Effect Of Chagan Decoction On Urine Metabolomics Of Immature Heat Syndrome Model Rats

Objective:Metabolomics liquid chromatography and mass spectrometry were used to study the changes of metabolites in urine of Chagan Decoction on immature heat syndrome model rats,and to preliminarily explore the therapeutic mechanism of Chagan decoction to promote immature heat.Methods:The rat model of fever was prepared by subcutaneous injection of 20% dry yeast suspension in the neck and back.The rat body temperature was measured once every 30 minutes by artificial anal temperature measurement method,which lasted for 15 hours.At the same time,the syndrome manifestations were observed,the immature heat syndrome period and mature heat syndrome period were distinguished,and the time point for urine sample collection was set.The administration group was given Chagan decoction for 4.5h,and the normal group was injected with equal dose of normal saline.The urine samples of normal group,model immature heat group and Chagan decoction immature heat high-dose group were detected by liquid chromatography and mass spectrometry,and the difference metabolites between groups were screened.The intergroup metabolites were identified by combining with database,and the metabolites related to fever were identified by literature search,and their metabolic pathways were further analyzed.Based on the metabolites,the therapeutic mechanism of Chagan decoction to promote ripens and immature fever was preliminatively discussed.Results:(1)The temperature change of the fever rat model was consistent with the process of human fever,and the syndrome manifestations were consistent with the clinical manifestations of immature fever and mature fever syndrome stage in Mongolian medicine,indicating the success of the model;The time point of immature heat syndrome of the model was defined as 4.5 hours ～ 7.5 hours.The duration of immature heat syndrome in Chagan decoction immature heat high-dose group was 4.5 hours to 6.5 hours;Therefore,the collection time point of urine samples in each group in the immature heat syndrome stage was set to 5 ～ 6 hours.Compared with the model immature fever group,the body temperature of rats in Chagan decoction immature fever high-dose group decreased significantly,and the immature fever period was shortened by about 60 minutes.(2)Under the positive and negative ion mode,a total of 599 different metabolites were found in the ZC and MXW groups,of which 483 were up-regulated and 116 were down-regulated.A total of 572 different metabolites were found in the MXW and CGW groups,among which109 were up-regulated and 460 down-regulated.There were 219 metabolites in urine samples of rats in the three groups,among which 5 differential metabolites related to fever were identified and confirmed,including glycine,citicoline,sphingomyelin(d18:1/24:1(15Z),sphingomyelin(d18:1/16:0)and uridine.Compared with the normal group,the above 5 metabolites in the immature heat group were significantly up-regulated.Compared with model immature heat group,the above 5 metabolites in Chagan decoction immature heat high-dose group were significantly down-regulated.Mainly involved in primary bile acid biosynthesis,porphyrin and chlorophyll metabolism,glutathione metabolism,glycine,serine and threonine metabolism,glycerol phospholipid metabolism,sphingolipid metabolism,pyrimidine metabolism,glyoxylic acid and dicarboxylic acid metabolism,aminoacyl-Tr NA biosynthesis and other metabolic pathways.Conclusion:(1)Through the analysis of urine samples of the three groups,five different metabolites related to fever were identified,which were glycine,cytidine choline,sphingomyelin(d18:1/24:1(15Z)),sphingomyelin(d18:1/16:0)and uridine.Chagan decoction may achieve its ripening effect by down-regulating these 5 metabolites.(2)The intervention pathway of Chagan decoction may balance the "three roots" of immature heat syndrome rats by adjusting the metabolic pathways of primary bile acid biosynthesis,porphyrin and chlorophyll metabolism,glutathione metabolism,glycine,serine and threonine metabolism,glycerol phospholipid metabolism,sphingolipid metabolism,pyrimidine metabolism,glyoxylic acid and dicarboxylic acid metabolism,and aminoacyl-Tr NA biosynthesis,etc.So as to give play to its ripening effect.