Melatonin as a chronobiotic/cytoprotector: its role in healthy aging

Preservation of sleep, a proper nutrition and adequate physical exercise are key elements for healthy aging. Aging causes sleep alterations, and in turn, sleep disturbances lead to numerous pathophysiological changes that accelerates the aging process. In the central nervous system, sleep loss impairs the clearance of waste molecules like amyloid-β or tau peptides. Melatonin, a molecule of unusual phylogenetic conservation present in all known aerobic organisms, is effective both as a chronobiotic and a cytoprotective agent to maintain a healthy aging. The late afternoon increase of melatonin “opens the sleep doors” every night and its therapeutic use to preserve slow wave sleep has been demonstrated. Melatonin reverses inflammaging via prevention of insulin resistance, suppression of inflammation and down regulation of proinflammatory cytokines. Melatonin increases the expression of α- and γ-secretase and decreases β-secretase expression. It also inhibits tau phosphorylation. Clinical data support the efficacy of melatonin to treat Alzheimer’s disease, particularly at the early stages of disease. From animal studies the cytoprotective effects of melatonin need high doses to become apparent (i.e. in the 40–100 mg/day range). The potentiality of melatonin as a nutraceutical is discussed.     

Antioxidant properties of melatonin: a pulse radiolysis study

Various one-electron oxidants such as OH*, tert-BuO*, CCl3OO*, Br2*- and N3*, generated pulse radiolytically in aqueous solutions at pH 7, were scavenged by melatonin to form two main absorption bands with lambda(max) = 335 nm and 500 nm. The assignment of the spectra and determination of extinction coefficients of the transients have been reported. Rate constants for the formation of these species ranged from 0.6-12.5x10(9) dm3 mol(-1) s(-1). These transients decayed by second order, as observed in the case of Br2*- and N3* radical reactions. Both the NO2* and NO* radicals react with the substrate with k = 0.37x10(7) and 3x10(7) dm3 mol(-1) s(-1), respectively. At pH approximately 2.5, the protonated form of the transient is formed due to the reaction of Br2*- radical with melatonin, pKa ( MelH* <=> Mel* + H+) = 4.7+/-0.1. Reduction potential of the couple (Mel*/MelH), determined both by cyclic voltammetric and pulse-radiolytic techniques, gave a value E(1)7 = 0.95+/-0.02 V vs. NHE. Repair of guanosine radical and regeneration of melatonin radicals by ascorbate and urate ions at pH 7 have been reported. Reactions of the reducing radicals e(aq)- and H* atoms with melatonin have been shown to occur at near diffusion rates.     

The organization of melatonin in lipid membranes

Melatonin is a hormone that has been shown to have protective effects in several diseases that are associated with cholesterol dysregulation, including cardiovascular disease, Alzheimer's disease, and certain types of cancers. We studied the interaction of melatonin with model membranes made of dimyristoylphosphatidylcholine (DMPC) at melatonin concentrations ranging from 0.5 mol% to 30 mol%. From 2-dimensional X-ray diffraction measurements, we find that melatonin induces a re-ordering of the lipid membrane that is strongly dependent on the melatonin concentration. At low melatonin concentrations, we observe the presence of melatonin-enriched patches in the membrane, which are significantly thinner than the lipid bilayer. The melatonin molecules were found to align parallel to the lipid tails in these patches. At high melatonin concentrations of 30 mol%, we observe a highly ordered melatonin structure that is uniform throughout the membrane, where the melatonin molecules align parallel to the bilayers and one melatonin molecule associates with 2 lipid molecules. Understanding the organization and interactions of melatonin in membranes, and how these are dependent on the concentration, may shed light into its anti-amyloidogenic, antioxidative and photoprotective properties and help develop a structural basis for these properties.     

从生物进化看褪黑素的功能意义

褪黑素在生物发布极为广泛,从低等原核生物到高５等脊椎动物,乃至植物体内均发现褪黑素。在漫长的生物进化过程中,它作为了种古老多效性的分子而保留至今。生物中门类浩繁,形态、功能千差万别,但体内的褪黑素却有相似的作用：（１）传递光暗信号,使生物的各种内源性节律与环境周期保持同步。（２）有铲清除生物体内的自由基。使机体免受氧化损伤。（３）褪黑素与钙调蛋白结合,调节细胞骨架的机能状态,以执行特定的功能、褪黑     

The use of melatonin in Alzheimer's disease

About 45% of Alzheimer's disease (AD) patients have disruptions in their sleep and sundowning agitation. Since melatonin secretion is greatly inhibited in AD patients we have used melatonin to treat sleep disorders in AD patients since 1995. In a first study [21] we reported, in 7 out of 10 dementia patients treated with melatonin (3 mg p.o. at bed time), a decreased sundowning. In a second study [22] we examined 14 AD patients who received 9 mg melatonin daily for 22 to 35 months, observing a significant improvement of sleep quality with stabilization of behavioral and cognitive parameters. In a third study [23] we reported two monozygotic twins with AD and similar cognitive impairment, one of them receiving 6 mg melatonin at bedtime daily for 3 years. Melatonin treatment improved sleep quality and suppressed sundowning. We now report the effect of melatonin (4-month-long treatment with 6 mg/day) in 45 AD patients with sleep disturbances. Melatonin improved sleep and suppressed sundowning, an effect seen regardless of the concomitant medication employed to treat cognitive or behavioral signs of AD. Melatonin treatment seems to constitute a selection therapy to ameliorate sundowning and to slow evolution of cognitive impairment in AD patients.     

The dim light melatonin onset, melatonin assays and biological rhythm research in humans

The most useful marker for human circadian phase position is the dim light melatonin onset (DLMO). This is optimally obtained by sampling blood or saliva in the evening at intervals of 30 min or less. Ambient light intensity should not exceed 30-50 lx. For many years, the DLMO was determined mainly with the 'gold standard' GCMS technique for measuring melatonin in human plasma. However, new and improved RIAs now provide the requisite sensitivity and accuracy (specificity) for detecting the time that low daytime levels begin to increase in the evening: the lower the operational threshold for the DLMO, the more reliable it is as a phase marker.     

Antioxidant and antiapoptotic properties of melatonin restore intestinal calcium absorption altered by menadione

The intestinal Ca²⁺ absorption is inhibited by menadione (MEN) through oxidative stress and apoptosis. The aim of this study was to elucidate whether the antioxidant and antiapoptotic properties of melatonin (MEL) could protect the gut against the oxidant MEN. For this purpose, 4-week-old chicks were divided into four groups: (1) controls, (2) treated i.p. with MEN (2.5 μmol/kg of b.w.), (3) treated i.p. with MEL (10 mg/kg of b.w.), and (4) treated with 10 mg MEL/kg of b.w after 2.5 μmol MEN/kg of b.w. Oxidative stress was assessed by determination of glutathione (GSH) and protein carbonyl contents as well as antioxidant enzyme activities. Apoptosis was assayed by the TUNEL technique, protein expression, and activity of caspase 3. The data show that MEL restores the intestinal Ca²⁺ absorption altered by MEN. In addition, MEL reversed the effects caused by MEN such as decrease in GSH levels, increase in the carbonyl content, alteration in mitochondrial membrane permeability, and enhancement of superoxide dismutase and catalase activities. Apoptosis triggered by MEN in the intestinal cells was arrested by MEL, as indicated by normalization of the mitochondrial membrane permeability, caspase 3 activity, and DNA fragmentation. In conclusion, MEL reverses the inhibition of intestinal Ca ²⁺ absorption produced by MEN counteracting oxidative stress and apoptosis. These findings suggest that MEL could be a potential drug of choice for the reversal of impaired intestinal Ca ²⁺ absorption in certain gut disorders that occur with oxidative stress and apoptosis.     

褪黑激素的研究进展

褪黑激素是一种在多个物种的多个组织中广泛存在并具有重要生理作用的激素。本文拟就褪黑激素在体内的分布,褪黑激素受体的分子结构、药理学特性、生物功能及调控模式作一简述。     

The profile of melatonin production in tumour-bearing rats

The pineal gland is involved in the regulation of tumour growth through the anticancer activity of melatonin, which presents immunomodulatory, anti-proliferative and anti-oxidant effects. In this study we measured melatonin content directly in the pineal gland, in an attempt to clarify the modulation of pineal melatonin secretory activity during tumour growth. Different groups of Walker 256 carcinosarcoma bearing rats were sacrificed at 12 different time points during 24h (12h:12h light/dark cycle) on different days during the tumour development (on the first, seventh and fourteenth day after tumour inoculation). Melatonin content in the pineal gland was determined by high-performance liquid chromatography with electrochemical detection. During tumour development the amount of melatonin secreted increased from 310.9 ng/mg of protein per day from control animals, to 918.1 ng/mg of protein per day 14 days after tumour implantation, and there were changes in the pineal production profile of melatonin. Cultured pineal glands obtained from tumour-bearing rats turned out to be less responsive to noradrenaline, suggesting the existence, in vivo, of putative factor(s) modulating pineal melatonin production. The results demonstrated that during tumour development there is a modification of pineal melatonin production daily profile, possibly contributing to cachexia, associated to changes in pineal gland response to noradrenaline stimulation.     

褪黑素受体与GABAA受体在褪黑素延长小鼠睡眠时间中的作用

目的：研究褪黑素受体和GABAA受体在褪黑素延长小鼠睡眠时间中的作用。方法：以翻正反射消失为睡眠开始的指标,至翻正反射恢复作为睡眠时间。观察不同受体激动剂或拮抗剂对褪黑素催眠作用的影响。结果：褪黑素3型受体拮抗剂盐酸哌唑嗪对褪黑素延长小鼠睡眠时间的作用无明显影响。GABA受体内源性激动剂GABA能明显增强褪黑素延长小鼠睡眠时间的作用,而GABAA受体上的印防己毒素结合位点的配基,即氯离子通道阻断剂印防己毒素能明显拮抗褪黑素的催眠作用,GABAA受体上的GABA结合位点的拮抗剂荷包牡丹碱则对褪黑素延长小鼠睡眠作用无明显影响。结论：褪黑素延长小鼠睡眠时间的作用与褪黑素3型受体无关,而与GABAA受体关系密切,其作用主要由印防己毒素结合位点介导。     

The use of melatonin for the treatment of insomnia

The pineal product melatonin is involved in the regulation of the sleep/wake cycle in humans. In blind individuals and in people travelling through time zones, melatonin rhythms are sometimes unsynchronized with the diel cycle, and nocturnal sleep may be disturbed. Low or distorted melatonin rhythms have repeatedly been reported in middle aged and elderly insomniacs. Melatonin administration effectively synchronized the sleep wake cycle in blind individuals and in subjects suffering from jet lag and advanced sleep onset in subjects suffering from delayed sleep phase syndrome. In elderly insomniacs, melatonin replacement therapy significantly decreased sleep latency, and/or increased sleep efficiency and decreased wake time after sleep onset. In addition, melatonin substitution facilitated benzodiazepine discontinuation in chronic users. These data show an association between melatonin rhythm disturbances and difficulties to promote or maintain sleep at night. Specific melatonin formulations may be useful to treat circadian-rhythm-related sleep disorders and age-related insomnia.     

The proposed use of melatonin in controlled sheep breeding

The regulation by melatonin of hypothalamic-pituitary events in the ewe to advance seasonal oestrous activity, with no undesirable effects upon fertility, and its induction of those seasonal responses associated with short days indicates an essential role for melatonin in controlled-breeding programs in major sheep-producing countries. The development of suitable controlled-release systems to provide a choice of practical methods of melatonin delivery under field conditions is discussed as also are geographical and breed factors in controlled breeding with melatonin.     

The oxidant/antioxidant network: role of melatonin

Melatonin is now known to be a multifaceted free radical scavenger and antioxidant. It detoxifies a variety of free radicals and reactive oxygen intermediates including the hydroxyl radical, peroxynitrite anion, singlet oxygen and nitric oxide. Additionally, it reportedly stimulates several antioxidative enzymes including glutathione peroxidase, glutathione reductase, glucose-6-phosphate dehydrogenase and superoxide dismutase; conversely, it inhibits a prooxidative enzyme, nitric oxide synthase. Melatonin also crosses all morphophysiological barriers, e.g., the blood-brain barrier, placenta, and distributes throughout the cell; these features increase the efficacy of melatonin as an antioxidant. Melatonin has been shown to markedly protect both membrane lipids and nuclear DNA from oxidative damage. In every experimental model in which melatonin has been tested, it has been found to resist macromolecular damage and the associated dysfunction associated with free radicals.     

The melatonin message: duration versus coincidence hypotheses

Whereas there is little doubt that melatonin is an important hormone which mediates the effects of the pineal gland, there is debate concerning the nature of the melatonin message which the animal interprets. This brief resume considers the two main features of the melatonin rhythm which the organism could "read" to determine whether it is in a long or a short day. The first scheme is what is referred to as the duration hypothesis. This hypothesis depends on the fact that the changing photoperiod likewise alters the duration of the daily melatonin peak and this signals the organism as to daylength and the appropriate endocrine adjustments are made. The second possibility depends on the synchronization of elevated melatonin levels with the sensitivity of a particular organ system to the melatonin peak; when this occurs the organ responds accordingly. Both the external and internal coincidence models are considered. The duration and coincidence models are fundamentally quite different. In the case of the former, the altered duration of the melatonin peak per se determines the response that will occur. In the case of the coincidence models, the elevated melatonin has a more passive role with the "decision" to respond being a function of end organ sensitivity. In the final analysis, it may be that organisms use a combination of absolute duration of the melatonin pulse, direction of change of the melatonin rhythm, and synchrony of peak melatonin with the increased sensitivity of the end organ before a response is forthcoming.     

The melatonin rhythm generating system: developmental aspects

Development of the melatonin rhythm generating system, including the suprachiasmatic nucleus, sympathetic innervation, and biochemistry of the pineal gland is reviewed. This system offers investigators an interesting opportunity to study the effects of drugs, hormones, and other factors on the developmental appearance of specific structures and biochemical parameters, and to determine the role, if any, each has in the development of an integrated neural system.     

褪黑素与缺血再灌注损伤

褪黑素(melatonin,MT)具有强效抗氧化作用,在肠、肝脏、心脏、脑等器官的缺血再灌注损伤实验中具有清除自由基、保护线粒体、抗细胞凋亡等保护性作用。本文综合褪黑素应用于缺血再灌注损伤的动物实验的近几年相关文献,总结并分析褪黑素在缺血再灌注损伤动物实验中的保护作用及其相关机制。     

褪黑素对盐碱复合胁迫下黄瓜幼苗光合特性和渗透调节物质含量的影响

为明确外源褪黑素(MT)对黄瓜盐碱胁迫的缓解效应,以‘新春四号’黄瓜为试材,采用复合盐碱(NaCl∶Na₂SO₄∶Na₂CO₃∶NaHCO₃=1∶9∶1∶9)模拟胁迫,测定外源根施MT和盐碱胁迫下黄瓜叶片光合特性和渗透调节物质含量。结果表明: 与正常生长黄瓜幼苗相比,在40 mmol·L^-1盐碱胁迫下,外源施加10 μmol·L^-1 MT能够显著增加黄瓜幼苗叶片的叶绿素、可溶性糖和可溶性蛋白质含量,提高净光合速率、气孔导度、蒸腾速率、光系统Ⅱ最大光化学效率、实际光化学效率、表观光合电子传递速率和光化学淬灭系数,而胞间CO₂浓度、非光化学淬灭系数、蔗糖、果糖、淀粉和脯氨酸含量减小了11.1%、13.8%、12.7%、27.5%、1.3%和32.8%,同时,碳同化关键酶核酮糖-1,5-二磷酸羧化/加氧酶、果糖-1,6-二磷酸酯酶活性显著升高,且Rubisco亚基(CsrbcS和CsrbcL)、CsFBA、CsRCA、CsFBPase、CsTK的mRNA水平均下调表达。综上,外源MT能够提高盐碱胁迫下黄瓜幼苗的叶绿素、渗透调节物质含量、光合化学效率和碳同化关键酶活性,增强幼苗光合能力,减轻复合盐碱胁迫对植株的伤害。研究结果可为抗盐碱栽培提供理论依据。     

Mechanism of melatonin action

Melatonin transduces the effect of photoperiod on the neuroendocrine system. Synthesis of melatonin in the pineal gland is well described, but the location of its target(s) and the mechanism of its action are little known. In attempt to localize melatonin target(s), the presence of high affinity binding sites in rat brain was determined. Such sites were detected in discrete brain areas, including the hypothalamus and anterior pituitary. Subcellular analysis indicated these binding sites were on plasma membranes, which suggests that melatonin modulates cell functions through intracellular second messengers. The effects of melatonin on second messengers were studied using the neonatal anterior pituitary, in which melatonin is known to inhibit the LHRH-induced release of LH. Studies on the effects of melatonin on second messenger indicated [corrected] that melatonin inhibits accumulation of cAMP and cGMP as well as synthesis of diacylglycerol and release of arachidonic acid. Time-course analysis indicates that inhibition by melatonin of the LHRH-induced release of LH increases following long preincubation. Since the effect of melatonin on LHRH-induced release of LH is prevented by dibutyryl cAMP, we conclude that melatonin might act by inhibiting production of cAMP.     

Antiglycation activity of melatonin

For the first time, it was found that the hormone melatonin exhibited antiglycation activity in vitro. It was shown that melatonin significantly slowed down the accumulation of fluorescent Schiff adducts formed as a result of BSA modification in the presence of high concentration of fructose. It was noted that, unlike the fructosylation reaction, melatonin did not affect the process of modification of BSA by methylglyoxal. We assume that melatonin is able to inhibit the development of the Maillard reaction but does not affect the process of BSA modification by reactive carbonyls.     

O-linked melatonin dimers as bivalent ligands targeting dimeric melatonin receptors

A series of dimeric melatonin analogues 3a-e obtained by connecting two melatonin molecules through the methoxy oxygen atoms with spacers spanning 16–24 atoms and the agomelatine dimer 7 were synthesized and characterized in 2-[¹²⁵-I]-iodomelatonin binding assays, bioluminescence resonance energy transfer (BRET) experiments, and in functional cAMP and β-arrestin recruitment assays at MT₁ and MT₂ receptors. The binding affinity of 3a-e generally increased with increasing linker length. Bivalent ligands 3a-e increased BRET signals of MT₁ dimers up to 3-fold compared to the monomeric control ligand indicating the simultaneous binding of the two pharmacophores to dimeric receptors. Bivalent ligands 3c and 7 exhibited important changes in functional properties on the G_i/cAMP pathway but not on the β-arrestin pathway compared to their monomeric counterparts. Interestingly, 3c (20 atoms spacer) shows inverse agonistic properties at MT₂ on the G_i/cAMP pathway. In conclusion, these findings indicate that O-linked melatonin dimers are promising tools to develop signaling pathway-based bivalent melatonin receptor ligands.