| OBJECTIVE:After the fingerprints getted by using NMR to detect the rat,urine,blood and lung tissue of small Qinglong Decoction syndrome, treated and analyzedby PCA or PLS-DA, the symbolitic compounds closely relating with small qinglongDecoction syndrome have been found out, which provide objective basis for clinicaldiagnosis and treatment.METHODS:（1）36male Wistar rats were randomly divided into the blankcontrol group (A)10rats, the model group (B) and small Qinglong Decoction group(C)with13rats in each group,4rats in one cage, feeded in the normal animal house andkeeped indoor temperature in (20±2)℃and relative humidity maintained for40-50%.We used traditional chinese medicine etiology theory to produce the Smallqinglong Decoction syndrome model:the model group and small Qinglong Decoctiongroup rats are placed in cold environment (control indoor temperature at10℃),feeded in frozen and in the environment of low temperature (0±4℃) about1H every day, given cold food and water,placed in ice water for swimming and takenout when rats,nasal tip was submerged, which continue for4weeks. The blankcontrol group were feeded in normal enviroment. We collected urine before and afterthe rats were modeled,4weeks later.And we took blood from the abdominal aorta andlung tissues and reserved（.2）The rats,urine,blood and lung tissue of the normal groupand model group were detected by NMR spectrum, from which the data coming arepretreated and integrated by MestRec and analyzed by PCA and PLS-DA by usingSIMCA-P+.Then,we can get the corresponding scatter diagram and diagram ofloading and find out the differences of metabolites by the comparison.Next,we useSPSS software to analyze metabolite differences with T-test or one-wayANOVA,finding material with statistically significant correlation in them andanalyzing dependency of metabolites with differences and small qinglong Decoctionsyndrome combining with the related literatures and subjects.RESULTS:（1）Rats,urine1HNMR results:the Results from comparison ofmodel group and the blank control group show that the contents of metabolites ofcreatinine, guanidinoacetic acid, trimethylamine, taurine, allantoin and othermetabolites in the model group have elevated (p<0.05),that the contents of metabolites of succinic acid, lactic acid,2-OG, glycine, methylnicotine metabolitehave decreased (p<0.05),that creatinine, trimethylamine, allantoin, taurine of smallqinglong Decoction group are still higher than the blank control group but have beenreduced than the model group, that lactic acid is lower than the blank control groupbut higher than the model group, that guanidinoacetic acid,2-OG, N-nornicotinechange have no statistical significance;（2）Rats,blood NMR results: the Results fromcomparison of model group and the normal group show that the contents ofmetabolites of creatine,3-hydroxybutyric acid, taurine, aspartic acid, citrulline andglutamine have increased (p<0.05), The contents of metabolites of lactic acid, aceticacid, proline, tyrosine and glycine have decreased (p<0.05). In small qinglongDecoction group, taurine is still higher than the blank control group but lower thanthe model group (p<0.05), tyrosine and proline are still lower than that of the blankcontrol group,but have increased compared with the model group,(p<0.05). Thecontents of metabolites of citrulline and glutamine are higher than the model group;（3）Rats,lung tissue NMR results: the Results from comparison of model group andthe normal group show that the contents of metabolites of leucine, isoleucine, valine,arginine, histidine, alanine and taurine have increased (p<0.05), which of lysine,threonine and methionine have decreased (p<0.05), that acetic acid has no statisticalsignificance. The small qinglong Decoction group compared with the blank controlgroup and model group shows that the contents of metabolites of valine, arginine,taurine, alanine and histidine are still higher than those in the model group, thecontents of taurine, valine and alanine are lower than those of the model group,thatarginine is higher than in model group, the contents of threonine and methionine arelower than the blank control group but higher than the model group.CONCLUSION:（1）It is feasible to use the traditional Chinese medicine theoryto produce model;（2）Research was carried out by using NMR technique for micromarker materials.The results are consistent with the methods of macroscophy andtradition.On one hand,it shows that the small qinglong decoction is effective,on theother hand,it shows that the research of TCM syndrome and type of TCM by using themetabonomics method is feasible;（3）From the point of view,the changes ofmolecular substance in small qinglong decoction syndrome in the body mainly have three aspects: energy metabolism, inflammation and immune function;（4）Throughthe comparison of the blood, lung tissue and rat urine,they show the consistency inpathological change,but blood is more sensitive and has the simplest approach, whichis more extensive in the application. |
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