| Objective To explore the effects of different fat-containing high-fat diets on blood lipids in rats,and provide experimental evidence and reference for the establishment of a rat model of dyslipidemia.Metabolomics analysis was used to study the metabolic mechanism of dyslipidemia rats and the effect of Gegen Qinlian Decoction on serum metabolomes in rats with dyslipidemia,to find potential biomarkers and analyze their metabolic pathways,from the perspective of metabolomics Gegen Qilian Decoction interferes with dyslipidemia in rats to achieve a possible mechanism of prevention and treatment of type 2 diabetes.Methods In order to simulate the dyslipidemia caused by poor dietary habits,SD rats were induced by high-fat diet with 36%and 60%fat supply respectively.The food intake,blood lipids,blood sugar and other biochemicals of rats under high-fat diet were observed at different time points.Dynamic changes in indicators and weight.After successful modeling,the rats in the high-fat group were randomly divided into the model group,the positive drug simvastatin group,and the Gegen Qinlian high,medium and low dose groups.Continue to the original condition,the model group was given normal saline,simvastatin was administered with 10mg·kg-1?15 times the amount of human?,and Gegen Qinlian Decoction?GQD?was given with 14.85,4.85,1.65g.kg-1was intragastrically administered at 10 ml/kg.The effects of GQD on blood lipids,blood glucose and body weight in rats were observed at different time points.Based on ultra-high performance liquid chromatography-four-stage time-of-flight tandem mass spectrometry?UHPLC/Q-TOF-MS?,the metabolic profile information of rat serum samples was collected using Mass Profiler Professional?MPP?,Simcap 14.1 and Graphpad prism 6.0 software combines the METLIN biological database with the human metabolite database HMDB for the screening and identification of endogenous biomarkers.The MetaboAnalyst 4.0 software combined with the SMPDB and KEGG databases for biomarker enrichment analysis and metabolic pathway analysis to explore the metabolic mechanism of rats with dyslipidemia induced by high-fat diet and the mechanism of intervention of Gegen Qinlian Decoction.Results1.The high fat diet of 36%fat was induced in rats for 12 weeks.The food intake of the rats decreased significantly in the first two weeks,and then gradually increased.After2 weeks,the body weight significantly exceeded the normal group,and there was no significant change in blood glucose;57%of the rats produced insulin resistance?IR?at the end of the 12th week,insulin resistant rats,non-insulin resistant rats compared with the normal group:IR group and non-IR The TC and HDL-C of the rats in the group were significantly lower than those in the normal group at the end of the 8th week?P<0.05,P<0.01?.The TG of the two groups was significantly lower than that of the normal group at 8 and 12 weeks?P<0.05?;LDL-C has no significant change.2.Compared with the changes in biochemical indicators,the plasma metabolome changes in the positive ion mode of the IR group and the normal group,including 9potential biomarkers including?-linolenic acid,arachidonic acid,propionic acid,and lysolecithin.Significant changes have occurred.Potential biomarkers are mainly involved in alpha-linolenic acid metabolism,arachidonic acid metabolism,glycerophospholipid metabolism,and unsaturated fatty acid biosynthesis metabolic pathways.3.The rats were induced by high-fat diet supplemented with 60%fat.After the model was established,the rats in the high-fat group were more than the normal group and preferred to the high-fat diet,and gradually decreased in the later stage.After 1 week,the body weight and Lee's index of the rats were significantly higher than that of the normal group?P<0.01?.After 4 weeks of modeling,the levels of TC and LDL-C in the high-fat group were significantly higher than those in the normal group?P<0.01?,HDL-C level was significantly decreased?P<0.01?;after 10 weeks of modeling,compared with the normal group,the TC and TG of the model group were significantly increased?P<0.01?,and HDL-C was significantly decreased?P<0.01?.Compared with the normal group,the TC,TG,and LDL-C in the model group were significantly increased?P<0.05,P<0.01?,and the HDL-C level was significantly decreased?P<0.05?.The blood glucose of the model group was significantly increased only at 10 weeks,and the blood glucose of the two groups did not significantly differ in other time periods.4.After 4 weeks of modeling,the rats had abnormal blood lipids.Simvastatin and Gegen Qinlian decoction were given high,medium and low doses.After 5 weeks of administration,simvastatin significantly decreased serum TC and TG levels in rats compared with the model group.Level.The high,medium and low dose groups of GQD significantly decreased serum TC and TG levels at 5,7 and 11 weeks?P<0.05,P<0.01?,and had little effect on HDL-C and LDL-C.After administration,the food intake of the GQD group gradually decreased,and the body weight change trend gradually became flat.After 7 weeks of administration,the body weight of the high dose group was significantly lower than that of the model group?P<0.05?.Because the blood glucose of the model group increased significantly at 10 weeks,Gegen Qinlian soup exerted significant hypoglycemic effect?P<0.01?.5.The optimal dose-high dose?GQD-H?of GQD lipid-lowering and hypoglycemic effect was selected.Serum metabolome analysis was performed on the normal group,model group and GQD-H group at the time of 5 weeks of administration.The results showed positive ions.In the model,23 potential biomarkers including?-linolenic acid,arachidonic acid,and lysolecithin were significantly changed.Potential biomarkers were mainly enriched in?-linolenic acid and linoleic acid metabolism,and long chains.Mitochondrial?-oxidation,plasmalogen synthesis of saturated fatty acids,mitochondrial?-oxidation of short-chain saturated fatty acids,arachidonic acid metabolism,and tryptophan metabolism.It mainly involves the biosynthesis of unsaturated fatty acids,?-linolenic acid metabolism,arachidonic acid,glycerophospholipid metabolism,tryptophan metabolism,and fatty acid biosynthesis metabolic pathways.Gegen Qilian Decoction can call back 16 biomarkers such as?-linolenic acid,arachidonic acid,lysolecithin,and arachidonic acid ethanolamine to different degrees and exert its pharmacological effects by regulating the above metabolic pathways.Conclusion1.In this study,36%fat-enriched high-fat diet had an unstable effect on blood lipids in rats,but it was proved that lipid metabolism disorder was strongly correlated with IR.Although biochemical indicators did not show significant changes,metabolites showed a significant change in metabolites in rats with a high-fat diet.The effect of 60%fat-enriched high-fat diet on blood lipids in rats is stable,and an ideal model of dyslipidemia can be established in a short period of time.2.Metabolome studies have shown that 36%high-fat diet involves phospholipids:LPC?16:1?,LPC?17:0?;unsaturated long-chain fatty acids:?-linolenic acid,hexadecenoic acid,arachidonic acid;acylcarnitine;Tetradecanoyl carnitine;anthracene derivative:propionic acid and other substances are significantly down-regulated.60%shigh-fat diets involve phospholipids:LPC?14:0/0:0?,LPC?20:5?,LPC?24:1?,and LysoPE?20:1?are significant in the model group.Downgrade,LysoPE?20:4?,LPC?22:4?are significantly up.Sphingomyelin:SM?D18:1/14:0?was significantly upregulated.Bile acid:7-ketodeoxycholic acid was significantly down-regulated,acylcarnitine:isobutyl L-carnitine was significantly down-regulated,and L-octanoylcarnitine and stearyl carnitine were significantly up-regulated.Unsaturated long-chain fatty acids:?-linolenic acid,docosahexaenoic acid are significantly down-regulated,arachidonic acid,octadecenoic acid,docosapentaenoic acid are significantly up-regulated,saturated long-chain fatty acids:stearic acid Significantly raised.The acylamino acid was significantly down-regulated,and the androstane 3?,17?-diol was significantly down-regulated.Endogenous fatty acid amides:arachidonic acid ethanolamine was significantly up-regulated,N-acetylethanolamine?NAE?was significantly down-regulated,and fatty acid ester:peanut ethyl acetate was significantly down-regulated.The acetaldehyde was significantly lowered.GQD has different degrees of call back to the above 16 substances.Both ratios of high-fat diet affect the metabolism of?-linolenic acid,arachidonic acid and glycerophospholipid in rats.This metabolic network may be an important regulatory network for dyslipidemia.3.Studies have shown that GQD has a significant lipid-lowering effect on rats with dyslipidemia induced by high-fat diet,its anti-pre-action mechanism and tryptophan metabolism,fatty acid biosynthesis,?-linolenic acid metabolism,arachidonic acid,Glycerol phospholipid metabolism is involved in metabolic pathways.4.The changes in potential biomarkers in this study also suggest that the lipid-metabolism disorder induced by high-fat diet is related to the endocannabinoid system and intestinal micro-ecological system,which deserves further investigation. |
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