| With the development of the economy,the dietary structure of the residents in China has changed,and the intake of lipids in the diet is increasing,which has become one of the main factors affecting the risk of hyperlipidemia and cardiovascular disease.Tartary buckwheat protein and buckwheat starch have the effect of improving hyperlipidemia,but the mechanism of lowering fat is not clear.In this study,a mouse model of metabolic disorder was established by high fat diet induction.The effect of tartary buckwheat protein and tartary buckwheat starch diet on the metabolism of blood lipids was studied.The plasma chromatography-mass spectrometry(liquid chromatography mass spectrometry)technique was used to analyze the plasma metabolism,and the lipid lowering mechanism of Tartary buckwheat Rhizoma Bletillae tartary buckwheat starch was preliminarily explored.(1)4 weeks male C57BL/6 mice were randomly divided into 4 groups: control group(low fat feed,LFD),model group(casein high fat feed,HF-CS),tartary buckwheat protein group(tartary buckwheat protein high fat feed,HF-BWP),tartary buckwheat starch intervention group(HF-BWS),15 rats in each group,and feed for 8 weeks.The experimental results showed that compared with the control group,the content of TG,TC and LDL-C increased significantly(P<0.05).The content of tartary buckwheat protein group was significantly lower than that of model group TG,TC and LDL-C,which were 85.6%,37.3% and 58.7%,respectively.The content of HDL-C was significantly increased by 32.5%.The TG and TC of tartary buckwheat starch group decreased by 54.6% and 16.7% respectively compared with the model group,indicating that tartary buckwheat protein and buckwheat starch had the effect of improving lipid metabolism disorder.(2)This study selects the appropriate mass spectrometry and chromatographic conditions,through the quality control sample(QC sample)to monitor the stability of the test process instrument in real time.It is found that the reproducibility of the retention time of the instrument is very good,the instrument analysis and the reliability of the data results are found.The retention time variation of the LC-MS(Thermo,Ultimate 3000 LC,Orbitrap Elite)data from the plasma QC samples showed that the retention time variation of the LC-MS spectrum data in the plasma QC samples was within + 0.03 min,indicating that the chromatogram mass spectrometry combined system was basically stable and the retention time of the chromatographic peaks between the QC samples was small.Based on the analysis of the above data reliability,it is shown that if the system detects the differences between the samples,the difference is derived from the difference of the sample metabolites rather than the analysis error.(3)The plasma metabolic profile of the mice in the control group,the model group and the buckwheat protein group was obtained by LC-MS technique,and the metabolic profile of the plasma of the model group deviated from the control group,and the tartary buckwheat protein group deviated from the model group.Compared with the model group,the buckwheat protein group was closer to the control group,indicating that the metabolic phenotype of tartary buckwheat protein group was closer to that of the control group.Multivariate statistical analysis found that there were 31 different metabolites between the control group and the model group.Compared with the control group,the plasma samples of the model group were decreased in the levels of arachidic acid,goosedeoxycholic acid,8,11,14-twenty carbon three eNIC acid,twenty-two carbon five enoic acid and adrenergic acid;LysoPC(14: 1(9Z)),LysoPC(14:0),citrulline,bovine The levels of sulfonylcholic acid and glutathione were increased.Compared with the model group,the content of five unsaturated fatty acids in the buckwheat protein group(8,11,14-carbon three enoic acid,twenty-two carbon five enoic acid,linoleic acid,adrenergic acid and stearic acid)increased significantly,palmitic acid,oleic acid,cortisol,oxydeoxycholic acid,LysoPC(14: 1(9Z)),LysoPC(14:0),LysoPC(15:),and LysoPC(15:).0),citrullinine and taurine decreased significantly.Combined with KEGG and MetPA database,it was found that the buckwheat protein mainly regulated the metabolic disorder of mice induced by high fat diet through amino acid metabolism,fatty acid metabolism,purine metabolism,and glucose metabolism pathway.(4)the multicomponent analysis showed that the plasma metabolic profile of the control group,the model group and the tartary buckwheat starch group was distinct,and it was found that there were 18 different metabolites between the tartary buckwheat starch group and the model group.Compared with the model group,the tartary buckwheat starch group palmitic acid,peanut four enoic acid,goosedeoxycholic acid,oxalate diacid,phenylacetone,and tartary buckwheat starch group were compared with the model group.The contents of L-lactic acid,oleic acid,LysoPC(14:0),citrulline,glycolic acid,acetoacetic acid and pyroglutamic acid decreased as the content of acid and citric acid increased.KEGG and MetPA database aided analysis found that tartary buckwheat starch improved lipid metabolism disorders mainly related to amino acid metabolism,fatty acid metabolism,purine metabolism,glucose metabolism,four metabolic pathways.(5)According to the multivariate analysis,the plasma metabolic profile of the control group,the model group,the buckwheat protein group and the tartary buckwheat starch group was obvious,and the distance between the buckwheat starch and the control group was closer than that of the tartary buckwheat contrast group.Therefore,the metabolic phenotypes of tartary buckwheat and tartary buckwheat starch groups were closer to the control group than those of the model group,and the buckwheat protein group had more obvious effect.Compared with the tartary buckwheat starch group,the tartary buckwheat protein group had higher content of CIS aconitic acid,citric acid,L-lactic acid,cholic acid,taurocholic acid,succinic acid,palmitic acid,oleic acid,oxycholic acid,LysoPC(15:0),LysoPC(14:1(9Z)),LysoPC(14:0),uric acid,stearic acid,cortisol,1-phosphoric acid sphingosine,and coke valley.The content of ammonia and citrullinine decreased.There were significant differences between tartary buckwheat protein and buckwheat starch in amino acid metabolism,fatty acid metabolism,purine metabolism,glycerol phospholipid metabolism and sphingolipid metabolism. |
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