Sodium cromoglycate inhibits absorption of the major soybean allergen, Gly m Bd 30K, in mice and human intestinal Caco-2 cells.
Our previous data showed that Gly m Bd 30K was absorbed from the gastrointestinal tract and circulated in blood in mice. This study was conducted to determine the mechanism and identify the inhibitor of such absorption. Using sandwich ELISA and immunoblotting, we found that intact Gly m Bd 30K was absorbed from apical to basolateral solutions and intracellularly accumulated by Caco-2 cells in a dose- and time-dependent manner. The absorption and intracellular accumulation of Gly m Bd 30K were significantly suppressed when Caco-2 cells were treated with sodium cromoglycate (SCG) (0-50 mmol/L) in a dose-dependent manner. In 24-d-old mice orally treated with SCG (10-1000 mg/kg body weight), plasma Gly m Bd 30K concentration decreased significantly 30-120 min after Gly m Bd 30K (2000 mg/kg body weight) administration. Moreover, inhibitors that suppress the clathrin-dependent endocytosis dansylcadaverine, the caveolae-dependent endocytosis nystatin and clathrin, and the caveolae-dependent endocytosis methyl-beta-cyclodextrin had inhibitory effects on the absorption and intracellular accumulation of Gly m Bd 30K by Caco-2 cells. These data indicate that Gly m Bd 30K is absorbed and intracellularly accumulated in Caco-2 cells via clathrin- or caveolae-dependent endocytosis. We propose that the absorption and intracellular accumulation of Gly m Bd 30K are inhibited by SCG via clathrin- or caveolae-dependent endocytosis.
Weangsripanaval
Thanakorn
T
Laboratory of Molecular Function of Food, Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Uji, Kyoto, Japan.
Murota
Kaeko
K
Murakami
Yoko
Y
Kominami
Masaru
M
Kusudo
Tatsuya
T
Moriyama
Tatsuya
T
Ogawa
Tadashi
T
Kawada
Teruo
T
D000469:Alkaline Phosphatase / Q000235:genetics; D000485:Allergens / Q000493:pharmacokinetics*; D000818:Animals; D052179:Antigens, Plant; D018938:Caco-2 Cells; D004205:Cromolyn Sodium / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D017097:Electric Impedance; D004705:Endocytosis; D006801:Humans; D007408:Intestinal Absorption / Q000187:drug effects*; D008353:Mannitol / Q000493:pharmacokinetics; D051379:Mice; D012333:RNA, Messenger / Q000032:analysis; D030262:Soybean Proteins / Q000493:pharmacokinetics*
D000485:Allergens; D052179:Antigens, Plant; C103236:Gly m BD 30K protein, Glycine max; D012333:RNA, Messenger; D030262:Soybean Proteins; D008353:Mannitol; D000469:Alkaline Phosphatase; D004205:Cromolyn Sodium