Leite Aline Zazeri, Rodrigues Nathália de Campos, Gonzaga Marina Ignácio, Paiolo João Carlos Cicogna, de Souza Carolina Arantes, Stefanutto Nadine Aparecida Vicentini, Omori Wellington Pine, Pinheiro Daniel Guariz, Brisotti João Luiz, Matheucci Junior Euclides, Mariano Vânia Sammartino, de Oliveira Gislane Lelis Vilela
Microbiome Study Group, School of Health Sciences Dr. Paulo Prata (FACISB), Barretos, Brazil.
QGene-Solutions and Logistics in Health, Sao Carlos, Brazil.
Front Immunol. 2017 Sep 15;8:1107. doi: 10.3389/fimmu.2017.01107. eCollection 2017.
Intestinal dysbiosis and metabolic endotoxemia have been associated with metabolic disorders, such as obesity, insulin resistance, and type 2 diabetes (T2D). The main goal of the present study was to evaluate the intestinal dysbiosis in Brazilian T2D patients and correlate these data with inflammatory cytokines and lipopolysaccharides (LPS) plasma concentrations. This study was approved by the Ethics Committees from Barretos Cancer Hospital and all individuals signed the informed consent form. Stool samples were required for DNA extraction, and the V3/V4 regions of bacterial 16S were sequenced using an Illumina platform. Peripheral blood was used to quantify inflammatory cytokines and plasma LPS concentrations, by CBA flex and ELISA, respectively. Statistical analyses were performed using Mann-Whitney and Spearman's tests. Analysis of variance, diversity indexes, and analysis of alpha- and beta-diversity were conducted using an annotated Operational Taxonomic Unit table. This study included 20 patients and 22 controls. We observed significant differences ( < 0.01) in the microbiota composition (beta-diversity) between patients and controls, suggesting intestinal dysbiosis in Brazilian T2D patients. The prevalent species found in patients' feces were the Gram-negatives , and . The proinflammatory interleukin-6 (IL-6) was significantly increased ( < 0.05) in patients' plasma and LPS levels were decreased. We find correlations between the proinflammatory interferon-gamma with Gram-negatives and species, and a positive correlation between the LPS levels and reads. The and species were associated with insulin resistance in previous studies. In this study, we suggested that the prevalence of Gram-negative species in the gut and the increased plasma IL-6 in patients could be linked to low-grade inflammation and insulin resistance. In conclusion, the and species could represent an intestinal microbiota signature, associated with T2D development. Furthermore, the identification of these Gram-negative bacteria, and the detection of inflammatory markers, such as increased IL-6, could be used as diabetes predictive markers in overweight, obese and in genetically predisposed individuals to develop T2D.
肠道菌群失调和代谢性内毒素血症与肥胖、胰岛素抵抗和2型糖尿病(T2D)等代谢紊乱有关。本研究的主要目的是评估巴西T2D患者的肠道菌群失调情况,并将这些数据与炎症细胞因子和脂多糖(LPS)血浆浓度相关联。本研究经巴雷托斯癌症医院伦理委员会批准,所有个体均签署了知情同意书。需要粪便样本进行DNA提取,并使用Illumina平台对细菌16S的V3/V4区域进行测序。分别通过CBA flex和ELISA法使用外周血来定量炎症细胞因子和血浆LPS浓度。使用Mann-Whitney检验和Spearman检验进行统计分析。使用带注释的操作分类单元表进行方差分析、多样性指数分析以及α-和β-多样性分析。本研究纳入了20例患者和22例对照。我们观察到患者和对照之间微生物群组成(β-多样性)存在显著差异(<0.01),提示巴西T2D患者存在肠道菌群失调。在患者粪便中发现的优势菌种为革兰氏阴性菌 、 和 。促炎白细胞介素-6(IL-6)在患者血浆中显著升高(<0.05),而LPS水平降低。我们发现促炎干扰素-γ与革兰氏阴性菌 和 菌种之间存在相关性,并且LPS水平与 读数之间存在正相关。在先前的研究中, 和 菌种与胰岛素抵抗有关。在本研究中,我们认为肠道中革兰氏阴性菌种的流行以及患者血浆中IL-6的升高可能与低度炎症和胰岛素抵抗有关。总之, 和 菌种可能代表一种与T2D发展相关的肠道微生物群特征。此外,鉴定这些革兰氏阴性菌以及检测炎症标志物,如升高的IL-6,可作为超重、肥胖以及有T2D遗传易感性个体患糖尿病的预测标志物。