Study On The Clinical Characteristics Of Intestinal Flora In Patients With Diabetic Nephropathy With Damp-heat Syndrome

Objective:The incidence of diabetes and diabetic kidney disease as a diabetic microvascular complication are also rising.Results of current epidemiological investigations indicate that chronic renal insufficiency due to diabetes has become one of the leading causes of kidney replacement therapy.Diabetic kidney disease is a kidney disease secondary to long-term blood glucose metabolism disorders in the body.The occurrence and development of this disease is affected by several factors.The pathogenesis is complex and has not been fully elucidated.In recent years,the intestinal microenvironment of patients with diabetic kidney disease has received increasing attention.Existing research shows that intestinal flora is closely related to diabetic kidney disease.Diabetic kidney disease patients with damaged glomeruli and tubules cannot completely excrete metabolic wastes such as indole sulphate and p-cresol sulphate,which directly cause intestinal microenvironment changes and microbiological disorders.At the same time,imbalance of the intestinal flora leads to increased permeability of the intestinal wall.A large amount of toxins such as lipopolysaccharides,cresols,and indoline-based molecules enter the blood circulation to promote the formation of micro inflammation in the body.Factors,leading to enlarged glomerular inflammation,necrosis of tubular epithelial cells,renal interstitial fibrosis,and aggravate the development of diabetic kidney disease.Diabetic kidney disease is closely related to the imbalance of intestinal flora.However,there are few reports on TCM syndrome types and clinical distribution of intestinal flora in diabetic kidney disease.This study takes the combination of disease and syndrome,and Uses high-throughput sequencing technology to explore the differences in the diversity,community composition,and species abundance of patients with damp-heat syndrome with diabetic kidney disease,non-damp-heat syndrome with diabetic kidney disease,type 2 diabetes,and healthy people To clarify the structural characteristics of intestinal flora in patients with damp-heat syndrome with diabetic kidney disease,in order to provide new ideas and methods for the prevention and treatment of diabetic kidney disease by integrated Chinese and western medicine.Method:Twenty patients with diabetic kidney disease damp-heat syndrome（SR）who were treated in the inpatient department of Dongzhimen Hospital of Beijing University of Traditional Chinese Medicine from October 2018 to November 2019 as the observation group,and another 20 patients with diabetic kidney disease non-damp-heat syndrome（FSR）,20 patients with simple type 2 diabetes（D）,and 20 healthy people（N）served as control groups.Fresh stools of each people were collected for gene DNA extraction,and the gene 16s V3-V4 region was amplified by polymerase chain reaction.Finally,a Miseq library was constructed and sequenced to obtain the biological information of the feces sample.Differences in diversity,colony composition,and species abundance between the observation and control groups were compared by biological analysis.Results:Comparison of results between the groups of diabetic kidney disease damp-heat syndrome group and healthy group:（1）Comparison of diversity:The chaol index,observed species index,PD whole tree index,and Shannon index of the diabetic kidney disease damp-heat syndrome group were higher than those of the healthy group.But the differences were not statistically significant（P>0.05）.（2）Species annotation differences:At the level of bacterial phylum:the abundance of Fusobacteria,Firmicutes and Bacteroidetes in the diabetic kidney disease damp-heat syndrome group is lower than that in the healthy group,and the abundance of Actinobacteria in the diabetic kidney disease damp-heat syndrome group is higher than that in the healthy group.The difference in Actinobacteria was statistically significant（P<0.05）.At the level of bacterial order:the abundance of Clostridiales,Bacteroidales,Fusobacteriales and Enterobacteriales in the diabetic kidney disease damp-heat syndrome group is lower than that in the healthy group,and the abundance of Bifidobacteriales and Lactobacillales in the diabetic kidney disease damp-heat syndrome group is higher than that in the healthy group.The difference in Bifidobacterium and Lactobacillus were statistically significant（P<0.05）.At the level of bacterial genus:Compared with the healthy group,the abundance of Blautia,Lachnospira,Clostridium_sensu_stricto₁,Roseburia and Veillonella were decreased,the abundance of Bifidobacterium,Lactobacillus,Neisseria,Ruminococcaceae UCG-007,Pseudomonas,Eubacterium nodatum_group were increased.The differences were statistically significant（P<0.05）.Comparison of results between the groups of diabetic kidney disease damp-heat syndrome group and type 2 diabetes group:（1）Comparison of diversity:The chao1 index,observed species index,PD whole tree index,and Shannon index of the diabetic kidney disease damp-heat syndrome group were higher than those of the type 2 diabetes group.But the differences were not statistically significant（P>0.05）.（2）Species annotation differences:At the level of bacterial phylum:the abundance of Fusobacteria and Firmicutes in the diabetic kidney disease damp-heat syndrome group is lower than that in the type 2 diabetes group,and the abundance of Actinobacteria in the diabetic kidney disease damp-heat syndrome group is higher than that in the type 2 diabetes group.The differences were not statistically significant（P>0.05）.At the level of bacterial order:the abundance of Fusobacteriales and Enterobacteriales in the diabetic kidney disease damp-heat syndrome group is lower than that in the type 2 diabetes group,and the abundance of Bifidobacteriales and Lactobacillales in the diabetic kidney disease damp-heat syndrome group is higher than that in the type 2 diabetes group.The differences were not statistically significant（P>0.05）.At the level of bacterial genus:Compared with the type 2 diabetes group,the abundance of Blautia was decreased and Prevotella₆,Prevotella₂,Neisseria,Ruminococcaceae UCG-014,Ruminococcaceae_UCG-002,Lachnospiraceae_NK4A136_group,Eubacterium_eligens_group were increased.The differences were statistically significant（P<0.05）.The abundance of Bifidobacterium in the diabetic kidney disease damp-heat syndrome group was increased,but the difference was not statistically significant（P>0.05）.Comparison of results between the groups of diabetic kidney disease damp-heat syndrome group and diabetic kidney disease non-damp-heat syndrome group:（1）Comparison of diversity:The chaol index,observed species index,PD whole tree index,and Shannon index of the diabetic kidney disease damp-heat syndrome group were higher than those of the diabetic kidney disease non-damp-heat syndrome group.But the differences were not statistically significant（P>0.05）.（2）Species annotation differences:At the level of bacterial phylum:the abundance of Proteobacteria and Bacteroidetes in the diabetic kidney disease damp-heat syndrome group is lower than that in the non-damp-heat syndrome group,and the abundance of Firmicutes and Actinobacteria in the diabetic kidney disease damp-heat syndrome group is higher than that in non-damp-heat syndrome group.The difference in Proteobacteria was statistically significant（P<0.05）.At the level of bacterial order:the abundance of Enterobacteriales,Bacteroidales and Clostridialesin the diabetic kidney disease damp-heat syndrome group is lower than that in the non-damp-heat syndrome group,and the abundance of Lactobacillales and Bifidobacteriales in the diabetic kidney disease damp-heat syndrome group is higher than that in non-damp-heat syndrome group.The difference in Enterobacteriales was statistically significant（P<0.05）.At the level of bacterial genus:Compared with the diabetic kidney disease non-damp-heat syndrome group,the abundance of Coprobacillus was increased and Klebsiella,Enterorhabdus and Proteus were decreased.The differences were statistically significant（P<0.05）.Conclusions:（1）Compared with the healthy group,the richness and uniformity of intestinal flora in diabetic kidney disease damp-heat syndrome group were increased.The composition of intestinal flora was not significant difference.At the level of bacterial phylum:The abundance of Firmicutes,Bacteroidetes and Fusobacteria in diabetic kidney disease damp-heat syndrome group were decreased,Actinobacteria was increased At the level of bacterial order;The abundance of Fusobacteriales,Clostridiales,Bacteroidales and Enterobacteriales in diabetic kidney disease damp-heat syndrome group were decreased,Lactobacillales and Bifidobacteriales were increased.At the level of bacterial genus:The abundance of Blautia,Lachnospira,Clostridium_sensu_stricto₁,Roseburia and Veillonella in diabetic kidney disease damp-heat syndrome group were decreased,Bifidobacterium,Lactobacillus,Neisseria,Ruminococcaceae_UCG-007,Pseudomonas and Eubacterium_nodatum_group were increased.（2）Compared with the type 2 diabetes group,the richness and uniformity of intestinal flora in diabetic kidney disease damp-heat syndrome group were increased.The composition of intestinal flora was not significant difference.At the level of bacterial phylum:The abundance of Firmicutes and Fusobacteria in diabetic kidney disease damp-heat syndrome group were decreased,Actinobacteria was increased.At the level of bacterial order:The abundance of Fusobacteriales and Enterobacteriales in diabetic kidney disease damp-heat syndrome group were decreased,Lactobacillales and Bifidobacteriales were increased.At the level of bacterial genus:The abundance of Blautia in diabetic kidney disease damp-heat syndrome group was decreased,Prevotella₆,Prevotella₂,Neisseria,Ruminococcaceae_UCG-014,Ruminococcaceae UCG-002,Lachnospiraceae NK4A136_group and Eubacterium_eligens_group were increased.（3）Compared with diabetic kidney disease non-damp-heat syndrome group,the richness and uniformity of intestinal flora in diabetic kidney disease damp-heat syndrome group were increased.The composition of intestinal flora was not significant difference.At the level of bacterial phylum:The abundance of Proteobacteria and Bacteroidetes in diabetic kidney disease damp-heat syndrome group were decreased,Firmicutes and Actinobacteria were increased.At the level of bacterial order:The abundance of Enterobacteriales,Bacteroidales and Clostridiales in diabetic kidney disease damp-heat syndrome group were decreased,Lactobacillales and Bifidobacteriales were increased.At the level of bacterial genus:The abundance of Coprobacillus in diabetic kidney disease damp-heat syndrome group was increased,Klebsiella,Enterorhabdus and Proteus were decreased.