| Objective To analyze the characteristics and differences of intestinal microflora between Tibetan and Han healthy people and to declare the relationship between intestinal microflora changing and insulin resistance in patients with type 2 diabetes mellitus(T2DM);to further observe the effect of fecal bacteria transplantation(FMT)on intestinal microflora and glucose metabolism in T2DM patients.Methods1.10 healthy Tibetan and 10 Han volunteers matched in age and sex were recruited for routine physical examination,dietary habits questionnaire survey was conducted to analyze the characteristics of dietary structure of the two groups,stool samples were collected in the morning,and the characteristics of intestinal flora of the two groups were analyzed by 16SrRNA high-throughput sequencing.The differences of intestinal microflora between Tibetan and Han population were compared and the influencing factors were discussed.2.46 patients with T2DM of Han nationality admitted from March 2017 to January 2018 were selected as the observation group,and 50 healthy subjects of Han nationality who underwent physical examination in the same period were selected as the control group.Fasting peripheral blood was collected to detect fasting blood glucose((FPG)),fasting insulin(FINS),serum total cholesterol(TC),triglyceride(TG),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C)and so on.Insulin resistance index(HOMA-IR)was calculated,and stool samples were collected in the morning.The characteristics of intestinal microflora in the two groups were analyzed by 16SrRNA high-throughput sequencing,and the structural differences of intestinal flora in the two groups were compared to explore the relationship between intestinal flora imbalance and insulin resistance in patients with T2DM.3.Five male T2DM patients of Han nationality with substandard blood glucose control(HbA1C?7.0%)from April to June 2018 were recruited as subjects and were treated with FMT by oral fecal paste capsule.FMT,follow-up was performed on days 1,2,7 and 8,respectively.Body weight,waistline,blood glucose,glycosylated hemoglobin(HbA1C),TG,FPG and FINS,were measured before and 6 weeks after FMT treatment.The changes of body weight,waist circumference,body mass index((BMI)),blood glucose,TC,TG,HDL-C,LDL-C,HOMA-IR,intestinal flora structure and hypoglycemic drug dose before and after FMT treatment were compared to evaluate the efficacy and safety of oral fecal paste FMT in the treatment of T2DMResults1.There were significant differences in the types and relative abundance of intestinal microflora between Tibetan and Han groups.At the level of biological taxonomy,the relative abundance of Bacteroides and Proteobacteria in Tibetan population was higher than that in Han population,The relative abundance of Firmicutes,Tenericutes and Actinobacteria was lower than that of Han population.At the biological strain level,the relative abundance of Prebotella in Bacteroides,Bifidobacteriales and Lactobacillus in Firmicutes in Tibetan population was lower than that in Han population.The Pearson correlation analysis showed that the abundance of Bacteroides in intestinal flora was positively correlated with the content of protein(livestock and poultry eggs)in dietary structure(Tibetan r=0.834,0.695;Han r=0.903,0.815,P<0.05).There was a positive correlation between the abundance of Prebotella and the content of fine grains(Han r=0.878,0.772,P<0.05).The abundance of Tenericutes was positively correlated with the content of vegetables and fruits(Han r=0.725,0.526,P<0.05).The abundance of Lactobacillus was positively correlated with the content of fine grain(Tibetan r=0.799,0.639;Han r=0.672,0.451,P<0.05).2.The abundance of Clostridium tenuiens and Enterococcus in T2DM group was significantly higher than that in healthy control group[(5.78±1.12)logN/g vs(5.21±1.01)logN/g,(4.61±0.89)logN/g vs(4.23±0.87)logN/g,P<0.05],The abundance of Bacteroides and E.coli was significantly lower than the control group[(6.64±1.07)logN/g vs(7.41±1.05)logN/g,(1.91±0.54)logN/g vs(2.18±0.55)logN/g,P<0.05],There was no significant difference in the abundance of Coccobacteria,Lactobacillus,Prebotella and Bifidobacteriales between the two groups(P>0.05);There were significant differences between TC,TG,HDL-C,LDL-C and HOMA-IR between the two groups[(5.77±1.13)mmol/L vs(4.02±0.58)mmol/L,(3.41±0.67)mmol/L vs(1.36±0.35)mmol/L,(1.21±0.46)mmol/L vs(1.63±0.43)mmol/L,(3.59±0.89)mmol/L vs(1.89±0.51)mmol/L,(1.02±0.23)vs(3.68±1.01),P<0.05];Pearson correlation analysis showed that Bacteroidetes abundance was negatively correlated with HOMA-IR index(r=-0.264,P<0.05),Enterococcus abundance is positively correlated with TG levels and negatively correlated with HDL-C levels(r=0.313,-0.247,P<0.05),Clostridium tenella abundance was negatively correlated with HDL-C level and positively correlated with HOMA-IR index(r=-0.221,0.314,P<0.05).3.Five Han subjects were followed up for 6 weeks after oral fecal paste FMT.No adverse reactions such as fever,nausea and diarrhea were found.After 6 weeks of FMT treatment,the ratio of B/F increased,the relative abundance of Lactobacillus increased,and the relative abundance of Clostridia decreased.The FPG,HbA1C and HMOA-IR of the subjects were improved compared with those before FMT[9.76±1.38)mmol/l vs(6.64±0.79)momol/l,(13.3±1.56)mmol/l vs(8.82±0.81)mmol/l,(9.46±1.22)%vs(6.22±2.05)%,(6.6±1.08)vs(4.89±0.82),P<0.05],Among them,the dose of hypoglycemic drugs decreased in 2 subjects(subjects 1 reduced the dose of insulin glargine by 12 u and discontinued oral acarbose 50mg/tid,and subjects 2 reduced the dose of insulin by 4 u).At the same time,it was observed that the TG,BMI and waist circumference of the subjects decreased[(3.62±0.58)mmol/l vs(2.98±0.34)mmol/l,(24.46±1.45)kg/m2 vs(24.02±1.47)kg/m2,(92.8±3.42)cm vs(86.4±3.78)cm,P<0.05].Conclusions1.There are differences in intestinal microflora between Tibetan and Han ethnic groups,mainly in the ratio of B/F,which is higher in Tibetan than in Han nationality,and the difference is related to different dietary structure.The relative abundance of Bacteroidetes in intestinal flora is positively correlated with the content of protein(livestock,poultry and eggs)in dietary structure,and the relative abundance of Prebotella is positively correlated with the content of fine grain.The relative abundance of Tenericutes was positively correlated with the content of vegetables and fruits,and the relative abundance of Lactobacillus was positively correlated with the content of fine grains.2.There were disorders of intestinal flora,insulin resistance and lipid metabolism in patients with T2DM,which were mainly characterized by increased relative abundance of Clostridium tenuiformis and Enterococcus,decreased relative abundance of Bacteroidetes and Escherichia coli,negative correlation between relative abundance of Bacteroidetes and HOMA-IR index,positive correlation between relative abundance of Enterococcus and TG level,negative correlation between Enterococcus relative abundance and HDL-C level.The relative abundance of Clostridium tender was negatively correlated with HDL-C level and positively correlated with HOMA-IR index.The imbalance of intestinal flora in patients with T2DM may affect lipid metabolism and aggravate insulin resistance through bile acid metabolism pathway,and ultimately promote the occurrence and development of diabetes.3.The disorder of intestinal flora in patients with T2DM was improved after FMT treatment,which showed that the ratio of B/F increased,the level of Lactobacillus increased and the level of Clostridia decreased compared with that before transplantation.FMT can also improve blood glucose control,blood lipids,BMI and waist circumference,increase insulin sensitivity and reduce the dose of hypoglycemic drugs in patients with T2DM.Oral fecal paste approach for FMT has the advantages of less trauma,strong maneuverability,safety and effectiveness,and is more easily accepted by patients. |
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