Immunomodulatory activity of enzymatically synthesized glycogen and its digested metabolite in a co-culture system consisting of differentiated Caco-2 cells and RAW264.7 macrophages

Michiko Yasuda, Takashi Furuyashiki, Toshiyuki Nakamura, Ryo Kakutani, Hiroki Takata, Hitoshi Ashida

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Previously, we developed enzymatically synthesized glycogen (ESG) from starch, and showed its immunomodulatory and dietary fiber-like activities. In this study, we investigated the metabolism of ESG and its immunomodulatory activity using differentiated Caco-2 cells as a model of the intestinal barrier. In a co-culture system consisting of differentiated Caco-2 cells and RAW264.7 macrophages, mRNA expression of IL-6, IL-8, IL-1β and BAFF cytokines was up-regulated in Caco-2 cells and IL-8 production in basolateral medium was induced after 24 h apical treatment with 5 mg ml-1 of ESG. The mRNA level of iNOS was also up-regulated in RAW264.7 macrophages. After characterization of the binding of anti-glycogen monoclonal antibodies (IV58B6 and ESG1A9) to ESG and its digested metabolite resistant glycogen (RG), an enzyme-linked immunosorbent assay (ELISA) system was developed to quantify ESG and RG. Using this system, we investigated the metabolism of ESG in differentiated Caco-2 cells. When ESG (7000 kDa, 5 mg ml-1) was added to the apical side of Caco-2 monolayers, ESG disappeared and RG (about 3000 kDa, 3.5 mg ml-1) appeared in the apical solution during a 24 h incubation. Neither ESG nor RG was detected in the basolateral solution. In addition, both ESG and RG were bound to TLR2 in Caco-2 cells. In conclusion, we suggest that ESG is metabolized to a RG-like structure in the intestine, and this metabolite activates the immune system via stimulation of the intestinal epithelium, although neither ESG nor its metabolite could permeate the intestinal cells under our experimental conditions. These results provide evidence for the beneficial function of ESG as a food ingredient.

Original languageEnglish
Pages (from-to)1387-1393
Number of pages7
JournalFood and Function
Volume4
Issue number9
DOIs
Publication statusPublished - Sep 2013
Externally publishedYes

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Caco-2 Cells
coculture
Coculture Techniques
Glycogen
glycogen
macrophages
Macrophages
metabolites
cells
interleukin-8
Interleukin-8
Messenger RNA
metabolism

ASJC Scopus subject areas

  • Food Science
  • Medicine(all)

Cite this

Immunomodulatory activity of enzymatically synthesized glycogen and its digested metabolite in a co-culture system consisting of differentiated Caco-2 cells and RAW264.7 macrophages. / Yasuda, Michiko; Furuyashiki, Takashi; Nakamura, Toshiyuki; Kakutani, Ryo; Takata, Hiroki; Ashida, Hitoshi.

In: Food and Function, Vol. 4, No. 9, 09.2013, p. 1387-1393.

Research output: Contribution to journalArticle

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