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Simultaneous invasive and noninvasive evaluations of baroreflex sensitivity with bolus phenylephrine technique.

Estimation of baroreflex sensitivity (BRS) is receiving increasing attention in clinical and experimental cardiology. Until recently, in most studies BRS has been assessed on the basis of invasive blood pressure measurement, which limits its use in large-scale studies and in clinical practice. The development of continuous noninvasive blood pressure monitoring has made it possible to assess BRS noninvasively. We compared central invasive and peripheral noninvasive techniques in the assessment of BRS during cardiac catheterization in 40 patients with possible coronary artery disease. The correlation between noninvasive and invasive BRS was high (r = 0.92; p < 0.001). However, the noninvasive method resulted in significantly higher BRS values than did the invasive method (7.1 +/- 6.5 msec/mm Hg vs 5.1 +/- 4.3 msec/mm Hg, respectively; p < 0.001) because of the smaller increase in systolic blood pressure after phenylephrine injection by the noninvasive technique than by the invasive technique (18.9 +/- 6.8 mm Hg vs 25.2 +/- 7.8 mm Hg, respectively; p < 0.01). The difference between noninvasive and invasive BRS correlated positively with invasive BRS (r = 0.54; p < 0.001) and inversely with age (r = -0.39; p < 0.01) and resting systolic blood pressure (r = -0.30, p < 0.05). A noninvasive BRS value of < 4.0 ms/mm Hg showed a sensitivity of 94%, a specificity of 91%, and an accuracy of 93% in identifying cases of reduced invasive BRS (< 3.0 msec/mm Hg). Our findings encourage the use of finger-cuff method in the assessment of BRS. However, noninvasive BRS values were slightly but significantly higher than invasive BRS values, a difference that should be taken into account when BRS is measured by the noninvasive approach.

Adult↗

Opioid receptors in rat cardiac sarcolemma: effect of phenylephrine and isoproterenol.

The present study demonstrates the presence of opioid receptors in the rat cardiac sarcolemma isolated by the hypotonic LiBr-shock procedure. Opioid binding was measured by using [3H]U69 593, [3H](2-D-penicillamine,5-D-penicillamine)enkephalin ([3H]DPDPE) or [3H][D-Ala2,MePhe4,Gly-(ol)5]enkephalin ([3H]DAGO) as selective radioligands for K, delta and mu opioid receptors, respectively. Both the K- and delta-selective ligands exhibited highly specific (75-86%) binding, saturable at a concentration of about 20 nM. No specific binding for the selective agonist DAGO was observed. A marked increase in both [3H]U69 593 and [3H]DPDPE binding was observed after incubation of the sarcolemma with the alpha-adrenoceptor agonist phenylephrine or with the beta-adrenoceptor agonist isoproterenol. These stimulatory effects were associated with an increase in the Bmax values, a decrease in the Kd values, and were completely antagonized by the respective antagonists phentolamine and propranolol.

Analgesics↗

Cytosolic calcium after carbon tetrachloride, 1,1-dichloroethylene, and phenylephrine exposure. Studies in rat hepatocytes with phosphorylase a and quin2.

Carbon tetrachloride (CCl4) and 1,1-dichloroethylene (DCE), both hepatotoxins, inhibit sequestration of Ca2+ by rat liver endoplasmic reticulum (ER) both in vivo and in vitro. It is possible that, as a result, cytosolic Ca2+ concentrations rise in liver cells. In experiments presented here, isolated hepatocytes were exposed to CCl4, DCE, and phenylephrine (PE), a non-hepatotoxic alpha 1-adrenergic agent that mobilizes Ca2+. Cytoplasmic Ca2+ concentrations were evaluated by two methods: indirectly by assaying the activity of glycogen phosphorylase a, and directly by monitoring the fluorescence of quin2. In primary hepatocyte cultures, CCl4, DCE, and PE exposure increased the activity of phosphorylase a at 5 min from 39 +/- 2 to 130 +/- 12, 80 +/- 13, and 97 +/- 10 nmoles PO4(3-)/mg protein/min respectively. In rat hepatocyte suspensions loaded with quin2 and exposed to CCl4, DCE, or PE, cytosolic Ca2+ concentrations were elevated within 20 sec to 0.83 +/- 0.13, 0.59 +/- 0.06 and 0.99 +/- 0.14 microM Ca2+ respectively. Basal Ca2+ levels in these cells averaged 0.25 +/- 0.03 microM. Thus, CCl4 and PE apparently increased cytosolic Ca2+ levels to approximately the same extent, whereas DCE was somewhat less effective. The durations of the effects of CCl4 and PE were examined further by determining their time courses of elevated phosphorylase a activity. In hepatocyte cultures, increased phosphorylase a activity persisted through at least 60 min following CCl4 exposure. In contrast, phosphorylase a activity returned to basal levels by 20 min after PE. Increases in cytoplasmic Ca2+ levels that are sustained rather than transient may distinguish these hepatotoxic chlorinated aliphatic hydrocarbons from non-toxic hormonal agents.

Aminoquinolines↗

Alpha-noradrenergic modulation of hypothalamic self-stimulation: studies employing clonidine, 1-phenylephrine and alpha-methyl-p-tyrosine.

An alpha-noradrenergic substrate of rewarding intracranial stimulation (ICS) has been hypothesized based on the observation that the alpha-antagonist phentolamine produced an inhibition of self-stimulation. The present experiment investigated the effects on hypothalamic self-stimulation of the alpha agonist clonidine in normal and in catecholamine-depleted rats. Using a shuttle-box technique that provides a rate-independent index of the rewarding and aversive components of ICS, it was demonstrated that clonidine produces a dose-dependent inhibition of reward that is clearly dissociable from any non-specific effects of the drug. The ineffectiveness of the peripheral alpha-agonist 1-phenylephrine indicates that the inhibition of reward produced by clonidine is mediated centrally. Clonidine and the catecholamine synthesis inhibitor alpha-methyl-p-tyrosine act together in a synergistic manner to greatly increase the magnitude and prolong the duration of the inhibition of reward while leaving the aversive component unaffected. These data are interpreted as supporting an alpha-noradrenergic basis of ICS reward while indicating that the aversive component of ICS is essentially independent of noradrenergic transmission.

Animals↗

Different effects of phenylephrine and clonidine on 86Rb efflux and on contraction in the rat caudal artery.

A large transitory efflux of 86Rb was induced by concentrations of phenylephrine (10(-6) and 10(-4)M) or clonidine (10(-4)M) which were able to produce phasic contraction. By contrast, a lower concentration of clonidine (10(-6) M), which could only induce a slower (tonic) contraction, only produced a weak but sustained 86Rb efflux. These results show qualitative and quantitative differences in the effects of the two alpha-agonists. They further support the view that phasic and tonic alpha-adrenergic responses may be related to different mechanisms.

Animals↗

Bronchopulmonary effects of phenylephrine and methoxamine in the guinea-pig. Interaction with bronchoconstrictor drugs.

The bronchopulmonary effects of phenylephrine (Phe) and methoxamine (Met) were investigated in vitro on isolated guinea-pig tracheas and lung strips and in vivo on pulmonary airway resistance (Raw) in conscious guinea-pigs. Phe, but not Met, relaxed the isolated trachea precontracted with acetylcholine (ACh); this effect was inhibited by propranolol and ascribed to beta-adrenergic stimulation. In the presence of propranolol, both Phe and Met contracted the isolated trachea (-log EC50 were 3.80 +/- 0.37 and 3.04 +/- 0.25 respectively (n = 5] and this effect was competitively antagonized by phentolamine. Phe and Met contracted the isolated lung strips more strongly than the trachea (-log EC50 were 5.14 +/- 0.23 and 4.30 +/- 0.14 respectively (n = 5]. In contrast with the latter, the maximum response was equivalent to that induced by ACh; this effect was also antagonized by phentolamine. In the conscious guinea-pig, Phe (100 and 300 micrograms/kg) and Met (1 and 3 mg/kg) had no effect on Raw but significantly reduced the bronchoconstriction induced by ACh (25 micrograms/kg), histamine (20 micrograms/kg) and serotonin (15 micrograms/kg); this protective effect was unmodified by propranolol (2 mg/kg), yohimbine (1 mg/kg) or piperoxan (0.3 mg/kg) but was significantly inhibited by prazosin (30 micrograms/kg) or AR- C239 (50 micrograms/kg). These results suggest that alpha-adrenergic vasoconstriction with subsequent shrinkage of the bronchial mucosa is responsible for the protective effect of Phe and Met against ACh-induced bronchoconstriction. In isolated lung strips, vasoconstriction would increase tension.

Acetylcholine↗

Effect of diltiazem upon contractile responses to phenylephrine, cirazoline, Sgd 101/75, St 587 and B-HT 920 in rabbit aorta and dog saphenous vein preparations.

Diltiazem (10 microM) did not significantly affect concentration-response curves to the full, relatively selective alpha 1-adrenoceptor agonists phenylephrine and cirazoline in rabbit aorta and dog saphenous vein preparations. The effects of these 2 agonists remained resistant to diltiazem even in tissues pretreated with phenoxybenzamine (0.03 or 0.1 microM, 20 min) to reduce the alpha-adrenoceptor reserve. Sgd 101/75 and St 587 were partial agonists in both vascular preparations. The concentration-response curves to these relatively selective alpha 1-adrenoceptor agonists were also unaffected, or only slightly attenuated, by diltiazem. B-HT 920 at low concentrations preferentially stimulated the dog saphenous vein preparation and only at high concentrations elicited small contractions of the rabbit aorta. The responses to B-HT 920 were mediated by alpha 2-adrenoceptors in the vein and by alpha 1-adrenoceptors in the aorta yet concentration-response curves to this agonist were significantly attenuated by diltiazem in both tissues. The results indicate that the resistance of certain alpha-adrenoceptor-mediated responses in vascular preparations to calcium entry blockers need not be associated with the presence of a significant receptor reserve and that calcium dependency of a response may be determined by the agonist.

Adrenergic alpha-Agonists↗

Effects of ring fluorination on the adrenergic properties of phenylephrine.

The adrenergic properties of 2-, 4- and 6-fluorophenylephrine (2-FPE, 4-FPE, 6-FPE) were compared to those of phenylephrine (PE). The order of affinities of these compounds for alpha 1-adrenoceptors as determined by displacement of [3H]prazosin and [3H]WB-4101 binding to brain membranes was the same as the order of potencies for eliciting two alpha 1-adrenergic metabolic responses in guinea-pig cerebral cortical synaptoneurosomes, namely the stimulation of phosphatidylinositol turnover and the potentiation of 2-chloroadenosine-induced accumulation of cyclic AMP. In all cases the order was 6-FPE greater than PE greater than 4-FPE greater than 2-FPE. The order of affinities for alpha 2-adrenoceptors as determined by displacement of binding of [3H]clonidine to brain membrane was 6-FPE greater than PE greater than or equal to 4-FPE = 2-FPE. In contrast, the order of potencies for inhibition of forskolin-stimulated adenylate cyclase activity in human platelet membranes via an alpha 2-adrenoceptor was 6-FPE approximately equal to PE greater than 4-FPE much greater than 2-FPE. The FPEs and PE were partial agonists compared to epinephrine in human platelets. The affinities of these compounds for beta-adrenoceptors as determined by displacement of binding of [3H]dihydroalprenolol to brain membranes are 2-FPE greater than PE greater than or equal to 4-FPE much greater than 6-FPE. The FPEs and PE had positive chronotropic and inotropic effects in isolated guinea-pig atria apparently through the activation of a beta-adrenoceptor, since pindolol blocked the response while prazosin did not. 6-FPE appeared less active than the other PEs in atria. In fat cell membranes, 2-FPE was more potent than PE in stimulating adenylate cyclase via a beta-adrenoceptor, while 4-FPE and 6-FPE were inactive. Both, 2-FPE and PE were partial agonists in fat cells compared to isoproterenol. Of the three FPEs, 6-FPE represents a more potent and more selective agonist for alpha-adrenoceptors compared to beta-adrenoceptors than PE, while 4-FPE and, in particular, 2-FPE are less potent and selective as alpha-agonists.

Animals↗

Influence of lithium on the positive inotropic effect of phenylephrine and isoprenaline in guinea-pig heart.

The influence of lithium on the positive inotropic effects mediated by alpha- and beta-adrenoceptor stimulation was studied in isolated guinea-pig ventricular preparations stimulated at 1 Hz. Lithium chloride (10(-3)-2 X 10(-2) M) shifted the concentration-response curve for the inotropic effect of phenylephrine to the right in a dose-dependent manner. [3H]Prazosin binding was not inhibited by 10(-2) M lithium. The antagonistic effect of lithium was almost completely prevented by the presence of 10(-2) M myoinositol, and was potentiated by 10(-4) M 2-2'-anhydro-2-C-(hydroxymethyl)-myo-inositol, an antagonist of myo-inositol. The positive inotropic effect of isoprenaline was completely unaffected by either 10(-2) M lithium, 10(-2) M myo-inositol, or 10(-4) M 2-2'-anhydro-2-C-(hydroxymethyl)-myo-inositol. Since it is known that lithium interferes with inositol phosphate metabolism, our results produce further indirect evidence of an involvement of inositol phosphates in the myocardial response to alpha-adrenoceptor, but not to beta-adrenoceptor, stimulation. Our findings also suggest that chronic lithium treatment could interfere with the adrenergic modulation of myocardial contractility in those physio-pathological conditions in which the role of myocardial alpha-adrenoceptors becomes predominant.

Animals↗

Phosphorylation of type-L pyruvate kinase in intact hepatocytes. Localisation of the phosphorylation site in response to both glucagon and the Ca2+-linked agonist phenylephrine.

Pyruvate kinase is one of the enzymes which can be phosphorylated by stimulation of the cell with either glucagon or Ca2+-linked hormones. Whether these two classes of hormones phosphorylate the same site on the enzyme is unclear. Our results demonstrate that isolation of [32P]phosphorylated type-L pyruvate kinase from glucagon-treated hepatocytes followed by aspartyl-prolyl cleavage yields a [32P]phosphorylated peptide of Mr 17,000. This fragment is also phosphorylated in response to the Ca2+-mediated agonist phenylephrine.

Adenylyl Cyclases↗

Augmentation in the kinetic characteristics of phenylephrine- and 5-hydroxytryptamine-induced contractions in the isolated rat aorta following eight weeks of STZ-diabetes.

Kinetic studies were conducted on the contractile response elicited by phenylephrine (PE) and 5-hydroxytryptamine (5-HT) activation of the alpha 1-adrenergic- and 5-HT2 receptor subtypes, respectively, in aortic rings isolated from streptozotocin (STZ)-diabetic and age-matched control rats. The maximal PE- and 5-HT-induced contractile responses were separated into distinct phasic and tonic components, and the tonic portion of the response was assessed by evaluation of the calculated maximal rate constant for onset of contraction (kobsmax; min-1). Statistical analysis revealed that the mean kobsmax values for PE alone (10 microM), 5-HT alone (10 microM) and mixtures of PE and 5-HT (10 microM each) were significantly greater in diabetic animals than in age-matched control animals. These increases in kobsmax resulted in significant diabetes-related increases in the rate and relative magnitude of response generation during the initial minutes of contraction. Such observations emphasize the importance of kinetic studies, and given the central role played by the aorta in cardiovascular homeostasis, suggest that altered aortic contractility may play a role in some aspects of diabetic vascular disease. Moreover, if these kinetic alterations reflect a more generalized feature of diabetic vasculature (e.g., resistance vessels), then it is conceivable that such changes may further exacerbate diabetic vasculopathy.

Animals↗

Effects of angiotensin II and phenylephrine on urinary endothelin in normal female volunteers.

Endothelin (ET) is a 21-amino acid peptide produced and secreted mainly by endothelial cells. Small amounts of ET are found in plasma, whereas large amounts are present in the urine. Despite the abundance of ET in the kidneys and urine, little is known about its regulation and clinical significance. The present study was designed to examine the effects of angiotensin II (Ang II) and phenylephrine (Phe) on the excretion of ET in normal female volunteers. Ang II and Phe were infused for 1 hour each and titrated to increase the mean arterial pressure by 20 mm Hg. There was a 60-minute recovery period before the second drug, and the order of the drugs was randomized. Infusion of Phe induced mild diuresis and natriuresis, which were associated with a significant increase in the excretion of ET. In addition, Phe significantly increased plasma atrial natriuretic factor (ANF). In contrast, infusion of equipressor doses of Ang II decreased urinary sodium excretion and did not significantly alter the excretion of ET. Moreover, Ang II induced only a small and nonsignificant increase in plasma ANF. These results demonstrate that (1) physiological doses of Ang II do not affect excretion of either ET or ANF; (2) Phe markedly increased the excretion of ET and ANF, independently of its effect on blood pressure; and (3) neither agent changed plasma ET, but Phe increased plasma ANF.

Adolescent↗

Both activators and inhibitors of protein kinase C promote the inhibition of phenylephrine-induced [Ca2+]i oscillations in single intact rat hepatocytes.

In single isolated rat hepatocytes Ca(2+)-mobilising hormones induce oscillations in cytosolic free Ca2+ ([Ca2+]i) in which the frequency of spiking depends on agonist dose, but the time course of individual spikes depends on the hormone species, rather than agonist concentration. We have previously presented data using sphingosine and staurosporine as evidence of a negative feedback role for protein kinase C (PKC) in the elongation of the falling phase of [Ca2+]i spikes. We show here that the principal effect of three specific PKC inhibitors, namely the bis-indolylmaleimide GF 109203X, the tetracyclic aromatic alkaloid chelerythrine, and a myristoylated PKC pseudosubstrate peptide, that act at different sites on the PKC molecule, is a reduction in, or a complete suppression of, the phenylephrine-induced [Ca2+]i oscillation frequency. These results resemble the effects of activators of PKC and modulators of diacylglycerol (DAG) metabolism. Furthermore, following phorbol ester-induced inhibition of the hepatocyte [Ca2+]i oscillator, the addition of all three of these PKC inhibitors further reduces the [Ca2+]i oscillation frequency, with high concentrations of chelerythrine being the only agent that overcomes this inhibition by phorbol ester. These paradoxical results point to the need for caution in interpreting the effects of protocols involving PKC activators and inhibitors in assessing the feedback control from PKC on cellular [Ca2+]i oscillations.

Alkaloids↗

High-performance liquid chromatographic determination of phenylephrine in human serum with coulometric detection.

A high-performance liquid chromatographic method for the determination of phenylephrine (PE) in human serum using coulometric detection is described. PE and internal standard, orciprenaline, were extracted from serum by solid-phase extraction and separation achieved on a coupled column system consisting of two C18 cartridge columns (250 x 4.6 mm I.D. coupled to a shorter 50 x 4.6 mm I.D. column) using a mobile phase of methanol-50 mM phosphate buffer (pH 3.2; 10:90) at 36 degrees C. Dual electrode coulometric detection was used in the "oxidative screen" mode. Calibration curves were linear over the range 0.3-4 ng/ml with a limit of quantification (LOQ) of 0.35 ng/ml. The method has a greater degree of sensitivity, precision and accuracy compared to previously published methods for PE and is suitable for use in pharmacokinetic and bioequivalence studies in humans.

Chromatography, High Pressure Liquid↗

Experimental design for the rapid selection of separation conditions for methyl and propyl parahydroxybenzoate, phenylephrine hydrochloride and chlorphenamine maleate by ion-pair liquid chromatography.

Methyl and propyl parahydroxybenzoate (MPHB, PPHB), phenylephrine hydrochloride (PE) and chlorphenamine maleate (CPM) are often combined as ingredients in cough-syrups. Due to distinct chemical structures, pKa values among other chemical properties are different. This may result in a particular chromatographic behaviour on ion-pair reversed-phase liquid chromatographic (LC) systems. A face-centred central composite design was applied to study the impact of four LC mobile phase parameters and parameter interactions on the retention of these four compounds. The mobile phase parameters studied were the concentration of methanol as organic modifier, the concentration of sodium dioctylsulphosuccinate (SDSS) as counter-ion, the concentration of dimethyloctylamine (DMOA) as competitive base and the pH. By means of the proposed design, mathematical regression models and response surface plots were calculated, which could predict the compounds' retention times with good statistical reliability. Adequate combination of the most relevant of these mobile phase parameters enabled complete chromatographic separations within short times of analysis.

Analysis of Variance↗

Induction of liver glycogenolysis by phenylephrine: comparison between superfusion and perfusion techniques.

Superfusion of liver tissue slices was performed by a technique specially developed for that purpose, and compared to the whole organ perfusion technique of Jakob and Diem in order to assess the merits of the former technique. Activation of liver glycogenolysis of phenylephrine has been examined by both techniques. Both techniques showed important similarities and some dissimilarities. It was concluded that the application of superfusion techniques requires careful quantitative assessment of each experimental detail for each physiological event examined.

Animals↗

Effect of phenylephrine bolus administration on left ventricular function during postural hypotension in anesthetized patients.

STUDY OBJECTIVE: To investigate the effect of intravenous (IV) phenylephrine (PHE) bolus administration on left ventricular function in patients who developed postural hypotension during isoflurane anesthesia in the head-up tilt (reverse Trendelenburg) position. DESIGN: Prospective "before-after" trial. SETTING: Operation theater of a university medical center. PATIENTS: 15 ASA physical status I and II patients without cardiovascular disorders. INTERVENTIONS: The anesthetized patients were tilted from a supine horizontal to a 30-degree reverse-Trendelenburg position. Once a steady state was achieved, PHE 3 micrograms/kg was administered as an IV bolus dose. MEASUREMENTS AND MAIN RESULTS: Transesophageal echocardiography was used to assess left ventricular function. We measured blood pressure (BP); heart rate; left ventricular end-systolic and end-diastolic area, diameter, and wall thickness; and ejection time at baseline and after tilt, immediately before and for a period of 3 minutes after PHE injection. We calculated fractional area change (FAC), mean velocity of circumferential fiber shortening (mVcf), and end-systolic wall stress. Head-up tilt caused a reduction of mean arterial pressure [from 68 to 54 mmHg (mean)], end-systolic and end-diastolic left ventricular area (from 9.7 to 6.5 cm2 and from 19.2 to 13.1 cm2, respectively) and end-systolic wall stress (from 56 to 33 10(3).dyne/cm2). FAC and mVcf remained unaltered. PHE administration restored baseline values or overcompensated the changes caused by tilt. FAC slightly decreased in response to PHE (from 0.51 to 0.43), end-systolic wall stress increased to 83 10(3).dyne/cm2, and mVcf remained unchanged. CONCLUSION: PHE bolus administration effectively restored BP and cardiac filling, which were reduced after head-up tilt, without causing a relevant impairment of left ventricular function or an increase in end-systolic wall stress above the upper normal limit.

Adult↗