Monochloramine-induced toxicity and dysregulation of intracellular Zn2+ in parietal cells of rabbit gastric glands.
Monochloramine (NH(2)Cl) is a potent, thiol-directed oxidant capable of oxidizing thiol (S-H) residues in a wide variety of proteins. Generated in the stomach by the interaction of bacterial and host products, monochloramine has been shown to dysregulate Ca(2+) homeostasis and disrupt mucosal integrity. In this report, we show that monochloramine also leads to disturbances in intracellular free zinc concentration ([Zn(2+)](i)) in the gastric gland of the rabbit and that the increased Zn(2+) within the cell causes an independent decrease in cell viability. Changes in [Zn(2+)](i) were measured by using the fluorescent reporter FluoZin-3, whereas cell viability was assessed by measuring the conversion of calcein-AM to fluorescent calcein, an assay that is not affected by intracellular oxidation state. Cell death was confirmed using propidium iodide and YO-PRO-1 dye uptake measurements. Our experiments demonstrate that [Zn(2+)](i) is increased in gastric glands exposed to NH(2)Cl and that elevated [Zn(2+)](i) decreases cell viability. Chelation of Zn(2+) with tetrakis-(2-pyridylmethyl) ethylenediamine decreases the toxicity of NH(2)Cl, but only when administered concurrently. These findings suggest that the toxic effect of thiol oxidants present during chronic gastritis is partially due to dysregulation of [Zn(2+)](i) early in the process and that zinc chelation can protect, but not rescue, gastric glands exposed to toxic doses of NH(2)Cl.
Kohler
Jonathan E
JE
Department of Surgery, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts 02115, USA.
Dubach
J Matthew
JM
Naik
Haley B
HB
Tai
Kaniza
K
Blass
Amy L
AL
Soybel
David I
DI
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D001205:Ascorbic Acid / Q000494:pharmacology; D015374:Biosensing Techniques; D002470:Cell Survival / Q000187:drug effects; D002614:Chelating Agents / Q000494:pharmacology; D002700:Chloramines / Q000633:toxicity*; D019610:Cytoprotection; D004229:Dithiothreitol / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005029:Ethylenediamines / Q000494:pharmacology; D005753:Gastric Mucosa / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006706:Homeostasis; D008856:Microscopy, Fluorescence; D016877:Oxidants / Q000633:toxicity*; D010084:Oxidation-Reduction; D010295:Parietal Cells, Gastric / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D011083:Polycyclic Compounds; D011817:Rabbits; D013997:Time Factors; D015032:Zinc / Q000378:metabolism*
D000975:Antioxidants; D002614:Chelating Agents; D002700:Chloramines; D005029:Ethylenediamines; C451182:FluoZin-3; D016877:Oxidants; D011083:Polycyclic Compounds; D015032:Zinc; C030816:chloramine; D001205:Ascorbic Acid; C044387:N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine; D004229:Dithiothreitol