| Purpose:The study is based on 16 S high-throughput sequencing technologies and liquid chromatography-mass spectrometry metabonomics technology,small molecule metabolites in the gut microbes and the breakthrough point,the metabonomics combined with gut microbes,patients with spleen deficiency and dyslipidemia were studied,to elucidate the therapeutic mechanism of Xiangsha Liujunzi decoction in preventing and treating dyslipidemia in spleen deficiency.It provides scientific basis for the traditional Chinese medicine to treat cardiovascular and cerebrovascular diseases from the spleen.Material and method:The study included 50 patients(disease group)with spleen deficiency and dyslipidemia in the outpatient department of the second affiliated hospital of Shenyang medical college from August 2018 to October 2018(disease group)and 30 healthy volunteers(healthy control group).The patients with spleen deficiency and dyslipidemia were treated with Xiangsha Liujunzi decoction for 4 weeks(treatment group).The serum lipid levels of the three groups were detected.The random number table method was used to extract15 subjects from the healthy control group,the disease group and the corresponding treatment group respectively(the disease group and the treatment group were the same population).The metabolites in the fecal samples of the three groups of subjects were detected by LC-MS metabolomics technology,and the differences of metabolites in the intestinal tract were analyzed.At the same time,16 S r DNA high-throughput sequencing technology(entrusted to Beijing orvisen gene technology co.,LTD.)was used to test the intestinal flora in stool samples of three groups of subjects,and the diversity and abundance of intestinal microbial flora and the different flora of subjects in different groups were analyzed.Results:1.Blood lipid level test results of the three groups of subjects In this study,a total of 50 patients with dyslipidemia were included,1 patient was excluded,and 30 healthy people were included.The serum lipid levels of the disease group were significantly higher than those of the healthy control group,while the serum levels of TC,TG and LDL-c were significantly higher and the serum HDL-c levels were significantly lower(P< 0.05).Compared with the disease group,serum levels of TC,TG and LDL-c were significantly reduced and serum HDL-c levels were significantly increased in the treatment group(P < 0.05).2.Metabolomics test results of three groups of subjects There were landmark metabolites between the healthy control group and the disease group,the disease group and the treatment group.After taking the intersection,it was found that there were 7 metabolites that changed together after the comparison between the two groups,which were Benzoic acid,(S)-naproxen,Eicosapentaenoic acid,dihom-gam-linolenic acid,11(Z),14(Z)-Eicosadienoic Acid,Adrenic Acid and Xanthine.3.Intestinal flora test results of three groups of subjects(1)OTU clustering results: a total of 637 otus were generated from 45 samples in the three groups.The healthy control group averaged 145 OTU per sample,the disease group averaged144 OUT per sample,and the treatment group averaged 152 OTU per sample,which was improved compared with the disease group.(2)analysis of intestinal flora Alpha diversity: the dilution curve,shannon-wiener curve and Specaccum curve all indicated that the sample size of this study was sufficient and the sequencing data was reasonable.Compared with the healthy control group,indexes of chao1,observed species,PD whole tree and shannon all decreased.Compared with the disease group,indexes of chao1,observed species,PD whole tree and shannon all increased.(3)analysis of species composition: at the level of phylum,the proportion of firmicutes and actinomycetes in the disease group decreased,and that of bacteroidetes and proteobacteria increased,compared with that of the healthy control group.Compared with the disease group,the proportion of firmicutes,actinomycetes and bacteroidetes increased and the proportion of proteobacteria decreased in the treatment group.The F/B ratio goes down significantly.At the genus level,compared with the healthy control group,the proportions of brauteria,coprobacter and bifidobacteria in the disease group decreased,while the proportions of eubacterium rectum,klebsiella,shigella escherichia coli and megalomonas were increased.Compared with the disease group,the proportions of fecal bacteria,bifidobacteria and bacteroidetes in the treatment group increased,while the proportions of brouttella,eubacterium rectum,klebsiella,shigella escherichia coli and megalomonas decreased.(4)analysis of the intestinal flora diversity: the healthy control group and the disease group were far from each other in the PCA map,the treatment group was far from the disease group,but close to the healthy control group,and the treatment group had little difference from the healthy control group.The matrix distance between the healthy control group and the treatment group was close,the similarity was high,and the Beta diversity showed a decreasing trend.However,the matrix distance between the disease group and the healthy control group and the treatment group was far,showing a significant difference in Beta diversity.(5)differential analysis of intestinal flora: there were 25 types of differential flora in the three samples,including 11 types of firmicutes,3 types of proteobacteria,2 types of actinomycetes,and Candidatus_Saccharimonas,Selenonmonadales,Negativicutes,and Acidaminococcaceae.The important differences in the healthy control group were probacteriaceae,phyllostaceae,streptococcaceae,lactobacilli,and bacillus.Selenonmonadales,Negativicutes,enterobacteriaceae and enterobacteriaceae were the important differential bacterial groups in the disease group.The important bacterial groups of the treatment group were Acidaminococcaceae and proteococcaceae.Conclusion:1.Xiangsha Liujunzi decoction may improve the serum levels of TC,TG,ldl-c and hdl-c in patients with spleen deficiency and dyslipidemia;2.Xiangsha Liujunzi decoction can improve the species abundance and diversity of intestinal flora of patients,regulate the levels of metabolites related to fatty acid synthesis and cholesterol metabolism,and treat the dyslipidemia. |
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