M.P. Terrón

Melatonin reduces body weight gain and increases nocturnal activity in male Wistar rats

AIM This study evaluated the effect of the administration of melatonin, the chief secretory product of the pineal gland, on the body weight in male Wistar rats. MAIN METHODS The animals were housed for 4months in cages equipped to log horizontal activity within a thermostatically-controlled chamber, under a 12h/12h light/dark photoperiod (lights on at 08:00h). After acclimatization, the animals were divided into two groups: (1) control animals, and (2) melatonin-treated animals. Melatonin was administered in tap water (20μg/ml), and fresh drinking fluid was changed twice weekly. Rats were fed a standard diet ad libitum. KEY FINDINGS Food and water intake, body weight, the amplitude of the activity/rest rhythm (motor activity), and blood melatonin and glucose concentrations were measured. The administration of melatonin did not influence either food or water intake or glucose levels relative to those found in the control animals. However, melatonin administration reduced body weight gain and increased nocturnal locomotor activity. The peak concentration of melatonin was found at night coinciding with the increase in nocturnal activity. SIGNIFICANCE The results show that exogenous melatonin reduces body weight gain without having marked effects on metabolism. This may be due in part to the increased nocturnal activity shown by the animals treated with the indoleamine.

Physiological concentrations of melatonin and corticosterone affect phagocytosis and oxidative metabolism of ring dove heterophils

A functional connection between the neuroendocrine and the immune systems has been established. Of particular interest is the finding that hormones such as melatonin and corticosterone are able to exert modulating effects on the immune function. Therefore, after determining the circadian rhythms of melatonin and corticosterone, we evaluated the in vitro effect of physiological concentrations of melatonin and corticosterone, separately and together, on the phagocytic function and superoxide anion levels of heterophils in ring dove (Streptopelia risoria). Trials were performed with concentrations corresponding to the nocturnal and diurnal levels reached by each of the hormones (50:300 pg/mL and 100:10 ng/mL for melatonin and corticosterone, diurnal:nocturnal, respectively). The phagocytes were incubated with the hormones both alone and concurrently. At the highest (nocturnal) concentration, melatonin augmented phagocytic function and at the same time inducing a fall in superoxide anion levels. At the highest (diurnal) concentration, corticosterone also enhanced phagocytic function, but without modifying the phagocyte oxidative metabolism. In the presence of both hormones, however, whether with nocturnal or diurnal concentrations, there was a greater increase in phagocytic function and a decrease in superoxide anion levels than was produced by either of the hormones alone. In conclusion, our findings suggest that melatonin and corticosterone may have an additive effect in the modulation of phagocytic function.

Melatonin and aging: in vitro effect of young and mature ring dove physiological concentrations of melatonin on the phagocytic function of heterophils from old ring dove.

We have studied the circadian rhythm of melatonin in the ring dove (Streptopelia risoria) for different age groups: young (1-1.5 years), mature (3-4 years) and old animals (>8 years). Melatonin levels were determined by radioimmunoassay. Results showed a significant decline in plasma melatonin levels in old animals when compared with the concentrations observed in the other two age groups, in which maximum (nocturnal) concentrations were 300 pg/ml and minimum (diurnal) concentrations were 50 pg/ml. We analyzed the in vitro effect of the physiological concentrations found in young and mature animals on the heterophils obtained from old animals, evaluating the capacity for ingestion and destruction of Candida albicans, and the oxidative metabolism associate to phagocytosis by determining the superoxide anion levels. Melatonin induced an increase in both the phagocytosis index and the candidicide capacity. This effect was dose-dependent. In relation with the oxidative metabolism, a decline in superoxide anion levels after incubation with both concentrations of the hormone was observed. Thus our results corroborate in this avian species the decline in plasma melatonin levels with advanced age, as well as the enhancing effect of physiological concentrations of melatonin on the phagocytic function.

Hydrogen peroxide increases the phagocytic function of human neutrophils by calcium mobilisation

We have studied the effect of exogenous administration of hydrogen peroxide (H2O2) on phagocytic activity of human neutrophils. The treatment of cells with increasing concentrations of H2O2 evoke a significant elevation of phagocytic function assayed as phagocytic index, percentage and efficiency; and was similar to that induced by the calcium mobilising agonist formyl-methionyl-leucyl-phenylalanine (fMLP). This stimulatory effect was reduced by pre-treatment of neutrophils with catalase and abolished in neutrophils loaded with the intracellular calcium quelator dimethyl BAPTA. In the absence of extracellular calcium, treatment of cells with H2O2 resulted in a increase in [Ca2+]i, indicating the release of calcium from intracellular stores. H2O2 abolished the typical calcium release stimulated by the physiological agonist fMLP, while depletion of agonist-sensitive calcium pools by fMLP was able to prevent H2O2-induced calcium release. We conclude that H2O2 induces calcium release from agonist-sensitive stores and consequently increase the phagocytosis process.

Phagocytosis of Candida albicans and Superoxide Anion Levels in Ring Dove (Streptopelia risoria) Heterophils: Effect of Melatonin

We observed in previous studies on avian heterophils that incubation with either physiological or pharmacological concentrations of the neurohormone melatonin increased the phagocytosis of inert particles (latex beads), and also provoked a decline in superoxide anion levels of those phagocytes. In the present study, we wanted to corroborate whether melatonin acts on the oxidative metabolism that accompanies the respiratory burst during phagocytosis by inducing a more effective phagocytic activity at the same time as exerting an antioxidant effect to eliminate and/or scavenge the free radicals left over after the destruction of the foreign material. To this end, we evaluated the ingestion and destruction of Candida albicans (live particles) by ring dove (Streptopelia risoria) heterophils after different times of incubation (30 and 60 min) with physiological concentrations of melatonin (50 pg/ml diurnal and 300 pg/ml nocturnal), as well as with a pharmacological concentration 23 × 106 pg/ml (100 µm) of the hormone. In parallel, using the same times of incubation, we evaluated the oxidative metabolism by determining the superoxide anion levels (O2−·). The results show that melatonin, at all the times and concentrations studied, increases both the phagocytosis index (number of C. albicans phagocytosed by 100 heterophils) and the candidicide power (percentage of C. albicans killed of those ingested by 100 heterophils). The effect was dose‐dependent. With respect to the oxidative metabolism accompanying the digestion and destruction, there was a decline in superoxide anion levels after incubation with all of the concentrations of the hormone studied. The effect was dose‐dependent and most pronounced at 60 min. These results thus corroborate the proposal that melatonin enhances the phagocytic function at the same time as neutralizing the oxidative stress derived from this immune function.

Effect of melatonin and tryptophan on humoral immunity in young and old ringdoves (Streptopelia risoria)

Melatonin is involved in the regulation of both cellular and humoral immunity. In the present study we have evaluated the effect of the oral administration of melatonin and its precursor, the amino acid tryptophan, on humoral immune response in ringdoves (Streptopelia risoria) from different age groups. Male and female ringdoves of 4-5years of age (young) and 12-14years of age (old) were used in this study. The animals received a single capsule of 300mg/kg b.w. of tryptophan (old animals) for 3 consecutive days 1h after lights on or a single oral dose (0.25 or 2.5mg/kg body weight/0.1ml per animal/day, young and old animals, respectively) of melatonin, for 3 consecutive days 1h before lights off. Blood samples were taken before beginning the treatment (basal values) and at the end of the treatment. Immunoglobulins, bactericidal and haemolytic activity were measured. Our results show that in old animals the humoral immune response was reduced with respect to the young. Both melatonin and tryptophan treatment increased the immunoglobulin concentration, with the nocturnal values being significantly higher than diurnal values and with a major effect in old animals. The bactericidal activity of the S. risoria serum against Staphylococcus aureus, after the treatment with melatonin or tryptophan, was increased at night with a greater effect in old animals. No significant differences were observed in the haemolytic activity of the serum in young animals, but there was an increase in old animals, with higher values at night after treatment with melatonin. In general, the oral administration of melatonin or tryptophan produced a stimulation of humoral immune response with greater effects in old ringdoves.

Melatonin, lipid peroxidation, and age in heterophils from the ring dove (Streptopelia risoria)

Numerous recent studies have shown the ability of physiological as well as all pharmacological concentrations of melatonin to prevent oxidative stress. We have found that incubating avian heterophils from young birds with a pharmacological concentration of 100 μM (23 × 106 pg/ml) melatonin reduced superoxide anion levels by modulating the activity of superoxide dismutase while also enhancing phagocytosis. There was also a decline in lipid peroxidation levels with both physiological and pharmacological concentrations of this indolamine. In the present work, we evaluated malonaldehyde (MDA) levels as an indicator of lipid peroxidation (both basal and antigen-induced) in young and old animals (ring doves) at different times of day (16:00 and 00:00) and with two incubation times (15 and 60 min). The lipid peroxidation was also measured in heterophils from old animals, incubated with the physiological concentrations of melatonin measured in young animals (50 and 300 pg/ml, diurnal and nocturnal, respectively). The results, expressed as nmol MDA/mg protein, show that MDA levels were higher in heterophils of old animals than in the young birds in all the experimental groups studied at both 16:00 and 00:00 (00:00 is the time at which the lowest peroxidation levels were obtained). Incubation with melatonin was found to reduce MDA levels, with the maximum reduction being after the 60 min incubation time and the nocturnal melatonin concentration. At both concentrations (diurnal and nocturnal), melatonin also counteracted the enhancement of MDA levels caused by latex beads, with the effect being greater at the longer incubation time. In conclusion, the results are further evidence of the antioxidant effect of melatonin even at physiological concentrations, and suggest its utility as a therapeutic agent in some pathological processes associated with age.

Systemic Inflammatory Load in Young and Old Ringdoves Is Modulated by Consumption of a Jerte Valley Cherry-Based Product

A chronic subclinical inflammatory status that coexists with immune dysfunction is commonly found in the elderly population. Consumption of foods rich in antioxidants (e.g., cherries) is an attractive strategy to reduce risk from chronic diseases. Based on previous studies showing the antioxidant effect of a Jerte Valley cherry derivative product in humans, the objective of this work was to evaluate the effect of the intake of a Jerte Valley cherry-based beverage on inflammatory load in both young and old ringdoves (Streptopelia risoria). To this purpose, circulating levels of pro-inflammatory and anti-inflammatory cytokines as well as serum levels of different acute-phase proteins were measured before and after a 10-day treatment with the Jerte Valley cherry-based beverage. Thus, the 10-day treatment with the cherry-based beverage modulated the balance of pro- and anti-inflammatory cytokines in both young and old ringdoves by down-regulating the levels of pro-inflammatory cytokines (interleukin [IL]-1β, tumor necrosis factor-α, and interferon-γ) and up-regulating the levels of anti-inflammatory cytokines (IL-4, IL-2, and IL-10). Moreover, the 10-day treatment with the Jerte Valley cherry-based product reduced the levels of several proteins involved in acute-phase responses, such as C-reactive protein, haptoglobin, α(2)-macroglobulin, and serum amyloid P component. On the other hand, old birds showed imbalanced levels of inflammatory markers toward a pro-inflammatory status, thereby underlining the fact that aging is usually accompanied by systemic inflammation and inflammation-related chronic diseases. To sum up, the data suggest a potential health benefit by consuming the cherry-based beverage, especially in aged populations, through their anti-inflammatory properties.

Tryptophan and Melatonin-Enriched Foodstuffs to Improve Antioxidant Status in Aging

Abstract As aging proceeds, the efficiency of antioxidant defense systems lowers, and the ability to remove deleterious reactive oxygen species and free radicals decreases. Consumption of foodstuffs containing high amounts of antioxidant compounds has been associated with the balance of the free radical/antioxidant status, which helps to minimize oxidative stress in the organism and to reduce the risk of suffering chronic diseases throughout life. Although polyphenols represent the archetype of the health-promoting effects of the Mediterranean diet, melatonin is currently revolutionizing the nutrition field by virtue of its biologic activities and its excellent bioavailability. It is known that increasing the levels of circulating melatonin by introducing, into the diet, foodstuffs rich in this compound (or its precursor, the amino acid tryptophan) enhances the individual’s antioxidant status and produces a widespread spectrum of physiologic effects. Therefore, the intake of tryptophan and/or melatonin-enriched foodstuffs may be a natural approach to prevent or combat important diseases related to oxidative stress.