Abdulla A.-B. Badawy

PRODUCTION OF TOLERANCE AND PHYSICAL DEPENDENCE IN THE RAT BY SIMPLE ADMINISTRATION OF MORPHINE IN DRINKING WATER

1 Rats are capable of consuming solutions of morphine sulphate in drinking water ad libitum in the absence of taste‐masking chemicals and without the need for scheduled provision or prior parenteral administration of the drug. 2 The success of this method depends on the initial provision of a 0.1 mg/ml solution of morphine sulphate. 3 When the drug concentration is increased to 0.4 mg/ml, the rats achieve an average daily intake of 50 mg/kg body wt. each. 4 Daily intake of morphine may be increased by at least about three fold by increasing the drug concentration to 1.2 mg/ml. 5 Oral morphine administration causes only a moderate loss in body weight. 6 Rats whose daily intake of the drug is 50 mg/kg exhibit tolerance to the analgesic action of morphine and show a drastic loss in body weight at 24 h after withdrawal and most of the behavioural symptoms of the naloxone‐precipitated withdrawal syndrome. 7 It is suggested that this simple method of morphine administration is suitable for further biochemical and behavioural studies of the actions of the drug.

Tryptophan metabolism in alcoholism

Acute and chronic alcohol (ethanol) intake and subsequent withdrawal exert major effects on tryptophan (Trp) metabolism and disposition in human subjects and experimental animals. In rats, activity of the rate-limiting enzyme of Trp degradation, liver Trp pyrrolase (TP), is enhanced by acute, but inhibited after chronic, ethanol administration, then enhanced during withdrawal. These changes lead to alterations in brain serotonin synthesis and turnover mediated by corresponding changes in circulating Trp availability to the brain. A low brain-serotonin concentration characterizes the alcohol-preferring C57BL/6J mouse strain and many alcohol-preferring rat lines. In this mouse strain, liver TP enhancement causes the serotonin decrease. In man, acute ethanol intake inhibits brain serotonin synthesis by activating liver TP. This may explain alcohol-induced depression, aggression and loss of control in susceptible individuals. Chronic alcohol intake in dependent subjects may be associated with liver TP inhibition and a consequent enhancement of brain serotonin synthesis, whereas subsequent withdrawal may induce the opposite effects. The excitotoxic Trp metabolite quinolinate may play a role in the behavioural disturbances of the alcohol-withdrawal syndrome. Some abstinent alcoholics may have a central serotonin deficiency, which they correct by liver TP inhibition through drinking. Further studies of the Trp and serotonin metabolic status in long-term abstinence in general and in relation to personality characteristics, alcoholism typology and genetic factors in particular may yield important information which should facilitate the development of more effective screening, and preventative and therapeutic strategies in this area of mental health.

Inhibition of rat liver tryptophan pyrrolase activity and elevation of brain tryptophan concentration by administration of antidepressants

Abstract The apoenzyme activity of rat liver tryptophan pyrrolase is decreased in vitro by 16–100% by concentrations of many antidepressants of 0.01–1 mM. Apo-(tryptophan pyrrolase) activity is also decreased by 37–86% at 2hr after administration of a 10 mg/kg dose of many antidepressants. This inhibition appears to be due to the prevention of the conjugation of the apoenzyme with its cofactor haem. Brain tryptophan concentration is elevated by 19–39% at 3.5 hr after administration of the above dose of antidepressants. Isocarboxazid is the only antidepressant tested that affects neither liver pyrrolase activity nor brain tryptophan concentration. The non-antidepressants chlorpromazine, β-flupenthixol, mefenamic acid and pargyline are also ineffective in both respects. The increase in brain tryptophan concentration caused by administration of mianserin, viloxazine, desipramine or tranylcypromine is associated with an accumulation of tryptophan in the liver and an increased availability of the circulating amino acid to the brain. It is suggested that antidepressants increase brain tryptophan concentration by inhibiting liver tryptophan pyrrolase activity. The results are briefly discussed in relation to the therapeutic effects of the drugs.

Inhibition of rat liver tryptophan pyrrolase activity and elevation of brain tryptophan concentration by acute administration of small doses of antidepressants.

1 Administration to rats of a 0.5 mg/kg dose of any of 19 antidepressants, but not that of many other drugs, causes a significant inhibition of the total enzyme and apoenzyme activities of liver tryptophan pyrrolase (of 24–48% and 37–65% respectively) and elevates brain tryptophan concentration by 13–66%. 2 When liver tryptophan pyrrolase activity is enhanced by pretreatment with cortisol or haematin, subsequent administration of a 0.5 mg/kg dose of some, but not other, antidepressants causes inhibition, which is weak (up to 38%). 3 This weak inhibition of the enhanced pyrrolase activity together with other pharmacological and physiological factors could explain the time lag between the start of antidepressant medication and the occurrence of a therapeutic response. 4 The cortisol‐induced and haematin‐activated pyrrolases respond differentially to inhibition by imipramine and amitriptyline, and this may explain the differential response to these two drugs of depressed patients in relation to urinary excretion of the noradrenaline metabolite 3‐methoxy‐4‐hydroxyphenylglycol. 5 The results are discussed in relation to the mechanism of action of antidepressants and the possible involvement of disturbed hepatic tryptophan metabolism in depressive illness.

Heterogeneity of serum tryptophan concentration and availability to the brain in patients with the chronic fatigue syndrome

We assessed the serotonin status of patients with the chronic fatigue syndrome (CFS). Tryptophan (Trp) availability to the brain, expressed as the ratio of concentration of serum Trp to the sum of those of its five competitors (CAA), and other parameters of Trp disposition were compared in 23 patients with the CFS and 42 healthy controls. The serum [free Trp]/[CAA] ratio was 43% higher in CFS patients, due to a 48% higher [free Trp]. [Total Trp] was also significantly higher (by 19%) in CFS patients, and, although the [total Trp]/[CAA] ratio did not differ significantly between the control and patient groups, the difference became significant when the results were co-varied with age and gender. [CAA] was not significantly different between groups, but was significantly lower in females, compared to males, of the CFS patient group. We have established normal ranges for Trp disposition parameters and propose criteria for defining the serotonin-biosynthetic status in humans. We have provisionally identified two subgroups of CFS patients, one with normal serotonin and the other with a high serotonin status. The relevance of our findings to, and their implications for, the pharmacological and other therapies of the chronic fatigue syndrome are discussed.

Effects of acute paroxetine administration on tryptophan metabolism and disposition in the rat.

1 The effects of acute oral administration of paroxetine on tryptophan metabolism and disposition were examined in the rat. 2 Basal liver tryptophan pyrrolase activity was inhibited by paroxetine in vitro and after oral administration. Maximum inhibition was caused by a 1 mg kg−1 dose. 3 Paroxetine administration also inhibited pyrrolase activity that had previously been enhanced by hormonal induction by cortisol or cofactor activation by haematin. The cortisol induction of the enzyme was, however, not inhibited by pretreatment of rats with paroxetine. 4 Paroxetine increased tryptophan availability to the brain, because of the above pyrrolase‐inhibitory mechanism. Cerebral 5‐hydroxytryptamine (5‐HT) synthesis was accordingly enhanced, though this was apparent only with doses of the drug of up to 1 mg kg−1. With larger doses, decreased 5‐HT turnover, probably as a result of 5‐HT uptake inhibition, was the more dominant feature. 5 Paroxetine lowered circulating corticosterone concentration, but did not influence those of albumin, non‐esterified fatty acids or glucose. 6 It is concluded that, in addition to inhibiting brain 5‐HT turnover, paroxetine also, in common with 20 other antidepressants, enhances 5‐HT synthesis by increasing brain tryptophan concentration secondarily to inhibition of liver tryptophan pyrrolase activity.

THE MECHANISM OF THE ANTAGONISM BY NALOXONE OF ACUTE ALCOHOL INTOXICATION

Naloxone lowers blood‐ethanol concentration and causes a simultaneous reversal of the disturbances in the redox states of the hepatic nicotinamide‐adenine dinucleotide (phosphate) couples in acutely‐ethanol‐intoxicated rats. It is suggested that these effects of naloxone form the basis of its antagonism of acute alcohol intoxication.

REVERSAL BY NALOXONE OF THE EFFECTS OF CHRONIC ADMINISTRATION OF DRUGS OF DEPENDENCE ON RAT LIVER AND BRAIN TRYPTOPHAN METABOLISM

1 Chronic administration of ethanol, morphine, nicotine or phenobarbitone has previously been shown to enhance rat brain 5‐hydroxytryptamine (5‐HT) synthesis by increasing the availability of circulating tryptophan to the brain secondarily to the NADPH‐mediated inhibition of liver tryptophan pyrrolase activity 2 Naloxone reverses the above enhancement of 5‐HT synthesis and the accompanying increase in tryptophan availability to the brain and the inhibition of liver tryptophan pyrrolase activity 3 It is suggested that naloxone exerts these effects by antagonizing the chronic drug‐induced increase in liver [NADPH] 4 Naloxone increases serum corticosterone concentration in rats chronically treated with the above four drugs of dependence. Possible explanations of this effect are discussed.

Mechanisms of elevation of rat brain tryptophan concentration by various doses of salicylate.

1 The roles of inhibition of liver tryptophan pyrrolase activity and of displacement of tryptophan from its binding sites on serum proteins have been investigated in relation to the increase in rat brain tryptophan concentration after administration of various doses of sodium salicylate. 2 The elevation of brain tryptophan concentration by sodium salicylate (0.5 mg/kg) was caused by inhibition of liver pyrrolase activity, whereas that by doses of the drug of 50 mg/kg and above was achieved mainly by tryptophan displacement. Both tryptophan displacement and pyrrolase inhibition caused the increase in brain tryptophan concentration by sodium salicylate at 10 mg/kg. 3 The smallest dose of salicylate capable of displacing serum‐protein‐bound tryptophan was 2.5 mg/kg.

The effects of lofepramine and desmethylimipramine on tryptophan metabolism and disposition in the rat

Acute and chronic administration of lofepramine and its major metabolite desmethylimipramine (DMI) to rats elevates brain tryptophan concentration, thereby enhancing cerebral 5-hydroxytryptamine (5-HT) synthesis, by increasing the availability of circulating tryptophan to the brain, secondarily to inhibition of liver tryptophan pyrrolase (tryptophan 2,3-dioxygenase, L-tryptophan:O2 oxidoreductase, decyclizing; EC 1.13.11.11) activity. The pyrrolase inhibition by lofepramine occurs independently of metabolism to DMI, because it can be demonstrated directly in vitro. Lofepramine also differs from DMI in its action profile on the above and related aspects of tryptophan metabolism and disposition. These results demonstrate that lofepramine influences tryptophan and 5-HT metabolism and disposition independently of its major metabolite DMI, and are discussed briefly in relation to the mechanism of action of antidepressants.