transforming-growth-factor-beta and Enterocolitis--Necrotizing

Necrotizing enterocolitis (NEC) yet remains a leading cause of morbidity and mortality in premature infants. The developmental deficiency of transforming growth factor-Beta (TGF-β) in the intestine is a risk factor for NEC in premature infants.We aimed to investigate the potential utility of serum TGF-β1 in the early diagnosis and severity assessment of NEC. This prospective case-control study was conducted on 102 VLBW neonates aging less than 32 weeks and weighing less than 1500 gm. They were divided into NEC group of 52 preterm neonates with symptoms and signs of NEC and 50 age and sex-matched neonates without NEC as a control group. All neonates underwent full medical history taking, clinical examination, radiological and laboratory investigations including CBC, CRP, fecal occult blood, and serum TGF-β1. Serum TGF-β1 was tested in NEC patients at the onset of symptoms and signs and 7 days later. Serum TGF-β1 was significantly lower in NEC patients at the onset of symptoms than the control group (P = 0.004) while after 7 days of onset serum TGF-β1 was significantly higher than at the onset of symptoms (P < 0.001). In NEC patients with stage I, TGF-β1 was significantly higher than in NEC patients with stage ≥II (P = 0.027).In conclusion serum TGF-β1 is downregulated in neonatal necrotizing enterocolitis and can be used as a useful biomarker for early diagnosis of NEC and to assess disease severity.

Topics: Case-Control Studies; Enterocolitis, Necrotizing; Female; Fetal Diseases; Humans; Infant; Infant, Newborn; Infant, Newborn, Diseases; Infant, Premature; Transforming Growth Factor beta; Transforming Growth Factor beta1

Necrotizing enterocolitis (NEC) is an inflammatory bowel necrosis of premature infants. Based on our recent findings of increased Smad7 expression in surgically resected bowel affected by NEC, we hypothesized that NEC macrophages undergo inflammatory activation because increased Smad7 expression renders these cells resistant to normal, gut-specific, transforming growth factor (TGF)-β-mediated suppression of inflammatory pathways.. We used surgically resected human NEC tissue, murine models of NEC-like injury, bone marrow-derived and intestinal macrophages, and RAW264.7 cells. Smad7 and IκB kinase-beta (IKK-β) were measured by quantitative PCR, western blots, and immunohistochemistry. Promoter activation was confirmed in luciferase reporter and chromatin immunoprecipitation assays.. NEC macrophages showed increased Smad7 expression, particularly in areas with severe tissue damage and high bacterial load. Lipopolysaccharide-induced Smad7 expression suppressed TGF-β signaling and augmented nuclear factor-kappa B (NF-κB) activation and cytokine production in macrophages. Smad7-mediated NF-κB activation was likely mediated via increased expression of IKK-β, which, further increased Smad7 expression in a feed-forward loop. We show that Smad7 induced IKK-β expression through direct binding to the IKK-β promoter and its transcriptional activation.. Smad7 expression in NEC macrophages interrupts TGF-β signaling and promotes NF-κB-mediated inflammatory signaling in these cells through increased expression of IKK-β.

Topics: Animals; Enterocolitis, Necrotizing; Humans; I-kappa B Kinase; Inflammation; Intestinal Mucosa; Lipopolysaccharides; Macrophages; Mice; Mice, Inbred C57BL; NF-kappa B; RAW 264.7 Cells; Signal Transduction; Smad7 Protein; Transforming Growth Factor beta

The goal was to identify cytokines associated with necrotizing enterocolitis (NEC). Based on our earlier reports of decreased tissue expression of transforming growth factor (TGF)-β, we hypothesized that infants with NEC also have low blood TGF-β levels. We further hypothesized that because fetal inflammation increases the risk of NEC, infants who develop NEC have elevated blood cytokine levels in early neonatal period.. Data on 104 extremely-low-birth-weight infants with NEC and 893 without NEC from 17 centers were analyzed. Clinical information was correlated with blood cytokine levels on postnatal day 1 (D1), D3, D7, D14, and D21.. Male gender, non-Caucasian/non-African American ethnicity, sepsis, lower blood TGF-β and interleukin (IL)-2 levels, and higher IL-8 levels were associated with NEC. The NEC group had lower TGF-β levels than controls since D1. The diagnosis of NEC was associated with elevated IL-1β, IL-6, IL-8, IL-10, monocyte chemoattractant protein-1/CC-motif ligand-2, macrophage inflammatory protein-1β/CC-motif ligand-3, and C-reactive protein.. Clinical characteristics, such as gender and ethnicity, and low blood TGF-β levels are associated with higher risk of NEC. Infants who developed NEC did not start with high blood levels of inflammatory cytokines, but these rose mainly after the onset of NEC.

Topics: Biomarkers; Cytokines; Enterocolitis, Necrotizing; False Positive Reactions; Female; Humans; Infant, Extremely Low Birth Weight; Infant, Newborn; Infant, Premature; Inflammation; Interleukin-2; Interleukin-8; Male; Reproducibility of Results; Risk; Transforming Growth Factor beta

Cronobacter sakazakii is a Gram-negative pathogen associated with the cases of necrotizing enterocolitis (NEC) that result from formula contamination. In a mouse model of NEC, we demonstrate that C. sakazakii infection results in epithelial damage by recruiting greater numbers of dendritic cells (DCs) than macrophages and neutrophils in the gut and suppresses DC maturation, which requires outer membrane protein A (OmpA) expression in C. sakazakii. Pretreatment of intestinal epithelial cell monolayers with supernatant from OmpA(+) C. sakazakii/DC culture markedly enhanced membrane permeability and enterocyte apoptosis, whereas OmpA(-) C. sakazakii/DC culture supernatant had no effect. Analysis of OmpA(+) C. sakazakii/DC coculture supernatant revealed significantly greater TGF-β production compared with the levels produced by OmpA(-) C. sakazakii infection. TGF-β levels were elevated in the intestinal tissue of mice infected with OmpA(+) C. sakazakii. Cocultures of CaCo-2 cells and DCs in a "double-layer" model followed by infection with OmpA(+) C. sakazakii significantly enhanced monolayer leakage by increasing TGF-β production. Elevated levels of inducible NO synthase (iNOS) were also observed in the double-layer infection model, and abrogation of iNOS expression prevented the C. sakazakii-induced CaCo-2 cell monolayer permeability despite the presence of DCs or OmpA(+) C. sakazakii/DC supernatant. Blocking TGF-β activity using a neutralizing Ab suppressed iNOS production and prevented apoptosis and monolayer leakage. Depletion of DCs in newborn mice protected against C. sakazakii-induced NEC, whereas adoptive transfer of DCs rendered the animals susceptible to infection. Therefore, C. sakazakii interaction with DCs in intestine enhances the destruction of the intestinal epithelium and the onset of NEC due to increased TGF-β production.

Topics: Animals; Bacterial Outer Membrane Proteins; Caco-2 Cells; Coculture Techniques; Cronobacter sakazakii; Dendritic Cells; Enterocolitis, Necrotizing; Humans; Intestinal Mucosa; Mice; Nitric Oxide Synthase Type II; Transforming Growth Factor beta

Treatment with transforming growth factor-beta(1) (TGF-beta(1)) has been shown to be effective in accelerating skin wound healing. Another approach to gain the beneficial effects of TGF-beta(1) on wound healing could be the activation of tissue stores of latent TGF-beta(1) with agents such as vitamin A. The aims of this study were to determine whether 1) vitamin A is effective in enhancing intestinal wound healing in vitro and 2) activation of TGF-beta(1) is increased during wound healing with vitamin A treatment. We used the intraluminal chemical induction model of necrotizing enterocolitis (NEC), which was adapted to the 1-wk-old piglet. Injured (NEC) and noninjured full-thickness ileum explants harvested from the piglets were cultured for 24 and 48 h in serum-free medium supplemented with all-trans retinol (ATR; 0, 2, 5, and 10 microM). All concentrations of ATR improved recovery of normal ileal wall cytoarchitecture of NEC explants, with maximal recovery observed with 2 microM ATR after 24 h of culture. Further recovery after 48 h was observed with 5 and 10 microM ATR but did not achieve the degree of healing observed with 2 microM ATR. There were no observable adverse effects of ATR on noninjured ileal explant morphology. Active TGF-beta(1) was identified only in the NEC explants incubated with ATR. The results of this study demonstrate that administration of vitamin A accelerates recovery of normal intestinal wall cytoarchitecture of injured ileum in vitro, without adversely affecting noninjured ileum. The increased activation of latent TGF-beta(1) may, in part, be responsible for the accelerated healing of injured ileum observed with vitamin A administration.

Topics: Animals; Enterocolitis, Necrotizing; Ileum; Sus scrofa; Tissue Culture Techniques; Transforming Growth Factor beta; Transforming Growth Factor beta1; Vitamin A; Wound Healing

Necrotizing enterocolitis (NEC) seems to result from the inflammatory response of an immature intestine. Human milk is protective against NEC via an unknown mechanism. We hypothesized that specific factors found in human milk would decrease stimulated IL-8 secretion in intestinal epithelial cells. HT29-cl19A and Caco2 cells were compared with the fetal human primary intestinal epithelial cell line H4 and temperature-sensitive conditionally immortalized fetal human intestinal (tsFHI) cells. Cells were pretreated with transforming growth factor-beta (TGF-beta), erythropoietin (Epo), IL-10, or epidermal growth factor (EGF) at physiologic concentrations before stimulation with tumor necrosis factor-alpha (TNF-alpha) or IL-1beta, and then IL-8 was measured by ELISA. The fetal cells produced significantly more IL-8 when stimulated by TNF-alpha or IL-1beta. There were also differences in the pattern of alteration of IL-8 secretion by human milk factors. In HT29-cl19A cells, IL-10 inhibited TNF-alpha-stimulated IL-8 secretion by 52%, and EGF increased secretion by 144%. In H4 cells, TGF-beta1 and Epo inhibited TNF-alpha-stimulated IL-8 secretion to control levels, and EGF increased secretion by 29%. IL-1beta-stimulated IL-8 secretion was inhibited 25% by TGF-beta1 in Caco2 cells and in H4 cells was inhibited by TGF-beta1, Epo, and TGF-beta2. TsFHI cells confirmed H4 cell results. Fetal human enterocytes have an exaggerated IL-8 secretion in response to TNF-alpha and IL-1beta. TGF-beta and Epo decrease this stimulated IL-8 secretion, which may partially explain the protective effect of human milk in NEC.

Topics: Age Factors; Caco-2 Cells; Enterocolitis, Necrotizing; Epidermal Growth Factor; Epithelial Cells; Erythropoietin; Fetus; HT29 Cells; Humans; In Vitro Techniques; Interleukin-1; Interleukin-10; Interleukin-8; Intestinal Mucosa; Milk, Human; Transforming Growth Factor beta