Microbiota Of Non-Alcoholic Fatty Liver Disease Complicated With Abnormal Glucose Metabolism Syndrome In Patients With Liver Depression And Spleen Deficiency

Background:As today’s global scope of the most common chronic liver disease,nonalcoholic fatty liver disease in clinic now the prevalence of diabetes mellitus is getting higher and higher,is day by day serious,need to be pay attention to health problems,but nonalcoholic fatty liver disease and its merger failed to clarify the related mechanism of sugar metabolism disorders,based on the existing research theory and 16 s r DNA technology unceasing development,Intestinal flora has become a new way to study abnormal glucose metabolism in nonalcoholic fatty liver disease.At the same time,through syndrome biological basic research to realize quantification of objective is an important research task,today’s Chinese medicine liver depression spleen deficiency as the main syndrome of nonalcoholic fatty liver disease,and diabetes type,also is in the initial stage of sugar,lipid metabolic disorder at the core of the card type,but the liver depression of nonalcoholic fatty liver disease patients with spleen deficiency of intestinal flora structure characteristics of the research is still inadequate,The effect of gdepression-spleen deficiency syndrome on the occurrence and development of non-alcoholic fatty liver disease combined with glucose metabolism disorder remains to be explored.At the same time,the specific structural characteristics of intestinal microflora disorder in patients with non-alcoholic fatty liver disease under different stages of glucose metabolism disorder and whether there are structural differences between each other remain unclear.Based on this,this study is carried out.Objective:Through this study,the gut microbiota of liver-depression and spleendeficiency syndrome in patients with non-alcoholic fatty liver was observed,and the characteristic structural basis of the gut microbiota of liver-depression and spleen-deficiency syndrome in patients with nonalcoholic fatty liver was explored.At the same time,based on the liver spleen deficiency of flora structure characteristics,study of nonalcoholic fatty liver disease in different stages of sugar metabolic abnormalities(prediabetes and diabetes),intestinal flora to nonalcoholic fatty liver abnormal glucose metabolism in patients with early changes of intestinal flora,explore the merger of sugar metabolism of nonalcoholic fatty liver disease in liver depression patients with abnormal spleen deficiency on common characteristics.Methods:1.There were 15 patients aged 18-65 years who were diagnosed with simple non-alcoholic fatty liver disease,and 15 healthy people with matching age were enrolled as the control group.Stool samples were collected and total DNA was extracted.High-throughput 16 S DNA sequencing was used to analyze the gut microbiota structure of the three groups.2.In addition,30 patients aged 18-65 years who were diagnosed with non-alcoholic fatty liver and conforming to the TCM syndrome differentiation diagnosis of Liver depression and spleen deficiency were enrolled.Based on the combination of abnormal glucose metabolism at different stages,the patients with non-alcoholic fatty liver were further divided into prediabetes group and type 2 diabetes group,with 15 patients in each group.At the same time,15 cases of simple non-alcoholic fatty liver disease and 15 cases of healthy control group were matched.Fecal samples were collected,total DNA was extracted,and the gut microbiota structural characteristics of these 4 groups were analyzed by 16 S DNA gene high-throughput sequencing.Results:1.In this study,compared with healthy people,the abundance of intestinal microflora in NAFLD patients significantly decreased,and the firmicutes/Bacteroides ratio decreased.The bacteria are Akkermansia,Mailhella,Catenibacterium,Eubacterium＿xylanophilum＿group,Eubacterium＿eligens＿group,Coprococcus＿2 and the abundance of Prevotellaceae＿UCG＿001 decreased significantly,while Succinivibrio,Clostridium＿innocuum＿group,Ruminococcus＿2,Eubacterium＿ruminantium＿group,Parabacteroid and the abundance of Prevotellaceae＿NK3B31＿group increased significantly.2.Compared with NAFLD dampness and heat accumulation syndrome,the relative abundance of Proteobacteria and Epsilonbacteraeota in the intestinal flora of NAFLD patients with liver depression and spleen deficiency was higher.The bacteria are Epulopiscium,Streptococcus,Gemella,Campylobacter,Actinomyces,Holdemania,Faecalibacterium,Prevotella,Ferruginibacter.The abundance of Llinsella and Ezakiella increased while that of Herbinix decreased.Moreover,the structural characteristics of microbiota in NAFLD syndrome of liver depression and spleen deficiency are more correlated with the disorder of glucose metabolism,which can promote the progression of NAFLD with abnormal glucose metabolism to some extent.3.Compared with patients with simple fatty liver disease,the abundance of microbacteria in NAFLD patients with abnormal glucose metabolism increased,the abundance of Patescibacteria decreased,and the Firmicutes/Bacteroidetes ratio decreased.Ferruginibacter,Peptoclostridium,Catenibacterium,Succinivibrio,Epulopiscium,The abundance of Prevotellaceae＿NK3B31＿group decreases while the abundance of Coprococcus＿2,Prevotellaceae＿UCG＿001,Akkermansia,Megamonas,Prevotella,Ruminococcaceae＿U decreases The abundance level of CG＿014 and Paraprevotella increased.The abundance of most butyrate bacteria decreased,but the abundance of bacteria related to the production of short-chain fatty acids increased.The abnormal aggravation of glucose metabolism in NAFLD patients may be caused by the fact that these bacteria promote the disorder of glucose and lipid metabolism by participating in the production of short-chain fatty acids,thus aggravating insulin resistance.4.Compared with NAFLD patients with diabetes,the abundance levels of Clostridium and Epsilonbacteraeota decreased significantly.The genera include Coprococcus＿2,Mailhella,Gemella,Lachnospiraceae,Campylobacter,Romboutsia,Prevotella,Alloprevotella,Dialister,Fusobac Terium,Lactobacillus,Ruminococcus＿1,Holdemanella,Turicibacter,Negativibacillus,Peptococcus,Psychrobacter,Eisenbergiella,Haemophilus,Roseburia,Parasutterella,Ruminococcaceae＿UCG＿014,Senegalimassilia,Ruminiclostridium＿6,Christensenellaceae＿R＿7＿group and Subdoligranulum decreased,while the abundance levels of Prevotellaceae＿UCG＿001 and Subdoligranulum increased.The abundance of some bacteria producing short-chain fatty acids in the intestinal flora of NAFLD patients with diabetes decreased,but bacteria closely related to chronic inflammation appeared,which was speculated to be associated with the progression of NAFLD patients with diabetes.Conclusions:Compared with healthy people,the structural characteristics of intestinal flora in NAFLD patients with liver depression and spleen deficiency syndrome were mainly proteobacteria abundance increased,microbacteria abundance decreased,Firmicutes/Bacteroides ratio decreased,Akkermansia and other bacteria decreased,and the abundance of Epulopiscium,Megamonas,Streptococcus and other bacteria increased,and the structure of the bacterial community was related to the abnormal development of glucose metabolism in NAFLD patients.In NAFLD patients with abnormal glucose metabolism,the abundance of microflora veruminata increased,the firmicutes/Bacteroides ratio decreased,and the abundance of Prevotellaceae＿UCG＿001,Prevotella,Megamonas and other bacteria closely related to the synthesis of short-chain fatty acids increased.But the abundance of Butyricproducing bacteria such as Faecalibacterium decreased;In NAFLD patients with diabetes,the species richness of the overall flora decreased significantly,the abundance of Clostridium decreased,and the abundance of Prevotella,Megamonas and other bacteria related to the synthesis of short-chain fatty acids decreased.The abundance of Peptoniphilus and Intestinibacter,which were related to chronic inflammation of diabetes,increased.For the increased abundance of Prevotella and Megamonas in NAFLD patients with liver depression and spleen deficiency syndrome,the abundance level of Prevotella and Megamonas continued to increase significantly in NAFLD patients with liver depression and spleen deficiency syndrome in prediabetes,and decreased in NAFLD patients with liver depression and spleen deficiency syndrome in diabetes.However,the abundance level of Prevotellaceae＿UCG＿001 in the same family as Prevotella continued to increase.