Tryptophan in alcoholism treatment I: kynurenine metabolites inhibit the rat liver mitochondrial low Km aldehyde dehydrogenase activity, elevate blood acetaldehyde concentration and induce aversion to alcohol.

AIMS: The aims were to provide proofs of mechanism and principle by establishing the ability of kynurenine metabolites to inhibit the liver mitochondrial low K(m) aldehyde dehydrogenase (ALDH) activity after administration and in vivo, and to induce aversion to alcohol. METHODS: Kynurenic acid (KA), 3-hydroxykynurenine (3-HK) and 3-hydroxyanthranilic acid (3-HAA) were administered to normal male Wistar rats and ALDH activity was determined both in vitro in liver homogenates and in vivo (by measuring blood acetaldehyde following ethanol administration). Alcohol consumption was studied in an aversion model in rats and in alcohol-preferring C57 mice. RESULTS: ALDH activity was significantly inhibited by all three metabolites by doses as small as 1 mg/kg body wt. Blood acetaldehyde accumulation after ethanol administration was strongly elevated by KA and 3-HK and to a lesser extent by 3-HAA. All three metabolites induced aversion to alcohol in rats and decreased alcohol preference in mice. CONCLUSIONS: The above kynurenine metabolites of tryptophan induce aversion to alcohol by inhibiting ALDH activity. An intellectual property covering the use of 3-HK and 3-HAA and derivatives thereof in the treatment of alcoholism by aversion awaits further development.

Tryptophan metabolite 3-hydroxyanthranilic acid selectively induces activated T cell death via intracellular GSH depletion.

Tryptophan-derived metabolites, initiated by indoleamine 2,3-dioxygenase (IDO), preferentially induce activated T cell death, which is an important mechanism in IDO-mediated T cell suppression. However, the mechanism of this phenomenon remains unclear. We found that 3-hydroxyanthranilic acid (3-HAA), the most potent metabolite, selectively eliminated activated T cells, which were stimulated with the bacterial superantigen staphylococcal enterotoxin A (SEA), but not resting T cells, by inducing apoptosis. We observed 3-HAA-induced depletion of intracellular glutathione (GSH) in activated T cells. When GSH levels were maintained by addition of N-acetylcysteine (NAC) and GSH, 3-HAA-mediated T cell death was completely inhibited. This was associated with extrusion of GSH from activated T cells without increased reactive oxygen species (ROS) formation. Finally, we showed that administration of 3-HAA in mice after allogeneic bone marrow transplantation reduced acute graft-versus-host disease (GVHD) lethality by inhibition of alloreactive T cell expansion through intracellular GSH depletion. Our data suggest that direct depletion of intracellular GSH is the major mechanism of 3-HAA-mediated activated T cell death.

Immunosuppressive effect of tryptophan metabolites in composite tissue allotransplantation.

BACKGROUND: Hand transplantations have intensified immunological research into composite tissue allotransplantation to induce tolerance. Pregnancy is a successful, natural model of immunological tolerance. The enzyme indoleamine 2,3-deoxygenase plays an important role by catabolizing the amino acid tryptophan. The resulting metabolites have been shown to be immunosuppressive. The effect of tryptophan metabolites has not been investigated in vascularized organ transplantation before. In this study, the authors applied to composite tissue allotransplantation what nature has developed for pregnancy, and examined the immunosuppressive effect of tryptophan metabolites in a model of hind limb transplantation. METHODS: Thirty-three allogeneic hind limb transplantations in the rat (Lewis --> Brown-Norway) were performed in three groups. Group A (n = 12) received no immunosuppression, group B (n = 13) received tryptophan metabolites (kynurenine and 3-hydroxyanthranilic acid) locally and systemically, and group C (n = 8) served as a control group receiving FK506. The timing of rejection was assessed by clinical observation. RESULTS: Rejection of the allogeneic hind limb occurred on average 6.58 days after transplantation in group A (no immunosuppression) and after 8.15 days in group B (tryptophan metabolites). Rejection was significantly delayed (log-rank test, p < 0.01). No rejection was seen with application of FK506 during the follow-up period of 21 days. CONCLUSIONS: For the first time, tryptophan metabolites have been applied in vascularized composite tissue allotransplantation and showed a significant immunosuppressive effect. These promising first results need further dose-effect and toxicological studies to increase the still limited immunosuppressive effect and define the clinical role these metabolites may play in the future.

[Effect of kynurenines on the actions of acetylcholine, oxotremorine, and nicotine]. / Vliianie kinureninov na éffekty atsetilkholina, oksotremorina i nikotina.

Kynurenine, 3-hydroxyanthranilic, anthranilic and nicotinic acids at concentrations 10(-4) and 10(-5) M potentiated contractions of the isolated rat rectum produced by acetylcholine. Contractile response of the phrenico-diaphragmatic preparation of young rats to acetylcholine was diminished by kynurenine, 3-hydroxykynurenine and by quinolinic acid. In rats of both sexes, 3-hydroxyanthranilic, anthranilic, picolinic and nicotinic acids in doses of 10 and 20 mg/kg shortened the latency of oxotremorine tremor. Quinolinic and nicotinic acids reduced the latency of nicotinic tremor and potentiated hypothermia.

[Effect of kynurenine and its metabolites on the vascular effects of serotonin, noradrenaline, and acetylcholine]. / Vliianie kinurenina i ego metabolitov na sosudistye éffekty serotonina, noradrenalina i atsetilkholina.

In female albino rats anesthetized with urethane, the depressor effect of serotonin was basically diminished by kynurenine, 3-hydroxyanthranilic acid (3-HAA), and anthranilic acid and was potentiated by quinolinic and picolinic acids (PA). All kynurenines were injected in a dose of 200 micrograms. 3-Hydroxykynurenine and nicotinic acid (NA) had a dual action. The pressor effect of noradrenaline was largely diminished by 3-HAA, PA and NA. Each of the rest kynurenines exerted an opposite action in different experiments. The pressor effect of acetylcholine was not affected by kynurenines.