In vitro study of transporters involved in intestinal absorption of inorganic arsenic.
Inorganic arsenic (iAs) [As(III)+As(V)] is a drinking water contaminant, and human exposure to these arsenic species has been linked with a wide range of health effects. The main path of exposure is the oral route, and the intestinal epithelium is the first physiological barrier that iAs must cross in order to be absorbed. However, there is a lack of information about intestinal iAs absorption. The aim of this study was to evaluate the participation of certain transporters [glucose transporters (GLUT and SGLT), organic anion transporting polypeptides (OATPs), aquaporins (AQPs), and phosphate transporters (NaPi and PiT)] in intestinal absorption of As(V) and As(III), using the Caco-2 cell line as a model of the intestinal epithelium. For this purpose, the effects of chemical inhibition and gene silencing of the transporters of interest on iAs uptake were evaluated, and also the differential expression of these transporters after treatment with iAs. The results show that chemical inhibition using rifamycin SV (OATP inhibitor), phloridzin (SGLT inhibitor), phloretin (GLUT and AQP inhibitor), and copper sulfate (AQP inhibitor) leads to a significant reduction in the apparent permeability and cellular retention of As(III). RT-qPCR indicates up-regulation of GLUT2, GLUT5, OATPB, AQP3, and AQP10 after exposure to As(III), while exposure to As(V) increases the expression of sodium-dependent phosphate transporters, especially NaPiIIb. Gene silencing of OATPB, AQP10, and GLUT5 for As(III) and NaPiIIb for As(V) significantly reduces uptake of the inorganic forms. These results indicate that these transporters may be involved in intestinal absorption of iAs.
Instituto de Agroquímica y Tecnología de Alimentos (CSIC) , Av. Agustín Escardino, 7, 46980 Paterna, Valencia, Spain.
Marta
M
Calatayud
Julio A
JA
Barrios
Dinoraz
D
Vélez
Vicenta
V
Devesa
D001151:Arsenic / Q000378:metabolism*; D018938:Caco-2 Cells; D002273:Carcinogens / Q000378:metabolism*; D019327:Copper Sulfate / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D020868:Gene Silencing; D006801:Humans; D007408:Intestinal Absorption; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D010539:Permeability / Q000187:drug effects; D010693:Phloretin / Q000494:pharmacology; D010695:Phlorhizin / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000235:genetics; D012294:Rifamycins / Q000494:pharmacology; D014874:Water Pollutants, Chemical / Q000378:metabolism*
D002273:Carcinogens; D026901:Membrane Transport Proteins; D034741:RNA, Small Interfering; D012294:Rifamycins; D014874:Water Pollutants, Chemical; D010695:Phlorhizin; C023808:rifamycin SV; D019327:Copper Sulfate; D001151:Arsenic; D010693:Phloretin