title: Secreted Metabolites of Bifidobacterium infantis and Lactobacillus acidophilus Protect Immature Human Enterocytes from IL-1β-Induced Inflammation: A Transcription Profiling Analysis.
abstract: Combination regimens of Bifidobacterium infantis and Lactobacillus acidophilus have been demonstrated to prevent necrotizing enterocolitis (NEC) in clinical trials. However, the molecular mechanisms responsible for this protective effect are not well understood. Additionally, conditioned media from individual cultures of these two probiotics show strain specific modulation of inflammation using in vitro human intestinal NEC models. Here we report a transcription profiling analysis of gene expression in immature human fetal intestinal epithelial cells (H4 cells) pretreated with conditioned media from B. infantis (BCM) or L. acidophilus (LCM) prior to IL-1β stimulation. Compared with control media, the two probiotic-conditioned media (PCM) treatments altered the expression of hundreds of genes involved in the immune response, apoptosis and cell survival, cell adhesion, the cell cycle, development and angiogenesis. In IL-1β-stimulated cells, PCM treatment decreased the upregulation of genes in the NF-κB activation pathway and downregulated genes associated with extracellular matrix (ECM) remodeling. Compared with LCM, BCM showed more significant modulatory effects on ECM remodeling, reflected by a lower p value. IL-6 and IL-8 production was significantly reduced in IL-1β-stimulated cells pretreated with PCM (p<0.05), which was consistent with their altered gene expression. Western blot analysis showed that compared with IL-1β stimulation alone, PCM treatment attenuated the decrease of cytoplasmic IκBα and NF-κB p65 levels as well as the increase of nuclear NF-κB p65 levels in the stimulated cells (p<0.05). In conclusion, PCM treatment exerted anti-inflammatory effects in immature human fetal enterocytes primarily by modulating genes in the NF-κB signaling and ECM remodeling pathways. Additionally, some components of these signaling pathways, particularly the ECM remodeling pathway, were more profoundly affected by BCM than LCM.
authors.affiliation: State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing, China; Mucosal Immunology and Biology Research Center, Massachusetts General Hospital for Children and Harvard Medical School, Charlestown, Massachusetts, United States of America.
authors.forename: Shuangshuang
authors.initials: S
authors.lastname: Guo
authors.affiliation: State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing, China.
authors.forename: Yuming
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts, United States of America.
authors.forename: Ayla
authors.initials: A
authors.lastname: Ergun
authors.affiliation: Section of Neonatology, Department of Pediatrics, The University of Chicago, Chicago, Illinois, United States of America.
authors.forename: Lei
authors.initials: L
authors.lastname: Lu
authors.affiliation: Mucosal Immunology and Biology Research Center, Massachusetts General Hospital for Children and Harvard Medical School, Charlestown, Massachusetts, United States of America.
authors.forename: W Allan
authors.initials: WA
authors.lastname: Walker
authors.affiliation: Mucosal Immunology and Biology Research Center, Massachusetts General Hospital for Children and Harvard Medical School, Charlestown, Massachusetts, United States of America.
authors.forename: Kriston
authors.initials: K
authors.lastname: Ganguli
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D001644:Bifidobacterium / Q000378:metabolism*; D002448:Cell Adhesion / Q000187:drug effects; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology; D020895:Enterocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D007249:Inflammation; D053583:Interleukin-1beta / Q000494:pharmacology*; D015850:Interleukin-6 / Q000032:analysis / Q000235:genetics; D016209:Interleukin-8 / Q000032:analysis / Q000235:genetics; D007779:Lactobacillus acidophilus / Q000378:metabolism*; D008954:Models, Biological; D000072000:NF-KappaB Inhibitor alpha; D051996:Transcription Factor RelA / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; D020672:I-kappa B Proteins; D053583:Interleukin-1beta; D015850:Interleukin-6; D016209:Interleukin-8; C000606204:NFKBIA protein, human; D051996:Transcription Factor RelA; D000072000:NF-KappaB Inhibitor alpha