Guanethidine induced sympathectomy in the adult rat. II. Functional effects following chronic administration

Guanethidine sulphate in doses of 5 and 40 mg/kg was administered intraperitoneally to adult rats daily for three months followed by discontinuation of administration for one day or three months. Following guanethidine 40 mg/kg a loss of approximately 95% of the nerve cells of the superior cervical ganglion was observed. No changes were observed after 5 mg/kg. The average mean arterial blood pressure (measured under chloralose-urethane anaesthesia) was lowered by 33 and 54 mmHg by guanethidine 40 mg/kg for three months followed by discontinuation for one day and three months respectively. The heart rate was unchanged. Guanethidine 5 mg/kg did not change any of these parameters. The increase in blood pressure and heart rate caused by intravenously administered noradrenaline was greatly enhanced following guanethidine 40 mg/kg, whereas only small changes were observed following 5 mg/kg. Following discontinuation for one day of 40 mg/kg for three months the potency ratios for the mean arterial blood pressure and the mean heart rate were 31 and 21 respectively, after discontinuation for three months 44 and 17 respectively. The potency ratios following guanethidine 5 mg/kg were 1.5 to 2.0. No changes were observed in the weight of the suprarenal glands following guanethidine. The present investigation has demonstrated an almost total chemical sympathectomy by guanethidine 40 mg/kg for three months and a persistent hypersensitivity to noradrenaline even after discontinuation of medication for three months.     

Guanethidine-induced acute hypersensitivity to noradrenaline in anaesthetized rats

The changes in mean arterial blood pressure, heart rate and the respective noradrenaline dose-response curves after guanethidine sulphate (0.25, 1.25, 5, 20, and 40 mg/kg given intravenously) were studied in chloralose-urethane anesthetized adult rats. After transient changes a dose-dependent maximum fall of the blood pressure was observed 15 minutes after the administration of guanethidine. A fall in heart rate was observed after guanethidine sulphate 0.25 mg/kg, whereas 1,25, 5, 20, and 40 mg/kg initially increased the heart rate followed by a decrease to subnormal levels with minimum values 6-24 hours after guanethidine. Normalization of blood pressure and heart rate occurred within 24 hours and 96 hours, respcetively. Maximum dose-dependent enhancement of changes in blood pressure by noradrenaline 0.5 mug/kg was obtained approximately 15 minutes after guanethidine. Following guanethidine 40 mg/kg the response was increased by a factor 2, and normalization occurred within 96 hours. Fifteen minutes after guanethidine, significant changes in the response of the heart rate to noradrenaline 0.5 mug/kg were obtained with guanethidine 1.25, 5, and 20 mg/kg, whereas fully developed hypersensitivity after 20 and 40 mg/kg did not occur until 6-24 hours later. Following guanethidine 40 mg/kg the response was increased by a factor of 3, and normalization was observed after 96 hours. The present investigation has demonstrated different time-dependent patterns of the effect of guanethidine on blood pressure and heart rate responses to noradrenaline.     

Guanethidine induced sympathectomy in the adult rat. I. Functional effects following subacute administration

Guanethidine sulphate 5 and 40 mg/kg was administered intraperitoneally to adult rats for 4, 8, 14, and 28 days followed by discontinuation for 1 day, after administration for 28 days and additionally for 8, 14, 29, and 60 days. Under chloralose-urethane anaesthesia the mean arterial blood pressure and the mean heart rate were determined and the response of these parameters to intravenous noradrenaline 3-1600 ng was recorded. The blood pressure was not significantly changed after guanethidine 5 mg/kg, but lowered by 40 mg/kg, the decrease being reversible on discontinuation. The response of both parameters to noradrenaline was increased by guanethidine depending on the dose. The hypersensitivity was partly reversible on discontinuation, but a significantly increased sensitivity of the heart rate to noradrenaline was observed 60 days after discontinuation of guanethidine 40 mg/kg for 28 days. Histologically a profound loss of nerve cells of the superior cervical ganglion was observed following guanethidine 40 mg/kg, whereas no change was observed after 5 mg/kg. The present investigation has demonstrated that guanethidine 5 mg/kg does not induce histological or permanent haemodynamic changes, whereas 40 mg/kg for 28 days result in an incomplete sympathectomy accompanied by a partially irreversible hypersensitivity to noradrenaline. There is no simple relation between the loss of ganglion cells and the haemodynamic changes, and hence the hypersensitivity to noradrenaline is only in part due to the destruction of the postganglionic sympathetic neurone obtained by long term administration of large doses of guanethidine.     

Augmentation of drug-induced blood pressure increases in rats by amobarbital

The effects of amobarbital anaesthesia on blood pressure responses to angiotensin and noradrenaline were determined in rats with chronically implanted venous and aortic catheters. Responses were tested twice on the same day in each rat, first in the conscious state and then during anaesthesia. Blood pressure rises produced by noradrenaline and angiotensin were larger when the rats were anaesthetized with amobarbital than when they were conscious. This enhancement was not characteristic of general anaesthesia since urethane, instead of enhancing, depressed the pressor responses.Blockade of acetylcholine receptors with atropine also increased the response to pressor drugs. However, amobarbital in anaesthetic doses did not affect the response to acetylcholine, indicating that removal of compensatory parasympathetic reflexes was not a major cause of its enhancing effect. The effects of amobarbital on arterial pressure and heart rate were similar to those of the ganglion blocking drug pentolinium. When pressor responses were increased by amobarbital, autonomic ganglia were depressed as indicated by the loss of the response to nicotine. These findings suggest that amobarbital enhances blood pressure responses to noradrenaline and angiotensin by blocking autonomic ganglia.     

THE EFFECT OF ALTHESIN, KETAMINE OR PENTOTHAL ON RENAL FUNCTION IN SALINE LOADED RATS

Rats were prepared with chronic cannulae in the carotid artery, jugular vein and urinary bladder; they were then kept 2 days to allow recovery from surgery. A steady-state continuous saline diuresis was established, then various anaesthetic agents were injected and changes in the urine flow, sodium and potassium excretion rates, GFR, ERPF, ERBF and blood pressure were measured. Five groups of rats were studied: (1) control animals given saline in place of any anaesthetic agent, all parameters measured remained constant; (2) althesin (1.2 mg/kg), all parameters remained constant except for the blood pressure which decreased slightly for 5 min; (3) althesin (12 mg/kg); (4) ketamine (50 mg/kg); (5) pentothal (50 mg/kg). The above anaesthetics altered all of the measured parameters except urine output. Various mechanisms for these anaesthetic agents are discussed. In althesin anaesthetized rats, all the retarded renal parameters recovered within 30 min. It is concluded that for the study of renal function, if a brief anaesthesia is needed, althesin is a more suitable anaesthetic agent.     

Hypothalamic alpha adrenergic receptors in cardiovascular regulation

Stereotaxic injection of noradrenaline (3–100 nmol) into the anterior hypothalamicpreoptic region of pentobarbital anaesthetized rats induced a fall in blood pressure and heart rate. The size of the effect was related to the size of the dose. Phenylephrine (70 nmol) and clonidine (1.5–100 nmol), two typical alpha adrenergic receptor stimulating agents, mimicked the noradrenaline induced depressor effects. Local pretreatment with phentolamine (106 nmol) abolished the fall in blood pressure and heart rate induced by noradrenaline (40 nmol) or clonidine (15 and 40 nmol). Noradrenaline (40 nmol) and clonidine (15 nmol) injected into the brain ventricles also caused a fall of both blood pressure and heart rate, but the size of these effects was significantly smaller than that after direct intrahypothalamic injection of the same amounts of these drugs.These results suggest that stimulation of alpha adrenergic receptors in the anterior hypothalamic-preoptic region of rats causes a fall in blood pressure and heart rate. The relevance of these data is discussed in view of the role of alpha adrenergic receptors in the hypothalamus in both cardiovascular regulation and thermoregulation.     

Modifications of the permeability of the blood-brain barrier and local cerebral metabolism in pentobarbital- and ketamine-anaesthetized rats

The state of deep surgical anaesthesia, induced by intraperitoneal injection of pentobarbital sodium (54 mg/kg) or ketamine hydrochloride (150 mg/kg) in the rat, was accompanied by a significant reduction in the permeability of the blood-brain barrier evaluated by calculating a unidirectional blood-to-brain constant (Ki) for the circulating tracer [14C]alpha-aminoisobutyric acid. Pentobarbital-induced anaesthesia was also characterized by a widespread and marked depression of local cerebral glucose utilization; on the contrary, when rats were anaesthetized with ketamine, cerebral glucose utilization increased in the striatum and hippocampus and decreased in the cerebellum and brain-stem. It is suggested, as a hypothesis, that two different mechanisms, depending on the kind of the anaesthetic drug used, may be involved in the changes in the permeability of the blood-brain barrier, observed in anaesthetized animals: (a) a neurogenic component; (b) a direct interaction of the anaesthetic with elements of the microvasculature.     

Effects of diethyl ether, halothane, ketamine and urethane on sympathetic activity in the rat

The present paper describes the effects of different general anaesthetics on plasma catecholamine (CA) concentrations taken as biochemical index of peripheral sympathetic activity. In chronically catheterized rats, diethyl ether, ketamine and urethane increased plasma adrenaline (A) and noradrenaline (NA) concentrations, indicating that these drugs stimulate both neurosympathetic and adrenomedullary functions. These effects appear to be centrally mediated, since ganglionic blockade or spinal transection completely counteracted the diethyl ether- and ketamine-induced increases in plasma CA levels. Halothane induced a transient decrease in circulating A and an increase in NA. These results support the concept that general anaesthetics may have different effects on sympathetic function. Arterial blood pressure and heart rate were also measured to look for possible correlations with peripheral sympathetic activity. The enhanced release of peripheral CAs seemed to be the determining factor for increasing blood pressure and heart rate with ketamine only. In the other instances the activation of the peripheral sympathetic system appeared to maintain homeostasis by counterbalancing the various depressive effects of anaesthetics on the cardiovascular system.     

Cardiovascular actions of phenoxybenzamine

Phenoxybenzamine increased the force of contraction and rate of the heart and lowered the blood pressure of dogs in pentobarbitone anaesthesia. This was preceded by depression of cardiac contractile force and a rise in blood pressure when phenoxybenzamine was injected following ganglion-blocking drugs or atropine. The positive inotropic and chronotropic and the hypertensive effects of phenoxybenzamine were prevented by pretreatment with reserpine or guanethidine.     

Effects of prostaglandin E2, propranolol and nitroglycerine with halothane, pethidine or pentobarbitone anaesthesia on arrhythmias and other responses to ligation of a coronary artery in rats

The effects of various cardiovascular drugs (prostaglandin E2 (PGE2), propranolol and nitroglycerine) and anaesthetic regimens (halothane, pethidine and pentobarbitone), upon the outcome of coronary artery ligation in acutely prepared rats were determined. Effects upon arrhythmias, blood pressure, heart rate, mortality, ECG and the size of the occluded zone were determined for each drug in the presence of each anaesthetic. PGE2 and nitroglycerine had no statistically significant effects on the outcome of ligation whatever the anaesthetic. Propranolol had limited antiarrhythmic actions. The anaesthetic used had major effects upon the outcome of ligation, regardless of the cardiovascular drugs administered. Pentobarbitone anaesthesia resulted in the highest mortality, and most arrhythmias. Pethidine-N2O anaesthesia was associated with fewer arrhythmias. Halothane-N2O anaesthesia markedly decreased the incidence and severity of arrhythmias, independent of the cardiovascular drug. It was concluded that the anaesthetic used can have a major influence on ligation-induced arrhythmias in acutely prepared anaesthetized rats.     

Action of depressor-I, a hypotensive phospholipid from bovine brain, on systemic and arterial blood pressures of various species.

Effects of "Depressor-I" (D-I), a new hypotensive phospholipid obtained from bovine brain lipid fraction, on systemic arterial blood pressure were investigated. The hypotensive activity of D-I in urethane anaesthetized rats was dose dependent and tachyphylaxis and/or sensitization were not observed. Increments of the respiration and the heart rate were observed with sharp falls in blood pressures following intravenous administration of D-I, in simultaneous recordings in anaesthetized rats. D-I elicited hypotension in all species of animals examined, and the sensitivities to D-I were much the same, however, there were two types in patterns of duration on responses and the durations were also dose dependent. D-I exhibited depressor-responses even in conscious rats, though responses were much smaller compared with those seen in anaesthetized rats. In a comparison of anaesthetic agents in rats, the highest hypotensive activity of D-I was observed with pentobarbital anaesthesia, a moderate response was seen with alpha-chloralose and the least response was seen with urethane. In spinal rats or those pretreated with reserpine or antagonists, such as atropine, diphenhydramine, propranolol and hexamethonium, D-I also elicited hypotension. These results suggest that "Depressor-I" does not elicit the depressor action via the stimulation of the central and the autonomic nervous systems but rather by a direct action on peripheral blood vessels.     

Comparative circulatory effects of adrenaline and noradrenaline during intra-osseous dental anaesthesia.

The effects of intra-osseous dental anaesthesia on heart rate and blood pressure was examined in 17 healthy volunteers. Adrenaline containing anaesthetic solutions were accompanied by increased heart rate and elevated systolic blood pressure which were not acceptable to a high proportion of subjects. Noradrenaline at the same concentration as adrenaline (1/80 000) usually decreased the heart rate and elevated the blood pressure. A few subjects experienced a feeling of weakness in the lower extremities when noradrenaline was injected. Endogenous catecholamine formation in these test conditions gave rise to insignificant circulatory effects.     

Effects of blood pressure changes on the catecholamine release in the locus coeruleus of cats anaesthetized with pentobarbital or chloralose

Summary Effects of carotid occlusion and drugs applied intravenously on the release of endogenous catecholamines in the locus coeruleus of cats anaesthetized with pentobarbital or chloralose were investigated. The locus coeruleus was superfused bilaterally with artificial cerebrospinal fluid through push-pull cannulae inserted stereotaxically. Dopamine, noradrenaline and in some experiments also adrenaline were determined radioenzymatically in the superfusate.Under pentobarbital anaesthesia, a bilateral carotid occlusion increased the release rate of noradrenaline in the locus coeruleus, while the release of dopamine was decreased. These changes were due to the fall of blood pressure in the carotid sinus caused by the occlusion. Loading of baroreceptors by elevating blood pressure with phenylephrine (10 g·kg^–1·min^–1, i.v. infusion) was accompanied by a decreased release of noradrenaline in the locus coeruleus. This decrease in noradrenaline release was not detected in the caudal aspect of the locus coeruleus. Under chloralose anaesthesia, phenylephrine diminished the release rate of noradrenaline to about the same extent as under pentobarbital anaesthesia. The release rate of adrenaline was also decreased. A prolonged infusion of phenylephrine led to a prolonged pressor response associated with a sustained decrease in the noradrenaline release rate. Intravenous injection of chlorisondamine (3 mg·kg^–1) did not change the release of noradrenaline, while dopamine release was reduced.It is concluded that the release of catecholamines in the locus coeruleus is influenced by signals originating from peripheral baroreceptors. The influences are similar under pentobarbital and chloralose anaesthesia. Noradrenergic neurons responding to haemodynamic signals are not uniformly distributed within the locus coeruleus. It is suggested that noradrenergic and possibly dopaminergic and adrenergic neurons of the locus coeruleus are involved in the baroreceptor reflex, thus contributing to central homeostasis of blood pressure.     

Differential effects of anesthetic agents on regional blood flow and central hemodynamic parameters in rats

Three commonly used anesthetic agents (pentobarbital, chloralose-urethane, and inactin) were studied in a rat model. The radiolabeled microsphere technique was used to evaluate rats anesthetized (no reaction to pain stimulus) with the three drugs as compared to awake unanesthetized animals. Of the three anesthetic agents studied, pentobarbital caused the smallest alteration in central hemodynamic parameters. Chloralose-urethane significantly lowered cardiac output (56%), stroke volume (35%), and minute work (51%). Chloralose-urethane also significantly increased total peripheral resistance (59%). Inactin at the concentration used in the present study had very little effect on cardiac index, heart rate, stroke volume, and minute work but significantly increased total peripheral resistance and mean arterial pressure. All three anesthetic agents reduced cerebral and skeletal muscle blood flows equally. While pentobarbital and chloralose-urethane significantly decreased renal blood flow (33%), inactin did not change flow to the kidney. It is concluded that anesthetic agents used in small animal experiments should be chosen carefully so that they do not influence blood flow to the organ being studied.     

Mechanism of action of guanethidine

The early hypotensive action of intravenous guanethidine in rabbits, rats and cats anaesthetized with urethane is reversed after pretreatment with iproniazid. The fall in blood pressure following injection of guanethidine in rabbits is reduced after previous administration of reserpine. Reserpine, like adrenalectomy and splenectomy, suppresses the early pressor effect of guanethidine in cats anaesthetized with chloralose. Guanethidine inhibits the action of tyramine and nicotine, but potentiates the effect of noradrenaline on isolated rabbit atria. Guanethidine is also a weak inhibitor of monoamine oxidase activity. The results are discussed and compared with those shown by reserpine. It is concluded that the early effects of guanethidine are mainly due to the release of endogenous catechol amines.     

Effects of the intravenously administered anaesthetics ketamine,propofol, and thiamylal on the cortical renal blood flow in rats

Intravenous anaesthetics such as ketamine, propofol, and thiamylal are widely used, although the direct effects of these anaesthetics on the renal blood flow (RBF) have not been well elucidated. In this study, we examined the effects of bolus and continuous administrations of ketamine, propofol, and thiamylal on cortical RBF and the effects of noradrenaline (NA) on RBF under continuous administration of these anaesthetics. We used laser Doppler flowmetry to measure the effects of bolus injection and continuous infusion of ketamine, propofol, and thiamylal on cortical RBF in male Wistar rats. We also examined the effects of the anaesthetics on mean arterial blood pressure (MAP) and heart rate (HR). Bolus injections of ketamine, propofol, or thiamylal (1-8 mg/kg each, n = 10) at clinically relevant concentrations did not affect MAP, HR, or RBF. Continuous administration of ketamine, propofol, or thiamylal (1-8 mg/kg/h each, n = 10) did not affect MAP, HR or RBF. Exogenous NA (2 microg/kg) caused an increase in MAP and a decrease in RBF and HR. In experiments with continuous infusions of propofol or thiamylal (1-8 mg/kg/h each, n = 10), similar results were observed without infusion of any anaesthetics. However, bolus injection of NA did not result in a decrease in RBF during continuous ketamine infusion (98.8 +/- 6.7% of control, n = 6, p < 0.05), while ketamine did not affect the NA-induced increase in MAP. In conclusion, bolus and continuous administrations of ketamine, propofol, and thiamylal did not affect the RBF. From our present findings, ketamine would be useful for maintaining the RBF.     

Differential effects of hypothermia upon blood acid-base state and blood gases in sodium pentobarbital and urethane anaesthetised rats.

1. The effects of two anaesthetics, sodium pentobarbital and urethane, and the effects of anaesthesia-associated hypothermia on acid-base status and blood gases were studied in rats without assisted ventilation. 2. Manipulation of conscious rats produces a progressive increase in arterial lactate associated with slight hyperventilation. 3. Sodium pentobarbital anaesthesia produces mild respiratory acidosis accompanied by increase in lactate arterial values. Urethane anaesthesia leads to partially compensated metabolic acidosis. 4. Hypothermia reduces metabolic acidosis and hypercapnia induced by sodium pentobarbital anaesthesia. No difference between hypothermic and normothermic values was observed in urethane anaesthesia.     

AN ANTIHYPERTENSIVE ACTION OF PROPRANOLOL IN DOCA/SALT-TREATED RATS

SUMMARY 1. The cardiovascular effects of acute and long-term (4–7 weeks) treatment with propranolol and guanethidine were observed in conscious, DOCA/salt-treated hypertensive rats. 2. During long-term treatment with a low dose of propranolol (0.2 mg/kg, subcutaneously, twice daily), the blood pressure is reduced to normotensive levels. 3. The subcutaneous injection of 0.2 mg/kg of propranolol lowers blood pressure and heart rate for 2–4 h at the start of long-term treatment, but these acute effects become less as treatment continues. 4. Long-term treatment with a high dose of propranolol (5 mg/kg, subcutaneously, twice daily) reduces heart rate but not blood pressure. 5. The acute effects of subcutaneous injection of 5 mg/kg of propranolol are a rise in blood pressure and a fall in heart rate; these effects are maintained throughout the period of long-term treatment. 6. Guanethidine (5 mg/kg, subcutaneously, once daily) produces a gradual fall in blood pressure. The acute depressor effect becomes less as treatment continues. 7. Observations were made on the responses to noradrenaline injections and thoraco-lumbar stimulation in pithed rats, pretreated 1 h beforehand with propranolol (0.2 or 5 mg/kg) or guanethidine (5 mg/kg). 8. Pressor responses to noradrenaline are slightly increased by all three pretreatments; the tachycardia produced by noradrenaline is increased by guanethidine and reduced by propranolol. 9. The pressor response to thoraco-lumbar stimulation is abolished by guanethidine and not affected by either dose of propranolol; the tachycardia is reduced by guanethidine and the high dose of propranolol (5 mg/kg) but not by the low dose. 10. The antihypertensive action of propranolol in DOCA/salt-treated rats can not be attributed to adrenergic neurone blockade or to a non-specific depressant effect, and does not appear to be due to a dose-dependent antagonistic effect on cardiac β-receptors. It is suggested that it is due to a central impairment of sympathetic activity.     

Differential influence of laboratory anaesthetic regimens upon renal and hepatosplanchnic haemodynamics in the rat

Renal blood flow in rats anaesthetized with the combination alphaxolone/alphadolone (3.90 mL min-1 (g tissue)-1) was significantly (P less than 0.05) greater than in rats anaesthetized with ketamine midazolam (3.24 mL min-1 (g tissue)-1, pentobarbitone (3.19 mL min-1 (g tissue)-1), fentanyl/fluanisone midazolam (2.84 mL min-1 (g tissue)-1) or urethane (1.99 mL min-1 (g tissue)-1). Renal blood flow in the urethane anaesthetized rats was significantly (P less than 0.05) lower than in animals anaesthetized with the other anaesthetic regimens, and is consistent with literature reports of a depressive effect of urethane anaesthesia upon xenobiotic renal clearance in the rat. Hepatosplanchnic blood flow was highest in the alphaxolone/alphadolone anaesthetized animals (71.7 mL min-1 kg-1), with the urethane anaesthetized animals demonstrating a significantly (P less than 0.05) lower (33.4 mL min-1 kg-1) blood flow. The fentanyl fluanisone/midazolam (65.4 mL min-1 kg-1), pentobarbitone (61.1 mL min-1 kg-1), and ketamine/midazolam (51.4 mL min-1 kg-1) regimens resulted in hepatosplanchnic blood flows of intermediate magnitude. The observed marked differential effects of the anaesthetic regimens upon renal and hepatosplanchnic blood flows may dramatically influence drug disposition in the experimental animal, and be of significance to laboratory pharmacokinetic studies in which anaesthesia is used.     

Isoflurane and other commonly used anaesthetics do not protect the isolated buffer perfused mouse heart from ischemia-reperfusion injury

1. Some anaesthetic agents such as barbiturates and opioids possess cardioprotective properties in rats, rabbits, dogs and pigs. The purpose of this study was to evaluate the effects of some commonly used anaesthetic agents (pentobarbital, isoflurane and a mixture of midazolam, fentanyl and fluanisone) on the tolerance of the isolated mouse heart to ischaemia-reperfusion injury. 2. The isolated, Langendorff-perfused hearts were subjected to 45 min of global ischaemia followed by 60 min of reperfusion. Left ventricular pressures, heart rate and coronary flow were measured and infarct size was determined using triphenyltetrazolium staining. 3. There were no differences in haemodynamic variables during reperfusion between groups. Infarct size was not influenced by the choice of anaesthesia. 4. None of the anaesthesia protocols exerted significant protective effects on the ischaemic-reperfused isolated mouse heart performance. In mice, isoflurane as well as pentobarbital, opioids and benzodiazepines may be safely used for anaesthesia without a risk of protective side-effects in isolated mouse heart studies.