Effects of absorption enhancers on cytoskeletal actin filaments in Caco-2 cell monolayers.
Histomorphological changes of actin filaments, intracellular levels of calcium ion, and amount of released lactate dehydrogenase (LDH) were examined, in order to elucidate the mechanism of action of three absorption enhancers, i.e., sodium caprate (Cap-Na), sodium deoxycholate (Deo-Na), and dipotassium glycyrrhizinate (Grz-K), using Caco-2 cell monolayers. The structure of actin filaments in microvilli was slightly modified by 0.5 %(w/v) Grz-K and was significantly changed by 0.2 %(w/v) Cap-Na, 0.05 %(w/v) Deo-Na, and 0.0015 %(w/v) ionomycin. All of the enhancers, except Grz-K, induced significantly histomorphological changes in the actin filaments on the middle depth and basal side of the cells. Furthermore, the altered structure of the actin filaments in the monolayers was restored after removal of the Cap-Na, Grz-K and ionomycin, but not Deo-Na. Intracellular levels of calcium ion increased in the following order: ionomycin = Cap-Na > Deo-Na. However, the intracellular calcium ion levels decreased by treatment with Grz-K. The changes in transepithelial electric resistance (TEER) at the initial stage of treatment with all enhancers correlated with intracellular calcium ion levels. These results suggest that one of the mechanisms by which these agents exert absorption-enhancing activity involves structural alterations in the cytoskeletal actin filaments which are provoked by changes in intracellular calcium ion levels. Only the monolayers which were treated with 0.05 %(w/v) Deo-Na released a significant amount of LDH and irreversibly altered the structure of actin filaments, thus indicating that Deo-Na might affect the actin filaments not only by increasing intracellular calcium ion level but also by other, presently unknown factors.
Sakai
M
M
Hisamitsu Pharmaceutical Co., Inc. Tsukuba Laboratories, Ibaraki, Japan.
Imai
T
T
Ohtake
H
H
Azuma
H
H
Otagiri
M
M
D000199:Actins / Q000737:chemistry / Q000187:drug effects*; D000890:Anti-Infective Agents / Q000009:adverse effects / Q000494:pharmacology; D018938:Caco-2 Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D002118:Calcium / Q000378:metabolism*; D002756:Cholagogues and Choleretics / Q000009:adverse effects / Q000494:pharmacology; D003599:Cytoskeleton; D003652:Decanoic Acids / Q000009:adverse effects / Q000494:pharmacology; D003840:Deoxycholic Acid / Q000009:adverse effects / Q000494:pharmacology; D019695:Glycyrrhizic Acid / Q000009:adverse effects / Q000494:pharmacology; D006801:Humans
D000199:Actins; D000890:Anti-Infective Agents; D002756:Cholagogues and Choleretics; D003652:Decanoic Acids; D003840:Deoxycholic Acid; C031071:decanoic acid; D019695:Glycyrrhizic Acid; D002118:Calcium