De Angelis Maria, Montemurno Eustacchio, Piccolo Maria, Vannini Lucia, Lauriero Gabriella, Maranzano Valentina, Gozzi Giorgia, Serrazanetti Diana, Dalfino Giuseppe, Gobbetti Marco, Gesualdo Loreto
Department of Soil, Plant and Food Sciences, University of Bari Aldo Moro, Bari, Italy.
Department of Emergency and Organ Transplantation, Nephrology Unit - University of Bari Aldo Moro, Bari, Italy.
PLoS One. 2014 Jun 12;9(6):e99006. doi: 10.1371/journal.pone.0099006. eCollection 2014.
This study aimed at investigating the fecal microbiota, and the fecal and urinary metabolome of non progressor (NP) and progressor (P) patients with immunoglobulin A nephropathy (IgAN). Three groups of volunteers were included in the study: (i) sixteen IgAN NP patients; (ii) sixteen IgAN P patients; and (iii) sixteen healthy control (HC) subjects, without known diseases. Selective media were used to determine the main cultivable bacterial groups. Bacterial tag-encoded FLX-titanium amplicon pyrosequencing of the 16S rDNA and 16S rRNA was carried out to determine total and metabolically active bacteria, respectively. Biochrom 30 series amino acid analyzer and gas-chromatography mass spectrometry/solid-phase microextraction (GC-MS/SPME) analyses were mainly carried out for metabolomic analyses. As estimated by rarefaction, Chao and Shannon diversity index, the lowest microbial diversity was found in P patients. Firmicutes increased in the fecal samples of NP and, especially, P patients due to the higher percentages of some genera/species of Ruminococcaceae, Lachnospiraceae, Eubacteriaceae and Streptococcaeae. With a few exceptions, species of Clostridium, Enterococcus and Lactobacillus genera were found at the highest levels in HC. Bacteroidaceae, Porphyromonadaceae, Prevotellaceae and Rikenellaceae families differed among NP, P and HC subjects. Sutterellaceae and Enterobacteriaceae species were almost the highest in the fecal samples of NP and/or P patients. Compared to HC subjects, Bifidobacterium species decreased in the fecal samples of NP and P. As shown by multivariate statistical analyses, the levels of metabolites (free amino acids and organic volatile compounds) from fecal and urinary samples markedly differentiated NP and, especially, P patients.
本研究旨在调查免疫球蛋白A肾病(IgAN)非进展型(NP)和进展型(P)患者的粪便微生物群以及粪便和尿液代谢组。研究纳入了三组志愿者:(i)16例IgAN NP患者;(ii)16例IgAN P患者;以及(iii)16名无已知疾病的健康对照(HC)受试者。使用选择性培养基来确定主要的可培养细菌群。分别对16S rDNA和16S rRNA进行细菌标签编码的FLX-钛扩增子焦磷酸测序,以确定总细菌和代谢活性细菌。主要采用Biochrom 30系列氨基酸分析仪和气相色谱-质谱联用/固相微萃取(GC-MS/SPME)分析进行代谢组学分析。通过稀疏度、Chao和香农多样性指数估计,P患者的微生物多样性最低。由于瘤胃球菌科、毛螺菌科、真杆菌科和链球菌科某些属/种的百分比更高,NP患者尤其是P患者粪便样本中的厚壁菌门增加。除少数例外,梭菌属、肠球菌属和乳杆菌属的物种在HC中含量最高。拟杆菌科、卟啉单胞菌科、普雷沃氏菌科和理研菌科在NP、P和HC受试者之间存在差异。萨特氏菌科和肠杆菌科物种在NP和/或P患者的粪便样本中几乎含量最高。与HC受试者相比,NP和P患者粪便样本中的双歧杆菌属物种减少。多变量统计分析表明,粪便和尿液样本中代谢物(游离氨基酸和有机挥发性化合物)的水平显著区分了NP患者,尤其是P患者。