To Investigate The Changes Of Intestinal Flora In Patients With Coronary Heart Disease And The Correlation Between Them Based On High Throughput Sequencing

BackgroundWith the improvement of living standards and changes in lifestyle,the incidence and prevalence rate of coronary heart disease increases year by year,and the trend of younger age is obvious,and the number of patients receiving treatment continues to increase.In the face of many patients with coronary heart disease,At present,people pay more attention to the treatment of diseases,pay less attention to the prevention before and after the onset of the disease,which leads to the further development of the disease and the burden on the society,the country,the family,and the individual.Therefore,prevention and control of coronary heart disease is of great significance.However,the specific pathogenesis of coronary heart disease is not clear,but most scholars study the risk factors of coronary heart disease,the risk factors can be divided into uncontrollable（old age,sex）and controllable（hypertension,diabetes,smoking,obesity,dyslipidemia）.In recent years,studies have shown that intestinal flora is associated with atherosclerosis,myocardial infarction,blood pressure,blood lipid,blood sugar,vitamin synthesis,etc.Its atherosclerosis,blood pressure,blood lipids,blood sugar and so on are risk factors of coronary heart disease.Therefore this paper aims to discuss the relationship between intestinal flora and coronary heart disease,and the correlation between intestinal flora and risk factors of coronary heart disease.It is used to study the metabolic pathway of risk factors which may lead to the occurrence and development of coronary heart disease.ObjectiveTo Investigate the Changes of Intestinal Flora in Patients with Coronary Heart Disease and the correlation between them based on high throughput sequencing.MethodFrom July 2016 to June 2017 in First Affiliated Hospital of Xinxiang Medical College Department of Cardiology and severe cardiac ICU were treated by ECG,myocardial enzyme markers,coronary angiography and coronary heart disease combined with clinical manifestations were diagnosed in 62 cases,male 38 cases,female 24 cases,age 29-75 years,the average age（54.67+9.45）,Subjects with coronary heart disease were divided into low-grade group（30 cases）and moderate-severe group（32 cases）according to the Gensini standard for evaluation of coronary heart disease.And the hospital physical examination of 60 healthy cases as the control group,there were 35 males and 25 females,aged 28-72 years old,the average age（53.7+7.25）.All subjects were excluded as follows:CHD group and control group were collected with the consent of the subjects.The fresh feces of the subjects were collected about 5 g in aseptic cryopreservation tube.After sealing it,it was sent to the heart disease key laboratory within 1 hour,and 2.00 g feces were taken,and the rest was stored in an ultra-low temperature refrigerator at-80 ℃.The DNA was extracted,and the concentration of DNA was detected.Fluorescence quantitative PCR was performed,and then High throughput sequencing of intestinal microbes was performed by Suzhou Jin Weizhi Science and Technology company for.There was no significant difference in age,sex,blood lipid,height,body weight,body mass index,fasting blood glucose and diet between chd group and control group（p >0.05）.Results1.Comparison of intestinal flora between coronary heart disease group and control group at phylum levelAt the phylum level of the CAD group and the control group contained the phylum of Bacteroidetes 、 Firmicutes 、 Proteobacteria 、 Fusobacterium 、 Actinobacteria 、Euryarchaeota and so on.Compared with the control group,Intestinal microflora in coronary heart disease group contain tenericutes,synergistetes,There was a significant change in the structure and composition of the phylum level.But Bacteroides phylum,Thickwall is still the dominant phylum.2.Comparison of intestinal microflora in each group of coronary heart diseaseCompared with the low degree group of coronary heart disease,the abundance of Bacteroides,Clostridium,Actinomycetes decreased,while the abundance of Thinchycetes and Proteus increased in moderate-severe group Coronary Heart Disease group,The difference was statistically significant.The results showed that the abundance of intestinal flora in coronary artery disease group was further changed with the increase of coronary artery stenosis.3.Comparison of of the intestinal flora at the level of the genus between the coronary heart disease group and the control groupThe intestinal flora of coronary heart disease group and control group wercompared and analyzed at the generic level,There are 99 different genera.The higheabundance of species in coronary heart disease group is followed by: Bacteroide（25.76%）、Faecalibacterium（7.86%）、Megamonas（5.38%）、Alistipes（4.22%）Escherichia-Shigella（4.05%）、Fusobacterium（3.72%）、Parabacteroides（3.46%）Roseburia（2.51%）、Prevotella₉（2.38%）、Bifidobacterium（0.92%）、Klebsiell（0.84%）;the higher abundance of species in control group is followed by: Bacteroide（50.12%）、Fusobacterium（3.721%）、Megasphaera（0.14%）、Roseburia（4.07%）Faecalibacterium（3.5%）、Prevotella₉（3.34%）、Phascolarctobacterium（3.32%）Bifidobacterium（2.07%）、Alistipes（2.04%）、Klebsiella（1.98%）、Parabacteroide（1.04%）、Escherichia-Shigella（0.23%）、Fusobacterium（0.022%）.99 genera in alsamples of coronary heart disease group and control group were analyzed for statisticadifferences.A new genus of bacteria appeared in coronary heart disease group,Thstructure ratio of genera has also changed.abundance of Faecalibacterium 、 Alistipes 、Escherichia-Shigella、Fusobacterium in coronary heart disease group were increased,whilabundance of Bacteroides 、Megasphaera、Roseburia、Prevotella₉、Bifidobacterium wer descend.It is new emerging genera of bacteria that Citrobacter、Weissella、Succinivibrio、Ruminiclostridium、Eisenbergiella、Enterococcus in coronary heart disease group.4.Comparison of intestinal flora at the generic level of each group of coronary heart diseasethe two groups in Coronary heart disease was compared each other,Abundance of Bacillus roseburia prevotella-9,fusobacterium parabacteroides,bifidobacterium,ruminococcus-2 descend,Abundance of Faecalibacterium 、 Alistipes 、 coprostanoligenes_group、Subdoligranulum、Dialister、Phascolarctobacterium increased,The difference is statistically significant.There was no significant difference in abundance of megasphaera,escherichia-shigella.5.Comparison of the level of the blood pressure,Hcy,folic acid and vitamin B12 in the coronary heart disease group with control group,Compared with the control group,The level of blood pressure,Hcy,folic acid and vitamin B12 in the coronary heart disease group were lower,the difference was statistically significant.6.Comparison of blood pressure,folic acid and vitamin b12 levels in patients with coronary heart diseaseCompared with the control group,the level of blood pressure homocysteine increased,folic acid and vitamin B 12 decreased in CHD group,the difference was statistically significant.7.Correlation of Bacteroidetes,Firmicutes,Bacteroides,Faecalibacterium,Roseburia、Prevotella、Bifidobacterium with Blood pressure,Homocysteine,folic Acid,Vitamin B12 in Coronary Heart Disease GroupFolic acid and vitamin B12 were negatively correlated with homocysteine in coronary heart disease group（r=-0.332、-0.378,P<0.05）.There was a positive correlation between folic acid,vitamin B12 and Bifidobacterium in coronary heart disease group（r=0.352 、0.257,P<0.05）.Folic acid,vitamin B12 were negatively correlated with blood pressure in coronary heart disease group,（r=-0.373、-0.401,P<0.05）.There was a positive correlation between homocysteine and blood pressure in coronary heart disease group（r=0.476,P<0.05）;Bacteroidetes,Roseburia and Bifidobacterium are negatively correlated with blood pressure（r=-0.247、-0.041、-0.387,P<0.05）.There is a positive correlation between blood pressure and the Firmicutes、Prevotella（r=0.455、0.386,P<0.05）.Conclusions1.There are changes in intestinal flora structure in patients with coronary heart disease,suggesting that intestinal flora was involved in the occurrence and development of coronary heart disease.2.The imbalance of intestinal flora leads to the increase of Firmicutes/ Bacteroidetes（F /B）ratio,the disorder of folic acid,vitamin B12 and so on,which affect the metabolic pathways of blood homocysteine and blood pressure and lead to the occurrence and development of atherosclerosis and coronary heart disease.It provides theoretical support for further study on the effect of probiotics on coronary heart disease.