Gut Dysbiosis and Detection of “Live Gut Bacteria” in Blood of Japanese Patients With Type 2 Diabetes

OBJECTIVE Mounting evidence indicates that the gut microbiota are an important modifier of obesity and diabetes. However, so far there is no information on gut microbiota and “live gut bacteria” in the systemic circulation of Japanese patients with type 2 diabetes. RESEARCH DESIGN AND METHODS Using a sensitive reverse transcription–quantitative PCR (RT-qPCR) method, we determined the composition of fecal gut microbiota in 50 Japanese patients with type 2 diabetes and 50 control subjects, and its association with various clinical parameters, including inflammatory markers. We also analyzed the presence of gut bacteria in blood samples. RESULTS The counts of the Clostridium coccoides group, Atopobium cluster, and Prevotella (obligate anaerobes) were significantly lower (P < 0.05), while the counts of total Lactobacillus (facultative anaerobes) were significantly higher (P < 0.05) in fecal samples of diabetic patients than in those of control subjects. Especially, the counts of Lactobacillus reuteri and Lactobacillus plantarum subgroups were significantly higher (P < 0.05). Gut bacteria were detected in blood at a significantly higher rate in diabetic patients than in control subjects (28% vs. 4%, P < 0.01), and most of these bacteria were Gram-positive. CONCLUSIONS This is the first report of gut dysbiosis in Japanese patients with type 2 diabetes as assessed by RT-qPCR. The high rate of gut bacteria in the circulation suggests translocation of bacteria from the gut to the bloodstream.

[1]  Kazumasa Matsumoto,et al.  Establishment of an Analytical System for the Human Fecal Microbiota, Based on Reverse Transcription-Quantitative PCR Targeting of Multicopy rRNA Molecules , 2009, Applied and Environmental Microbiology.

[2]  S. Hazen,et al.  Intestinal microbial metabolism of phosphatidylcholine and cardiovascular risk. , 2013, The New England journal of medicine.

[3]  E. N. Bergman Energy contributions of volatile fatty acids from the gastrointestinal tract in various species. , 1990, Physiological reviews.

[4]  J. Ferrières,et al.  Metabolic Endotoxemia Initiates Obesity and Insulin Resistance , 2007, Diabetes.

[5]  M. Hattori,et al.  Bifidobacteria can protect from enteropathogenic infection through production of acetate , 2011, Nature.

[6]  Zeneng Wang,et al.  Intestinal Microbial Metabolism of Phosphatidylcholine and Cardiovascular Risk , 2013 .

[7]  F. Bushman,et al.  Intestinal microbiota metabolism of L-carnitine, a nutrient in red meat, promotes atherosclerosis , 2013, Nature Medicine.

[8]  Patrice D Cani,et al.  Diabetes, obesity and gut microbiota. , 2013, Best practice & research. Clinical gastroenterology.

[9]  F. Liew,et al.  Negative regulation of Toll-like receptor-mediated immune responses , 2005, Nature Reviews Immunology.

[10]  A. M. Habib,et al.  Short-Chain Fatty Acids Stimulate Glucagon-Like Peptide-1 Secretion via the G-Protein–Coupled Receptor FFAR2 , 2012, Diabetes.

[11]  Patrice D Cani,et al.  The role of the gut microbiota in energy metabolism and metabolic disease. , 2009, Current pharmaceutical design.

[12]  J. Shaw,et al.  IDF diabetes atlas: global estimates of the prevalence of diabetes for 2011 and 2030. , 2011, Diabetes research and clinical practice.

[13]  S. Rane,et al.  Beneficial Metabolic Effects of a Probiotic via Butyrate-induced GLP-1 Hormone Secretion* , 2013, The Journal of Biological Chemistry.

[14]  Toshiaki Shimizu,et al.  Bacterial rRNA-Targeted Reverse Transcription-PCR Used To Identify Pathogens Responsible for Fever with Neutropenia , 2010, Journal of Clinical Microbiology.

[15]  J. Gordon,et al.  Human nutrition, the gut microbiome and the immune system , 2011, Nature.

[16]  Chih-Hsin Tang,et al.  Lipoteichoic acid enhances IL-6 production in human synovial fibroblasts via TLR2 receptor, PKCdelta and c-Src dependent pathways. , 2010, Biochemical pharmacology.

[17]  Fredrik H. Karlsson,et al.  Symptomatic atherosclerosis is associated with an altered gut metagenome , 2012, Nature Communications.

[18]  P. Kubes,et al.  Lipoteichoic Acid Induces Unique Inflammatory Responses when Compared to Other Toll-Like Receptor 2 Ligands , 2009, PloS one.

[19]  J. Moreno-Navarrete,et al.  Serum lipopolysaccharide-binding protein as a marker of atherosclerosis. , 2013, Atherosclerosis.

[20]  S. Sasaki,et al.  Comparison of relative validity of food group intakes estimated by comprehensive and brief-type self-administered diet history questionnaires against 16 d dietary records in Japanese adults , 2011, Public Health Nutrition.

[21]  S. Sørensen,et al.  Gut Microbiota in Human Adults with Type 2 Diabetes Differs from Non-Diabetic Adults , 2010, PloS one.

[22]  R. Knight,et al.  Diversity, stability and resilience of the human gut microbiota , 2012, Nature.

[23]  D. Kobayashi,et al.  Changes of the Intestinal Microbiota, Short Chain Fatty Acids, and Fecal pH in Patients with Colorectal Cancer , 2013, Digestive Diseases and Sciences.

[24]  Patrice D Cani,et al.  Involvement of gut microbiota in the development of low-grade inflammation and type 2 diabetes associated with obesity , 2012, Gut microbes.

[25]  H. Tilg,et al.  Obesity and the microbiota. , 2009, Gastroenterology.

[26]  S. Sasaki,et al.  Self-administered diet history questionnaire developed for health education: a relative validation of the test-version by comparison with 3-day diet record in women. , 1998, Journal of epidemiology.

[27]  T. Asahara,et al.  Sensitive Quantitative Detection of Commensal Bacteria by rRNA-Targeted Reverse Transcription-PCR , 2006, Applied and Environmental Microbiology.

[28]  R. Bibiloni,et al.  Changes in Gut Microbiota Control Metabolic Endotoxemia-Induced Inflammation in High-Fat Diet–Induced Obesity and Diabetes in Mice , 2008, Diabetes.

[29]  Qiang Feng,et al.  A metagenome-wide association study of gut microbiota in type 2 diabetes , 2012, Nature.

[30]  T. Muta,et al.  Essential roles of CD14 and lipopolysaccharide-binding protein for activation of toll-like receptor (TLR)2 as well as TLR4 Reconstitution of TLR2- and TLR4-activation by distinguishable ligands in LPS preparations. , 2001, European journal of biochemistry.

[31]  E. Mardis,et al.  An obesity-associated gut microbiome with increased capacity for energy harvest , 2006, Nature.

[32]  J B L Hoekstra,et al.  The therapeutic potential of manipulating gut microbiota in obesity and type 2 diabetes mellitus , 2012, Diabetes, obesity & metabolism.

[33]  V. Tremaroli,et al.  Functional interactions between the gut microbiota and host metabolism , 2012, Nature.

[34]  J. Doré,et al.  Involvement of tissue bacteria in the onset of diabetes in humans: evidence for a concept , 2011, Diabetologia.

[35]  Fiona Powrie,et al.  Microbiota, Disease, and Back to Health: A Metastable Journey , 2012, Science Translational Medicine.

[36]  P. Sansonetti,et al.  Intestinal mucosal adherence and translocation of commensal bacteria at the early onset of type 2 diabetes: molecular mechanisms and probiotic treatment , 2011, EMBO molecular medicine.

[37]  T. Asahara,et al.  Sensitive Quantitative Detection of Commensal Bacteria by rRNA-Targeted Reverse Transcription-PCR , 2007, Applied and Environmental Microbiology.