| objective:This study used a non-targeted metabolomics method based on ultra-high performance liquid chromatography-time-of-flight mass spectrometry(UPLC-QTOF-MS)to study the therapeutic effect of aurantio-obtusin on hyperlipidemia rats,looking for hyperlipidemia-related metabolites with the help of chemometrics,and exploreing the intervene effects of aurantio-obtusin on potential metabolites,to initially clarify the mechanism of aurantio-obtusin and to provide rational theoretical basis for clinical practice.Methods:The rats were randomly divided into 2 groups:the control group(10 rats):fed with normal food;the hyperlipidemia model group(30 rats):fed with high-fat food.7 days later,blood samples were taken from tail veins in both groups to test their biochemical indexes and to confirm the effect of modeling.Afterwards,30 hyperlipidemia model rats were randomly divided into 3 groups,namely model group(10),aurantio-obtusin group(10)and atorvastatin group(10).Atorvastatin group was given intragastric administration of atorvastatin 10mg/(kg·d),and aurantio-obtusin group was given aurantio-obtusin 3.6mg/(kg·d).Control group and model group were fed with high-fat food for 30 days,and then the rats were processed to collect liver tissue and plasma.Plasma was used to detect biochemical indicators.The liver tissue was preprocessed and analyzed with UPLC-Q-TOF-MS.The obtained data were integrated with the instrument's own software.SMICA-P14.1 and MATLAB software were imported for principal component analysis(PCA),partial least squares Discriminant analysis(PLS-DA)and random forest analysis.Variable importance projection index(VIP)and single factor t test were applied to analyze the substance.Finally,biomarker was confirmed,and its metabolic pathway was analyzed with software.Results:(1)Biochemical indicators were detected after 7 days of high-fat feeding.It was found that compared with the control group(p<0.05),the total values of cholesterol(TC)and low-density lipoprotein(LDL-C)in the model group were significantly increased,indicating that the model was successful.(2)The UPLC-QTOF-MS technology was used to test liver tissue,and the obtained data through integrated analysis showed that the retention time of the sample and its relative standard deviation(RSD)of the content met the analysis requirements.The other sample data was combined with LC-MS database,HMDB database and online database to identify the substances in rats.A total of 13 hyperlipidemia-related biomarkers were obtained with VIP value and t test.After the intervention of aurantio-obtusin,9,10-epoxyoctadecenoic acid,flavin mononucleotide,D-glucuronic acid,L-glutamic acid,L-histidine,and hippuric acid in rat liver tissue decreased,among which 9,10-epoxy octadecenoic acid,flavin mononucleotide,and D-glucuronide had an obvious drop.These substances involved various metabolism,including lipid metabolism,energy metabolism,fatty acid metabolism and amino acid metabolism.Therefore,this study on the mechanism of aurantio-obtusinhad to the treatment of hyperlipidemia had clinical values.Conclusion:This study confirmed the successful modeling of hyperlipidemia rats by detecting the biochemical indicators of rat plasma.Then the method of UPLC-QTOF-MS metabolomics was used to study the metabolic changes of rat liver that was caused by hyperlipidemia.The protective mechanism of aurantio-obtusinon about liver was clarified,and these performances may involve energy metabolism,lipid metabolism,fatty acid metabolism and amino acid metabolism. |
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