| Migraine is one of the four chronic diseases that are internationally recognized as the most disabling,which seriously affects the quality of life of patients.Western medicine cannot treat diseases well due to its side effects and drug resistance.Traditional Chinese medicine has the characteristics of multi-target and multi-effect comprehensive regulation.Therefore,it is a popular research to find effective therapeutic drugs from traditional Chinese medicine.Among them,Tianma Gouteng has a significant effect on chronic migraine and has been widely used in clinical treatment.But what is the pathogenesis of chronic migraine,and what is the mechanism of action of Tianma Gouteng on the treatment of chronic migraine,which is still unclear.At present,metabolomics technology has been widely used in the research of disease and drug mechanism.In addition,many diseases and drug interventions can cause changes in the structure of intestinal flora,which provides a new idea for us to explain the clinical effects of traditional Chinese medicine compound.In view of the complex pathogenesis of chronic migraine and the significant effect of Tianma Gouteng on chronic migraine,the main contents of this research are as follows: 1.Intervention of Tianma Gouteng on the model of chronic migraine induced by nitroglycerin in ratsIn order to verify whether a rat model of chronic migraine induced by nitroglycerin is established and to confirm the therapeutic effect on water extracts of Tianma Gouteng,this chapter was conducted by measuring body weight,thermal pain threshold and plasma NO,CGRP,ET,5-HT levels.After modeling,the rats gained weight slowly,the pain threshold decreased,plasma NO,CGRP,ET levels increased,5-HT content decreased,and these characters tends to be normal after administration of Tianma Gouteng.It is initially proved that the model is established successfully and the drug has certain curative effect.However,the mechanism of disease occurrence and drug treatment is still unclear,and it needs to be further elaborated by metabolomics research.2.Metabolomics study on Terapeutic mechanism of Tianma Gouteng on chronic migraine ratsTo elucidate the pathogenesis of nitroglycerin-induced chronic migraine model and the mechanism of action of Tianma Gouteng on chronic migraine in rats.In this experiment,UPLC-QTOF-MS/MS technique was used to study the plasma endogenous metabolites in rats,and analyzed the regulation of biomarkers after administration.Using QI and UNIFI software and qualitative analysis of the data through database matching,40 differential metabolites were discovered,and Tianma Gouteng could regulate 34 components.Using Meataboanalyst 3.0,KEGG database enrichment,mapped related signaling pathways(linoleic acid metabolism,tryptophan metabolism,arginine metabolism,...),providing a basis for the pathogenesis study of chronic migraine from the metabolic level.It is speculated that Tianma Gouteng may play a role in regulating these metabolic pathways.However,some studies have reported that the bioavailability of Tianma Gouteng is relatively low,and the componds are not completely obsorbed into the blood.Because the drug stays in the gastrointestinal tract for a long time,it may cause the change of intestinal flora structure,thus participating in the therapy process of Tianma Gouteng.3.Study on the Terapeutic mechanism of Tianma Gouteng by regulating intestinal flora disorderTo further clarify whether the mechanism of action of Tianma Gouteng on chronic migraine in rats is related to intestinal flora.16 S rRNA sequencing was performed to proved that the intestinal flora level of the disease model was dysregulated.The abundance level of each microbe after the intervention of Tianma Gouteng was getting normal.And through 16 S rRNA functional prediction analysis,these bacteria are mainly involved in amino acid metabolism,energy metabolism,etc.,consistent with the results obtained by plasma metabolomics,Pearson correlation analysis also shows that intestinal bacteria and plasma biomarkers are related.However,these conclusions are only speculated from the experimental results,whether there is a correlation,and experiments are needed.3.Validation analysis of correlation between intestinal flora and metabolomicsIn order to prove that Tianma Gouteng medicine affects the structure of intestinal flora is also one of the ways to exert drug efficacy,we conducted two parts of experiments,using UPLC-QTOF-MS/MS technology to establish a rapid analysis method,first determined the metabolic profile of the intestinal contents and the endogenous differential components prove that the changes in plasma metabolic components may be caused by the absorption of components in the intestine into the blood;then the experiment of co-incubation of feces in vitro is carried out,and the metabolism of intestinal bacteria is inhibited by antibiotics,and Tianma Gouteng could not play a regulatory role on the components in the feces of the model rats;while the samples without antibiotics were added,the content of some biomarkers tends to be normal after co-incubation,indicating that the drugs pass through the intestinal flora to the inside of the intestines to regulate endogenous components by changing the structure of intestinal flora.In this experiment,combined with metabolomics technology and intestinal flora research,nitroglycerin-induced chronic migraine model and the mechanism of treatment of Tianma Gouteng medicine were studied.The correlation analysis between the two was carried out,and the treatment mechanism of Tianma Gouteng was explained from a new perspective.This research also provides a new research idea and theoretical basis for the clinical treatment of chronic migraine and the mechanism of action of Tianma Gouteng. |
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