Study On The Curative Effect Mechanism Of Yimai Granules Against Atherosclerosis

Purpose:This study aims to explore the therapeutic mechanism of Yimai Granules in the treatment of atherosclerosis through bioinformatics,metabolomics high-throughput sequencing technology,and metagenomic techniques,and to find changes in related intestinal flora and metabolites.And the correlation between them provides a reliable treatment basis for Yimai Granule in the clinical treatment of atherosclerosis.Material and method:1 Study on the therapeutic mechanism of Yimai Granule:The genes for the potential regulation of all drugs of Yimai Granules were predicted by bioinformatics technology.The TCMBATMAN database was used to query the name of the decoction tablets of Yimai Granules,and the main components of each drug were obtained.The target of the main components of the drug was searched by DRUGBANK,and then mapped to the human gene by uniprot;the Disgenet and TCD databases were used;The related genes of the disease,and cross-matching the genes with the main components of Yimai granules,speculate that Yimai granules can be used to treat the target of AS;use STRING database to construct the background network,calculate the node degree of each target,according to the node Sorting the above targets,and finally obtaining the potential molecular targets of Yimai particles acting on AS,that is,according to the composition of Yimai granules,the target gene of all the drugs in the drug is mapped to the target gene to determine the important target.gene.SPF grade C57BL/6J mice were selected,56 days old,56 days old,as normal group(ZC group),given normal diet + normal saline;SPF ApoE-/-mice were selected,56 days old,60 days,Randomly divided into 5 groups,12 in each group.The model group was given high-fat and high-cholesterol diet + normal saline;Yimai granules high-dose group(YF-G group)was given high-fat and high-cholesterol diet + high-dose intragastric administration;Yimai granules in the middle dose group(YF-Z)Group)high-fat high-cholesterol diet + Chinese medicine medium-dose intragastric administration;Yimai granules low-dose group(YF-D group)to high-fat and high-cholesterol diet + Chinese medicine low-dose intragastricadministration;western medicine positive drug control group(XY group)12 Only high-fat,high-cholesterol diet + atorvastatin.Continuous treatment for 12 weeks.After treatment,blood was collected from the abdominal aorta,and blood lipids,TNF-?,IL-6 and plasma viscosity and whole blood high,medium and low viscosity,erythrocyte sedimentation rate,and red blood cell aggregation index were measured.At the same time,myocardial vascular tissue was selected for histopathological observation,and the aorta of the mouse was selected to detect CES1,HMGCR,PPARD,PPARG and other proteins.2 Metabolomics study of Yimai Granules in the treatment of atherosclerotic diseases:SPF-class C57BL/6J mice were selected,aged 28-56 days,males,and 6 rats,as normal group(ZC group),given normal diet + normal saline.SPF-class ApoE-/-mice were selected,aged 28-56 days,males,24 rats,randomly divided into 4 groups,6 in each group.The model group(MX group)was given high-fat and high-cholesterol diet + normal saline,western medicine positive drug control group(XY group)to high-fat and high-cholesterol diet +atorvastatin group,Chinese herbal extract positive group(HP group)High-fat and high-cholesterol diet + quercetin,Yimai granule treatment group(YF group)was given high-fat and high-cholesterol diet + high-dose Chinese medicine for continuous treatment for12 weeks.After treatment,non-targeted metabolomics tests were performed on the feces of each group to explore the metabolomic changes and the changes in biological functions of Yimai Granules in the treatment of atherosclerotic diseases.3 Using 16 SrDNA to analyze the effect of Yimai Granule on intestinal flora of atherosclerotic mice:SPF-class C57BL/6J mice were selected,aged 28-56 days,males,and 3 rats,as normal group(ZC group),given normal diet + normal saline.SPF-class ApoE-/-mice were selected,aged 28-56 days,males,12 rats,randomly divided into 4 groups,3 in each group.Model group(MX group)was given high-fat and high-cholesterol diet + normal saline;Western medicine positive drug control group(HP group)was given high-fat and high-cholesterol diet+ quercetin;Yimai granule treatment group(YF group)was given high-fat High cholesterol diet + Yimai Granules group,continued treatment for 12 weeks.After the treatment,the mice were sacrificed and the small intestine samples were collected,and the total DNA was extracted,and the 16 SrDNA of the intestinal flora of the mice was sequenced by PCR toanalyze the diversity of the intestinal flora and the difference of species in each group.Redundant analysis of gut microbiota results and metabolomics was conducted to explore the correlation between the two.Results:1 Study on the mechanism of Yimai granules:A total of 235 potential overlapping gene targets were obtained from Yimai Granules,and a total of 18 genes associated with AS pathogenesis.According to the Yimai granules,there are 4 overlapping gene targets in each group,which are CES1,HMGCR,PPARD and PPARG.Furthermore,the LXR-? and ABCA1 genes were further predicted,and it is speculated that Yimai granule may play a therapeutic role through the signaling pathway of PPAR-?/LXR-?/ABCA1.Yimai Granule can effectively regulate the levels of TG,TC,LDL-C,TNF-?,IL-6,blood viscosity and blood aggregation in AS mice.The blood lipids,TNF-?,IL-6 of Yimai Granules,The levels of blood viscosity and blood aggregation were close to normal levels,and the difference was statistically significant(P<0.05)compared with the MX group.The effect was comparable or even better compared with the XY group.In addition to effectively down-regulating the expression of CES1 and up-regulating the expression of HMGCR,PPARD and PPARG,Yimai granules can also up-regulate the expression of LXR-?/ABCA1 protein downstream of PPARG,and finally achieve the therapeutic effect on AS.The concentration of Yimai particles was proportional(P<0.05).The expression of CES1,PPARD,PPARG and LXR-? was significantly improved in the YF-D and YF-Z groups.Compared with the MX group,the difference was statistically significant(P<0.05).The expression of CES1,HMGCR,PPARD,PPARG,LXR-? and ABCA1 protein was significantly improved in YF-G group,and the difference was statistically significant(P<0.05).Under LIGHT,the vascular damage of XY group and YF group was repaired.The thickness of each part of YF-D group,YF-Z group and YF-G group was uneven,no plaque was found in the lumen,and there were fewer cells between cells.Inflammatory cell infiltration,the least in the YF-G group,with thickening of the intima,but thinner than theMX group,the intima thickness of YF-G is thinner,the muscle layer of YF-D group and YF-Z group is higher.The dose group was loose and a small amount of myofilament was seen.2 Metabolomics study of Yimai Granules in the treatment of atherosclerotic diseases:Fourteen metabolites were identified that Yimai Granules were able to improve the significant differences involved in AS mice,including known 4-nitrocinnamate,humic acid,isobutyl acetate,1-testosterone tetrahydrogen Eight metabolites such as pyridyl ether,docetaxel,vitamin k,shofar,and ergothin have been significantly regulated after treatment,positive ions are down-regulated,and negative ion-mediated metabolic pathways include amino acid metabolism,acarids.Metabolism of compounds and polyketides,biodegradation and metabolism of xenobiotics,lipid metabolism,and the like.3 Using 16 SrDNA to analyze the effect of Yimai Granule on intestinal flora of atherosclerotic mice:The results of PCoA showed that Yimai Granule could regulate the diversity of intestinal flora in AS mice.Compared with MX group,it was mainly composed of thick-walled bacteria and Bacteroidetes at the level of the door.The genus was mainly composed of Pseudomonas and Labiata,and the difference was statistically significant(P<0.05).Compared with ZC,MX and HP groups,the levels of YF anaerobic bacteria,intestinal bacteria and Helicobacter pylori were significantly different(P<0.05).4 Correlation analysis between intestinal flora and metabolitesRda results showed that the spirochete had a positive effect on all sample metabolites,and negatively correlated with Bacteroides and Proteus,while Bacteroides and Proteus were negative for all sample metabolites.Positive effects,the two showed a positive correlation;the number of spirochetes and Bacteroides gates were sensitive to the effects of metabolites,and the sensitive metabolites were: organic heterocyclic compounds,benzene ring compounds,phenylpropanoids and polyketides,Organic acids and their derivatives,lipids and lipid-like molecules and organic oxygen compounds.Conclusion:1 Yimai Granule can effectively down-regulate the expression of CES1,up-regulate theexpression of HMGCR,PPARD and PPARG,and can effectively regulate the blood lipid,blood viscosity and inflammatory factor level of AS mice by up-regulating the expression of LXR-?/ABCA1 protein downstream of PPARG.Etc.,play a role in the treatment of AS.2 Yimai Granules can improve 14 significant differences in metabolites involved in AS mice,including known 4-nitrocinnamate,humic acid,isobutyl acetate,1-testosterone tetrahydropyridinium Eight metabolites,such as norbornene,docetaxel,vitamin k,shofar,and ergot,showed significant regulation after treatment,positive ion down-regulation,and negative ion up-regulation.Yimai Granule can play a role in the treatment of AS by regulating amino acid metabolism,metabolism of terpenoids and polyketides,biodegradation and metabolism of xenobiotics,and lipid metabolism.3 Yimai Granule can affect intestinal flora by improving thick-walled bacteria,Bacteroides,Zymomonas,Labiata,Anaerobacter,Enterobacter,and Helicobacter Species,structure,flora diversity and metabolites play a role in the treatment of AS.At the same time,Enterobacter(Spirulina,Bacteroides and Proteus)and potential biomarkers(organic heterocyclic compounds,benzene ring compounds,phenylpropanoids and polyketides,organic acids and their derivatives,lipids)Classes and lipid-like molecules and organic oxygen compounds have strong correlations.