Effect of hydrogen peroxide on interleukin-8 synthesis and death of Caco-2 cells.
Intestinal epithelial cells can secrete interleukin-8 (IL-8), among other substances in response to different stimuli, which plays an important role in mucosal immune response. Above a certain concentration range, hydrogen peroxide causes cell death by necrosis or apoptosis. We investigated the time- and dose-dependent induction of IL-8 by hydrogen peroxide in the human colon adenocarcinoma cell line Caco-2. In addition, the changes of transepithelial electrical resistance and cell death induction in response to hydrogen peroxide were studied. Nonfilter-grown and filter-grown Caco-2 cells were employed in our experiments. Interleukin-8 synthesis was measured by ELISA. Necrosis was determined by DAPI staining of cells, apoptosis by measuring caspase-3 enzyme activity or annexin V staining. In nonfilter-grown Caco-2 cells, 1 mM of hydrogen peroxide induced the highest level of IL-8 production 24 hr after treatment. In filter-grown Caco-2 cells, IL-8 was produced only on the apical side in response to 1 mM of hydrogen peroxide. This level was 10-fold lower than that measured in nonfilter-grown Caco-2 cells 24 hr after the treatment. In filter-grown Caco-2 cells 10 mM hydrogen peroxide induced the highest IL-8 level on the apical as well as basolateral side. Transepithelial electrical resistance decreased markedly upon application of 40 mM hydrogen peroxide. Late effect of hydrogen peroxide was observed in nonfilter-grown Caco-2 cells, as 1 mM hydrogen peroxide caused necrosis after 24 hr while early-necrosis induction occurred in filter-grown cells exposed to 40 mM of hydrogen peroxide after 1 hr. Filter-grown Caco-2 cells were less sensitive to hydrogen peroxide than the nonfilter-grown ones.
Németh
Edina
E
Department of Biology, Central Food Research Institute, Budapest, Hungary. nemeth.edina@aotk.szie.hu
Halász
Anna
A
Baráth
Agnes
A
Domokos
Mónika
M
Gálfi
Péter
P
D018938:Caco-2 Cells; D053148:Caspase 3 / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D017097:Electric Impedance; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D016209:Interleukin-8 / Q000096:biosynthesis*; D009336:Necrosis / Q000473:pathology; D018384:Oxidative Stress / Q000187:drug effects
D016209:Interleukin-8; D006861:Hydrogen Peroxide; D053148:Caspase 3