The expression of epidermal growth factor and transforming growth factor‐α mRNA in the small intestine of suckling rats: organ culture study

Dvořák, Bohuslav; Kolínská, J; McWilliam, Debra L.; Williams, Catherine S.; Higdon, Travis A.; Zákostelecká, Marie; Koldovský, Otakar

doi:10.1016/s0014-5793(98)01050-3

Cited by 19 publications

(3 citation statements)

References 47 publications

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“…Single-stranded cDNA was reverse-transcribed as previously described in detail [27]. The amounts of total RNA used in the RT reactions were calculated from the absorbency at 260 nm, and verified by densitometry of the 28S ribosomal RNA band separated on denaturing agarose gels (by Gel Doc 1000 Documentation System with Molecular Analyst/PC software, BIO-RAD, Hercules, CA).…”

Section: Methodsmentioning

confidence: 99%

Active Transport of Bile Acids Decreases Mucin 2 in Neonatal Ileum: Implications for Development of Necrotizing Enterocolitis

et al. 2011

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Necrotizing enterocolitis (NEC) is the most common gastrointestinal emergency of premature infants, but its etiology remains unclear. We have previously shown that mucin 2 (Muc2) positive goblet cells are significantly decreased in NEC. We have also shown that ileal bile acids (BAs) are significantly increased during the development of this disease. Because BAs can affect mucins, we hypothesized that elevated ileal BAs contribute to decreased Muc2 in experimental NEC. The role of Muc2 in NEC was evaluated in Winnie +/+ mice, a strain that produces aberrant Muc2. Muc2 and trefoil factor 3 (Tff3) were assessed in neonatal rats subjected to the NEC protocol when bile acids were removed, and in ileal explants from newborn and older rats cultured with and without BAs. Further, the role of active transport of BAs was determined using neonatal rats given the apical sodium dependent bile acid transporter (Asbt) inhibitor SC-435 and in neonatal Asbt knockout mice subjected to the NEC protocol. Mice with aberrant Muc2 had significantly greater incidence and severity of NEC. Using both in vivo and ex vivo techniques, we determined that BAs decrease Muc2 positive cells in neonatal but not older ileum. However, Tff3 positive cells are not decreased by BAs. In addition, active transport of BAs is required for BAs to decrease Muc2 in immature ileum. These data show that functional Muc2 plays a critical role in the prevention of NEC and BAs can potentiate the decreased Muc2 in disease development. Further, BAs have a more profound effect on Muc2 in immature versus older ileum, which may explain at least in part why NEC occurs almost exclusively in premature infants.

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Section: Methodsmentioning

confidence: 99%

Active Transport of Bile Acids Decreases Mucin 2 in Neonatal Ileum: Implications for Development of Necrotizing Enterocolitis

et al. 2011

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“…RT real‐time PCR assays were performed to specifically quantify rat interleukin‐18, interleukin‐12, and interleukin‐10 steady state mRNA concentrations. Single‐stranded complementary DNA was reverse transcribed from 1 μg of total RNA in a 10‐μL reaction mixture, as previously described in detail (31). The amounts of total RNA used in the RT reactions were calculated from the absorbency at 260 nm and verified by densitometry of the 28S ribosomal RNA band separated on denaturing agarose gels (by Gel Doc 1000 Documentation System with Molecular Analyst/PC software; BIO‐RAD, Hercules, CA, U.S.A.).…”

Section: Methodsmentioning

confidence: 99%

“…The RT competitive-PCR assay was used to quantify intestinal Sp1 and STAT5 mRNA concentrations as previously described in detail (31,35). The sequences of primers used were as follows: Rat Sp1 sense primer (5Ј-AGAACCGC-ACAGTCTCTGGT-3Ј corresponding to nucleotides 470-489) and antisense primer (5Ј-GGGACAGCTTGCTGGAGTAG-3Ј corresponding to nucleotides 748-767) (36); rat Stat5 sense primer (5Ј-CTGCAGTCCTGGTGTGAGAA-3Ј corresponding to nucleotides 933-952) and antisense primer (5Ј-GATGAT-GGTTGCCTTCACCT-3Ј corresponding to nucleotides 1213-1232) (37).…”

Section: Competitive Polymerase Chain Reactionmentioning

confidence: 99%

Ileal Cytokine Dysregulation in Experimental Necrotizing Enterocolitis Is Reduced by Epidermal Growth Factor

Halpern,

Dominguez,

Dvorakova

et al. 2003

J. pediatr. gastroenterol. nutr.

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BackgroundNecrotizing enterocolitis (NEC) is the most common gastrointestinal disease of premature infants. We have shown in previous studies that proinflammatory interleukin‐18 and interleukin‐12 are up‐regulated in the ileum of rats with experimental NEC and that epidermal growth factor (EGF) reduces the development of disease. Here we investigated whether the protective effects of EGF are a result of changes in ileal interleukin‐18, interleukin‐12 and/or antiinflammatory interleukin‐10.MethodsNewborn rats were artificially fed with either growth‐factor–free rat milk substitute (RMS) or RMS supplemented with 500 ng/mL EGF (RMS + EGF) and NEC was induced via exposure to asphyxia and cold stress. Cytokine expression and localization were assessed using reverse‐transcription real‐time polymerase chain reaction and immunohistology/confocal microscopy.ResultsEnteral administration of EGF (RMS + EGF) decreased overproduction of interleukin‐18 and increased interleukin‐10 production in the ileum. Furthermore, increased interleukin‐10 production was associated with up‐regulation of the transcription factor Sp1 in RMS + EGF rats.ConclusionsThese data suggest that EGF may reduce NEC via increased interleukin‐10 and decreased interleukin‐18 and that EGF‐mediated up‐regulation of Sp1 may account for the increased interleukin‐10.

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Growth Factors in the Gastrointestinal Tract

Kuemmerle¹,

Barnard²,

McHugh³

2012

Physiology of the Gastrointestinal Tract

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The expression of epidermal growth factor and transforming growth factor‐α mRNA in the small intestine of suckling rats: organ culture study

Cited by 19 publications

References 47 publications

Active Transport of Bile Acids Decreases Mucin 2 in Neonatal Ileum: Implications for Development of Necrotizing Enterocolitis

Active Transport of Bile Acids Decreases Mucin 2 in Neonatal Ileum: Implications for Development of Necrotizing Enterocolitis

Ileal Cytokine Dysregulation in Experimental Necrotizing Enterocolitis Is Reduced by Epidermal Growth Factor

Growth Factors in the Gastrointestinal Tract

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