Based On Proteomics To Study The Property And Flavor Of Chinese Cold And Hot Natural Properties

It has always been the hot and difficult spots in the study of Cold and Hot natural properties in Traditional Chinese Medicine.The modernization of Traditional Chinese Medicine(TCM)calls for modern scientific methods to research the pharmacology and mechanism of functions of TCM in molecular level.Following modern science and technology combined with TCM,theory of property of Chinese herbs has been rising to new height.Study on cold and heat property traditional Chinese medicines energy metabolism and expression of liver protein in normal rats,and made cold and heat property traditional Chinese medicines in property and flavor-effect-pharmacological effects difference sand similarities.This research could provide a new visual angle and experimental reference for the study on the modernization of theory of Chinese Cold and Hot Natural Properties.Therefore,we carried out the work as the followings:1.We comprehended the nature and flavor theory of TCM by lots of literature domestic and abroad;through collection the literatures on proteomies as well as cold and heat syndrome animal model determined the study method and mode of the nature and flavor theory,and then provided a solid literature foundation of our study.2.Proteomics has recently become a hotspot in life science.It is applied to do some properties theory of Chinese medicinal herbs research.Proteomics is an important component of functional genomics.Therefore,the core of life sciences transfers from genomics to proteomics.Isobaric tags for relative and absolute quantitation(iTRAQ)is now the key technique in proteome research.ITRAQ is a new quantitative proteomics technology,which has proved to be efficient,reproducible,and able to simultaneously perform quantitative protein analysis of eight samples.Many such studies have shown that iTRAQ is provides a robust platform for showing differential protein expression under distinct physiological or pathological conditions.Here we used a proven technique to study the liver,serum,hypothalamus,liver microsomal proteomics and explored the depth and breadth of Chinese medicinal herbs research.Our study provides new data for Chinese medicinal herbs research.Further research may be able to explain"New hypothesis of scientific connotation of traditional Chinese Medicine".3.Using typical cold property of drug,hot property of drug flat Chinese medicine in animal body several physiological and biochemical indexes in combination with pathological histology,From the liver,spleen,lung,hypothalamus,kidney pathological changes to explore Chinese medicine medicinal properties,and doing comparative analysis for the physiological and biochemical characteristics of these changes on common of cold and hot.Warm-heat herbs pathological changes on blood stasis due to cold syndrome can be obviously improved and cold-cool herbs pathological changes on blood stasis due to cold syndrome has increasing trend.cold herbs pathological changes on heat-syndrome can be obviously improved and warm-heat herbs pathological changes on heat-syndrome has increasing trend.The hepatic ATP,Na+-K+-ATPase and Ca2+-Mg2+-ATPase in heat herbs group increased significantly,the hepatic ATP,Na+-K+-ATPase and Ca2+-Mg2+-ATPase in the cold-cool herbs group reduced significantly.Preliminary clear theory of traditional Chinese medicine "cure cold to hot,hot to cold " point of view,and clear principles for the treatment of cold and heat syndrome,as to provide theoretical basis for clinical medicine of traditional Chinese medicine;Warm-heat herbs and cold-cool herbs influence the hepatic energy metabolism.4.Adopt Sprague Dawley rat,the rats were anesthetized via an intraperitoneal injection of 1%pentobarbital sodium(0.15 ml per 100g body weight).Tissue proteins are separated by differential centrifugation,BCA protein quantitation by enzyme linked immunosorbent assay(ELISA),using iTRAQ technology combined with the 2D-LC-MS/MS analysis method identification of liver proteins.In liver,serum,hypothalamus and liver,85 enzymes were significantly alteredin responses to Cold traditional Chinese medicine(Rhizoma Coptidis(RC),Scutellaria baicalensis Georgi(SB),Cortex Phellodendriare(CP)),Kegg passway analysis of differentially expressed proteins were mainly distributed in the insulin signal transduction pathway,starch and sucrose metabolism,fatty acid biosynthesis,steroid synthesis,tricarboxylic acid cycle,three calcium signal transduction pathway,Wnt signaling pathway,long term potentiation and so on,reduce the body's energy production,reduce energy use.Pharmacological studies indicate that it also has antitumour,anti-infective,coronary heart disease functions.127enzymes were significantly altered in responses to warm-heat natural herbs(adix Aconiti Lateralis Preparata,Rhizoma Zingiberis,Pricklyash Peel),Kegg passway analysis of differentially expressed proteins were mainly distributed in Sugar metabolism,phosphate pentose pathway,amino acid metabolism,fatty acid metabolism,linoleic acid metabolism,peanut arachidonic acid metabolism,vitamin A metabolism,tricarboxylic acid cycle increase the body's energy generation,improve the utilization of energy of the body.Pharmacological studies indicate that it plays a role in liver cancer,aging,myocardial infarction.The results of the study for "Chinese medicine has the property(gas)and flavor,the characteristics of characteristics of property(gas)and flavor.Flavor of a drug was related with specific efficacy.Property of a drug affects the body's energy metabolism and substance metabolism by different pass ways.Influencing therapeutic effects of drugs or lead to side-effect associated with biological effects." Nature and flavor of medicines theory provides a scientific basis.At the same time as the evaluation of traditional Chinese medicine provides a new method for Cold and Hot natural properties.5.To facilitate analysis of protein-protein interaction data,STRING integrates the information about the interactions from metabolic pathways,crystal structures,binding experiments and drug-target relationships.Inferred information obtained from phenotypic effects,text mining and chemical structure similarity was used to predict the relationships between Chinese medicinal herbs research and protein targets.Using STRING and Cytoscape to facilitate analysis of protein-protein interaction data.In liver,serum,hypothalamus and liver microsomes,cold and heat of traditional Chinese Medicine were used to predict 30 associated proteins.6.One of the most widely used non-prescription pain and fever reducers is acetaminophen(N-acetyl-p-aminophenol,APAP).When kept within a normal therapeutic range,APAP is safe and has no serious side effects.However,acute overdoses can oftentimes result in serious-and sometimes fatal-hepatotoxicity.Rhizoma Dioscoreae bulibferae(RDB)is commonly used in clinical medicine.It has been used in many kinds of traditional Chinese medicine(TCM).But the toxicity of RDB,easily lead to hepatotoxicity.To study the synergistic protective effect of Rhizoma Coptidis(RC),Scutellaria baicalensis Georgi(SB)with Cortex Phellodendriare(CP)on RDB or APAP caused liver toxicity in rats.Observed pathologic histological and ultra structure changes,evaluation of transaminase and antioxidant enzymes.Rat liver toxicity model was successful,Liver cell were degeneration,swelling,piecemeal necrosis in central lobular parenchymal,cells that extended into the hepatic lobule,infiltration by inflammatory cells with steatosis.Indexes of liver function(Alt,AST and ALP)were significantly increased(P<0.05 or P<0.01).RDB or APAP caused rats liver toxicity,synergistic protective effect of RC,SB,and CP on RDB or APAP liver toxicity in rats.SB and CP exerted the strongest hepato-protective effects against liver hepatotoxicity.SB,CP and RC were observed to exhibit hepatoprotective effect as demonstrated by significant decrease in serum the activities of alanine aminotransferase(ALT),aspartate aminotransferase(AST)and alkaline phosphatase(ALP),and MAD level in liver tissue(P<0.001),increased the GSH content in liver tissue(P<0.001),and by preventing liver histopathologic changes in rats.The combination of RC,SB and CP reduced the ALT,ALP and AST level in serum,improved the liver histopathological changes of liver injury rats through synergistically increasing GSH contentand antioxidantability of liver.