Evidence for a peripheral component in the sympatholytic actions of clonidine and guanfacine in man

Summary The time courses of the changes in plasma growth hormone and noradrenaline concentrations in response to 15 min infusions of clonidine 0.2 mgs and guanfacine 2 mgs, were studied in six normal volunteers, in a double-blind, randomised, crossover study. Plasma noradrenaline fell within 15 min of the commencement of drug administration, by 36±14% after clonidine (p<0.05) and by 32±11% (p<0.05) after guanfacine. Plasma growth hormone was not significantly elevated until the 30th minute to 12.0±4.7 lU/ml (p<0.05) after clonidine and 14.7±11.5 lU/ml (p<0.05) after guanfacine, having been undetectable prior to both drugs. The reduction in plasma noradrenaline by these ₂-adrenergic agonists, prior to activation of central adrenoceptors as detected by changes in plasma growth hormone, is evidence for a peripheral component in their sympatholytic effect.     

Evidence for a central alpha-sympathomimetic action of clonidine in the rat.

The antihypertensive effects of clonidine (0.15 mg kg-1, i.p.) were studied in conscious DOCA/saline hypertensive rats having chronically implanted arterial cannulae. The response to clonidine was markedly reduced by simultaneously administered desipramine (3 mg kg-1, i.p.), antagonized dose-dependently by piperoxan (2-10 mg kg-1, i.v.) and prevented by pretreatment with phentolamine (0.2 mg, i.c.v.). Pretreatment with 6-hydroxydopamine (3 x 250 mu-g, i.c.v.), haloperidol (1 mg kg-1, i.p.), p-chloro-N-methylamphetamine (3.5 mg kg-1, i.p.) or 5,6-dihydroxytryptamine (50 mu-g and 25 mu-g, i.c.v.) did not significantly modify the antihypertensive response. It is concluded that the antihypertensive response to clonidine is mediated via stimulation of central alpha-adrenoceptors and is independent of central dopaminergic receptors and intact central serotoninergic neurons. The necessity for intact central noradrenergic neurons remains uncertain.     

In vivo production of steroids with central depressant actions by the ovary of the rat

1. In addition to progesterone and 20-dihydroprogesterone, the following unconjugated steroids were isolated from ovarian tissue and ovarian venous blood of mature, non-pregnant rats: pregnenolone, 3betaOH-5alpha-pregnan-20-one, 3alphaOH-5alpha-pregnan-20-one, 20alphaOH-5alpha-pregnan-3-one and 5alpha-pregnane-3alpha, 20alpha-diol.2. Their concentration in the ovarian tissue and their secretion rates into the ovarian venous blood was of the same order of magnitude as that of progesterone.3. In the evening of the day of pro-oestrus significant increases in the ovarian contents of the following steroids were observed: progesterone, +140%; 3alphaOH-5alpha-pregnan-20-one, +320%; 20alphaOH-5alpha-pregnan-3-one, +195%; sum of pregnenolone and 3betaOH-5alpha-pregnan-20-one, +275%.4. At the same time the ovarian progeterone secretion rate was increased by 365%, that of 3alphaOH-5alpha-pregnan-20-one by 190%.5. A possible physiological role of the ovarian allopregnane derivatives as central depressant agents is discussed.     

Importance of central noradrenergic and serotonergic pathways in the cardiovascular actions of rilmenidine and clonidine.

We examined the role of central monoamine neurotransmitters noradrenaline and serotonin in the cardiovascular actions of the alpha 2-adrenoceptor agonist rilmenidine in the conscious rabbit and compared it to clonidine. Rilmenidine and clonidine were equieffective in producing a maximum 24% reduction in blood pressure and heart rate when given intracisternally (i.c.), but rilmenidine was approximately 20-30 times less potent than clonidine. The effective i.c. doses of both drugs were 25-30 times lower than those required peripherally (i.v.). Comparison of the time course of an equieffective dose of rilmenidine and clonidine showed that the time to reach maximum effect was 10 min for both drugs but time to recovery was greater for rilmenidine (2.5-3 h vs. 1.5 h for clonidine). Destruction of central noradrenergic neuron pathways with i.c. 6-hydroxydopamine attenuated the hypotensive and bradycardic actions of both rilmenidine and clonidine at 2, 4, and 8 weeks after treatment. The maximum observed attenuation of the hypotension was at 8 weeks (40% of control for rilmenidine and 29% for clonidine), while for the bradycardia this was at 2 weeks (9 and 2% of control, respectively) with some recovery by 8 weeks (29 and 50% of control). Destruction of the central serotonergic neurons with i.c. 5,6-dihydroxytryptamine also attenuated the hypotension and bradycardia to rilmenidine and clonidine but was slower in onset, needing the full 8 weeks to reach its maximum effect. At this time the hypotension and bradycardia to rilmenidine and clonidine were reduced to between 35 and 48% of control.(ABSTRACT TRUNCATED AT 250 WORDS)     

The cardiovascular actions of clonidine and neuropeptide-Y in the ventrolateral medulla of the rat.

1. The cardiovascular responses to neuropeptide-Y (NPY) (25 and 50 pmol) and clonidine (10 and 20 nmol) were examined following microinjection into the rostral ventrolateral medulla (RVLM) and the caudal ventrolateral medulla (CVLM). Mean arterial pressure (MAP) and heart rate (HR) were measured in anaesthetized rats, pre- and post-injection. 2. The alpha 2-adrenoceptor agonist clonidine (10 and 20 nmol) reduced MAP and HR significantly when microinjected into the CVLM and RVLM. 3. NPY (25 and 50 pmol) microinjected into the CVLM decreased MAP and HR. However, in the RVLM neither dose had a significant cardiovascular effect. 4. The possibility of a functional interaction between the adrenergic system and NPY was examined by co-administration of clonidine and NPY in doses that gave submaximal blood pressure responses. In the CVLM this produced hypotension and bradycardia which was similar in magnitude to the sum of their individual responses, indicating that in this area their actions appear to be independent. 5. In the RVLM, where NPY has no significant cardiovascular effects, co-administration with clonidine, did not alter the response to clonidine. 6. It appears that in the areas investigated, there is no functional interaction between NPY and clonidine.     

Cyclic adenosine 3',5'-monophosphate in rat cerebral cortical slices: effects of methoxamine and clonidine.

In incubated slices of cerebral cortex from Sprague-Dawley rats, methoxamine and clonidine have no effect on basal levels of cyclic AMP. Methoxamine effectively inhibits the noradrenaline-stimulated formation of cyclic AMP. The inhibitory constant for methoxamine was 12.6 mumol/l. In the presence of 100 mumol/l adenosine, methoxamine does not inhibit the activity of noradrenaline, but is capable to activate alpha-adrenergic receptors leading to enhanced formation of cyclic AMP. The mechanism by which adenosine alters adrenergic receptors to become methoxamine-sensitive is not known. Clonidine inhibits the effect of noradrenaline alone or in combination with adenosine on the cyclic-AMP-generating system. It does not, as reported earlier, enhance the activity of submaximal concentrations of the beta-adrenergic agonist isoproterenol. These data do not support the concept of adrenergic receptors which require both, alpha- and beta-stimulation for maximal activation of adenylate cyclase.     

Differences between central and peripheral rat alpha-adrenoceptors revealed using binuclear ligands

We have used two homologous series of binuclear ligands, diacridines and diquinolines, and the radioligand receptor assay to compare the topology of alpha 1- and alpha 2-adrenoceptors in rat cerebral cortex and kidney membranes. While the chain length-dependence of affinity of the diacridines, as well as that of the diquinolines, for the alpha 1-adrenoceptors of these central and peripheral tissues are similar, we find marked differences in affinity profiles for interaction with central and peripheral alpha 2-adrenoceptors. In the context of our previously proposed model for the binding of diacridines and diquinolines to alpha-adrenoceptors the results suggest that the surface features of central and peripheral alpha 2-adrenoceptors differ in the area surrounding the noradrenaline binding site. This difference may prove to be of therapeutic relevance.     

Role of central dopaminergic mechanisms in piribedil and clonidine induced hypothermia in the rat

The colonic temperature of rats maintained at different ambient temperatures (6, 20 and 30°C) was measured following intraperitoneal injection of piribedil and clonidine. Piribedil caused a dose-dependent fall in temperature, most prominent at 6°C, which was attenuated by pretreatment with pimozide but not with phenoxybenzamine or propranolol.Intraperitoneal clonidine also produced dose-dependent hypothermia but this was not related to ambient temperature, since hypothermia was as great at 20°C as at 6°C. The clonidine induced fall was abolished by phenoxybenzamine but not by pimozide pretreatment.Selective depletion of brain dopamine, achieved by intracisternal 6-hydroxydopamine following intraperitoneal desmethylimipramine, significantly enhanced the hypothermic effect of piribedil at 6°C. By contrast the effect of clonidine was unchanged.It is suggested that the hypothermia induced by piribedil is due to stimulation of central dopaminergic receptors, while the effect of clonidine is probably mediated via adrenergic mechanisms.     

Acute effects of clonidine on central and peripheral haemodynamics and plasma renin activity

Summary After the intravenous administration of clonidine normo- and hypertensive patients were studied by measurement of central, peripheral and renal hemodynamics, and plasma renin activity. The patients were examined either at rest, or while supine after tilting to 30° for 10 min. In most patients 300 µg of clonidine lowered both the blood and pulse pressures, heart rate, stroke volume and cardiac index, as well as the central venous pressure. In general the peripheral vascular resistance did not change, although it fell in some hypertensive patients. Renal vascular resistance diminished and renal blood flow and glomerular filtration rate sometimes rose, but more frequently remained unchanged. — The results suggest a centrally mediated action of clonidine, possibly on the sympathetic system as they can be interpreted as evidence of inhibition of sympathetic responses. The failure of the peripheral renin level to rise supports this hypothesis.Synonyms: Catapres^®; investigational code number: ST 155; generic name: DCAI (dichloro-amino-imidazoline).Work done with the aid of the Deutsche Forschungsgemeinschaft and of Boehringer Sohn, Ingelheim.     

Pre- and postjunctional actions of endothelin in the rat iris sphincter preparation

Effects of endothelins (ETs) were studied in the rat iris sphincter preparation. Three peptides (ET1, ET-2 and ET-3) caused contractile responses, and the rank order of agonist potency was: ET-1 = ET-2 > ET-3. The concentration-response curve to ET-1 was shifted to the right by the ET_A receptor antagonist cyclo [d-Asp-l-Pro-d-Val-l-Leu-d-Trp] (BQ-123: 10^–7 M), the pA₂ value of which was 7.41 ± 0.09 (n = 4).ET-1 and ET-3, at the concentration of 10^–9 M, potentiated cholinergic contractions evoked by electrical field stimulation (5 and 20 Hz) without affecting the postjunctional sensitivity to carbachol. This potentiating effect was not influenced by BQ-123 (10^–6 M). The ET-evoked percentage increase in the stimulation-induced contraction observed at 5 Hz was significantly greater than that at 20 Hz. A release of immunoreactive ET was detected when the preparation was stimulated at 20 Hz (1.81 ± 0.36 pg/sphincter n = 6). ET release evoked by 20 Hz stimulation was completely abolished by tetrodotoxin (10^–7 M).In conclusion, ET interacts with two different receptor types, ET_A and non-ET_A receptors (probably ET_B) which exist post- and presynaptically at cholinergic neuroeffector junctions of the rat iris preparation. Stimulation of ET_A receptor results in a direct muscle contraction and non-ET_A receptor activation facilitates the acetylcholine output from cholinergic nerve endings. It is suggested that ET released from a tetrodotoxin-sensitive site is involved in the modulation of acetylcholine release in the rat iris sphincter preparation. Correspondence to: I. Takayanagi at the above address     

Effects of clonidine and guanethidine on peripheral sympathetic nerve function in the pithed rat

Clonidine, in low intravenous doses, inhibited the increased heart rate of pithed rats caused by peripheral sympathetic nerve stimulation. The magnitude of this effect was greatest at low frequencies of nerve stimulation, responses to high frequencies being little affected by the drug. In contrast, guanethidine reduced cardiac responses to both low and high rates of nerve stimulation. The difference between the depressant effects of the two drugs on responses to various frequencies of sympathetic nerve traffic may contribute to the differences known to occur between their properties as hypotensive agents.     

Relationship between central actions of bradykinin and prostaglandins in the conscious rat

Intraventricular injection of bradykinin produced a dose-dependent increase in the mean arterial blood pressure of conscious rats. With 5 nmol of bradykinin, a dual pressor response was observed, which was associated with a biphasic behavioral change. With repeated hourly injections of bradykinin, tachyphylaxis developed to the pressor and central nervous system (CNS) stimulating effect. Indomethacin, given intraventricularly, reduced the hypertension and the behavioral excitation caused by bradykinin in a dose-dependent manner. When prostaglandin E₂ was injected into the cerebral ventricles, it induced hypertension and behavioral sedation similar to the secondary response to bradykinin. These results suggest that bradykinin has a dual action on the CNS, and this is mediated by prostaglandin-related systems in the brain.     

Tofupill lacks peripheral estrogen-like actions in the rat reproductive tract

OBJECTIVE: The objective of this study was to evaluate the estrogenic effect of phytoestrogens contained in a commercial food supplement (Tofupill) on the reproductive tract of ovariectomized rats. METHODS: Food supplement (3.4 or 10.2 mg/kg) and conjugated equine estrogens (CEE, 31 or 100 microg/kg) were orally administered, daily during 14 days to ovariectomized rats. At the end of treatment, the following determinations were done: dry and wet uterine weight, vaginal epithelium condition, and uterine serotonin-induced contractile response. A group treated with 17beta-estradiol was included as control for serotonin-induced contractile response. RESULTS: Food supplement did not display clear estrogenic effects on vaginal epithelium, uterine weight or myometrial sensitivity to serotonin, whereas high doses of conjugated equine estrogens showed estrogenic action. CONCLUSIONS: The present data showed that Tofupill displayed a lower estrogenic effect than conjugated equine estrogens, which are one of the most commonly used hormone replacement therapy for postmenopausal women. However, further studies are needed to evaluate the risk associated to the use of Tofupill as an alternative to hormone replacement therapy for postmenopausal women.     

Investigation of the actions of chloroethylclonidine in rat aorta.

1. The interaction between chloroethylclonidine (CEC) and noradrenaline (NA) has been examined at alpha-adrenoceptors mediating contractions of rat aorta. 2. In rat aorta, the competitive antagonist prazosin, over the concentration-range 0.01-10 microM, produced concentration-dependent shifts in the contractile potency of NA, so that there was no component of the NA contraction resistant to prazosin. 3. The irreversible alpha 1-adrenoceptor antagonists, phenoxybenzamine (PBZ) (1-10 microM) and benextramine (10 microM) produced shifts in potency of NA and reduced the maximum response in a concentration-dependent manner. 4. The irreversible alpha 1-adrenoceptor antagonist, CEC (100 microM), produced a non-parallel shift in the NA concentration-response curve so that low concentrations of NA produced relatively small contractions but relatively high concentrations produced further contractions, so that the maximum response was not significantly reduced. 5. The combination of CEC pretreatment and subsequent prazosin (0.1 microM) produced a parallel shift in the potency of NA. However, prazosin (10 microM) failed to produce any further effect on the response to high concentrations of NA following CEC pretreatment. Hence, a component of the contraction to NA in the presence of CEC was resistant to subsequent prazosin. Likewise, this component was resistant to a combination of prazosin (10 microM) and yohimbine (10 microM). 6. Receptor protection experiments were carried out in which tissues were exposed to NA (100 microM), yohimbine (10 microM) or prazosin (0.1 microM) prior to and during exposure to CEC. Receptor protection with NA, yohimbine or prazosin (0.1 microM), followed by washout prevented the shift in potency of NA produced by CEC. 7. Further experiments examined the effects of prazosin (10 microM) on responses to NA following receptor protection with NA (100 microM), yohimbine (10 microM), prazosin (10 microM), or xylazine (100 microM).(ABSTRACT TRUNCATED AT 250 WORDS)     

Investigation of the actions and antagonist activity of some polyamine analogues in vivo

1. The ability of three putative polyamine antagonists to antagonize behavioural changes induced by spermine was assessed.
2. Injection of an excitotoxic dose of spermine (100 μg, i.c.v.) in mice results in the development of a characteristic behavioural profile, which has two temporally distinct phases. The early events include clonic convulsions, and the later, more general excitation, includes tremor and culminates in the development of a fatal tonic convulsion.
3. Co-administration of arcaine (25 μg, i.c.v.) potentiated the early phase effects after spermine injection, but antagonized the development of spermine-induced tonic convulsions. A larger dose of arcaine (50 μg, i.c.v.) given alone resulted in the development of spermine-like body tremor and convulsions. It therefore appears that arcaine is not a pure polyamine antagonist in vivo, but may be a partial agonist.
4. Similarly, 1,10-diaminodecane appeared to act as a partial agonist in vivo, although it was less potent than arcaine.
5. In contrast, diethylenetriamine (DET) effectively inhibited the development of the early effects of spermine, but was ineffective against the spermine-induced CNS excitation and tonic convulsions.
6. It is concluded that none of the putative polyamine antagonists tested behaved as effective polyamine antagonists in vivo, although each produced some antagonism.

     

Cholinergic false transmitters: physiological and biochemical actions in central and peripheral systems

Three N-ethyl substituted analogs of acetylcholine (ACh) were evaluated for potential use as false neurotransmitters to decrease cholinergic transmission. This evaluation included (1) the elevation of arterial blood pressure upon central administration, (2) depression of blood pressure upon intravenous injection and (3) interactions with central muscarinic and peripheral nicotinic receptors. With respect to the central pressor response, ACh, acetylmonoethylcholine (AMECh) and acetyldiethylcholine (ADECh) were full agonists of decreasing potency; acetyltriethylcholine (ATECh) was a partial agonist of considerably lower potency. The duration of response paralleled potency. With respect to the peripheral depressor response, ACh and AMECh were full agonists of equal potency, and ADECh and ATECh were partial agonists of at least 100-fold lower potency. In terms of their affinity for central muscarinic receptors (brainstem and cerebral cortex), the following series was obtained: ACh greater than AMECh much greater than ADECh = ATECh. All of the agents had a greater affinity for muscarinic receptors in the brainstem compared to cortex. Acetylcholine and AMECh recognized multiple receptor binding conformations; the binding of ADECh and ATECh indicated interaction with a single set of equivalent sites. The affinity for nicotinic ACh receptors from the Torpedo electric organ was ACh = AMECh much greater than ADECh; ATECh had little affinity for these receptors. Acetylcholine, AMECh and ADECh stimulated the binding of [3H]phencyclidine to the ion channel of nicotinic receptor (potency series = ACh greater than AMECh = ADECh); ATECh was inactive. Acetylcholine, AMECh and ADECh also induced receptor conversion to a desensitized conformation; ATECh did not.(ABSTRACT TRUNCATED AT 250 WORDS)