Study On The Compatibility Effect And Material Basis Of Coptidis Rhizoma-Euodiae Fructus Herb Pair Against Sepsis

Sepsis is a major global health problem,which is defined as life-threatening organ dysfunction caused by a host’s dysfunctional response to infection.As research progresses,it was found that the gut is particularly important in the development of sepsis.The combination of Coptidis Rhizoma-Euodiae Fructus herb pair(CR-EF)has a long history.Modern pharmacological studies have shown that it has various pharmacological effects,such as anti-inflammation,antiulcerogenic,protection of the gastrointestinal tract and repair of the intestinal mucosal barrier.Therefore,this thesis will carry out research on the material basis of CR-EF,explore the compatibility effect and mechanism,and provide a theoretical basis for CR-EF against intestinal mucosal lesions of sepsis.In order to provide a scientific basis for its further development.This thesis is divided into 3 chapters;the research contents are as follows:Chapter 1.Literature research Section 1.Advances in the study of intestinal mucosal injury in sepsisThe intestinal mucosa barrier in normal body prevents pathogenic bacteria and toxins from crossing the intestinal mucosa to invade the blood circulation and peripheral organs,thus maintaining the stability of the internal environment of the organism.Local inflammation reactions during sepsis can cause damage of the intestinal mucosa barrier,which can accelerate the development of sepsis and even endanger patient’s life.Therefore,this section summarizes the progress of intestinal mucosal injury in sepsis,and also summarizes the pathways of action of TCM in the treatment of intestinal mucosal barrier injury,in order to provide ideas for subsequent research.Section 2.Advances in the Study of Coptidis Rhizoma-Euodiae Fructus herb pairDrug pair is one of the common forms of clinical prescriptions.Through the combination of two drugs,it can play effect-enhancing or toxicity-reducing.CR-EF combination has a long history.It is commonly used in clinics as Zuojin pills,which comes from Danxi’s Mastery of Medicine.In this section,we summarize the chemical composition of CR-EF,and summarize its synergistic and antidotal effects,and review its modern pharmacological effects to lay the foundation for this thesis.Section 3.Research ideas and program designThis section aims to clarify the research ideas,experimental design,and overall framework of the research content.Focusing on the CR-EF,by replicating the mouse model of LPS-induced sepsis,the following key issues are planned to be solved:(1)explaining the chemical composition of CR-EF;(2)evaluation of the protective effect of CR-EF drug pair on mice with sepsis;(3)Uncovering the mechanism of CR-EF drugs against sepsis.Chapter 2.Chemical constituents of Coptidis Rhizoma-Euodiae Fructus herb pair and its protective effect on sepsis mice Section 1.Qualitative analysis of chemical composition of Coptidis Rhizoma-Euodiae Fructus herb pair based on UPLC-Q-TOF/MS methodIn this section,the UPLC-Q-TOF/MS method was used for the qualitative analysis of the chemical constituents in CR-EF drug pair,and a total of 17 compounds were identified as 9,10-dihydroxy columbamine,rutaecarpine,2-hydroxyjatrorrhizine,demethylene coptisine,8-oxoepiberberine,jatrorrhizine,groenlandicine,columbamine,epiberberine,coptisine,dehydroevodiamine,13-methylberberine,palmatine,berberine,umbelliferone,limonin,evodiamine.Including 15 alkaloids,one limonoid,and one coumarins.Five of them from Euodiae Fructus,12 from Coptidis Rhizoma.In order to provide a theoretical basis for the further study of pharmacological mechanism.Section 2.Study on the extraction process of Coptidis Rhizoma-Euodiae Fructus herb pair based on analytic hierarchy process and multi-index orthogonal testIn this section,the extraction process of the CR-EF drug pair was investigated by using hierarchical analysis,multi-indicator comprehensive scoring method combined with orthogonal test,and seven indicative ingredients(coptisine,epiberberine,jatrorrhizine,berberine hydrochloride,palmatine chloride,evodiamine and rutaecarpine)were used as inspection indicators.The results show that the optimal processing parameters were as follows: the solid-liquid ratio of 1:8 g/m L,extracting three times,each for one and a half hours.The optimized process of CR-EF extraction has been verified to be reasonable,stable,and reproducible,which could provide reference for further development and utilization of the formula.Section 3.Evaluation of the protective effect of Coptidis Rhizoma-Euodiae Fructus herb pair on septic miceIn this section,we used intraperitoneal injection of LPS to successfully induce a mouse model of sepsis,and treated with different doses of CR-EF drug pair(low dose:0.78 g/kg,medium dose: 1.56 g/kg,and high dose: 3.12 g/kg),CR(1.30 g/kg)and EF(0.26 g/kg)were administered for intervention.The results showed that MD groups(i.e.,human clinical equivalent dose)could improve the survival rate of septic mice,significantly reduce the serum inflammatory factors,and also improve inflammatory infiltration of intestinal tissue,and increase the expression of tight junction proteins.Compared with the other dose groups and the single drug CR group and EF group,Among them,the best improvement was achieved by the medium dose group of CREF drug pair.Chapter 3.Study on the mechanism of anti-sepsis action of Coptidis Rhizoma-Euodiae Fructus herb pair Section 1.Mechanism of Coptidis Rhizoma-Euodiae Fructus herb pair against sepsis based on metabolomicsIn this section,we study the mechanism of the anti-sepsis action of CR-EF drug pair based on a metabolomic approach of UPLC-Q-TOF/MS.A total of 23 differential metabolites(11 in positive ion mode and 12 in negative ion mode)were selected,and relevant metabolic pathways were constructed.After analysis,it was found that CR-EF drug pair may exert its anti-septic effect by promoting glycerophospholipids metabolism to normal and inhibiting the excessive activation of the arachidonic acid metabolic pathway.Compared with CR and EF,CR-EF drug pair interferes with multiple metabolic pathways by regulating the expression of multiple differential metabolites,leading to better treatment effects.Section 2.Mechanism of Coptidis Rhizoma-Euodiae Fructus herb pair against sepsis based on transcriptomeIn this section,the colon tissues of mouse in control group,sepsis model group and CR-EF drug pair middle dose group were sequenced by transcriptome technique.Then the differentially expressed genes were analyzed by cluster analysis,GO analysis and KEGG pathway enrichment analysis were performed on the selected 109 differentially expressed genes.The results showed that these differentially expressed genes were mainly enriched in HIF-1 signaling pathways,TNF signaling pathways and IL-17 signaling pathways and so on.It showed that CR-EF may exert the mechanism of action against sepsis by regulating the expression of related genes and inhibiting the above-mentioned inflammation-related signaling pathways.Section 3.Correlation analysis of metabolic and transcriptomic data reveals mechanism of Coptidis Rhizoma-Euodiae Fructus herb pair against sepsisIn this section,the combined analysis of metabolomics and transcriptome was used to explore the anti-sepsis mechanism of CR-EF herb pair.It was found that there was a strong correlation between selected metabolites and differentially expressed genes.CREF reverses differentially expressed genes in mice caused by sepsis.It plays an important role in preventing intestinal mucosal damage of sepsis by regulating genes closely related to inflammation.Section 4.Study on the mechanism of anti-sepsis effect of Coptidis Rhizoma-Euodiae Fructus herb pair based on intestinal floraIn this section,16 S r DNA sequencing technology was used to study the structure of intestinal flora in the sepsis mouse model and the mice treated with CR-EF,CR,and EF.The results showed that sepsis reduced the diversity of gut flora in mice,increased the abundance of harmful bacteria,and decreased the abundance of beneficial bacteria.Each administration group inhibited the growth of harmful bacteria and promoted the growth of beneficial bacteria to a certain extent,with CR-EF group playing a significant role.It alters and rebalances the composition of intestinal flora in septic mice,which has a protective effect on septic mice.