Gut Microbiome And Metabolomics For Preventive Effect Of Proanthocyanidin On Dyslipidemia In Mice

Proanthocyanidins widely exist in nature and have antioxidant activities and effects on disturbance of lipid metabolism.Proanthocyanidin B₂ is the dimer form of proanthocyanidin and the main oligomer of foodborne proanthocyanidin.With the rise of social and economic level,diet also tends to be rich in high-fat meat products,and hyperlipidemia and other symptoms caused by hyperlipidemia are common.The intervention of abnormal lipid metabolism has become one of the research hotspots.However,few studies have reported the improvement of hyperlipidemia by proanthocyanidins B₂.In this paper,the hyperlipidemic model of mice induced by hyperlipidemic diet was established,and the intervention effect of dietary supplementation of procyanidins B₂ on hyperlipidemic model mice was evaluated,and the mechanism of procyanidins B₂ regulating hyperlipidemic disorder was explored from two aspects of intestinal flora and metabolomics.The research results are as follows:?1?C57BL/6 mice aged 5 weeks were randomly divided into three groups?n=9?:control group?CON?,high-fat group?HF?,procyanidin B₂ group?PB₂?,The indexes of blood lipid,liver lipase activity,antioxidant and behavior of mice were determined.Procyanidin B₂can significantly lower abdomen fat index,blood lipid index TC,TG and AI were significantly reduced?P<0.05?,significantly improve the blood HDL-C and enzyme activity in the liver tissue lipoprotein enzymes,hepatic lipase activity and total fat enzyme activity,and significantly enhance the antioxidant activity?T-AOC,SOD,CAT and GSH index?,inhibit oxidative damage product MDA content?P<0.05?,suggesting that the procyanidin B₂has high intervention fat prandial dyslipidemia and oxidative stress.In addition,animal behavior experiments showed that proanthocyanid B₂ significantly improved the central region distance and time of high-fat diet mice in the open field,as well as the target quadrant distance and residence time in the Morris water maze experiment,indicating that proanthocyanid B₂ could improve the learning and memory ability of high-fat diet mice.?2?Illumina high-throughput sequencing technology was used to analyze the changes of intestinal flora structure in mice.Alpha diversity Sobs and Shannon index dilution curves and difference test plots showed that there were significant differences among control group,high-fat group group and procyanidins B₂.Venn diagram of species composition showed that52 OUT were unique in the control group,10 in the high-fat group,and 12 in the proanthocyanidin B2 group.It was found that proanthocyanidins B₂ could increase the ratio of Bacteroidetes/Firmicutes in the intestinal flora of high-fat model mice,and increase the relative abundance of Lactobacillaceae,Peptostreptococcaceae,Bacteroidales-s24-7-group and other flora.Analysis of the difference between the level of subordinate classification and species reveals that proanthocyanin B₂ reduces the relative abundance of bacteria such as escherichia-shigella and Ruminococcaceae in the intestinal flora of high-fat model mice and increases the relative abundance of bacteria such as Romboutsia and fissicatena_group.In addition,the analysis of short-chain fatty acids in mice feces showed that proanthocyanidin B₂could significantly increase the contents of butyric acid in mice feces with high fat.The results showed that proanthocyanidin B₂ could interfere with the intestinal microflora structure caused by high-fat diet.?3?LC-MS technique was used for data collection of plasma and urine samples of mice,and then SIMCA-P and metabolomics database?HMDB,KEGG and Mtlin,etc.?were used for comparison and analysis.Screening the three groups of different metabolites found 14kinds of plasma?arachidonic acid,citric acid,l-phenylalanine,taurine,etc?and 13 kinds of urine?aminoadipic acid,acetylcarnitine,xanthurenic acid,etc?.Among them,the high-fat diet caused the decrease of the content of metabolites such as dicarboxytetraenoic acid,palmitic acid,linoleic acid,arachidonic acid,citric acid,taurine and amino-adipic acid in mice,and the increase of the content of metabolites such as arachidonic acid,citric acid and taurine after the intervention of procyanidin B₂.Path analysis showed that procyanidin B₂ intervention lipid metabolic disorder induced by high fat diet mainly involved in plasma in 5 kinds of metabolic pathways?fatty acid metabolism,glyoxylic acid and dicarboxylic acid metabolism,primary bile acid biosynthesis,the Krebs cycle,biosynthesis of aromatic amino acids?and the urine have 4 kinds of metabolic pathways?bile acid synthesis,lysine biosynthesis,cyanide amino acid metabolism,taurine and taurine metabolism?.Proanthocyanidin B₂ can significantly interfere with the abnormal blood lipid metabolism and oxidative stress induced by high-fat diet in mice,and the intervention effect of proanthocyanidin B₂ may be related to the regulation of multiple metabolic pathways such as the regulation of intestinal flora structure and fatty acid metabolism,bile acid metabolism and tricarboxylic acid cycle.