Endothelin (ET)-1 is known to inhibit active NaK transport by as much as 50% in kidney tubule and other tissues. The presence of low levels of ET-1 in aqueous humor combined with the potential for release of ET-1 from ciliary processes suggests that the lens could be exposed to ET-1 in vivo. In this study, experiments were conducted to examine the influence of ET-1 on active NaK transport in porcine lens.
1. The uptake of 14C-ascorbic acid by the iris-ciliary body in vitro was examined in the rabbit, guinea pig and rat. 2. It was observed that iris-ciliary body from the rabbit and guinea pig, but not the rat, accumulated 14C-ascorbate to levels exceeding that in the bathing medium. 3. In all three species, the uptake of 14C-ascorbate was diminished by cold temperature; the degree of uptake at 0 degrees C was similar in the rabbit, guinea pig and rat iris-ciliary body. 4. Chromatographic examination of the 14C accumulated by the rabbit and guinea pig tissue demonstrated that the label remains almost exclusively as 14C-ascorbate.
This study evaluated poly(ADP-ribose) polymerase (PARP) inhibition as a new therapeutic approach for peripheral diabetic neuropathy using clinically relevant animal model and endpoints, and nitrotyrosine (NT), TNF-alpha, and nitrite/nitrate as potential biomarkers of the disease. Control and streptozotocin-diabetic rats were maintained with or without treatment with orally active PARP inhibitor 10-(4-methyl-piperazin-1-ylmethyl)-2H-7-oxa-1,2-diaza-benzo[de]anthracen-3-one (GPI-15,427), 30 mg kg(-1) d(-1), for 10 wk after first 2 wk without treatment. Therapeutic efficacy was evaluated by poly(ADP-ribosyl)ated protein expression (Western blot analysis), motor and sensory nerve conduction velocities, and tibial nerve morphometry. Sciatic nerve and spinal cord NT, TNF-alpha, and nitrite/nitrate concentrations were measured by ELISA. NT localization in peripheral nervous system was evaluated by double-label fluorescent immunohistochemistry. A PARP inhibitor treatment counteracted diabetes-induced motor and sensory nerve conduction slowing, axonal atrophy of large myelinated fibers, and increase in sciatic nerve and spinal cord NT and TNF-alpha concentrations. Sciatic nerve NT and TNF-alpha concentrations inversely correlated with motor and sensory nerve conduction velocities and myelin thickness, whereas nitrite/nitrate concentrations were indistinguishable between control and diabetic groups. NT accumulation was identified in endothelial and Schwann cells of the peripheral nerve, neurons, astrocytes, and oligodendrocytes of the spinal cord, and neurons and glial cells of the dorsal root ganglia. The findings identify PARP as a compelling drug target for prevention and treatment of both functional and structural manifestations of peripheral diabetic neuropathy and provide rationale for detailed evaluation of NT and TNF-alpha as potential biomarkers of its presence, severity, and progression.
The effect of tryptophan and three major metabolites of tryptophan on the rabbit lens electrolyte balance was examined. Even at 3 x 10(-3) M tryptophan, tryptamine and xanthurenic acid had no effect on lens sodium, potassium and calcium content after a 20 hr incubation. Small but significant changes in lens sodium and potassium content were induced by 20 hr incubation in 2 x 10(-3) M 3-hydroxy-DL-kynurenine and 10(-3) M hydroxyanthranilic acid. Lens Na, K-ATPase activity was unaffected by 3 x 10(-3) M tryptophan. Tryptamine reduced, to a small extent, Na,K-ATPase activity at 3 x 10(-3) M, but was without effect at 10(-3) M. 3-hydroxy-DL-kynurenine (2 x 10(-3) M) and 3-hydroxyanthranilic acid (10(-3) M) inhibited Na,K-ATPase activity by 27% and 30% respectively. Only 3-hydroxy-DL-kynurenine had a demonstrable effect on 86Rb uptake. It is concluded that, in contrast to findings in the amphibian lens, metabolites of tryptophan have minimal or no detrimental effects upon rabbit lens electrolyte balance under the in vitro conditions of these experiments. However, this does not rule out a long term effect of these compounds should they accumulate in the lens.
The lens epithelium is thought to conduct Na-K transport for the entire lens cell mass. Lens fibers have a poor ion transport capacity. The authors tested whether different Na,K-ATPase polypeptides are expressed in the two cell types and whether both cells have the machinery needed for ongoing Na,K-ATPase expression as judged by the presence of mRNA for the Na,K-ATPase alpha subunit.
