based on network pharmacology and experimental validation.Methods1.A network pharmacological study of Tianma Gouteng Decoction in the treatment of epilepsy: a search of the TCMSP and TCMID databases and references for the active ingredients of Tianma Gouteng Decoction.TCMSP,Swiss Target Prediction,and Pharm Mapper databases were used to detect the potential targets of the active components,and the epilepsy genes were queried in the OMIM,Drug Bank and Genecards databases.The String database was used to construct using Cytoscape3.9.1software.the protein interaction network,and the GO functional enrichment and the KEGG pathway were analyzed in Metascape database.The network was constructed using Cytoscape3.9.1 software to predict the possible targets and molecular mechanisms.2.A part of the mice was gavaged with Tianma Gouteng Decoction for 15 days after moulding,and the blank and moulded mice were gavaged with the same dose of saline.The brains of the mice were stained with HE for pathological histological analysis,WB for protein expression of NF-κB signaling pathway,q RT-PCR and ELISA for inflammatory factors.ResultsPart I: The network pharmacology study of Tianma Gouteng Decoction for epilepsy obtained 106 active ingredients,301 potential drug targets and 2160 epilepsy disease targets through screening.A total of 101 drug-disease intersection targets were obtained based on the Venny diagram,and core targets such as AKT1,IL-6,TP53,CASP3 and TNF were obtained from the PPI network analysis,with the core targets mainly related to inflammation.Based on the Kyoto Encyclopedia of Genes and Genomes(KEGG)study,it was shown that Tianma Gouteng Decoction for epilepsy mainly includes: NF-κB signaling pathway,tumour signaling pathway,HIF-1 signalling pathway and JAKSTAT signaling pathway.The main active substances quercetin,β-sitosterol and kaempferol dock well with AKT1.Part II:(1)HE staining showed that neuronal cells in the cortical hippocampus of mice in the blank group were regular and had large,round,dark blue nuclei located in a transparent cytoplasm.The neurons of the mice in the epileptic modelling group were crinkled and deeply stained and irregular,and the number of normal neuronal cells in the mice in the drug intervention group was increased.(2)The results of the water maze experiment showed that on the third and fourth day,the mice in the drugintervention group began to have a shorter time on the platform compared to the epilepsy-modelling group,and the time when the mice in the drug-intervention group entered the platform latency for the first time was also shorter than that in the epilepsymodelling group.(3)The results of q RT-PCR and ELISA showed that the levels of AKT1,IL-6 and TNF-α in the drug intervention group were lower than those in the epilepsy modeling group and were statistically significant(P < 0.05).(4)WB results showed a decrease in NF-κB protein expression levels in the drug intervention group compared with the epilepsy modeling group(P<0.05).ConclusionThrough the network pharmacology,it was found that Tianma Gouteng Decoction was effective in treating epilepsy by using multiple elements,multiple targets and multiple pathways.It can reduce the concentration of inflammatory factors AKT1,IL-6,TNF and control the protein expression of NF-κB signaling pathway,which can be used to treat epilepsy.In summary,we can conclude that the predicted results of network pharmacology have been validated by animal studies.