Nitric oxide generation mediates lipid A-induced oxidant injury in renal proximal tubules.
In previous studies, we found that lipid A, the biologically active component of lipopolysaccharide, triggers a rapid release of intracellular calcium, the activation of nitric oxide synthase (NOS), and nitric oxide (NO) production in rat proximal tubules. This pathway leads ultimately to cell death [as measured by the release of lactate dehydrogenase (LDH)], initiated by early generation of NO. In the present studies we found that lipid A produces a time- and concentration-dependent increase in lipid peroxidation [malondialdehyde (MDA) formation] prior to cell death. Furthermore, preventing lipid peroxidation protected against cell death. Lipid A (50 micro;g/ml) produced significant MDA formation in 30 min. The addition of two antioxidants 5 min prior to lipid A completely inhibited MDA formation and LDH release at 90 min. Preincubation with 5 mm GSH also significantly reduced MDA formation. The involvement of NOS activation in lipid A-induced lipid peroxidation was established when an NOS inhibitor and an inhibitor of intracellular calcium release completely blocked MDA formation. In addition, superoxide generation was significantly increased in the presence of lipid A, and the involvement of superoxide was established when superoxide dismutase protected against oxidant injury. The iron chelators deferoxamine (also a scavenger of peroxynitrite) and diethylenetriaminepentaacetic acid prevented lipid A-induced lipid peroxidation and cell death, indicating a role for iron and peroxynitrite. The addition of an NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-3-oxide-1-oxyl, prior to lipid A also completely protected tubule cells from lipid peroxidation and subsequent cell death. These results indicate that lipid A-stimulated NO generation in the rat proximal tubule initiates oxidant injury.
Traylor
L A
LA
Department of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock, Arkansas, 72205, USA.
Mayeux
P R
PR
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002614:Chelating Agents / Q000494:pharmacology; D004790:Enzyme Induction; D007501:Iron; D007674:Kidney Diseases / Q000139:chemically induced*; D007687:Kidney Tubules, Proximal / Q000187:drug effects / Q000503:physiopathology*; D008050:Lipid A / Q000633:toxicity*; D008054:Lipid Peroxides / Q000378:metabolism; D008297:Male; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000096:biosynthesis; D010101:Oxygen Consumption / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction; D013481:Superoxides / Q000378:metabolism
D000975:Antioxidants; D002614:Chelating Agents; D008050:Lipid A; D008054:Lipid Peroxides; D013481:Superoxides; D009569:Nitric Oxide; D007501:Iron; D019001:Nitric Oxide Synthase; D002118:Calcium