| Epimedium is the traditional Chinese tonic medicine.Clinical research found that raw Epimedium and processed products with different efficacy,raw products used for rheumatism and treatment of osteoporosis,processed with suet oil used for Kidney deficiency syndrome.The processing mechanism of epimedium in the early stage of the research group has been explained from the viewpoints of chemical composition and absorption of metabolic changes.However,the role of traditional Chinese medicine in the human body is a complex system of biological processes,only from the above two aspects can not fully explain its processing mechanism.In order to evaluate the intervention effect of epimedium on kidney yang deficiency from the whole level,and to explain the difference of the effect of epimedium on the kidney and yang deficiency,in this study,we used metabolomics method and multivariate statistical methods to study the changes of endogenous metabolites in urine and plasma after taking Herba Epimedii and its different processed products.We searched for the identification of biological markers of kidney yang deficiency,from the overall level of its evaluation of the role of ?warming kidney and enhancing yang?,investigating its metabolic pathways.Reveal epimedium warming kidney and enhancing yang efficiency mechanism from a new perspective,to clarify the processing mechanism.1.In this paper,we selected the epimedium brevieornum Maxim as the research object,and prepared the emulsions of suet oil and epimedium,heating products,suet oil processed prodects extract.Continuous intraperitoneal injection of hydrocortisone 10 days,we established the rat kidney yang deficiency model and given the model group rats with the above liquid.Then we observed the body weight,body temperature,survival status,HPAT axis correlation index and thymus,adrenal gland,hypothalamus,pituitary,testicular pathology of the six groups(normal group,model group,suet oil group,epimedium extract group,epimedium heating processed extract group,epimedium suet oil processed extract group),We found that intraperitoneal injection of hydrocortisone after 10 days,the model was successfully established.After administration of Epimedium and its different prcessed products extracts,each group of rats kidney yang deficiency have a certain reduction.Among them,the application of suet oil processed epimedium on the treatment of kidney yang deficiency,the effect is the best,followed by heating processed products,Epimedium products the most times,the treatment of suet oil is not significant.Suggesting that oral administration of suet oil processed Epimedium can achieve a therapeutic effect by better regulating the dysfunction of the HPAT axis.2.In this paper,GC-MS and NMR were used to detect the type and concentration of endogenous metabolites in urine and plasma of six groups respectively,and the principal component analysis(PCA)model was established.PCA model showed that the model group and the blank group were well distributed in the PCA model after intraperitoneal injection of hydrocortisone,indicating successful modeling.Feeding the rats with above extract respectively,overlap area ratio distribution of the model group in the four-administered group decreased in PCA model,the sample distribution areas gradually biased to the normal group,and the ratio of the overlapping area gradually increased normal group.The results showed that the contents of endogenous metabolites in the urine and plasma of the epimedium extract group,epimedium heating processed extract group and epimedium suet oil processed extract group were gradually higher than those in the model group and the overall difference gradually increases,and compared with the blank group,the gap is getting smaller and smaller,which suggesting that emulsions of suet oil,epimedium,heating products,suet oil processed prodects extract of the treatment of kidney yang deficiency more and more strong.3.The multivariate statistical analysis of the endogenous metabolites in the blank and model groups resulted in 41 accurate metabolites: Ribitol,Tyramine,Galacturonic acid,Gulonate,3-hydroxyproline,Ethanolamine,Formate,Glycerophosphorylcholine,Indole-3-lactate,Phenylacetyglycine,m-Hydroxyphenylacetate,Alanine,Citrate,Glyceraldehyde,Malonate,Trigonelline,Oxoproline,Mannose,Ornithine,Citric acid,Tryptophan,Sucrose,2,4,6-trimethyl-Pyridine,Glycerol,Threonic acid,Cholesterol,3-Hydroxybuyarate,Acetoacetate,Creatine,Glutamate,Glutamine,Glycero,Glycine,Histidine,Lactate,Lysine,Methoinine,Phenylalanine,Pyruvate,VLDL,?-Glucose.Comparison of the above differences in metabolites in the six groups found that intraperitoneal injection of hydrocortisone modeling,the metabolites abnormal increase / decrease,and given Epimedium and its processed products,the content of each metabolite have a certain callback.The overall regulation of the effect of surt oil processed epimedium > heating epimedium > epimedium > suet oil,suggesting that suet oil and epimedium have a certain synergistic effect,through the regulation of the relevant pathways to play a role in warming kidney and enhancing yang.Therefore,the "oil" and "heating" these two key links in the processing of epimedium is an essential part of the process.4.The above 41 differential metabolites were introduced into the KEGG and MetaboAnalyst 3.0 databases to obtain a total of 35 metabolic pathways associated with kidney yang deficiency.By means of statistical analysis,nine pathways were obtained: Nitrogen metabolism,Aminoacyl-tRNA biosynthesis,D-Glutamine and D-glutamate metabolism,Alanine,aspartate and glutamate metabolism,Methane metabolism,Glycine,serine and threonine metabolism,Glyoxylate and dicarboxylate metabolism,Phenylalanine,tyrosine and tryptophan biosynthesis,Synthesis and degradation of ketone bodies,involving Glycan Biosynthesis and Metabilish,Lipid Metabolism,Metablism of Terpenoids and Polytetides,Xenobiotics Biodegradation and Metabolism,Carbohydrate metabolism,Amino Acid Metabolism,Energy Metabolism,Nucleotide Metabolism,Metabolism of Cofactors and Vitamins,Biosynthesis of Other Secondary Metabolism and Metabolism of Other Amino Acid.In summary,this paper studies the necessity of "heating" and "suet oil" these two key links is an essential part of the process in the processing of epimedium to links on the role of epimedium which can enhance the efficacy of ?warming kidney and enhancing yang? through the pharmacodynamics,metabolomics study of epimedium processing.In this study,the overall macro animal pharmacodynamic evaluation,the endogenous metabolites microscopic changes within these two levels,multiple perspectives enriches the processing mechanism of Epimedium,also provided new ideas and methods to traditional Chinese medicine. |
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