Pectin from Prunus domestica L. induces proliferation of IEC-6 cells through the alteration of cell-surface heparan sulfate on differentiated Caco-2 cells in co-culture

• Published in: Glycoconjugate journal Vol. 32; no. 3-4; pp. 153 - 159
• Main Authors: Nishida, Mitsutaka, Murata, Kazuma, Oshima, Kazuya, Itoh, Chihiro, Kitaguchi, Kohji, Kanamaru, Yoshihiro, Yabe, Tomio
• Format: Journal Article
• Language: English
• Published: New York Springer US 01.05.2015; Springer Nature B.V
• Subjects: Animals; Biochemistry; Biomedical and Life Sciences; Caco-2 Cells - drug effects; Cell Differentiation - drug effects; Cell Line - drug effects; Cell Proliferation - drug effects; Cellular biology; Coculture Techniques; Dietary fiber; Epithelial Cells - cytology; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Heparitin Sulfate - chemistry; Heparitin Sulfate - metabolism; Humans; Intestinal Mucosa - cytology; Intestinal Mucosa - metabolism; Life Sciences; Original Article; Pathology; Pectins - pharmacology; Prunus domestica - chemistry; Rats; Studies; Surface Plasmon Resonance; Wnt Proteins - genetics; Wnt Proteins - metabolism; Wnt3A Protein - secretion; Heparan sulfate; Pectin; IEC-6 cell; Wnt3a; Differentiated Caco-2 cell
• ISSN: 0282-0080; 1573-4986
• DOI: 10.1007/s10719-015-9588-4

Dietary fiber intake provides various physiological and metabolic effects for human health. Pectin, a water-soluble dietary fiber, induces morphological changes of the small intestine in vivo . However, the molecular mechanisms underlying pectin-derived morphological alterations have not been elucidated. Previously, we found that pectin purified from Prunus domestica L. altered the sulfated structure of cell-surface heparan sulfate (HS) on differentiated Caco-2 cells via fibronectin and α5β1 integrin. In this study, we investigated the biological significance of the effect of pectin on HS in differentiated Caco-2 cells. An in vitro intestinal epithelium model was constructed by co-culture of differentiated Caco-2 cells and rat IEC-6 cells, which were used as models of intestinal epithelium and intestinal crypt cells, respectively. We found that pectin-treated differentiated Caco-2 cells promoted growth of IEC-6 cells. Real-time RT-PCR analysis and western blotting showed that relative mRNA and protein expression levels of Wnt3a were upregulated by pectin treatment in differentiated Caco-2 cells. Analysis by surface plasmon resonance spectroscopy demonstrated that pectin-induced structural alteration of HS markedly decreased the interaction with Wnt3a. However, depression in the secretion of Wnt3a from Caco-2 cells by anti-Wnt3a antibody did not affect the proliferation of IEC-6 cells in co-culture system. These observations indicated that pectin altered the sulfated structure of cell-surface HS to promote secretion of Wnt3a from differentiated Caco-2 cells and Wnt3a indirectly stimulated the proliferation of IEC-6 cells.