Endogenous Monoamine Oxidase Inhibitor Research Articles

Possible synergies between isatin, an endogenous MAO inhibitor, and antiparkinsonian agents on the dopamine release from striatum of freely moving rats.

Isatin is an endogenous indole that inhibits monoamine oxidase (MAO). When exogenously administered, it increases the striatal dopamine and acetylcholine levels and presents neuroprotective effects in the brain. Previous studies show that intrastriatal administration of isatin increased the in vivo dopamine release from striatum in a concentration-dependent form. In the present work, we investigated the effects of combined administration of isatin together with other substances actually used in antiparkinsonian pharmacotherapy on in vivo dopamine overflow. For this, we co-administered isatin with the MAO inhibitors selegiline and clorgyline, l-DOPA, the catechol-o-methyl-transferase (COMT) inhibitors tropolone and dinitrocatechol, with the dopaminergic agonist ropinirole, and with the psychostimulant caffeine, in order to evaluate possible synergies between these substances to increase the dopamine extracellular levels in freely moving rats. Intrastriatal administration of isatin (10 mM, 60 min) significantly increased dopamine release to 1164 ± 152%, compared to the baseline. Co-administration of isatin together with selegiline (1 mM) or clorgyline (1 mM) alone or in combinations showed a similar profile to increase in vivo dopamine release. Intrastriatal infusion of isatin together with antiparkinsonian drugs l-DOPA (25 μM), tropolone (1 mM), dinitrocatechol (100 μM), amantadine (5 mM) and caffeine (5 mM) significantly elevated extracellular dopamine levels more than any single drug, showing a good neurochemical synergy by improving the effect of isatin on the extracellular dopamine levels in the striatum. Infusion of isatin + ropinirole (5 mM) did not change the isatin-induced increase in dopamine overflow. These results could be useful to carry out further investigations with a possible clinical application.

Effect of monoamine oxidase inhibitors on ischaemia/reperfusion-induced acute kidney injury in rats

Increases in renal sympathetic nerve activity during ischaemia and renal venous norepinephrine levels after reperfusion play important roles in the development of ischaemia/reperfusion-induced acute kidney injury. In the present study, we examined the effect of isatin, an endogenous monoamine oxidase inhibitor, on renal venous norepinephrine levels, superoxide production after reperfusion, and ischaemia/reperfusion-induced acute kidney injury. Ischaemia/reperfusion-induced acute kidney injury was accomplished by clamping the left renal artery and vein for 45min, followed by reperfusion, 2 weeks after contralateral nephrectomy. Renal superoxide production and norepinephrine overflow were elevated and significant renal tissue damage was observed following ischaemia/reperfusion injury. Intravenous injection of isatin (10mg/kg) at 5min before ischaemia increased the renal venous plasma norepinephrine level after reperfusion and aggravated ischaemia/reperfusion-induced renal dysfunction and histological damage. The excessive superoxide production after reperfusion was significantly suppressed by isatin administration, indicating that the inhibition of oxidative deamination effectively suppressed superoxide production. These data suggest that the exacerbation effect of isatin is associated, at least in part, with increased norepinephrine levels but not with superoxide production. To the best of our knowledge, this is the first report of isatin involvement in the pathogenesis and/or development of acute kidney injury.

The relation between isatin and selegiline by the alteration of serum isatin, an endogenous monoamine oxidase inhibitor, induced by selegiline hydrochloride in Parkinson's disease

Background: Monoamine oxidase (MAO) is an enzyme that metabolizes a wide range of monoamines and play an important role in the control of neurotransmitter concentrations. However, the detailed CNS functions of an endogenous MAO inhibitor, isatin, were not determined. Isatin was found to be a potent MAO inhibitor that is more active against MAO-B than MAO-A. Exogenously administered isatin increased dopamine (DA) levels in the rat striatum. Furthermore isatin improves motor dysfunction and the DA levels in the striatum in rat model of Parkinson’s disease(Ogata et al., 2003). We showed the low concentration in the serum isatin was found in Parkinson's disease patients as compared with controls.

Analysis of the serum isatin, an endogenous monoamine oxidase inhibitor, in aging and Alzheimer's disease

Background: Isatin, an endogenous monoamine oxidase (MAO)-A and -B inhibitor, distributes in all organs. Isatin makes monoamine in rat brain increased. MAO plays an important role in the metabolic regulation as metabolic enzyme of monoamine which is neurotransmitter substance. The MAO activity in both brain and platelet increase according to age. Several reports indicate that both A and B forms of MAO are altered in Alzheimer’s disease and Parkinson's disease.

Enzymatic detection and quantification assay of isatin, a putative stress biomarker in blood.

Isatin is an endogenous inhibitor of monoamine oxidase B and is found in human blood and tissue. Increased levels of isatin have been linked to stress and anxiety in rodents and humans; however, the metabolism of isatin in humans is largely unknown. We have developed a fluorescence-based enzymatic assay that can quantify isatin in blood samples. A phase extraction of isatin followed by a second phase extraction combined with an enzymatic reaction performed by an isatin hydrolase is used to extract and quantify isatin in whole blood samples. This results in a purity of more than 95% estimated from RP-HPLC. The hydrophobic molecule isatin is in equilibrium between an organic and aqueous phase; however, conversion by isatin hydrolase to the hydrophilic product isatinate traps it in the aqueous phase, making this step highly specific for isatin. The described protocol also offers a novel method for fast and efficient removal of isatin from any type of sample. The isolated isatinate is converted chemically to anthranilate that allows fluorescent detection and quantification. Pig plasma isatin levels are quantified to a mean of 458 nM ± 91 nM. Biophysical characterization of the isatin hydrolase shows enzymatic functionality between pH 6 and 9 and at temperatures up to 50 °C. Isatin hydrolase is highly selective for manganese ions with a dissociation constant determined to be 9.5 μM. We deliver proof-of-concept for the enzymatic quantification of isatin in blood and provide a straightforward method for further investigation of isatin as a biomarker in human health.

1-Methyl-1,2,3,4-tetrahydroisoquinoline, an Endogenous Neuroprotectant and MAO Inhibitor with Antidepressant-Like Properties in the Rat

Oxidative stress is a major contributing factor in a range of brain pathologies and in the etiology of depression. 1-Methyl-1,2,3,4-tetrahydroisoquinoline (1MeTIQ) is an endogenous substance which is present in the mammalian brain and exhibits neuroprotective, and monoamine oxidase (MAO)-inhibiting properties. In the present study, in order to investigate the potential role of 1MeTIQ as an antidepressant, we tested antidepressant-like effects of 1MeTIQ in comparison with desipramine (a classic antidepressant) in the forced swimming test (FST), and using HPLC methodology, we measured the concentrations of monoamines (dopamine, noradrenaline, serotonin) and the rate of their metabolism. 1MeTIQ given alone as well as in combination with desipramine produced an antidepressant-like effect and decreased the immobility time in the FST. Neurochemical data have shown that 1MeTIQ like desipramine, activated the noradrenergic system. However, the mechanism of action of 1MeTIQ is broader than the actions of desipramine, and 1MeTIQ inhibits the MAO-dependent oxidation of dopamine and serotonin in all investigated structures. We can conclude that 1MeTIQ exhibits antidepressant-like activity in the FST in the rat. The mechanism of its antidepressant action differs from desipramine and seems to be mostly associated with the inhibition of the catabolism of monoamines and their increased concentrations in the brain. 1MeTIQ seems to be very beneficial from the clinical point of view as a reversible MAO inhibitor with a significant antidepressant effects.

Immobilization stress increases endogenous monoamine oxidase (MAO) inhibitor in rat liver

Although the physiological role of endogenous monoamine oxidase (MAO) inhibitor still remains unclear, the present study examined whether or not immobilization stress (IMMO) induce MAO inhibitor. An endogenous inhibitor of MAO was separated by gel filtration from 105,000 g supernate in rat liver cytosol following IMMO. The molecular weight of this inhibitor was estimated to be 500–600 by gel filtration. This inhibitor was proved to be heat-stable resistant to protease treatment. IMMO for 2 h significantly decreased MAO. These results suggest that this inhibitor is induced by IMMO. MAO activity in rat liver might be regulated by the level of this inhibitor.

Isatin, an endogenous monoamine oxidase inhibitor, triggers a dose- and time-dependent switch from apoptosis to necrosis in human neuroblastoma cells

Isatin is an endogenous indole that is increased in stress, inhibits monoamine oxidase (MAO) B and improves bradykinesia and striatal dopamine levels in rat models of Parkinson's disease. Consequently, it has been suggested that isatin might be a possible treatment for Parkinson's disease although little is known about its effects on neural cell growth and survival. The aim of this study was to investigate the survival of dopaminergic human neuroblastoma (SH-SY5Y) cells following treatment with increasing concentrations of isatin. SH-SY5Y cells were exposed to isatin for defined time points, after which cell survival was determined by MTT assay. A combination of Annexin V binding and propidium iodide (PI) exclusion was used to distinguish apoptosis from necrosis in flow cytometry experiments and FACS profiles of permeabilised PI-labelled cells were employed for the assessment of cell cycle distribution. Isatin treatment (1–400 μM) for 24 h induced a significant dose-dependent increase in MTT metabolism by SH-SY5Y cells in culture, but this was not due to an increase in cell division. At the higher concentrations (200–400 μm) isatin triggered cell death, although MTT metabolism was still increased in the culture, suggesting that surviving cells were hypermetabolic. Following a longer (48 h) exposure, isatin was found to cause cell death in a dose-dependent manner; at lower concentrations (50 μM), the predominant mode of cell death was apoptosis while at the highest concentration (400 μm) increasing numbers of necrotic cells were also evident. Thus, in dopaminergic SH-SY5Y cells isatin induces cell death in dose- and time-dependent manner. This death occurred as a continuum of survival, apoptosis and necrosis. Our results re-emphasise that caution should be exercised when considering high doses of isatin as a putative anti-Parkinson's disease therapeutic.

Inhibition of brain mitochondrial monoamine oxidases by the endogenous compound 5-hydroxyoxindole

5-Hydroxyoxindole is a recently identified endogenous compound. Its physiological role remains unclear but certain evidence exists, that it may share some regulatory properties with isatin, a known endogenous inhibitor of monoamine oxidase (MAO) type B (MAO-B). In this study several oxidized indoles were tested for their in vitro inhibition of MAO type A (MAO-A) and B of rat brain non-synaptic mitochondria. 5-Hydroxyoxindole was less potent MAO-A inhibitor ( ic 50 56.8 μM) than isatin (31.8 μM) and especially 5-hydroxyisatin (6.5 μM), but it was the only highly selective MAO-A inhibitor among the all compounds studied ( ic 50 MAO-A: ic 50 MAO-B=0.044). Thus, the in vitro data suggest that MAO-A may represent potential target for 5-hydroxyoxindole.

Comparative Study of the Effects of Isatin, an Endogenous MAO-Inhibitor, and Selegiline on Bradykinesia and Dopamine Levels in a Rat Model of Parkinson’s Disease Induced by the Japanese Encephalitis Virus

We previously reported that exogenously administered isatin, an endogenous monoamine oxidase (MAO) inhibitor, significantly increased acetylcholine (ACh) and dopamine (DA) levels in the rat striatum. Selegiline [(−)-deprenil] was developed as a MAO-B inhibitor more than 30 years ago and widely used in the treatment of Parkinson’s disease. Effects of isatin or selegiline were investigated in Japanese encephalitis virus (JEV)-induced post-encephalitic parkinsonism rats by a pole test for detecting motor activity and by the determination of biogenic amine levels. Motor activity of JEV-induced rats receiving isatin (100 mg/kg per day for 1 week, i.p.) or selegiline (0.2 mg/kg per day for 1 week, i.p.) was significantly improved compared with that of untreated JEV-infected rats. Both isatin and selegiline prevented the decrease in striatal DA levels in JEV-rats. The increased turnover of DA (DOPAC/DA) induced by JEV was significantly inhibited by isatin, but not by selegiline. These results suggested that exogenously administered isatin and selegiline can improve JEV-induced parkinsonism by increasing DA concentrations in the striatum.

Isatin, an endogenous MAO inhibitor, improves bradykinesia and dopamine levels in a rat model of Parkinson's disease induced by Japanese encephalitis virus

Isatin, an endogenous monoamine oxidase (MAO) inhibitor, has an important role in the control of neurotransmitter concentration. We previously reported that exogenously administered isatin significantly increased acetylcholine (ACh) and dopamine (DA) levels in the rat striatum. In order to test the possibility of treating Parkinson's disease by isatin, we evaluated DA levels in the striatum and bradykinesia using a rat model of Parkinson's disease induced by the Japanese encephalitis virus (JEV). We have already reported that in adult Fischer rats infected with JEV at day 13, there was a marked decrease of tyrosine hydroxylase-positive neurons in the bilateral substantia nigra after 12 weeks. Effects of isatin were investigated in JEV-induced post-encephalitic parkinsonism rats by a pole test and high performance liquid chromatograph (HPLC) with an electrochemical detector (ECD). Isatin (100 mg/kg per day for 1 week, intraperitoneal injection) improved the bradykinesia observed in the JEV-induced parkinsonism rats. Dopamine (DA) concentrations in the JEV-infected rats were profoundly reduced in the striatum as compared with controls. Isatin also increased DA in the striatum of parkinsonism rats. These results suggest that isatin could be a possible treatment for Parkinson's disease as well as for post-encephalitic parkinsonism.

Effects of isatin on rotational behavior and DA levels in caudate putamen in Parkinsonian rats

Isatin was a potent endogenous monoamine oxidase (MAO) inhibitor that is more active against MAO-B than MAO-A. The acute effects of isatin on apomorphine (APO)-induced rotations were evaluated in Parkinsonian rats induced by 6-hydroxydopamine (6-OHDA) lesion. Furthermore, the effects of isatin on DA release in caudate putamen (CPu) of model and normal rats were monitored using fast cyclic voltammetry (FCV). The contents of monoamine transmitters and their metabolites in CPu of model and normal rats were also analyzed by high performance liquid chromatography with electrochemical detection after administration of isatin. Here we show that isatin (100 mg/kg, i.p.) apparently inhibited APO-induced rotations of Parkinsonian rats to 39.1±3.7% of the control ( n=12), while it had no apparent effects on electrical stimuli-induced DA release either in normal rats or in model rats. In addition, the content of 5-hydroxytryptamine but not DA was increased in both normal rats and model rats after isatin (100 mg/kg, i.p.) was administered ( P<0.01, n=6). The content of 5-hydroxyindole acetic acid was not changed. These results suggest that isatin cannot increase DA levels in rat CPu. Therefore, the effects of isatin on APO-induced rotations of our Parkinsonian rats could not attribute to its inhibition of DA catabolism as a MAO inhibitor.

Pharmacological role of isatin, an endogenous MAO inhibitor

Isatin (indole-2,3-dione) has been found in mammalian tissues as one of major components of tribulin, a postulated endogenous marker of stress and anxiety. I previously identified isatin as an endogenous inhibitor of monoamine oxidase (MAO) in the human urine and the brain of stroke-prone spontaneously hypertensive rats (SHRSP) using GC-MS. A single dose of isatin significantly increased norepinephrine (NE) and 5-hydroxytryptamine (5-HT) concentrations measured 2 h later in the various brain regions of normotensive Wistar Kyoto rats (WKY). Striatal acetylcholine (ACh) and dopamine (DA) levels significantly increased 2 h after the administration of isatin. Perfused through a microdialysis probe, isatin also produced a significant and concentration-dependent increase in the ACh and DA concentration in the perfusate from the rat striatum. In the patients with Parkinson's disease, urinary isatin concentrations tended to increase according to the severity of disease, as classified by the Hoehn and Yahr criteria. Isatin significantly increased striatal DA levels in a rat model of Parkinson's disease. Isatin may play a role in the regulation of the brain levels of ACh and DA. Furthermore, isatin has a wide spectrum of biological properties: (a) a marker of stress and anxiety, (b) an inhibitor of a number of enzymes, (c) an anti-seizure agent, (d) an inhibitor of benzodiazepin receptors and ANP binding to its receptors.

Effects of isatin, an endogenous MAO inhibitor, on dopamine (DA) and acetylcholine (ACh) concentrations in rats

Isatin (indole-2,3-dione), an endogenous inhibitor of monoamine oxidase (MAO), has several physiological properties for stress and anxiety. We previously identified isatin in the brain of stroke-prone spontaneously hypertensive rats (SHRSP) using gas-chromatography mass spectrometry. This study elucidated the effects of isatin on the ACh and DA levels of brain tissues in rats. Furthermore, we evaluated the effect of isatin on DA levels in a rat model of Parkinson's disease induced by Japanese encephalitis virus. Striatal ACh and DA levels significantly increased at 2 hours after isatin (50-200 mg/kg, i.p.) administration. Perfused through a microdialysis probe, isatin (10(-6)-10(-4) M) also produced a significant and concentration-dependent increase in the ACh and DA concentrations in the perfusate from the rat striatum. Furthermore, urinary isatin concentrations in patients with Parkinson's disease tend to increase according to the severity of disease. Isatin (100 mg/kg, i.p.) significantly increased striatal DA levels in a rat model of Parkinson's disease. These results suggest that urinary isatin may become a diagnostic marker for the clinical severity of Parkinson's disease and that endogenous isatin, a new biological modulator, may play a role in the regulation of the brain levels of ACh by increasing the level of DA under stress.

The relationship between circadian patterns of salivary cortisol and endogenous inhibitor of monoamine oxidase a

The circadian pattern of free cortisol, measured in saliva, was monitored in normal healthy adults (N = 41) for the first half hour immediately after awakening and in a smaller group (N = 8) at timed intervals throughout the day. The endogenous inhibitor of monoamine oxidase A (MAO-AI) was measured in the same saliva samples in order to explore the relationship between circadian activation of the hypothalamic-pituitary-adrenocortical (HPA) axis and MAO-AI. A marked elevation of salivary cortisol was recorded in the first half hour immediately after awakening resulting in a two to three fold increase from the first awakening level. By contrast MAO-AI was highest immediately upon awakening and fell subsequently. Hence the cortisol response to awakening is preceded by heightened MAO-AI. Moreover those subjects who showed more persistently elevated MAO-AI were characterised by a more pronounced cortisol response. An association between MAO-AI and cortisol was also manifest in the diurnal pattern recorded at timed intervals throughout the day. The decline of salivary cortisol from the morning acrophase to the evening nadir was paralleled by MAO-AI. Both patterns of decline were significant (P < 0.01). Taken together with previously reported psychological stress studies these findings suggest a possible relationship between MAO-AI and HPA activity.

Determination of Isatin, an Endogenous Monoamine Oxidase Inhibitor, in Urine and Tissues of Rats by HPLC

1. We have previously identified isatin as one of the endogenous monoamine oxidase (MAO) inhibitors in the urine and the brain of stroke-prone spontaneously hypertensive rats (SHRSP), using gas chromatography-mass spectrometry (GC-MS). 2. In this study, we attempted to develop a convenient assay to determine isatin using high performance liquid chromatography with an ultraviolet detector (HPLC-UV). The standard curve for authentic isatin was linear at a range from 2 to 20 nmol per ml. The coefficient of variance was within 3% for both intra-assay and inter-assay. The sensitivity was 20 pmol per 10 μl of urine sample. 3. Isatin concentration correlated significantly and positively with endogenous MAO activity (tribulin-like activity) in both urine ( r=0.924, P<0.001) and kidney extracts ( r=0.862, P<0.01). There was a significant difference in urinary isatin between Wistar Kyoto rats (WKY) and SHRSP. Oral administration of isatin increased urinary isatin concentration and systolic blood pressure in WKY. 4. Determination of isatin using HPLC-UV may be useful for elucidating role of isatin in various conditions of stress and disease.

Does isatin interact with rat brain monoamine oxidases in vivo?

Isatin is a reversible endogenous monoamine oxidase (MAO) inhibitor found in the brain. The influence of isatin on the degree of irreversible MAO inhibition by phenelzine and pargyline has been studied in vitro and in vivo experiments. In vitro 10 and 100 μM isatin reduced irreversible inhibition of MAO A and MAO B assayed after mitochondria wash. Pretreatment of rats with `anxiogenic dose' of isatin (10 mg/kg) protected neither MAO A nor MAO B against subsequently administered phenelzine (10 mg/kg). `Anticonvulsant' dose of isatin (80 mg/kg) reduced phenelzine-dependent inhibition of MAO B but not MAO A. Data obtained suggest that the anticonvulsant effect of isatin does not result from the inhibition of MAO A and the increase of monoamine level in the brain reported in few laboratories is apparently stipulated by a mechanism that does not involve MAO A inhibition.

Urinary cortisol and endogenous monoamine oxidase inhibitor(s), but not isatin, are raised in anticipation of stress and/or arousal in normal individuals

Cortisol, isatin and monoamine oxidase (MAO) inhibitory activity (tribulin) distinguished by its A and B components were measured in spot-urine samples provided by 20 students prior to an assessed oral seminar presentation. Stress arousal checklists (SACL) were completed on five occasions at timed intervals before and after the presentation over a 3 1/2-hour period. Significant changes in stress and arousal (p ?t 0.000) were reported during this period; levels reached their peak at the time of presentation and fell to their lowest 30 minutes after the presentation. Urinary cortisol concentration was found to correlate with MAO-A inhibitory activity (r = 0.444, p = 0.05) and with the elevation in perceived stress (but not arousal) in terms of peak levels compared to post-presentation base levels (r = 0.478, p ?t 0.05). Both MAO-A and B inhibitory activities correlated with the elevation in arousal (but not stress) in terms of peak levels, experienced during the presentation, compared to post-presentation base levels (respectively: r = 0.6477, p ?t 0.005; r = 0.6068, p ?t 0.01). Additionally, there was a difference between absolute peak arousal (but not stress) experienced by those subjects who had previously exhibited high levels of MAO-B inhibitory activity compared to those with low levels (p ?t 0.05, two-tailed). Isatin did not correlate with any of the measured parameters.

Urinary cortisol and endogenous monoamine oxidase inhibitor(s), but not isatin, are raised in anticipation of stress and/or arousal in normal individuals

Urinary cortisol and endogenous monoamine oxidase inhibitor(s), but not isatin, are raised in anticipation of stress and/or arousal in normal individuals

P-749 - Effect of isatin, an endogenous monoamine oxidase (MAO) inhibitor, on acetylcholine release from rat striatum.

P-749 - Effect of isatin, an endogenous monoamine oxidase (MAO) inhibitor, on acetylcholine release from rat striatum.