| Objective: The purpose of this study is to detect the structure and diversity of intestinal microflora in patients with coronary heart disease and healthy people in Guangxi by 16 s r RNA high-throughput sequencing,and to explore the differences of intestinal microflora in patients with coronary heart disease with different TCM syndrome types,so as to observe the changes of intestinal microflora which may play an important role in the occurrence and development of coronary heart disease at the level of intestinal microorganisms.Methods: The patients with coronary heart disease with phlegm and blood stasis syndrome,qi deficiency and blood stasis syndrome,phlegm blocking heart pulse syndrome and heart and spleen deficiency syndrome from April2020 to December 2020 were collected,and the subjects who were excluded from other diseases by physical examination and laboratory examination in the same period were selected as healthy control group with 10 cases in each group.The clinical data of the subjects were collected,and the levels of blood total cholesterol(TC),triglyceride(TG),low density lipoprotein(LDL-C),high density lipoprotein(HDL-C)and fasting blood glucose(GLU);were detected.The intestinal flora abundance,structure and species diversity of each group of fecal samples were compared by high throughput sequencing technique.Results: 1 Baseline data: there was no statistical difference in age,sex,BMI,blood lipid and fasting blood glucose among each group,and the study was comparable.2 Intestinal flora results: A total of 3993386 pairs of Reads,double-terminal Reads were sequenced and filtered to produce 3915266 Clean tags.Each sample produced at least 77376 Clean tags,samples with an average sequence number of 78305.A total of 10 phylums,15 classes,21 orders,44 families and 150 genera were detected.There were 291 OTUs in five groups of samples.The results showed that there were not only most of the same colonies in the five groups of samples,but also their own unique colonies at the same time.3 Microflora diversity: Alpha diversity analysis of microflora: Chao1 index and Ace index were statistically significant,suggesting that there were significant differences in flora abundance among the five groups.However,Shannons index and Simpson index showed that there was no statistical difference in community diversity among the five groups.4 Beta diversity analysis: each group of samples have a certain clustering position,indicating that the same group of samples have a certain similarity,the heart and spleen deficiency syndrome group and the healthy control group have the greatest discreteness,indicating that there are significant differences in the composition of intestinal flora between the two groups.The species accumulation curve,dilution curve and grade abundance curve suggested that the adequacy,sequencing depth and species richness of the samples collected in this experiment were higher.5 The results of different taxonomic levels are as follows: 1)At the phylum level,the abundance of bacteroidetes and fusobacteria in the healthy control group was significantly higher than that in the coronary heart disease group(P < 0.05),Firmicutes:the heart and spleen deficiency syndrome group was higher than the healthy control group,and the difference was statistically significant(P < 0.05).Comparison of firmicutes in different TCM syndrome types of coronary heart disease: heart and spleen deficiency syndrome group > phlegm blocking heart pulse syndrome group > qi deficiency and blood stasis syndrome group >phlegm and blood stasis syndrome group,heart and spleen deficiency syndrome group and phlegm and blood stasis syndrome group,there was statistical significance(P < 0.05).2)at the class and order level,the abundance of the dominant bacteria in the five groups was different,but there was no statistical significance(P > 0.05).3)At the family level,the abundance of prevotellaceae and ruminococcaceae were significant difference between the healthy control group and the heart and spleen deficiency syndrome group(P < 0.05).Bifidobacteriaceae: healthy control group > phlegm blocking heart pulse syndrome group > phlegm and blood stasis syndrome group > qi deficiency and blood stasis syndrome group > heart and spleen deficiency syndrome group,there were statistical differences among the three groups(P< 0.05).4)At the genus level,through the analysis of the significant difference of community species among the groups,the relative abundance of different bacteria among the groups was as follows: Rothia:healthy control group >phlegm blocking heart pulse syndrome group > phlegm and blood stasis syndrome group > qi deficiency and blood stasis syndrome group > heart and spleen deficiency syndrome.Bacteroides and Bifidobacterium: healthy control group > phlegm and blood stasis syndrome group > qi deficiency and blood stasis syndrome group > phlegm blocking heart pulse syndrome group > heart and spleen deficiency syndrome group.Ruminococcace: heart and spleen deficiency syndrome group > qi deficiency and blood stasis syndrome group > phlegm blocking heart pulse syndrome group > phlegm and blood stasis syndrome group > healthy control syndrome group.Escherichia-Shigella: heart and spleen deficiency syndrome group > phlegm and blood stasis syndrome group > qi deficiency and blood stasis syndrome group > phlegm blocking heart pulse syndrome group > healthy control group.Conclusions: 1compared with healthy people,the difference of intestinal flora between patients with coronary heart disease and healthy people was reflected in the abundance of microflora at phylum,family and genus level.2compared with the intestinal flora of patients with coronary heart disease of different TCM syndrome types,the number of dominant bacteria in patients with deficiency of both heart and spleen syndrome decreased more significantly than the other three syndrome types.3 Prevotellaceae,Ruminococcace and Escherichia-Shigella may be the markers of intestinal microorganisms in patients with heart and spleen deficiency syndrome of coronary heart disease. |
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