Eva Fulep

Prevention of pericyte loss by trolox in diabetic rat retina.

Prolonged hyperglycemia results in a number of diabetic complications, including retinopathy. Pericyte degeneration is one of the earliest histological changes observed in the development of diabetic retinopathy. Increased free radicals generated under hyperglycemia could damage the retina, which abounds in polyunsaturated fatty acids. In the current study, a severalfold increase in thiobarbituric acid-reactive substances was found in rat retina cultured in hyperglycemic medium, which decreased significantly when trolox, an amphipathic antioxidant, was included in the medium. To examine the contribution of oxidative stress in vivo, diabetic rats were fed trolox (0.4% in the diet) during the course of the experiments. After 5 mo of hyperglycemia, whole mounts of retinal vessels were prepared and endothelial cells (E) and pericytes (P) were counted. The ratio of E/P in the retinas obtained from normal rats, diabetic rats, and diabetic rats fed trolox were 1.74 +/- 0.186, 3.78 +/- 0.47, and 2.32 +/- 0.24, respectively. A significant restoration of pericytes by trolox suggests the involvement of oxidative injury during pericyte loss in diabetic retinopathy.

Modulation of immune responses by anabolic androgenic steroids

Anabolic androgenic steroids (AS) have recently been placed on the Food and Drug Administrations (FDAs) list of controlled substances, because of the adverse effects seen in athletes taking accelerated dosages in attempts to enhance performance. Reported deleterious effects on abusers include sterility, gynecomastia in males, acne, balding, psychological changes, and increased risks of heart disease and liver neoplasia. Considering the roles of the immune and neuroendocrine systems and their interactions in many of these pathologies, it is important to determine the effects of these derivitized androgens on this connection. Little is known in this respect. We therefore determined the effects of anabolic steroids on certain immune responses and their effects on the extrapituitary production of corticotropin by lymphocytes. We present evidence that (1) both 17-beta and 17-alpha esterified AS, nandrolone decanoate and oxymethenelone, respectively, significantly inhibited production of antibody to sheep red blood cells in a murine abuse model; (2) the control androgens testosterone and dehydroepian-drosterone (DHEA) or sesame seed oil vehicle had no significant effects on antibody production; (3) nandrolone decanoate and oxymethenelone directly induced the production of the inflammatory cytokines IL-1 beta and TNF-alpha from human peripheral blood lymphocytes but had no effect on IL-2 or IL-10 production; (4) control androgens had no direct cytokine inducing effect; (5) nandrolone decanoate significantly inhibited IFN production in human WISH and murine L-929 cells; and (6) nandrolone decanoate significantly inhibited the production of corticotropin in human peripheral blood lymphocytes following viral infection. These data indicate that high doses of anabolic steroids can have significant effects on immune responses and extrapituitary production of corticotropin. Furthermore, the mouse model should provide an effective means by which to study other deleterious effects of anabolic steroid abuse in humans.

Calcitonin gene-related peptide dilates the pregnant rat uterine vascular bed via guanylate cyclase, ATP- and Ca-sensitive potassium channels and gap junctions.

Summary We studied the mechanism of calcitonin gene-related peptide (CGRP)-induced vasorelaxation in isolated uterine vascular beds of pregnant rats. The vascular beds were perfused in situ with Krebs buffer containing dextran and indomethacin, an inhibitor of cyclooxygenase. Baseline perfusion pressure was maintained with norepinephrine. When applied as a bolus, CGRP caused a decreased perfusion pressure in uterine vascular beds that was dose-dependent and equal in both mid-pregnant and late-pregnant rats. The non-selective inhibitor of nitric oxide synthase (NOS), Nω-nitro-L-arginine methyl ester (L-NAME), did not significantly affect CGRP-induced vasodilatation in vascular beds of either group. CGRP-induced vasodilatation was not influenced by preincubation with the inhibitors of adenylate cyclase (SQ 22536 or MDL 12330A), but was significantly attenuated by the selective inhibitor of soluble guanylate cyclase (ODQ). The vasorelaxant effect of CGRP was not significantly influenced by the inhibitor of voltage-gated potassium (KV) channels (4-aminopyridin), but was significantly attenuated by an inhibitor of calcium-regulated potassium (KCa) channels (tetraethylammonium) and by an inhibitor of adenosine triphosphate-sensitive potassium (KATP) channels (glibenclamide). The gap junction uncoupling agent (carbenoxolone) also significantly attenuated the CGRP-induced decrease in perfusion pressure. We conclude that vasorelaxation induced by CGRP in the pregnant rat uterine vascular bed is not dependent on endothelial nitric oxide. In the uterine circulation of late-pregnant rats, the CGRP effect involves activation of soluble guanylate cyclase, but not adenylate cyclase, and does involve KCa and KATP channels and gap junctions.

Responses of isolated perfused uterine vascular beds of nonpregnant and pregnant rats to endogenous and exogenous nitric oxide

The responses to endothelial vasodilators and exogenous nitric oxide (NO) were characterized in intact isolated uterine vascular beds of nonpregnant, midpregnant and late-pregnant rats perfused with Krebs buffer (37 degrees C, 5% CO(2) in air, pH approximately 7.4) containing 2% dextran and indomethacin. Phenylephrine increased perfusion pressure in the vascular beds equally in all three groups. In the presence of phenylephrine, N(omega)-nitro-L-arginine methyl ester (L-NAME) significantly augmented perfusion pressure in the order: nonpregnant<midpregnant<late-pregnant uterine vascular bed. Acetylcholine and bradykinin-induced attenuation of perfusion pressure did not depend on gestational age. The decrease in perfusion pressure induced by acetylcholine was nonsignificantly attenuated by L-NAME in vascular beds from pregnant rats. The attenuation induced by bradykinin reached significant level in the vascular beds from midpregnant rats. The diethylamine (DEA)/NO-induced decrease in perfusion pressure was not influenced by L-NAME in any group. These data demonstrate the augmentation of basal release of NO associated with progression of pregnancy, while the responses to endothelial vasodilators do not depend on gestational age and are not abolished by inhibition of NO synthase, suggesting involvement of nonprostanoid non-NO factor in the control of uterine circulation.

Effect of fluoxetine on contractile activity of pregnant rat uterine rings.

OBJECTIVE We sought to compare the effects of fluoxetine, imipramine, and nortriptyline on spontaneous and serotonin-activated contractile activity of the uterine rings from midterm and term pregnant rats. STUDY DESIGN Uterine rings from timed-pregnant Sprague-Dawley rats on day 14 (midgestation) and day 22 (term gestation) were used for isometric tension recording. Responses to cumulative concentrations of fluoxetine, imipramine, nortriptyline, and serotonin in the absence and presence of the monoamine reuptake inhibitors were studied. RESULTS Neither of the monoamine reuptake inhibitors significantly influenced spontaneous contractile activity, whereas the concentration-dependent increase in activity induced by serotonin was inhibited in rings from both midterm and term pregnant rats. CONCLUSIONS The reported increase in preterm delivery in women receiving fluoxetine during the third trimester cannot be explained by a direct effect on uterine contractility.