| Baoyuan Decoction is composed of ginseng,astragalus,licorice and cinnamon.Ginseng and astragalus are valuable traditional Chinese medicines for nourishing health.They have the functions of reinforcing vital energy and better for harmonious in our body.In Zhang's Yitong Zuji,this is the first prescription for invigorating qi.The efficacy advantages of Baoyuan Decoction mostly come from the summary of clinical experience,but the mechanism research of each disease is mostly blank,lack of experimental and scientific basis.Metabonomics is different from the study of drug metabolism.It does not focus on the metabolic changes of a specific component in the body.It is a holistic study of the types,contents and changes of metabolic products before and after disturbance of the target organism.Its application to the study of traditional Chinese medicine,especially complex traditional Chinese medicine prescriptions,can serve as a modern pharmacology and mechanism of traditional Chinese medicine.In this study,the anti-fatigue and anti-hypoxia mechanisms of Baoyuan Decoction were studied by gas chromatography-mass spectrometry(GC-MS)and liquid chromatography-mass spectrometry(LC-MS).Metabolic markers were searched and metabolic pathways were plotted.One,Study on the Anti-fatigue Effect of Baoyuan Decoction Based on Metabonomics.In order to explore the anti-fatigue effect of Baoyuan Decoction,the exhaustive swimming model of rats was established.After obtaining serum and urine samples,GC-MS and LC-MS analysis were carried out,and metabolomic techniques were used to find metabolic markers to elucidate the metabolic mechanism.The LC-MS method of lactic acid in serum was established to replace the traditional ELISA method.Lactic acid was detected by using CAPCELL PAK ADME(50 mm x 2.0 mm,5 ?m)column with a mobile phase of water(0.1% formic acid)(A)-methanol(0.1% formic acid)(B)= 98-2.The RSD of peak area precision was 4.53%,The correlation coefficients was 0.999,and the recovery of sample was 96.47%.This method is proved to be simpler with good reproducibility.It provides a new method and idea for the determination of lactic acid in serum.The method was used to analyze and compare the lactic acid of rats in different drug-administered groups,and to study the effect of Baoyuan Decoction on lactic acid changes.We also established a mixed-mode retention mechanism(reversed-phase and cation-exchange)to determinate the amino acids.In this method,eighteen amino acids can be retained on a CAPCELL PAK CR 1:4(150 mm x 2.0 mm,5 ?m)column with a mobile phase of 0.1% formic acid(A)-acetonitrile(0.1% formic acid)(B)-50 mmol/L ammonium formate(C)using a gradient elution program.The RSDs of peak area precision ranged from 0.73% to 5.85%.The correlation coefficients of eighteen compounds were greater than 0.996.And the recoveries of serum and urine samples ranged from 92.2% to 113.6%.It provides a new method and idea for the determination of amino acids in biological samples,using simple mobile phase without the ion-pair and derivatization.Using this method,the difference of amino acids in serum and urine of different groups was analyzed,and the different amino acids was obtained.The method was used to analyze and compare the amino acids of rats in different drug-administered groups,and to study the effect of Baoyuan Decoction on amino acid changes.According to the results of GC-MS and LC-MS analysis,19 marker metabolites were found,including lactic acid,glycerol,tryptophan,methionine,valine,leucine,phenylalanine,tyrosine,proline,glycolic acid,hippuric acid,isocitric acid,citric acid,mannitol,gluconic acid,alanine,glutamic acid,glycine and lysine,involving 6 main metabolites.Metabolic pathways: 1.Phenylalanine,tyrosine and tryptophan biosynthesis 2.Valine,leucine and isoleucine biosynthesis 3.Phenylalanine metabolism 4.Glyoxylate and dicarboxylate metabolism 5.Alanine,aspartate and glutamate metabolism 6.Citrate cycle.Baoyuan Decoction can affect the body's energy metabolism and play an anti-fatigue role.The results showed that Baoyuan Decoction could effectively prolong the exhaustive swimming time and restore the weight of rats.Baoyuan Decoction can affect the body's energy metabolism and excitatory neurotransmitters,eliminate the accumulation of lactic acid,and play an anti-fatigue role.Two,Study on the Anti-hypoxia Effect of Baoyuan Decoction Based on Metabonomics.In order to explore the anti-hypoxia effect of Baoyuan Decoction and establish mice models of atmospheric pressure airtight hypoxia and hematologic hypoxia by sodium nitrite injection.The survival time of mice under hypoxia was compared.After obtaining serum and urine samples,GC-MS analysis were carried out,and metabolomic techniques were used to find metabolic markers to elucidate the metabolic mechanism.Baoyuan Decoction can effectively prolong the survival time of pressure airtight hypoxia,but no significant prolongation of the survival time under hematologic hypoxia.The serum samples of atmospheric pressure airtight hypoxia mice were analyzed by GC-MS,and 22 marker metabolites were found by metabolomic analysis.They were alanine,lactic acid,1-phenylethylamine,glycine,isoleucine,aminobutyric acid,dimethylglycine,serine,ethanolamine,tyramine,tryptamine,ethylamine and threonine.2,3-butanediol,succinic acid,threonic acid,glycerol phosphoric acid,N-acetyl-L-lysine,butylamine,erythrose,fucose,inositol-1-phosphoric acid.Metabolic pathways: 1.Glycine,serine and threonine metabolism 2.Methane metabolism 3.Aminoacyl-tRNA biosynthesis 4.Valine,leucine and isoleucine biosynthesis.The anti-fatigue mechanism of Baoyuan Decoction are described.In conclusion,Baoyuan Decoction has the ability of anti-fatigue and anti-hypoxia under normal pressure.Its mechanism involves the interaction of multiple metabolic pathways,especially energy metabolism,which is of great significance in clinical application.This experiment provides a new idea and experimental basis for the study of traditional Chinese medicine prescriptions through the statistical analysis of metabonomics. |
PDF Full Text Request