ABSTRACT
BACKGROUND: Little is known about factors which trigger and/or contribute to hereditary angioedema or ACE-inhibitor-mediated angioedema including variations in bradykinin type 2 receptor (B2R) expression and activity. METHODS: Protein and mRNA expression of B2R and the increase of intracellular calcium (iCa) in response to bradykinin were monitored in porcine and murine endothelial cells in response to NO donors or bradykinin. B2R protein expression was evaluated in skin, heart, and lung of (i) mice with endothelial-specific overexpression of eNOS (eNOS(tg) ), (ii) in eNOS(-/-) mice and (iii) in C57BL/6 mice treated with the NO donor pentaerythritol tetranitrate (PETN), the NOS inhibitor l-nitroarginine (L-NA), plasma pool C1-INH, and the B2R antagonist icatibant. Aortic reactivity to bradykinin was investigated including eNOS(-/-) mice. RESULTS: B2R protein and mRNA expression remained unchanged in cells subjected to L-NA, NO donors, and bradykinin in a time- and concentration-dependent manner. Likewise, increases of iCa in murine brain endothelial cells remained unchanged. B2R protein levels were similar in eNOS(tg) and eNOS(-/-) as compared to transgene-negative littermates. Likewise, treatment of C57BL/6 mice with PETN, L-NA, C1-INH or icatibant did not change B2R protein expression. In aortic rings of C57BL/6 mice, bradykinin induced B2R-dependent constrictions which were attenuated by endothelial NO and abolished by diclofenac indicating the functional importance of B2R-induced activation of endothelial NO synthase and cyclooxygenase. CONCLUSION: These data suggest that alterations of B2R protein expression induced by NO, bradykinin, C1-INH, or icatibant unlikely contribute to bradykinin-induced angioedema. This finding does not rule out a role for NO in bradykinin-induced extravasation and/or angioedema.
Subject(s)
Bradykinin B2 Receptor Antagonists/pharmacology , Bradykinin/analogs & derivatives , Bradykinin/metabolism , Complement C1 Inhibitor Protein/pharmacology , Nitric Oxide/metabolism , Receptor, Bradykinin B2/metabolism , Animals , Bradykinin/pharmacology , Calcium Signaling/drug effects , Cells, Cultured , Endothelial Cells/drug effects , Endothelial Cells/metabolism , Gene Expression Regulation/drug effects , Male , Mice , Mice, Knockout , Mice, Transgenic , Receptor, Bradykinin B2/genetics , SwineABSTRACT
BACKGROUND AND PURPOSE: The regulation of vascular soluble guanylyl cyclase (sGC) expression by nitric oxide (NO) is still under discussion. In vitro, NO has been shown to downregulate the expression of sGC but it is unclear if this mechanism is operative in vivo and occurs during nitrate treatment. EXPERIMENTAL APPROACH: We investigated whether high dose isosorbide mononitrate (ISMN) or pentaerythrityl tetranitrate (PETN) treatment changes vascular sGC expression and activity in vivo. New Zealand White rabbits received a standard diet, 2 or 200 mg ISMN kg(-1) d(-1) for 16 weeks, and C57BL/6 mice received a standard diet, 6, 60 or 300 mg PETN kg(-1) d(-1) for four weeks. Absorption was checked by measuring the plasma levels of the drug/metabolite. KEY RESULTS: Western blots of rabbit aortic rings showed similar protein levels of sGC alpha1- (P=0.2790) and beta1-subunits (P=0.6900) in all groups. Likewise, ANOVA showed that there was no difference in the expression of sGC in lungs of PETN-treated mice (P=0.0961 for alpha1 and P=0.3709 for beta1). The activities of isolated sGC in response to SNAP (1 microM-1 mM) were identical in aortae of ISMN-treated rabbits (P=0.0775) and lungs of PETN-treated mice (P=0.6348). The aortic relaxation response to SNAP slightly decreased at high ISMN but not at high PETN. CONCLUSIONS AND IMPLICATIONS: These data refute the hypothesis that therapeutic treatment with long acting NO donors has a significant impact on the regulation of vascular sGC expression and activity in vivo.
Subject(s)
Guanylate Cyclase/drug effects , Isosorbide Dinitrate/analogs & derivatives , Nitric Oxide Donors/pharmacology , Pentaerythritol Tetranitrate/pharmacology , Receptors, Cytoplasmic and Nuclear/drug effects , Administration, Oral , Analysis of Variance , Animals , Aorta, Thoracic/drug effects , Aorta, Thoracic/metabolism , Blotting, Western , Dose-Response Relationship, Drug , Gene Expression Regulation, Enzymologic/drug effects , Guanylate Cyclase/metabolism , Isosorbide Dinitrate/administration & dosage , Isosorbide Dinitrate/pharmacokinetics , Isosorbide Dinitrate/pharmacology , Lung/drug effects , Lung/metabolism , Male , Mice , Mice, Inbred C57BL , Nitric Oxide Donors/administration & dosage , Nitric Oxide Donors/pharmacokinetics , Pentaerythritol Tetranitrate/administration & dosage , Pentaerythritol Tetranitrate/pharmacokinetics , Protein Subunits/metabolism , Rabbits , Receptors, Cytoplasmic and Nuclear/metabolism , Soluble Guanylyl Cyclase , Vasodilator Agents/administration & dosage , Vasodilator Agents/pharmacokinetics , Vasodilator Agents/pharmacologyABSTRACT
Angioedema is an underestimated clinical problem. Many cases are nonallergic reactions, e.g. bradykinin-induced angioedema caused by genetic defects and angiotensin-converting enzyme (ACE) inhibitors. This difference is crucial for successful therapy, in particular when complete emergency care is not available. Five important forms of nonallergic angioedema can be distinguished: hereditary (HAE), acquired (AAE), renin-angiotensin-aldosterone system (RAAS)-blocker-induced (RAE), pseudoallergic angioedema (PAE) and idiopathic angioedema (IAE). Some angioedema are present in the larynx and may cause death. A vast majority of nonallergic angioedema are RAE, particularly those caused by ACE inhibitors. It appears important to emphasize that in patients with complete intolerance to RAAS-blockers, cessation of RAAS-blockers is likely to be associated with increased cardiovascular risk. Currently, there is no published algorithm for diagnosis and treatment. Angioedema is usually treated by a conservative clinical approach using artificial ventilation, glucocorticoids and antihistamines. Today, a plasma pool C1-esterase inhibitor (C1-INH) concentrate is the therapy of choice in HAE. The current pharmacotherapy of nonallergic angioedema is not satisfactory, thus requiring the identification of effective agents in clinical trials. Recently, several new drugs were developed: a recombinant C1-INH, a kallikrein inhibitor (ecallantide) and a specific bradykinin-B2-receptor antagonist (icatibant). According to currently available reports, these drugs may improve the treatment of kinin-induced angioedema.
Subject(s)
Angioedema , Bradykinin/metabolism , Angioedema/diagnosis , Angioedema/drug therapy , Angioedema/physiopathology , Complement C1 Inhibitor Protein/metabolism , Complement C1 Inhibitor Protein/therapeutic use , Humans , Recombinant Proteins/therapeutic useABSTRACT
This study was designed to investigate the dose-rate dependent effects of ionising radiation on endothelium- and NO-mediated reactivity of aorta and coronary vessels. Rats were exposed to acute ((137)Cs, 9 x 10(-4) Gy s(-1), 18 min) and chronic ((137)Cs, 2.8 x 10(-7) Gy s(-1), 41 days) radiation in 1 Gy dose. Acute irradiation transiently increased coronary flow in eNOS-activity-dependent manner on day 3 after exposure. In striking contrast, chronic irradiation caused a significant depression of coronary flow even on day 90 after irradiation and abolished the effects of NO-synthase inhibitor N-nitro-L-arginine methyl ester (10 micromol l(-1)). Furthermore, low intensity radiation strongly diminished the vasodilator properties of NO-donor sodium nitroprusside (5 micromol l(-1)). A similar pattern was observed in aortic rings. Endothelium-dependent vasodilation was increased on days 3 and 10 after acute irradiation, but strongly inhibited following chronic exposure for the entire post-radiation period. This was accompanied by a diminished vasodilator response to NO-donor on days 3, 10 and 30 of post-radiation but not on day 90. The data suggest that ionising radiation in 1 Gy induces changes of aortic and coronary vessels reactivity depending on the dose-rate and the interval after exposure.
Subject(s)
Aorta/physiology , Aorta/radiation effects , Blood Flow Velocity/physiology , Coronary Vessels/physiology , Coronary Vessels/radiation effects , Vasoconstriction/physiology , Vasoconstriction/radiation effects , Animals , Blood Flow Velocity/radiation effects , Dose-Response Relationship, Radiation , Male , Radiation Dosage , Radiation, Ionizing , RatsABSTRACT
Endothelium- and NO-dependent relaxation of aortic segments and basal coronary flow and response of coronary vessels to exogenous NO in the Langendorff-perfused rat hearts were examined on the 3rd, 10th, 30th and 90th days following whole body irradiation in 1 Gy dose with different dose rate. NO-mediated elevation of coronary flow and increased aortic endothelium-dependent vasodilation were found at the early stage after acute irradiation (137Cs, 9 x 10(-4) Gy/s), while vascular reactivity to exogenous NO was not changed. Chronic irradiation (137Cs, 2.3 x 10(-7) Gy/s) significantly impaired endothelium-dependent relaxation within whole experimental period and decreased NO component of basal coronary flow at the 3rd, 10th and 90th day. It was associated with attenuated reactivity to NO-donor in aorta one month after irradiation and in the coronary vessels--shortly after irradiation. In the delayed period endothelial dysfunction and the diminution of NO-dependent coronary flow were mediated by selective impairment of NO synthesis/release. Decreased survival rate of the lethally-irradiated Nwnitro-L-arginine methyl ester-treated animals revealed radiosensitizing properties of NO synthase inhibition. The data obtained indicate the involvement of NO in the alterations of vascular reactivity depending on the dose rate and on the interval after the irradiation.
