title: Experimental exposure of arsenic in cultured rat intestinal epithelial cells and cell line: toxicological consequences.
abstract: Arsenic is a naturally occurring metalloid and the drinking water contamination by inorganic arsenic remains a major public health problem. The trivalent arsenic (arsenite) is more toxic than the pentavalent form (arsenate), and is known to cause gastrointestinal toxicity. Specific immortal cell lines are considered to be suitable for toxicity screening and testing of chemicals as they are easy to handle and possess most of the biochemical pathways present in the corresponding cells present in vivo. The present study was designed to evaluate and compare the in vitro toxicity of arsenite on rat intestinal epithelial cell line (IEC-6) and primary cultures of rat intestinal epithelial cells (IEC). To evaluate in vitro toxicity, cultures of IEC and IEC-6 cells were assessed for viability, morphometric analysis, membrane transport enzymes and structural constituents for membrane damage, dehydrogenase activity test for respiratory and energy producing processes and esterase activity test for intra and extra cellular degradation, following the post exposures to arsenite (0-20 ppm). Significantly similar concentration-dependent changes in these toxicity-screening parameters in IEC and IEC-6 were observed. Highest tested concentration of arsenite (20 ppm) was found to be detrimental in both IEC and IEC-6. Furthermore, to evaluate arsenite toxicity in epithelial cells of rat intestine, intestinal loops were filled with arsenite solutions and incubated for 30 min in situ. In situ studies also showed a significant arsenite concentration-dependent decline in epithelial cell membrane transport enzyme activities and total hexose and sialic acid contents. Concomitant release of membrane enzymes, hexose and sialic acid in the intestinal luminal fluid following higher arsenite exposures further indicated partial membrane damage. Similar morphological changes in IEC and IEC-6 were also evident. These findings also suggest that IEC-6 cell lines are suitable for initial screening of gastrointestinal cellular toxicity caused by arsenite.
authors.affiliation: Industrial Toxicology Research Centre, Biomembrane Toxicology Division, Mahatma Gandhi Marg, P.O. Box 80, Lucknow 226001, India. upretirk@rediffmail.com
authors.forename: Raj K
authors.initials: RK
authors.lastname: Upreti
authors.forename: A
authors.initials: A
authors.lastname: Kannan
authors.forename: A B
authors.initials: AB
authors.lastname: Pant
mesh_terms: D000818:Animals; D001151:Arsenic / Q000633:toxicity*; D018053:Arsenites / Q000633:toxicity; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002461:Cell Line, Transformed; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology; D048430:Cell Shape / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000187:drug effects* / Q000648:ultrastructure; D004950:Esterases / Q000378:metabolism; D007202:Indicators and Reagents; D007413:Intestinal Mucosa / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D007700:Kinetics; D008297:Male; D010088:Oxidoreductases / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013778:Tetrazolium Salts; D013844:Thiazoles
chemical_list: D018053:Arsenites; D007202:Indicators and Reagents; D013778:Tetrazolium Salts; D013844:Thiazoles; D010088:Oxidoreductases; D004950:Esterases; C022616:thiazolyl blue; D001151:Arsenic