Study Of Small Qinglong Decoction Syndrome Essence Based On GC/MS And LC/MS Of Metabonomics

OBJECTIVE:After the fingerprints getted by using GC/MS and LC/MS to detectin rats urine, blood and lung, spleen, liver and kidney tissue of small Qinglong Decoction syndrome, treated and analyzed by PLS-DA, the symbolitic compounds closely relating with small qinglong Decoction syndrome have been found out, which provide objective basis for clinical diagnosis and treatment.METHODS:(1)36 male Wistar rats were random Ly divided into the blank control group(A) 10 rats, the model group(B) and small Qinglong Decoction group(C) for each 13 animals,4 rats in one cage,feeded in general animal room,the indoor temperature was maintained at(20 ± 2) ℃, relative humidity of 40-50%.Combined with traditional chinese medicine etiology drink produced within the theoretical cold outside(small Qinglong Decoction syndrome) card model, will be placed in the model group and small Qinglong Decoction syndrome rats in a cold environment(control room temperature at about 10 ℃) in the fed daily at low temperatures(0 ± 4 ℃ about) freeze 1 hour, and the rats were placed in ice water to swim to the surface drowned rat nose remove the rats for 4 weeks, while eating into the cold(freezing feed and ice). The control group received no treatment. We collected urine before and after the rats were modeled, 4 weeks later.And we took blood from the abdominal aorta and lung,spleen, liver and kidney tissues and reserved.(2)The rats, urine,blood and lung spleen, liver and kidney tissues of the normal group and model group were detected by GC/MS and LC/MS,from which the data coming are pretreated and integrated and analyzed by PLS-DA by using SIMCA-P+.After comparing find the differences metabolites, then use spss software to identify the differences in metabolites t test or ANOVA analysis to identify statistically significant material, combined with analysis of the relevant literature and issues metabolic differences matter and small Qinglong Decoction syndrome permit correlation.RESULTS:(1)GC/MS detected 13 kinds of markers in blood, containing lactic acid, 3- hydroxy butyric acid, urea, succinic acid, glycine, serine, lysine, arachidonic acid, glutaric acid, pyran galactose glucoside, sorbitol, valine.Seven kinds of markers were detected in the urine,glycine, glucose, inositol, proline, fumaric acid, alanine, urea.Six kinds markers were detected of lung tissue, palmitic acid, stearic acid, fumaric acid, inositol and 2- butanone acid, glycine. Twelve markers were detected of liver tissue fumaric acid, lactic acid, stearic acid, inositol and glucose, valine, isoleucine, alanine, glycine, glutamic acid, aspartic acid, threonine.10 markers were detected in the kidney tissues, lactic acid, urea, leucine, valine, alanine, glycine, serine and threonine, aspartic acid, proline.(2)LC/MS detected six kinds of markers in the blood, histidine, hemolytic phosphatidyl ethanolamine, choline, lysine, lecithin.Three kinds of urine markers, amino acids, glucoside acid, 4- hydroxy benzene pyruvic acid.Four kinds of markers were detected of lung tissue,spermine, spermine, pancreatic acyl- isoleucine, guanosine, liver tissue five markers aspartic acid, lecithin, choline, guanidine butyric acid, 4-4- hydroxy benzene pyruvic acid. Four markers were detected in the kidney tissue,4- hydroxy benzene pyruvic acid, linoleic acid, 3, 5- two br- L- tyrosine, amino acids. Four markers were detected spleen tissue, glycine, phenylalanine, phosphatidyl ethanolamine, phosphoric acid guanidine acetate.(3)Small molecules analysis results show that the technical inspection of the GC/MS analysis results show that the small qinglong decoction syndrome syndrome essence is mainly associated with energy metabolism, inflammation and immunity. LC/MS to detect the related markers, concentrated expression in phospholipid metabolism, inflammation, immune, and inhibition of the Krebs cycle. Analysis of small qinglong decoction syndrome from the Angle of the metabolic syndrome essence, small qinglong decoction can affect phospholipid metabolism, inflammation, immune and the Krebs cycle to achieve anti-inflammatory, enhance immunity, adjust the energy metabolism within the treatment such as cold drink type of asthma, in conclusion, the small qinglong decoction syndrome syndrome essence and phospholipids metabolic abnormalities, inflammation, immune, and inhibition of the Krebs cycle these four aspects.CONCLUSION:(1)It is feasible to use the traditional Chinese medicine theory to produce model;(2) The use of metabolomics technology changes caused by the changing nature of microscopic markers study results are in agreement with the macro, the traditional method, on the one hand that the model is successful,the small qinglong decoction is effective, is another example of research using metabolomics methods syndromes indeed feasible, and for the Chinese classic recipe research provides new methods and ideas;(3)Small molecules angle shows the small qinglong decoction certificate variation caused by the body there are four main areas: phospholipid metabolism, inhibition of the citric acid cycle, inflammation and immune function;(4) Small qinglong decoction group of rats, after taking small qinglong decoction, the body can get better, characterized by sneezing and coughing reduced, increased drinking and eating, etc., that small qinglong decoction outside in the cold drink asthma does have effect;(5)By rat urine, blood and markers of comparison between organizations, show the consistency in the aspect of pathological changes, but more sensitive than blood, pretreatment method is the most simple, is more extensive in application.