The effects of lipopolysaccharide-induced reactive oxygen species were blunted by calcium oxalate in renal tubular epithelial cells.

BACKGROUND/AIM: Previously we demonstrated that calcium oxalate (CaOx) in LLC-PK1 cells and oxalate in MDCK cells induce tubular damage and greater glycosaminoglycan synthesis. We test the hypothesis that reactive oxygen species (ROS) and prostaglandins mediate these effects. METHODS: LLC-PK1 and MDCK cells were exposed to graded concentrations of CaOx, oxalate or both. Glycosaminoglycan synthesis was analyzed through metabolic labeling and gel electrophoresis. Cell permeability and lipid peroxidation were assessed by lactate dehydrogenase release and malondialdehyde levels. Hydrogen peroxide and superoxide anion were analyzed using 2',7'-dichlorofluorescein and luminol. Cyclooxygenase-2 expression and prostaglandin E2 production were assessed by RT-PCR and ELISA, respectively. RESULTS: In LLC-PK1 cells exposed to CaOx, we observed increased cell permeability, no induction of ROS or lipid peroxidation, inability to produce lipopolysaccharide-induced ROS and increases in prostaglandin E2. Indomethacin used alone increased glycosaminoglycan synthesis but did not potentiate CaOx-induced effects. In MDCK cells exposed to oxalate we observed increased cell permeability, ROS production only at higher concentrations and inability to produce lipopolysaccharide-induced ROS. Indomethacin alone had no effect but increased oxalate-induced glycosaminoglycan synthesis. CONCLUSIONS: Prostaglandins modulate endogenous production of glycosaminoglycans in LLC-PK1 cells, as well as regulate oxalate-induced glycosaminoglycan synthesis in MDCK cells. Rather than increasing, CaOx and oxalate blunted lipopolysaccharide-induced ROS production. We could speculate that patients with recurrent nephrolithiasis may lose antimicrobial protection induced by ROS during infections.

Endogenous accumulation of IFN-gamma in IFN-gamma-R(-/-) mice increases resistance to B16F10-Nex2 murine melanoma: a model for direct IFN-gamma anti-tumor cytotoxicity in vitro and in vivo.

Syngeneic IFN-gamma(-/-) and IRF-1(-/-) mice are very sensitive to B16F10-Nex2 murine melanoma cells implanted subcutaneously. In contrast, IFN-gamma-R(-/-) (GRKO) mice are remarkably resistant to tumor development. Only 0-30% of these animals, challenged with a high dose of melanoma cells (5 x 10(5)), developed tumors at a late stage. The hypothesis of interferon gamma (IFN-gamma) accumulation and consequent cytotoxicity to implanted tumor cells was confirmed in vitro and ex vivo. IFN-gamma reduced tumor-cell growth in vitro in 60-81%, added alone or with LPS. Splenocytes and peritoneal macrophages from naïve GRKO mice activated with anti-CD3 and interleukin-12 (IL-12), respectively, accumulated IFN-gamma at levels 10-fold those of the wild-type. Supernatants of IL-12-activated macrophages from GRKO mice were toxic to B16F10-Nex2 cells, an effect reversible by anti-IFN-gamma antibody treatment. IL-12-activated macrophages from iNOS(-/-) mice were still highly cytotoxic to B16F10-Nex2 cells, but IL-12-activated macrophages from IFN-gamma-deficient mice were not inhibitory. In vivo, a single injection of anti-IFN-gamma antibody 18 h after tumor-cell challenge in GRKO mice rendered all animals susceptible to B16F10-Nex2 melanoma development. No tumors developed in the untreated GRKO mice during up to 45 days of observation. This model can be useful in understanding immune responses that involve IFN-gamma as a direct cytotoxic factor.

Synthesis of disubstituted tetrahydrocarbazoles with potential antidepressive activity.

A new series of disubstituted tetrahydrocarbazoles were synthesized. They are tested for antidepressive activity by the Porsolt's forced swimming test (one of the acute stress methods) and by the prevention of reserpine induced hypothermia and ptosis in mice. 3-Morpholino-1-[N-(6-methoxy-1,2,3,4- tetrahydrocarbozolyl)2-propanol, fumarate (XI) was demonstrated to be the most promising compound of this series. Besides, this series did not present the most common adverse effects of the conventional tricyclic-antidepressants (loss of locomotor coordination, ataxia and anticholinergic activity).