Analysis of the Mechanism of Gegen Decoction in Improving Cerebral Hypoxia and Alleviating Epileptic Seizure Based on Network Pharmacology,Molecular Docking,Metabolomics and Experimental Verification
10.19378/j.issn.1003-9783.2024.09.012
- VernacularTitle:基于网络药理学、分子对接、代谢组学及实验验证分析葛根汤改善大脑缺氧减轻癫痫发作的机制
- Author:
Zige WANG
1
;
Runxue REN
;
Wentao FAN
Author Information
1. 陕西中医药大学,陕西 咸阳 712056
- Keywords:
Epilepsy;
hypoxia;
Gegen Decoction;
network pharmacology;
metabolomics;
molecular docking;
experimental verification;
rats
- From:
Traditional Chinese Drug Research & Clinical Pharmacology
2024;35(9):1389-1400
- CountryChina
- Language:Chinese
-
Abstract:
Objective Network pharmacology,molecular docking,metabolomics and experimental verification were used to investigate the effect of Gegen Decoction on epileptic seizure under cerebral hypoxia.Methods The core targets of Gegen Decoction,epilepsy and cerebral hypoxia were screened by network pharmacology,and the correlation between ligand and receptor was verified by molecular docking.Further animal experiments were carried out to verify the effect of Gegen Decoction on the rat model with epilepsy accompanied by cerebral hypoxia,which was established by lithium chlorine-pilocarpine method combined with unilateral common carotid artery ligation and placing in a closed chamber with oxygen content of 10%-15%.Sixty rats were randomly divided into blank group,model group,phenytoin sodium(26 g·kg-1)group,high-,medium-and low-dose(10.62,5.31,2.655 g·kg-1)Gegen Decoction groups.After treatment,the number of epileptic seizures rats in each group was counted,and the recovery of nerve function and epileptic seizures were evaluated.HE staining was used to observe the morphological changes of the CA1 region of hippocampus in rats.The expression of interleukin-6(IL-6)and interleukin-17(IL-17)were detected by immunohistochemistry.The changes of brain neurotransmitters were analyzed by targeted metabolomics.Results Network pharmacological results suggested that 128 active components and 228 targets were obtained.A PPI network map of drug-disease was constructed by STRING.The results showed that STAT3,IL-6 and AKT1 were the core targets.GO enrichment for biological processes(BP)involves responses to external irritant,oxygen levels,hypoxia and metabolic processes of reactive oxygen species.Cell components(CC)mainly involve in the impact on cell membrane and protein kinase complex,while molecular function(MF)involves in influence on the binding of ubiquitin-like protein and ubiquitin-like protein ligase.The key pathways of Gegen Decoction in the treatment of hypoxic-induced epilepsy in the brain,which were analyzed by KEGG,involved lipid and atherosclerosis signaling pathway,IL-17 signaling pathway,Th-17 signaling pathway and so on.The experimental results showed that the number of seizures in the Gegen Decoction group was significantly decreased compared with that in the model group(P＜0.01),the morphology and structure of hippocampal cells were improved to varying degrees,and cell vacuolation was reduced.The expression of IL-17A and IL-6 in all treatment groups was lower than those in model group,and there was statistical significance in medium-dose Gegen Decoction group(P＜0.01,P＜0.05).Brain metabolomics test showed that compared with model group,monoamine neurotransmitters including 5-hydroxytryptophan(5-HTP)and norepinephrine(NE)were increased in high-dose Gegen Decoction group,while tryptophan metabolite kynurenine(Kyn)was decreased.Conclusion Gegen Decoction can reduce the levels of serum inflammatory factors IL-6 and IL-17A in epileptic rats under hypoxia.Moreover,Gegen Decoction can reduce epileptic seizures by improving hypoxia-induced inflammatory response,increasing the content of monoamine neurotransmitter(5-HTP and NE),as well as reducing the content of tryptophan metabolite Kyn and the toxin of tryptophan metabolism pathway.
