• Asian-Australasian Journal of Animal Sciences
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• v.22 no.8
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• pp.1160-1166
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• 2009

This study was conducted according to the single-factor design principle to investigate in vitro the effects of different glucagon-like peptide-2 (GLP-2) concentrations (0, $1{\times}10^{-11}$, $1{\times}10^{-10}$, $1{\times}10^{-9}$, $1{\times}10^{-8}$ and $1{\times}10^{-7}$ mol/L) on the morphology, proliferation and enzyme activity of intestinal enterocyte cells of 28-d-old weaned piglets. These cells were primary cultured in 4 pieces of 24-well cell culture plate. After having been grown for 48 h in culture media with hGLP-2, the ileal enterocyte cells of 28-d-old weaned piglets exhibited the typical characteristics of simple columnar epithelium. Compared with the control groups, the quantities of treated cells significantly increased (p<0.05) and their corresponding absorption values in 540 nm (MTT OD) also significantly increased (p<0.01). Likewise, lactic acid concentration, total protein content and protein retention significantly increased (p<0.05). $Na^{+}$, $K^{+}$-ATP enzyme activity was more active (p<0.05), although the activity of alkaline phosphatase, lactic acid dehydrogenase and creatine phosphokinase in culture media significantly decreased (p<0.01). To summarize, the results indicated that GLP-2 in vitro is capable of promoting the proliferation of intestinal enterocyte cells of 28-d weaned piglets, restraining their apoptosis and maintaining the integrity of their morphology.

• Proceedings of the Korean Nutrition Society Conference
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• 1995.11b
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• pp.11-34
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• 1995

Growth hormone (GH) plays a key role in regulating postnatal growth and can stimulate growth of animals by acting directly on specific receptors on the plasma membrane of tissues or indirectly through stimulating insulin-like growth factor (IGF)-I synthesis and secretion by the liver and other tissues. IGF-I and IGF-Ⅱ are polypeptides with structural similarity with proinsulin that stimulate cell proliferation by endocrine, paracrine and autocrine mechanisms. The initial event in the metabolic action of IGFs on target cells appears to be their binding to specific receptors on the plasma membrane. Current evidence indicates that the mitogenic actions of both IGFs are mediated primarily by binding to the type I IGF receptors, and that IGF action is also mediated by interactions with IGF-binding proteins (IGFBPs). Six distinct IGFBPs have been identified that are characterized by cell-specific interaction, transcriptional and post-translational regulation by many different effectors, and the ability to either potentiate or inhibit IGF actions. Nutritional deficiencies can have their devastating consequence during growth. Although IGF-I is the major mediator of GH's action on somatic growth, nutritional status of an organism is a critical regulator of IGF-I and IGFBPs. Various nutrient deficiencies result in decreased serum IGF-I levels and altered IGFBP levels, but the blood levels of GH are generally unchanged or elevated in malnutrition. Effects of protein, energy, vitamin C and D, and zinc on serum IGF and IGFBP levels and tissue mRNA levels were reviewed in the text. Multiple factors are involved in the regulation of intestinal epithelial cell growth and differentiation. Among these factors the nutritional status of individuals is the most important. The intestinal epithelium is an important site for mitogenic action of the IGFs in vivo, with exogenous IGF-I stimulating mucosal hyperplasia. Therefore, the IGF system appears to provide and important mechanism linking nutrition and the proliferation of intestinal epithelial cells. In order to study the detailed mechanisms by which intestinal mucosa is regulated, we have utilized IEC-6 cells, an intestinal epithelial cell line and Caco-2 cells, a human colon adenocarcinoma cell line. Like intestinal crypt cells analyzed in vivo or freshly isolated intestinal epithelial cells, IEC-6 cells and Caco-2 cells possess abundant quatities of both type Ⅰ and type Ⅱ IGF receptors. Exogenous IGFs stimulate, whereas addition of IGFBP-2 inhibits IEC-6 cell proliferation. To investigate whether endogenously secreted IGFBP-2 inhibit proliferation, IEC-6 cells were transfected with a full-length rat IGFBP-2 cDNA anti-sense expression construct. IEC-6 cells transfected with anti-sense IGFBP-2 protein in medium. These cells grew at a rate faster than the control cells indicating that endogenous IGFBP-2 inhibits proliferation of IEC-6 cells, probably by sequestering IGFs. IEC-6 cells express many characteristics of enterocyte, but do not undergo differentiation. On the other hand, Caco-2 cells undergo a spontaneous enterocyte differentiation. On the other hand, Caco-2 cells undergo a spontaneous enterocyte differentiation after reaching confluency. We have demonstrated that Caco-2 cells produce IGF-Ⅱ, IGFBP-2, IGFBP-3, and an as yet unidentified 31,000 Mr IGFBP, and that both mRNA and peptide secretion of IGFBP-2 and IGFBP-3 increased, but IGFBP-4 mRNA and protein secretion decreased after the cells reached confluency. These changes occurred in parallel to and were coincident with differentiation of the cells, as measured by expression of sucrase-isomaltase. In addition, Caco-2 cell clones forced to overexpress IGFBP-4 by transfection with a rat IGFBP-4 cDNA construct exhibited a significantly slower growth rate under serum-free conditions and had increased expression of sucrase-isomaltase compared with vector control cells. These results indicate that IGFBP-4 inhibits proliferation and stimulates differentiation of Caco-2 cells, probably by inhibiting the mitogenic actions of IGFs.

• Korean Journal of Veterinary Research
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• v.34 no.4
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• pp.805-813
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• 1994

Immunohistochemical study on the intestinal tissues obtained from the 21 pigs of the 14 terms in Korea in which the clinical and epidemiological features had indicated the possible outbreaks of porcine epidemic diarrhea(PED) was performed using the indirect immunofluorescence test and/or the immunoperoxidase method in order to detect PED viral antigens in the infected cells of the intestines, and histopathological features were described as well. By immunohistochemical analysis, PED viral antigens were detected in the epithelial cells covering the small intestinal villi and recognized slightly in the cells lining the colonic surface epithelium as well. Occasional fluorescence was also seen in a few intestinal crypt epithelium. On light microscopy, the piglets with PED showed marked villous atrophy and fusion, and severe enterocyte degeneration and desquamation. On the other hand, the older pigs more than 4 week old age was mild villous atrophy and fusion, severe villous epithelial cell proliferation, and moderate mononuclear cell infiltration.

• Journal of Microbiology and Biotechnology
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• v.13 no.2
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• pp.276-281
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• 2003

The adhesion of probiotic bacteria to the intestinal mucosa is one of the desirable properties for their colonization in the intestinal tract, where these bacteria constantly compete with other bacteria. The adhesion of different strains of bifidobacteria to Caco-2 cells was compared. Among the strains examined, BGN-4 showed the highest adhesion level and the greatest cell surface hydrophobicity (CSH). No close relationship was found between the adhesion and CSH of the strains. Upon protease and heat treatment, the adhesion of the BGN-4 to the Caco-2 cells decreased significantly. The cells grown at $42^{\circ}C$ showed a lower CSH and self-aggregation levels than cells grown at $37^{\circ}C$. The treatment of EGTA did not have any effect on the adhesion. The degree of adhesion did not differ among the experimental groups in which galactose, mannose, or fucose were added in the adhesion assay mixture. The results suggest that the adhesion of the Bifidobacterium to the epithelial cells may be affected by the composition and structure of the cell membrane and interacting surfaces.

• Molecules and Cells
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• v.41 no.4
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• pp.290-300
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• 2018

Using an in vitro model of intestinal organoids derived from intestinal crypts, we examined effects of indole-3-carbinol (I3C), a phytochemical that has anticancer and aryl hydrocarbon receptor (AhR)-activating abilities and thus is sold as a dietary supplement, on the development of intestinal organoids and investigated the underlying mechanisms. I3C inhibited the in vitro development of mouse intestinal organoids. Addition of ${\alpha}$-naphthoflavone, an AhR antagonist or AhR siRNA transfection, suppressed I3C function, suggesting that I3C-mediated interference with organoid development is AhR-dependent. I3C increased the expression of Muc2 and lysozyme, lineage-specific genes for goblet cells and Paneth cells, respectively, but inhibits the expression of IAP, a marker gene for enterocytes. In the intestines of mice treated with I3C, the number of goblet cells was reduced, but the number of Paneth cells and the depth and length of crypts and villi were not changed. I3C increased the level of active nonphosphorylated ${\beta}$-catenin, but suppressed the Notch signal. As a result, expression of Hes1, a Notch target gene and a transcriptional repressor that plays a key role in enterocyte differentiation, was reduced, whereas expression of Math1, involved in the differentiation of secretory lineages, was increased. These results provide direct evidence for the role of AhR in the regulation of the development of intestinal stem cells and indicate that such regulation is likely mediated by regulation of Wnt and Notch signals.

• Korean Journal of Veterinary Pathology
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• v.3 no.1
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• pp.35-44
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• 1999

The morphologic changes of small intestinal epithelium in pigs diagnosed as porcine epidemic diarrhea(PED} by virus isolation and immunohistochemistry were studied through light microscope and transmissible electron microscope. On semi-thin section, the histologic findings showed severe villous atrophy and fusion with hyperplasia of cuboidal epithelium in the villi, inflammatory cell infiltration in lamina propria, and increased mitotic figures in the crypt. The structural changes were mostly restricted to the cytoplasm of affected absorptive epithelium of villi. 3 types of epithelial changes were found; degenerated virus-affected cells, undifferentiated cuboidal cells, and normal columnar cells. On electron microscopy, round to spherical viral particles of 50∼l00nm in diameter were found within the dilated vesicles and endoplasmic reticulums of degenerated cells, which had decreased their cytoplasmic electron density due to dilated and missing organelles(e.g. mitochondria, ERs, etc.). Microvilli were shortened and sparse, leaving denuded terminal web of the villous epithelial cells. Fat globules were often found within slightly degenerated enterocytes. On the tip of villi, severely damaged cells were exfoliated and replaced by undifferentiated cuboidal cells We found distinct ultrastructural changes in the jejunal epithelium confirming PED virus infection is involved in malabsorptive diarrhea.

• Journal of Plant Biotechnology
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• v.36 no.3
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• pp.268-274
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• 2009

Porphyromonas gingivalis, the gram-negative anaerobic oral bacterium, initiates periodontal disease by binding to saliva-coated oral surface. The cholera toxin B subunit (CTB) genetically linked to FimA1 (1-200 aa) or FimA2 (201-337 aa) of the P. gingivalis fimbrial antigen were introduced into Solanum tuberosum cells by Agrobacterium tumefaciens-mediated transformation method. The integration of CTB-FimA1 or CTB-FimA2 fusion genes were confirmed in the chromosome of transformed leaves by genomic DNA PCR amplification method. Synthesis and assembly of the CTB-FimA fusion proteins into oligomeric structures with pentamer size was detected in transformed tuber extracts by immunoblot analysis. The binding activities of CTB-FimA fusion proteins to intestinal epithelial cell membrane receptors were confirmed by GM1-ganglioside enzyme-linked immunosorbent assay (GM1-ELISA). The ELISA showed that the expression levels of the CTB-FimA1 or CTB-FimA2 fusion proteins were 0.0019, 0.002% of the total soluble protein in transgenic tuber tissues, respectively The synthesis of CTB-FimA monomers and their assembly into biologically active oligomers in transformed potato tuber tissues demonstrates the feasibility of using edible plants for the production of enterocyte targeted fimbrial antigens that could elicit mucosal immune responses.