Biomedical subjects

Satoru Fukuda

Publications and source records attributed to Satoru Fukuda.

18 recordsLinked to original sources

PubMed2006

Protamine sulfate causes endothelium-independent vasorelaxation via inducible nitric oxide synthase pathway.

PURPOSE: The precise mechanism of systemic hypotension frequently observed with the use of protamine is unclear. Although it has been reported that protamine stimulates the release of nitric oxide (NO) from endothelium NO synthase (eNOS), the association with inducible NOS (iNOS) remains unknown, despite the induction of iNOS by lipopolysaccharides (LPS) and/or inflammatory cytokines during cardiopulmonary bypass (CPB). The purpose of this study was to determine whether protamine stimulates the release of NO from iNOS induced by LPS. METHODS: We performed prospective and controlled functional examinations with isolated endothelium-denuded thoracic aortas from 21 male Wister rats. Aortic strips were mounted in Krebs solution and treated with LPS (1 microg x mL(-1)) for six hours to induce iNOS. Changes in tension caused by L-arginine (a substrate of NOS), protamine or a heparin-protamine complex (heparin: protamine = 1 unit: 10 microg) were measured in strips pre-contracted by phenylephrine. RESULTS: No drug relaxed the strips before LPS-treatment, but each drug relaxed the strips in a dose-dependent manner after LPS-treatment (P < 0.05). Aminoguanidine (an iNOS inhibitor) and methylene blue (a guanylyl cyclase inhibitor) inhibited the relaxations. CONCLUSION: These results indicate that protamine and the heparin-protamine complex stimulated the release of NO from iNOS. As iNOS is induced during CPB, protamine or a heparin-protamine complex might cause systemic hypotension, at least in part, by stimulating iNOS.

Animals↗

PubMed2006

Excitatory and inhibitory actions of isoflurane on the cholinergic ascending arousal system of the rat.

BACKGROUND: The cholinergic arousal systems are known to critically regulate the state of consciousness. The aim of this study was to determine the effect of isoflurane on the inhibitory or excitatory neurotransmitters efflux in important nuclei within the cholinergic arousal system using in vivo intracerebral microdialysis. METHODS: The efflux of glutamate, gamma-aminobutyric acid (GABA), or acetylcholine in the posterior hypothalamus (PH), the basal forebrain (BF), and the somatosensory cortex (S1BF) of rats was detected using intracerebral microdialysis under an awake condition and at 0.5-2.0 minimum alveolar concentration (MAC) isoflurane anesthesia. The intrabasalis perfusion of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) and N-methyl-D-aspartate on the cortical acetylcholine effluxes was also examined under both conditions. RESULTS: Isoflurane had no influence on the glutamate and GABA efflux in the PH, whereas in the BF, it dose-dependently increased glutamate efflux and decreased GABA efflux. A transient increase in glutamate efflux at 1.0 MAC and a decrease in GABA at 0.5-1.5 MAC were observed in the S1BF. Isoflurane dose-dependently decreased acetylcholine efflux in the S1BF. Perfusion of the BF with AMPA increased acetylcholine efflux in the S1BF with electroencephalographic activation during 0.75 MAC isoflurane anesthesia, suggesting an inhibitory action of isoflurane on AMPA receptors in the BF. However, N-methyl-D-aspartate had no effect on these parameters. CONCLUSION: Isoflurane induces both excitatory and inhibitory actions in the cholinergic arousal system. The predominant inhibitory action of isoflurane over its excitatory action at the BF would result in the decrease in the acetylcholine efflux in the S1BF.

Acetylcholine↗

PubMed2006

Orexins increase cortical acetylcholine release and electroencephalographic activation through orexin-1 receptor in the rat basal forebrain during isoflurane anesthesia.

BACKGROUND: Cholinergic arousal system plays an important role in the maintenance of consciousness. The authors investigated whether the intrabasalis injection of orexin-A or orexin-B and the electrically stimulated pedunculopontine tegmentum nuclei (PPTg: the origin of cholinergic ascending pathways) may alter acetylcholine efflux and electroencephalographic activity in the somatosensory cortex in relation to the orexinergic system in isoflurane-anesthetized rats. METHODS: Either orexin-A (10, 30, or 100 pmol) or orexin-B (10, 30, or 100 pmol) (n = 6 each) was injected into the basal forebrain while the electroencephalogram was measured during 1.0 minimum alveolar concentration (1.2%) isoflurane anesthesia. Injection of Ringer's solution was used as a control. The PPTg was electrically stimulated twice with the following conditions: 1-s stimulus train (0.2 ms, 100 Hz, 400 microA) per min for 20 min. Twenty minutes before the second PPTg stimulation, Ringer's solution or 20 microg SB334867, an orexin-1 receptor antagonist (n = 5 each) was injected into the basal forebrain. RESULTS: Injection of orexin-A (30 and 100 pmol) and orexin-B (100 pmol) significantly increased the acetylcholine efflux in the somatosensory cortex (P < 0.05). Injection of orexin-A (10, 30, 100 pmol) and orexin-B (30, 100 pmol) changed the burst and suppression patterns to arousal electroencephalogram. Compared with orexin-B, injection of a lower dose of orexin-A induced increase in the acetylcholine efflux and arousal electroencephalogram. SB334867 significantly attenuated the increases in the acetylcholine efflux and electroencephalographic activation evoked by PPTg stimulation. CONCLUSION: The authors demonstrated that orexin-A was more potent than orexin-B in producing alteration of cholinergic basal forebrain neuronal activity and that the cortical activation induced by the PPTg stimulation against isoflurane anesthesia may be mediated through the orexin-1 receptors in the basal forebrain.

Acetylcholine↗

PubMed2006

Lymphocytoma cutis due to pierced earrings.

We report two cases of lymphocytoma cutis caused by pierced earrings, with the results of patch tests and X-ray microanalyses on electron microscopy. We detected zinc through scanning electron microscopy from the specimen of Case 1 and gold and titanium through transmission electron microscopy from Case 2. This is the first report to demonstrate the presence of metal fragments in the lesion, which may suggest the remanence of metal for 20 years, bringing about persistent allergic reaction.

Adult↗

PubMed2006

[Progress in perioperative neuromonitoring: preface and comments].

The aims of the neuromonitoring during perioperative period are 1) to detect the brain, spinal cord and peripheral nerve ischemia immediately and hence prevention from neuronal injury, 2) to measure the depth of anesthesia, especially sedative effect of anesthetics and 3) to evaluate brain death. For the detection of brain and spinal cord ischemia during operation, we have to bear in mind 1) whether we truly monitor the possible ischemic risk sites in the brain and spinal cord; 2) whether we know the false-negative and false-positive ratio of the neuromonitoring with account being taken of the effect of anesthetics, the patient's own pathologies, the body temperature etc. If we detect the abnormal change through neuromonitoring, we should immediately warn the surgeon to improve his subsequent procedures and confirm his decision to prevent neuronal damage. To understand these reviews, we have to know the fundamental basic knowledge such as International 10-20 System on EEG monitoring, the expression of negative wave and positive wave in evoked potential, the meaning of near-field potential and far-field potential, temporal dispersion and so on. In this issue, the recent progress in neuromonitoring is described by the specialists in each fields. I hope the anesthesiologists understand the significance of neuromonitoring and apply these techniques for the patient undergoing the neurological operation during perioperative periods.

Brain↗

PubMed2006

[Somatosensory evoked potential].

Somatosensory evoked potential (SEP) has been widely used for monitoring the abnormal nerve conduction in various diseases. In non-anesthetized patients, Abeta fibers are electrically stimulated during SEP measurements. In anesthesiological field, it is used as a short latency somatosensory potential (SSEP), because its latency and amplitude are relatively constant. To detect the conduction abnormality from the upper extremities to the brain, median nerve stimulation is used. For the detection of spinal cord abnormality during operation, posterior tibial nerve stimulation is often used. It is important to know the origin of the wave appearing in SSEP to find the lesion in the nervous system. SSEP has been used in scoliosis surgery, carotid endarterectomy, thoracoabodominal aortic surgery and cervical operations to detect brain and spinal ischemia. In an intensive care unit, it is used for the diagnosis of brain death or ischemia and other neuronal diseases such as Guillain-Barre syndrome and polyneuritis etc. In pain clinic, laser evoked potential (LEP) has been recently introduced for the analysis of the mechanisms of nerve and spinal cord diseases. Using the LEP, pain mechanism would be clarified. During SSEP measurements, it is necessary for the anesthesiologists, intensivists and pain clinicians to understand the effect of anesthetic drugs and hypothermia on SSEP.

Anesthetics↗

PubMed2005

Craniofacial anomalies of the cultured mouse embryo induced by inhibition of sonic hedgehog signaling: an animal model of holoprosencephaly.

The pathogenesis of holoprosencephaly is multifactorial, and blockage of Sonic hedgehog signaling is one of the most important causative factors in animal models and human cases. In this study, the authors analyzed facial anomalies of mouse embryos, which were cultured in vitro and exposed to cyclopamine, an alkaloid blocker of Sonic hedgehog signaling. When cultured with cyclopamine for embryonic day 8.5 to 10.5, the whole body size was smaller than normal, and the distance and angle between the nasal placodes were remarkably reduced. Extension of the cranial surface vessels also was noted. No cyclopia was observed. Migration of the cranial neural crest cells seemed to be intact. Expressions of Patched-1 and Gli-1, downstream genes of Sonic hedgehog signaling, also were down-regulated in in situ hybridization and real-time reverse transcriptase-polymerase chain reaction analyses. The authors consider that these facial anomalies represent milder phenotypes of holoprosencephaly.

Animals↗

PubMed2005

[The management of the difficult pediatric airway].

BACKGROUND: We are faced sometimes with the difficult pediatric airway due to congenital abnormalities. However, there has been no systematic examination for the management of the difficult pediatric airway. METHODS: We retrospectively examined the incidence of difficult airway in 13,557 pediatric patients who had undergone general anesthesia with tracheal intubation. The difficulties of the intubation were classified into grade 1 to 4; grade 1: intubated one time, grade 2: two times, grade 3: three times or more, grade 4: changed to another way. We defined grade 3 and 4 as "difficult airway". RESULTS: Twenty-five patients (0.17%) are "difficult airway" among 13,557 patients in which 21 patients (0.15%) are classified as grade 3, and 4 patients (0.02%) are grade 4. The difficulties were significantly different among the syndromes (P< 0.001). The rate of the incidence in the difficulty is high in Treacher Collins syndrome, arthrogryposis multiprex congenita and first and second brachial arch syndrome, but few is in Pierre-Robin syndrome, Crouzon syndrome and Apert syndrome which are known to accompany difficult airway. CONCLUSIONS: We demonstrated that the incidence of difficult airway is different among the syndromes.

Anesthesia, General↗

PubMed2005

[Comparison of semitransparent colored labels with opaque colored labels for prevention of adverse drug administration].

BACKGROUND: We made a semitransparent color label to supplement a demerit of an opaque color label for prevention of adverse drug administration, and evaluated whether a semitransparent color label is superior to an opaque color label. METHODS: We prepared a total of 16 syringes (8 colors; two syriges of each color) in the opaque (NC) group and in the semitransparent (CL) group. Each ten subjects were asked to pick up the same drug label alternately in each group, and we measured the time and the number of syringes until the examinee can pick up the five correct syringes. We also examined the adhesiveness of the label to the syringe for six hours in each group. RESULTS: The time and syringe number until the examinee could pick up five correct syringes were 24.6+/-4.6 seconds and 16.2+/-2.7 in NC group (P= 0.0004) and 10.9+/-3.7 seconds and 6.5+/-1.7 in CL group, respectively (n-10, each, P <0.0001). In CL group the label adhered to the syringes tightly for six hours, whereas all the labels in NC group were detached (P<0.0001). CONCLUSIONS: The semitransparent color label is superior to the opaque color label in discrimination and adhesion.

Anesthetics↗

PubMed2004

Unique roles of G protein-coupled histamine H2 and gastrin receptors in growth and differentiation of gastric mucosa.

Disruption of histamine H2 receptor and gastrin receptor had different effects growth of gastric mucosa: hypertrophy and atrophy, respectively. To clarify the roles of gastrin and histamine H2 receptors in gastric mucosa, mice deficient in both (double-null mice) were generated and analyzed. Double-null mice exhibited atrophy of gastric mucosae, marked hypergastrinemia and higher gastric pH than gastrin receptor-null mice, which were unresponsive even to carbachol. Comparison of gastric mucosae from 10-week-old wild-type, histamine H2 receptor-null, gastrin receptor-null and double-null mice revealed unique roles of these receptors in gastric mucosal homeostasis. While small parietal cells and increases in the number and mucin contents of mucous neck cells were secondary to impaired acid production, the histamine H2 receptor was responsible for chief cell maturation in terms of pepsinogen expression and type III mucin. In double-null and gastrin receptor-null mice, despite gastric mucosal atrophy, surface mucous cells were significantly increased, in contrast to gastrin-null mice. Thus, it is conceivable that gastrin-gene product(s) other than gastrin-17, in the stimulated state, may exert proliferative actions on surface mucous cells independently of the histamine H2 receptor. These findings provide evidence that different G-protein coupled-receptors affect differentiation into different cell lineages derived from common stem cells in gastric mucosa.

Animals↗

PubMed2004

Nitric oxide generated by nNOS in the macula densa regulates the afferent arteriolar diameter in rat kidney.

Nitric oxide (NO) derived from neuronal NO synthase (nNOS) in the macula densa is a modulator of tubuloglomerular feedback. However, little is known about the regulation of the afferent arteriolar diameter by NO from the macula densa in salt-sensitive hypertension. We investigated the relationship between nNOS in the macula densa and the afferent arteriolar diameter in deoxycorticosterone acetate (DOCA)-salt hypertensive rats treated with angiotensin converting enzyme (ACE) inhibitor or thiazide for 5 weeks. DOCA rats had reduced nNOS expression in the macula densa compared to controls and reduction of NO production evaluated with 4,5-diaminofluorescein diacetate in the juxtaglomerular apparatus (JGA). Treatment with ACE inhibitor and thiazide increased nNOS and NO production in the JGA. The diameter of the afferent arteriole observed by SEM using microvascular casts was smaller in DOCA rats compared to control rats, and ACE inhibitor or thiazide dilated the afferent arteriole with a positive correlation to nNOS immunoreactivity in the macula densa. In conclusion, the afferent arteriolar diameter might be regulated by NO derived from nNOS in the macula densa in DOCA-salt hypertensive rats.

Animals↗

PubMed2004

[Spinal cord protection during descending thoracic and thoracoabdominal aortic surgery].

The incidence of postoperative paralysis after thoracic and thoracoabdominal aortic operations has decreased, but is still high in comparison with other operations. The analysis of the mechanism involved in the ischemic tolerance of the spinal cord could contribute to the protection of the spinal cord from ischemia. The identification of the Adamkiewicz artery and the predictive factors for postoperative paralysis in the preoperative period, the use of motor evoked potential, several adjuncts to keep the spinal cord circulation, the stabilization of the hemodynamics with good oxygenation, and hypothermia contribute to the prevention of the spinal cord ischemia. The anesthetics appropriate for the monitoring of the motor-evoked potential are propofol and fentanyl with or without ketamine. Among the anesthetic drugs, narcotics might exacerbate the motor function after the spinal cord ischemia. The analgesic drugs which do not aggravate the spinal cord dysfunction would be expected. Good cooperation of surgeons and anesthesiologists greatly contributes to the finding of the spinal cord ischemia during this operation.

Anesthetics↗

PubMed2003

Deactivation of norepinephrine by peroxynitrite as a new pathogenesis in the hypotension of septic shock.

BACKGROUND: Vascular hyporeactivity to catecholamines limits successful treatment of hypotension in septic shock. Large amounts of nitric oxide (NO) and superoxide anion (O(2)(-1).) are produced in response to bacterial endotoxins and/or inflammatory cytokines. NO reacts with O(2)(-1). to form the potentially toxic NO metabolite, peroxynitrite (ONOO(-1)). The purpose of this study was to investigate whether ONOO(-1) decreases the vasocontractile activity of norepinephrine. METHODS: Norepinephrine was treated with ONOO(-1) or 3-morpholinosydonimine-N-ethyl-carbamine (SIN-1; an ONOO(-1) producer) in a 5 x 10(-2) m sodium phosphate buffer solution at pH 7.4, and absorbance of the product was measured spectrophotometrically at 295 and 370 nm. Norepinephrine pretreated with ONOO(-1) was administered to isolated rat thoracic aortas to observe contractions in functional experiments. The rate constant between norepinephrine and ONOO(-1) was determined via a competition assay with cysteine in functional experiments. Norepinephrine pretreated with ONOO(-1) was injected intravenously into anesthetized rats to measure blood pressure. RESULTS: Norepinephrine pretreated with ONOO(-1) was confirmed spectrally as oxidized norepinephrine. Norepinephrine pretreated with ONOO(-1) decreased its vasocontractile force in an ONOO(-1) (10(-6), up to 3 x 10(-4) m) concentration-dependent manner (EC(50) = 5.1 x 10(-5) m). The decrease in its force was lower at pretreatment with ONOO(-1) in a lower pH buffer. A rate constant for the ONOO(-1)-norepinephrine reaction was 6 x 10(2) m/s. Norepinephrine (10(-7) m) incubated with SIN-1 (10(-3) m) decreased its vasocontractile force in an incubation time-dependent manner. Administration of norepinephrine pretreated with ONOO(-1) to anesthetized rats caused no significant change in arterial blood pressure. CONCLUSIONS: These results indicate that norepinephrine was oxidized and deactivated by ONOO(-1). This deactivation may, at least in part, account for the hyporeactivities of vasocontraction to norepinephrine in septic shock.

Anesthesia↗

PubMed2003

Peroxynitrite decreases dopamine's vasoconstrictive activity.

UNLABELLED: Peroxynitrite (ONOO(-1)) reacts with dopamine to form an oxidized derivative. To investigate the vasoconstrictive activity of this derivative, we performed functional examinations with dopamine treated with ONOO(-1) or 3-morpholinosydonimine-N-ethyl-carbamine (SIN-1; an ONOO(-1) producer) on isolated strips of rat thoracic aorta. To exclude the direct effect of ONOO(-1), the strips were pretreated with methylene blue, a guanylyl cyclase inhibitor. Dopamine induced concentration-dependent contraction, but dopamine pretreated with ONOO(-1) decreased the contraction in an ONOO(-1)-concentration-dependent manner. Both maximum contractions and 50% effective concentration values for dopamine-induced vasocontraction were significantly decreased by pretreatment with ONOO(-1). Dopamine incubated with SIN-1 also decreased the contraction, the decrease being dependent on the incubation time. ONOO(-1) formation is a favored reaction and occurs easily when cellular production of both nitric oxide and superoxide increases, as in septic shock. These results may, at least in part, account for dopamine's limitation as a vasoconstrictor in septic shock. IMPLICATIONS: Peroxynitrite (ONOO(-1)) reacts with dopamine to form an oxidized derivative. We investigated the vasoconstrictive activity of this derivative with functional examinations using rat thoracic aorta and found the activity decreased. As ONOO(-1) formation increases in septic shock, our results may account for dopamine's limitation as a vasoconstrictor in septic shock.

Animals↗

PubMed2003

Orexin a elicits arousal electroencephalography without sympathetic cardiovascular activation in isoflurane-anesthetized rats.

UNLABELLED: We studied the effects of intracerebroventricular injection of the novel neuropeptide orexin A on electroencephalogram (EEG) and autonomic nervous system activity in rats under isoflurane anesthesia. The administration of orexin A changed burst suppression patterns to arousal patterns on the EEG at 1.0 minimum alveolar anesthetic concentration (MAC) isoflurane and decreased the burst suppression ratio at 1.5 MAC isoflurane. However, orexin A did not influence the heart rate or mean arterial blood pressure at either isoflurane concentration. These findings demonstrated that orexin A elicited anesthetic arousal under isoflurane anesthesia in terms of EEG pattern without sympathetic cardiovascular activation in the rat. IMPLICATIONS: The novel neuropeptides orexins induce arousal associated with activation of the sympathetic nervous system in conscious rats. It is not known whether orexins affect the electroencephalogram (EEG), autonomic nerve activity, or both under anesthesia. Orexin A induced EEG arousal without sympathetic cardiovascular activation in the isoflurane-anesthetized rat. Orexin A might influence the depth of anesthesia.

Anesthesia, Inhalation↗

PubMed2002

Cell density and growth-dependent down-regulation of both intracellular calcium responses to agonist stimuli and expression of smooth-surfaced endoplasmic reticulum in MC3T3-E1 osteoblast-like cells.

A two-dimensional intracellular Ca(2+) ([Ca(2+)](i)) imaging system was used to examine the relationship between [Ca(2+)](i) handling and the proliferation of MC3T3-E1 osteoblast-like cells. The resting [Ca(2+)](i) level in densely cultured cells was 1.5 times higher than the [Ca(2+)](i) level in sparsely cultured cells or in other cell types (mouse fibroblasts, rat vascular smooth muscle cells, and bovine endothelial cells). A high resting [Ca(2+)](i) level may be specific for MC3T3-E1 cells. MC3T3-E1 cells were stimulated with ATP (10 microM), caffeine (10 mM), thapsigargin (1 microM), or ionomycin (10 microM), and the effect on the [Ca(2+)](i) level of MC3T3-E1 cells was studied. The percentage of responding cells and the degree of [Ca(2+)](i) elevation were high in the sparsely cultured cells and low in densely cultured cells. The rank order for the percentage of responding cells and magnitude of the Ca(2+) response to the stimuli was ionomycin > thapsigargin = ATP > caffeine and suggests the existence of differences among the various [Ca(2+)](i) channels. All Ca(2+) responses in the sparsely cultured MC3T3-E1 cells, unlike in other cell types, disappeared after the cells reached confluence. Heptanol treatment of densely cultured cells restored the Ca(2+) response, suggesting that cell-cell contact is involved with the confluence-dependent disappearance of the Ca(2+) response. Immunohistological analysis of type 1 inositol trisphosphate receptors and electron microscopy showed distinct expression of inositol trisphosphate receptor proteins and smooth-surfaced endoplasmic reticulum in sparsely cultured cells but reduced levels in densely cultured cells. These results indicate that the underlying basis of confluence-dependent [Ca(2+)](i) regulation is down-regulation of smooth-surfaced endoplasmic reticulum by cell-cell contacts.

3T3 Cells↗

PubMed2002

Isoflurane ameliorates the posthypoxic deoxygenation of the rat brain--the role of cell adhesion molecules of polymorphonuclear leukocytes.

One of the serious problems that occurs after cardiopulmonary resuscitation is brain posthypoxic/ischemic deoxygenation. However, there has been no report concerning the effect of isoflurane (ISO) on the brain oxygenation during hypoxia-reoxygenation in relation to cell adhesion molecules (CD11b) in polymorphonuclear leukocyte. Rats were anesthetized with a low concentration of ISO (0.5 MAC: low ISO) or high concentration of ISO (1.5 MAC: high ISO) and brain oxygenation was detected by near infrared spectroscopy during 10-min hypoxia (5% O(2)) and a subsequent 120-min reoxygenation period. Hypoxia induced a decrease in oxyhemoglobin (HbO(2)) and an increase in deoxyhemoglobin (Hb). Reoxygenation induced a significant decrease in total hemoglobin (tHb) and HbO(2) with low ISO, but not with high ISO. The changes in Hb were minimal during reoxygenation in both groups. CD11b increased during reoxygenation with low ISO anesthetization, but not with high ISO. A significant negative correlation was observed between CD11b and two of the measured oxyparameters, HbO(2) and tHb, during reoxygenation at low ISO, but not at high ISO. These findings suggest that brain deoxygenation during hypoxia-reoxygenation is partly related to the expression of CD11b. We conclude that ISO modifies the brain circulation at least in part through attenuating the expression of CD11b during hypoxia-reoxygenation.

Anesthetics, Inhalation↗

PubMed2002

Modification of alpha1 -adrenoceptors by peroxynitrite as a possible mechanism of systemic hypotension in sepsis.

OBJECTIVE: It is well known that nitric oxide synthase is induced by endotoxin or inflammatory cytokines, and consequently large amounts of nitric oxide cause vascular hyporeactivity to vasoconstrictor agents and myocardial dysfunction, hence hypotension. However, there is considerable controversy as to whether these pathologic cardiovascular features are mediated directly by nitric oxide or also through the formation of secondary reaction products such as peroxynitrite (ONOO-1). Our objective was to investigate inhibitory effects of ONOO-1 on alpha1-adrenoceptors. DESIGN: Prospective, controlled, in vitro, laboratory study. SETTING: Laboratory of a health sciences university. SUBJECTS: Chinese hamster ovary cells that expressed the human recombinant alpha1a-, alpha1b-, or alpha1d-adrenoceptors, rat aorta strips. INTERVENTIONS: Binding experiments of [3H]prazosin were done in the Chinese hamster ovary cell membranes pretreated with 100 microM to 3 mM ONOO-1. Displacement experiments with noradrenaline or 3-nitro-l-tyrosine also were conducted. Mobilization of intracellular Ca2+ evoked by 1 nM to 10 microM noradrenaline was monitored in a fluorescence spectrophotometer with dual excitation at 340 nm/380 nm and emission at 500 nm in fura-2/AM-loaded Chinese hamster ovary cells. Contractile force produced by noradrenaline was monitored in rat aorta strips that have alpha1a- and alpha1d-adrenoceptors, pretreated with 1 mM ONOO-1. Either 0.3 N NaOH or the decomposed ONOO-1 was used as the control. MEASUREMENTS AND MAIN RESULTS: The specific binding of [3H]prazosin to alpha1a- and alpha1d-adrenoceptor was inhibited by ONOO-1 in a concentration-dependent manner. We found that 3 mM ONOO-1 decreased maximum binding sites by 40% to 50% in alpha1a- and alpha1d-adrenoceptors. Binding affinities for prazosin and noradrenaline were not affected by 1 mM ONOO-1 in all subtypes. We found that 3-nitro-l-tyrosine did not affect the prazosin binding to three adrenoceptor subtypes. Noradrenaline increased intracellular Ca2+ concentration ([Ca2+]i) concentration-dependently, which was inhibited by ONOO-1 in alpha1a- and alpha1d-adrenoceptors. ONOO-1 had no effect on alpha1b-adrenoceptor. Contractile force produced by noradrenaline decreased significantly in aorta strips pretreated with ONOO-1. CONCLUSION: ONOO-1 reduces the binding capacity of alpha1a- and alpha1d- but not alpha1b-adrenoceptors without changing the affinities. Treatment with ONOO-1 attenuates noradrenaline-stimulated increase in [Ca2+]i in alpha1a- and alpha1d-adrenoceptors but not in alpha1b-adrenoceptor. ONOO-1 also weakens noradrenaline-induced contractions in rat aorta that has alpha1a- and alpha1d-adrenoceptors. Cardiovascular hyporeactivity to catecholamines in septic shock may be caused in part by the inactivation of alpha-adrenoceptors by ONOO-1.

Adrenergic alpha-Antagonists↗