Moses A. Akanmu

Neuropharmacological effects of oleamide in male and female mice.

Oleamide, a fatty acid amide accumulates selectively in the cerebrospinal fluid of sleep deprived cats and rats. Oleamide has been reported to have effects on a wide range of receptors and neurotransmitter systems especially the centrally acting ones for example, dopamine acetylcholine, serotonin, gamma aminobutyric acid (GABA), cannabinoid and vanilloid among others. This suggests a wide range of central nervous system effects of the compound. The effects of intraperitoneal administered oleamide on Novelty-induced behaviours, learning and memory and forced swimming-induced depression were studied. The relative effects of the compound on the male and female mice were also noted. Oleamide dose-dependently reduced (p<0.05) novelty induced rearing, grooming and locomotion. The effects on the all NIBs started within the first 10 min of the test and the peak of the effects was observed during the third 10 min period of the test. Effect of oleamide on short-term working memory was significantly (p<0.05) affected only with the dose of 5mg/kg while the other dose of 10mg/kg had no effect. In the forced swimming test, acute triple intraperitoneal administration of oleamide at 10mg/kg induced a significant reduction in the immobility duration in mice signifying an antidepressant effect. Sex differences in the effects of oleamide (10mg/kg, i.p.) were clearly evident in active behaviours in FST. These results confirm the multiplicity of central nervous system receptors and neurotransmitters that oleamide interacts with hence its numerous and diverse neuropharmacological effects. Most importantly, the present study suggests that oleamide has antidepressant-like property.

A novel melatonin derivative modulates sleep–wake cycle in rats

The aim of this study was to evaluate, in comparison with melatonin (MLT), the effects of a novel 2-melatonin derivative, having a 2-trifluoromethyl group (MLTD) on the sleep-wake cycle in freely behaving rats. Doses of MLTD (100, 300 and 500 nmol/100 microl saline) or MLT (500 nmol/100 microl saline) replaced the i.c.v. infusion of saline during the diurnal infusion period (7:00-17:00). Diurnal infusion of MLTD significantly (P < 0.05) increased NREM sleep at doses of 300 and 500 nmol during the second 4-h time interval of the light period and the lowest dose showed delayed effects on NREM sleep in the third 4-h time interval. However, REM sleep was only increased significantly at the dose of 500 nmol. The infusion of MLT did not have effects on REM sleep but significantly increased NREM sleep. These findings support the notion that MLTD diurnal infusion into the third ventricle produces soporific effects dose-dependently and more potently than MLT. Thus, MLTD may play an important role in studies of sleep.

Effects of ketamine and N-methyl-d-aspartate on fluoxetine-induced antidepressant-related behavior using the forced swimming test

This study investigated the effects of ketamine on fluoxetine-induced antidepressant behavior using the forced swimming test (FST) in mice. In order to understand the possible role of N-methyl-d-aspartate (NMDA) neurotransmission in the antidepressant effect of fluoxetine, different groups of mice (n=10) were administered with acute ketamine (3mg/kg, i.p.), acute NMDA (75mg/kg and 150mg/kg, i.p.) and a 21-day chronic ketamine (15mg/kg, i.p./day) were administered prior to the administration of fluoxetine (20mg/kg, i.p.) in the mice. Antidepressant related behavior (immobility score) was measured using the forced swimming test. The results showed that the acute ketamine and fluoxetine alone treatments elicited a significant (p<0.05) reduction in immobility score compared with saline control. Furthermore, pre-treatment with acute ketamine significantly enhanced by the fluoxetine-induced decrease in immobility score. In contrast, pre-treatment with NMDA (150mg/kg) significantly (p<0.05) reversed fluoxetine-induced decrease in immobility score. On the other hand, chronic administration of ketamine significantly elicited an increase in immobility score as well as reversed the reduction induced by fluoxetine. Similarly, NMDA administration at both 75mg/kg and 150mg/kg increased immobility score in chronically administered ketamine groups. Furthermore, chronic administration of ketamine, followed by NMDA (75mg/kg) and fluoxetine significantly elevated the immobility score when compared with the group that received NMDA and fluoxetine but not chronically treated with ketamine. It can be suggested) that facilitation of NMDA transmission blocked fluoxetine-induced reduction in immobility score, while down-regulation of NMDA transmission is associated with increase in fluoxetine-induced antidepressant-related behavior in mice. Down-regulation of the NMDA transmission is proposed as an essential component of mechanism of suppression of depression related behaviors by fluoxetine. Modulation of NMDA transmission is suggested to be relevant in the mechanism of action of fluoxetine.

Analgesic, learning and memory and anxiolytic effects of insulin in mice

Insulin is a polypeptide hormone that is present in mammals and its main function is the maintenance of adequate blood sugar level. Insulin receptors are widely but unevenly distributed in the brain. Insulin has been reported to be involved in the regulation of neurotransmitters release. It has also been linked to the pathogenesis of neurodegenerative disorders such as Alzheimers and Parkinsons diseases. Although there is abundant literature on the study of biochemical and molecular properties of insulin, there has been no literature on its central behavioural effects on anxiety and pain relief among other behavioural effects. This study therefore investigates whether insulin has any anxiolytic and other CNS effects. This experiment was carried out in mice using animal behavioural models including a hot plate analgesic test, holeboard and elevated plus maze for anxiolytic test. A Y-maze was used for the locomotor activity and spontaneous alternation investigations. Mice were administered intraperitoneally with insulin at different doses of 0.5, 1.0 and 2.0IU/kg. The results obtained showed that insulin has no analgesic activity, however, it caused significant central inhibitory effects by decreasing both locomotor activity in both holeboard and Y-maze models and also decreased the exploratory behaviour in holeboard at doses administered dose-dependently indicating its sedative effects. In elevated plus maze, insulin had no effects on percentage of open arm entries at all doses but had a significant effect on percentage of open arm duration at the dose of 1.0IU/kg only. Insulin administration at lower doses (0.5 and 1.0IU/kg, i.p.) had no effect on spatial working memory, however, it had significant spatial working memory impairment at the dose of 2.0IU/kg, i.p. in mice. The study showed that insulin has several neuropharmacological effects at doses used.

Neuropeptide-Y Y2-receptor agonist, PYY3-36 promotes non-rapid eye movement sleep in rat

PYY3-36 is a major component of the gut-brain axis and peripheral administration has been reported to exert significant effects on feeding, brain function and is more selective for neuropeptide Y2 receptor. Therefore, we investigated the effects of nocturnal intraperitoneal administration of single doses of PYY3-36 (30 and 100 microg/kg i.p.) on food intake, water intake and the sleep-wake cycle in rats. Sleep recordings were carried out in male Sprague-Dawley rats implanted with cortical electroencephalogram (EEG) and neck electromyogram (EMG) electrodes. The EEG, EMG, food intake and water intake were assessed. The electrographic recordings obtained were scored visually as rapid eye movement (REM) sleep, non-REM (NREM) sleep and wakefulness. PYY3-36 administration 15 min prior to dark onset significantly (p<0.05) increased non-rapid eye movement (NREM) sleep and decreased wakefulness. Analysis of the dark-period at 4-h time intervals showed that nocturnal administration of PYY3-36 (30 and 100 microg/kg) significantly suppressed wakefulness and increased non-REM sleep during the first 4-h time interval. Time spent in wakefulness was significantly decreased after administration of PYY3-36 (30 and 100 microg/kg) when compared with administration of vehicle. In addition, PYY3-36 (30 and 100 microg/kg i.p.) induced an increase in the time spent in NREM sleep. The nocturnal intraperitoneal administration of the lower dose of PYY3-36 (30 microg/kg) also significantly decreased food intake [F (2,23)=4.90, p<0.05] but had no effect on water intake. These findings suggest that PYY3-36 may play an important role in the enhancement of NREM sleep and feeding behavior.

Psychoneuropharmacological activities and chemical composition of essential oil of fresh fruits of Piper guineense (Piperaceae) in mice

ETHNOPHARMACOLOGICAL RELEVANCE Piper guineense Schum & Thonn (Piperaceae) is a medicinal plant used in the Southern States of Nigeria to treat fever, mental disorders and febrile convulsions. AIMS OF THE STUDY This study aims at determining the chemical composition and the central nervous system (CNS) activities of the essential oil obtained from the plant׳s fresh fruits in order to rationalize its folkloric use. MATERIALS AND METHODS Essential oil of P. guineense (EOPG) obtained by hydrodistillation was analysed by GC/MS. EOPG (50-200mg/kg, i.p.) was evaluated for behavioural, hypothermic, sedative, muscle relaxant, anti-psychotic and anticonvulsant activities using standard procedures. RESULTS AND DISCUSSION Analysis of the oil reveals 44 compounds of which 30 compounds constituting 84.7% were identified. The oil was characterized by sesquiterpenoids (64.4%) while only four monoterpeneoids (21.3%) were found present in the oil. Major compounds identified were β-sesquiphellandrene (20.9%), linalool (6.1%), limonene (5.8%), Z-β-bisabolene (5.4%) and α-pinene (5.3%). The EOPG (50-200mg/kg, i.p.) caused significant (p<0.01) inhibition on rearing {F(4,20)=43}, locomotor {F(4,20)=22} activity and decreased head dips in hole board {F(4,20)=7} indicating CNS depressant effect; decreased rectal temperature {F(4,20)=7-16}, signifying hypothermic activity; decreased ketamine-induced sleep latency {F(4,20)=7.8} and prolonged total sleeping time {F(4,20)=8.8}, indicating sedative effect; reduced muscular tone on the hind-limb grip test {F(4,20)=22}, inclined board {F(4,20)=4-49} and rota rod {F(4,20)=13-106}, implying muscle relaxant activity; induced catalepsy {F(4,20)=47-136}, inhibited apomorphine-induced climbing behaviour {F(4,20)=9} and inhibited apomorphine-induced locomotor {F(4,20)=16}, suggesting anti-psychotic effect; and protected mice against pentylenetetrazole-induced convulsions, indicating anticonvulsant potential. CONCLUSION The most abundant component of the fresh fruits essential oil of P. guineense was β-sesquiphellandrene (20.9%); and the oil possesses CNS depressant, hypothermic, sedative, muscle relaxant, antipsychotic and anticonvulsant activities, thus providing scientific basis for its ethnomedicinal applications.

Hypnotic, anticonvulsant and anxiolytic effects of 1-nitro-2-phenylethane isolated from the essential oil of Dennettia tripetala in mice.

This study investigated the hypnotic, anti-convulsant and anxiolytic effects of 1-nitro-2-phenylethane (BPNE) obtained from the oil of Dennettia tripetala G. Baker (Annonaceae) and established its mechanism of action. The essential oil (EO) from the leaf, fruit and seed was obtained by hydrodistillation, followed by isolation of BPNE purified to 99.2% by accelerated gradient chromatography on silica, and identified by NMR and GC-MS. The pure BPNE and EO of the dried seed (93.6%) were comparatively evaluated for hypnotic, anticonvulsant and anxiolytic effects in mice. The acute toxicity of BPNE was determined and the LD50 was 490 mg/kg, intrapritonealy. The hypnotic activities of the EO and BPNE (50-400 mg/kg, i.p.) were assessed by loss of righting reflex, while sodium pentobarbitone (PBS) and diazepam (DZM) were used as positive controls. The anticonvulsant and anxiolytic effects of the EO and BPNE were evaluated in mice. Both BPNE and EO at doses ≥100 mg/kg induced spontaneous hypnosis with loss of righting reflex, significantly decreased sleep latency (SL) and also increased total sleeping time (TST) dose-dependently. They had comparable activity with NAP in TST. The BPNE exhibited higher hypnotic potency than EO at the same dose levels. The EO and BPNE offered comparable dose-related protections against PTZ- and strychnine-induced convulsions. Flumazenil (2 mg/kg) blocked the hypnotic and anticonvulsant (PTZ-convulsions) effects of both EO and BPNE. The essential oil at 5-20 mg/kg dose levels significantly (p<0.05) increased the percentage time spent and number of entries into the open arms. While at the same dose range BPNE significantly (p<0.05) increased the percentage time spent and the number of entries into the open arms respectively. The study concluded that 1-nitro-2-phenylethane exhibited dose dependent significant hypnotic, anticonvulsant and anxiolytic effects and it is the compound largely responsible for the neuropharmacological effects of the oil.

Effect of dehusked Garcinia kola seeds on the overall pharmacokinetics of quinine in healthy Nigerian volunteers.

We investigated the effect of concurrent ingestion of Garcinia kola seed on the pharmacokinetics of quinine. In a randomized crossover study, 24 healthy Nigerian volunteers were assigned into 2 groups (A and B; n = 12 per group) on the basis of G. kola dose orally ingested. Each subject received 600 mg quinine sulfate before and after ingesting 12.5 g of G. kola once daily for 7 days (group A) or 12.5 g twice daily for 6 days and once on the seventh day (group B). Blood samples were collected and analyzed for plasma quinine and its metabolite (3‐hydroxyquinine) using a validated high performance liquid chromatography method. Concurrent administration of quinine with G. kola reduced quinine tmax by 48% (group A), mean Cmax by 19% and 26% in groups A and B, respectively, and slight reduction in mean AUC0–∞ of quinine in both groups. 3‐hydroxyquinine Cmax also reduced by 29% and 32%; AUC0–∞ by 13% and 9%, respectively. The point estimates of the T/R ratio of the geometric means for all Cmax obtained and only the AUC0–∞ at a higher dose of G. kola were outside the 80%–125% bioequivalence range. In conclusion, an herb–drug interaction was noted with concurrent quinine and G. kola administration.

Hypotensive and vasorelaxant effects of (E) – Methyl isoeugenol: A naturally occurring food flavour

Application of naturally occurring (E) - methyl isoeugenol (MIE) as food flavour has been widely accepted despite the growing concerns over cardiovascular issue. Hence, we sought to investigate hypotensive property of MIE and the involvement of central and/or peripheral mechanism (s). Variation in mean arterial pressure (MAP), heart rate (HR), systolic blood pressure (SBP), baroreflex sensitivity of normotensive rats and vascular reactivity were recorded. MIE (1.11, 2.25 or 4.50mg/kg, iv) elicited dose-related decrease in MAP (-16.9±1.13; -19.0±4.18 or -27.2±3.65mmHg, respectively) and an increase in HR (17.4±1.79; 24.4±5.11 or 29.9±6.62 bmp, respectively). MIE 25 or 50mg/kg (p.o) reduced the SBP (-13.6±4.18 or -16.6±5.60mmHg, respectively) without altering baroreflex sensitivity. The hypotensive effect of MIE remained unaltered by WAY100635 (antagonist of 5-HT1A) and L-NAME (NO synthase inhibitor). Intracerebroventricular injection of MIE did not change MAP. MIE elicited endothelium independent vasorelaxation (endothelium-intact vessels, Emax 92.5±1.75%; Endothelium-denuded vessels, Emax 91.4±2.79%). MIE blocked CaCl2 or BAY K8644 (L-type voltage gated calcium channel activator)-induced vascular contractions. Our findings showed evidence of hypotensive and vasorelaxation effects of MIE with involvement of calcium channel.

Neuropharmacological profile and chemical analysis of fresh rhizome essential oil of Curcuma longa (turmeric) cultivated in Southwest Nigeria

Graphical abstract