Antioxidative properties of organotellurium compounds in cell systems.

The protective/antioxidative properties of diaryl tellurides were demonstrated in cellular systems of increasing complexity. In the presence of glutathione, bis(4-hydroxyphenyl) telluride (1a), bis(4-aminophenyl) telluride (1d) and bis(2-carboxyphenyl) telluride (1h) reduced by more than 50% t-butyl hydroperoxide-induced cell death in lung fibroblast cultures at concentrations below 2 microM. Bis(2,6-dimethyl-4-hydroxyphenyl) telluride (2b) reduced by more than 50% leukocyte-mediated and phorbol-12-myristate-13-acetate-stimulated damage to Caco-2 cells at 0.1 microM concentration. As judged by their abilities to reduce formation of thiobarbituric acid reactive substances at concentrations close to 1 microM, diaryl tellurides 1a, 1d and 2b protected rat kidney tissue against oxidative damage caused by anoxia and reoxygenation. The organotellurium compounds also offered protection after systemic administration. In the presence of diaryl telluride 2b (0.1-1 microM), the ischemia/reperfusion-induced vascular permeability increase in the hamster cheek pouch was significantly reduced as compared with the control. Some of the most active organotellurium cell protectants were evaluated for their ability to inhibit formation of the inflammatory mediators leukotriene B4 and interleukin-1beta. An inhibitory effect on the secretion of these species was seen for compounds 1a and 2b at or above 10 microM concentrations. The protective effects of diaryl tellurides against t-butyl hydroperoxide-induced cell injury can be ascribed mainly to the peroxide-decomposing, glutathione peroxidase-like capacity of the compounds. The chain-breaking, electron- or hydrogen atom-donating ability of diaryl tellurides seems to be the main reason for their protection against leukocyte-mediated cell damage in Caco-2 cells and in the oxidatively challenged rat kidney and hamster cheek pouch.

Intestinal inflammation and barrier function in HLA-B27/beta 2-microglobulin transgenic rats.

BACKGROUND: Since intestinal inflammation is correlated with impaired barrier functions, transgenic HLA-B27/human beta 2-microglobulin rats that spontaneously develop intestinal inflammation were used to investigate whether onset of inflammation or impaired barrier function was the initial event. METHODS: During the age period of 9-14 weeks, transgenic and non-transgenic (control) rats were gavaged weekly with the marker molecules, 51Cr-ethylenediaminetetraacetic acid, 1-deamino-8-D-arginine vasopressin, and albumin, which were quantified in blood or urine. RESULTS: At 12 weeks of age the first signs of inflammation appeared with decreased body weight gain, decreased urine production, and onset of diarrhea. By 14-15 weeks of age all transgenic rats had developed intestinal inflammation, as confirmed by histology and increased myeloperoxidase content, whereas no inflammation was observed in controls. Intestinal passage of the markers did, however, not differ between transgenic and control rats over the studied period. CONCLUSIONS: The results suggest that intestinal inflammation precedes altered intestinal barrier function in this inflammation model.

Topical anticolitic efficacy and selectivity of the glucocorticoid budesonide in a new model of acetic acid-induced acute colitis in the rat.

AIMS: To study the effect of local or parenteral administration of the glucocorticoid budesonide in the acetic acid-induced colitis model in the rat. METHODS: Colitis was induced in an exteriorized colonic segment by administration of 4% acetic acid for 15 s. Four days later, this colonic segment with colitis was examined using a morphological scoring system, and measurements of myeloperoxidase activity and of plasma exudation into the colonic segment. The experimental colitis showed morphological similarities to human ulcerative colitis, with 3-fold increase in myeloperoxidase activity and 6-fold increase in the plasma exudation. Budesonide in different doses administered for 3 days, starting one day after acetic acid instillation, prevented the development of colitis in a dose-dependent manner. The best effect of budesonide on the morphological score was achieved after local treatment at a dose of 10(-5) M twice daily (76% reduction compared with a control colitis group) and parenteral treatment with 0.75 mg/kg (80% reduction). These doses also normalized myeloperoxidase activity and significantly reduced the plasma exudation. The systemic effects of the drug were most pronounced in the group treated with parenteral budesonide. This group showed the greatest reduction in body weight and a significant reduction of the weight of adrenal glands and spleen (as compared to controls). Thymus weight in animals treated systemically was significantly lower than in locally treated animals. In the group treated with local budesonide the weight of adrenals was reduced. However, the weights of spleen and thymus were not reduced and the reduction of the body weight was even less than in the control group. CONCLUSION: Local treatment with budesonide at a dose of 10(-5) M (0.17 mg/kg if completely absorbed, but only 0.03 mg/kg with 15% bioavailability on colonic application) was as effective as parenteral treatment at a dose of 0.75 mg/kg in the attenuation of acetic acid-induced colitis in the rat, but resulted in minor systemic side-effects.

Modification of postischemic increase of leukocyte adhesion and vascular permeability in the hamster by Iloprost.

This study of the postischemic events in the hamster cheek pouch showed that there is an increase in number of leukocytes adhering to the venular endothelium after reperfusion. It also showed that the stable prostacyclin analogue Iloprost could counteract both the postischemic increase in leukocyte adhesion and the postischemic increase in vascular permeability to macromolecules. The hamsters were anesthetized and the cheek pouch was everted and prepared for intravital microscopy. Temporary ischemia (30 min) was obtained using an expandable cuff placed around the proximal part of the cheek pouch. Fluorescein labelled dextran (FITC-dextran, Mw 150,000) was used as a tracer of macromolecular leakage from the postcapillary venules. Iloprost, given either topically (0.1 nM) or as an intravenous infusion (40 ng/kg/min), reduced the postischemic permeability increase (P less than 0.05) but did not affect the hemodynamics or the permeability response induced by histamine. It is proposed that these effects could be due to inhibition of leukocyte activation by Iloprost, indicating that these cells could play a role in the permeability increase during reperfusion after ischemia.

Superoxide dismutase as an inhibitor of postischemic microvascular permeability increase in the hamster.

The purpose was to elucidate the involvement of superoxide radical (O2-.) in the postischemic increase in the vascular permeability in the hamster cheek pouch. Cheek pouches of anesthetized hamsters were everted, prepared for intravital microscopy, and superfused with a bicarbonate buffered saline solution. Local ischemia for 30 min was obtained using a cuff placed around the proximal part of the cheek pouch. The vascular permeability in the postcapillary venules was quantified as leakage of intravenously injected fluorescein labeled dextran (FITC-dextran, Mw 150,000), using intravital microscopy and fluorimetry. There was a significant and reversible permeability increase after the reperfusion started. In the first series of experiments, combined intravenous infusion and topical application of human recombinant extracellular superoxide dismutase C (EC-SOD C) reduced the postischemic permeability response by 80%. Bovine CuZn-SOD given in exactly the same way reduced the response by 60%. In the second series of experiments, inactivated EC-SOD C was given to the control animals and active EC-SOD C was given to the treated animals. The topical treatment was excluded. Only active EC-SOD C reduced significantly the postischemic permeability increase when present during the ischemic period. Treatment with mannitol (i.v.) did not alter the postischemic response. Since active EC-SOD C and CuZn-SOD but not inactivated EC-SOD C effectively inhibited the response, we suggest that the superoxide anion is involved in the mediation of the postischemic permeability increase in the hamster.

Leukotriene B4-induced permeability increase in postcapillary venules and its inhibition by three different antiinflammatory drugs.

Leukotriene B4 (LTB4) is a potent chemotactic and chemokinetic substance that causes leukocyte accumulation and increases vascular permeability. We have used the hamster cheek pouch model to study the effect of some different antiinflammatory drugs on the LTB4-induced microvascular permeability. Intravascular fluorescein-labeled dextran (mol wt 150,000) was used as a tracer of macromolecular leakage, which was measured as the number of venular leaky sites and as efflux of the vascular tracer. Topical application of LTB4 (10(-8) M, 5 min) resulted in a reversible increase of the vascular permeability and could be repeated with 40-min intervals without any significant reduction in permeability response. The LTB4-induced response could be inhibited by budesonide (a glucocorticoid) given locally, by iloprost (a prostacyclin analog) given intravenously and by a combined local and intravenous treatment with terbutaline (a beta 2-receptor agonist) but not by local terbutaline treatment only.

Macromolecular permeability increase following incomplete ischemia in the hamster cheek pouch and its inhibition by terbutaline.

The microvascular permeability increase during the reperfusion after incomplete ischemia in the everted hamster cheek pouch preparation was measured by intravital microscopy. FITC-dextran 1500,000 was given i.v. as tracer of macromolecular leakage. Incomplete ischemia of the left cheek pouch was obtained by occlusion of the ipsilateral carotid artery, its external branch and the contralateral external carotid artery. Reperfusion after thirty minutes of incomplete ischemia resulted in a fully reversible increase in the number of FITC-dextran leakage sites (leaks) from 0 to 68 +/- 18 in 7 hamsters. The response to a second period of ischemia, starting one hour after the first ischemia period, was significantly smaller (9 +/- 5 leaks). Local treatment with terbutaline (10(-6) M), a beta 2-receptor agonist, abolished the permeability response to ischemia almost completely (0.5 +/- 0.3 leaks, n = 6). Local application of histamine (2 X 10(-6) M) at the end of the experiments gave a response that was in accordance with that seen in hamsters not subjected to ischemia. These results were compared with those obtained from previous studies in our laboratory with a complete ischemia model and it appears as these two methods, incomplete and complete ischemia, give similar results.

Terbutaline and budesonide as inhibitors of postischaemic permeability increase.

A temporary ischaemia with total circulatory arrest of the hamster cheek pouch was obtained by clamping the neck of the everted cheek pouch. The macromolecular permeability increase in postcapillary venules was quantified as the leakage of fluorescein-labelled dextran using intravital microscopy and a fluorometer simultaneously. At reperfusion after 30 min ischaemia, there was a significant and reversible permeability increase. This response could be totally prevented by topical administration of either terbutaline, a selective beta 2-receptor agonist, or budesonide, a glucocorticoid, but it was not significantly impeded by the antihistamine mepyramine. The study shows that, in conformity with the situation in inflammation, the postischaemic permeability increase at reperfusion after ischaemia can be blocked by either beta 2-stimulation or glucocorticoids. Furthermore, it indicates that histamine, a common inflammatory mediator, is not responsible for the postischaemic permeability increase.

The hamster cheek pouch--an experimental model to study postischemic macromolecular permeability.

In order to study permeability changes after local ischemia the cheek pouch of the hamster was everted for intravital microscopy. Macromolecular permeability was quantified as leakage of fluoroscein labelled dextran (Mw 70,000). Temporary ischemia with total circulatory arrest of the whole cheek pouch was obtained by a cuff mounted around the everted cheek pouch where it leaves the mouth of the hamster. At reperfusion after repeated ischemic periods of 5 min no leaks were seen. A prolongation of the ischemic period to 30 min resulted in a temporary increase in the number of postcapillary venular leaks. When the occlusion was repeated the response was significantly smaller. An occlusion period of 60 min gave no further increase in macromolecular permeability. Locally applied histamine was used to test the reactivity of the cheek pouch. It was concluded that the model is suited for studies of permeability increase after local ischemia.