Effect of Melatonin and Caffeine Interaction On Caffeine Induced Oxidative Stress and Sleep Disorders
Effect of Melatonin and Caffeine Interaction On Caffeine Induced Oxidative Stress and Sleep Disorders
Effect of Melatonin and Caffeine Interaction On Caffeine Induced Oxidative Stress and Sleep Disorders
25(2010) 17 – 24
www.physocnigeria.org/njps
Summary: Effect of interaction of melatonin and caffeine on caffeine induced oxidative stress and sleep disorders was
studied. Fifteen wistar rats were randomly assigned into three study groups. The animals in group 1 (the control) received a
placebo of 10.0 ml distilled water via gastric intubation. The hosts in groups 2 and 3 were treated with 100 mg caffeine/ kg,
or melatonin/ kg, respectively, in a total volume of 10.0 ml vehicle. The experiment lasted for 30 days. One day after the
final exposure, the animals were euthanized by inhalation of overdose of chloroform. Blood was collected by cardiac
puncture. Serum was obtained by centrifugation (6000 Xg, 30 mins), and used for serum total protein and serum blood urea
nitrogen levels. The brain of each rat was also harvested and processed into whole homogenate, frozen in liquid nitrogen
(N2), and maintained at -80oC until used for total brain cholesterol and tryptophan levels. The results showed that
interaction of melatonin and caffeine enhanced protein synthesis; stimulated gonadotrophin release, and could be used as
oral contraceptive for women, and may be beneficial in the treatment of impotence (androgen depression), leading to
improved reproductive and sex life; stimulated tryptophan metabolism, which prevents vitamin B6 deficiency, anemia,
negative nitrogen balance, tissue wasting and accumulation of xanthurenic acid, which promotes sleep; and could be
beneficial in the treatment of hyper cholesterolemia, thereby preventing coronary heart disease, and post menopausal
osteoporosis.
Key words: melatonin, caffeine, oxidative stress, sleeps disorders and free radicals.
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blood vessels (Eteng et al, 1997). Caffeine and other et al, 1994; Lissoni et al, 1994; Sandyk, 1994; Chan
methylxanthines occupy adenosine receptors and then et al, 1995; Lissoni et al, 1995). It also lowers total
block the action of the neurotransmitter. Adenosine cholesterol and LDL levels in rats (Chan et al, 1995);
receptors are linked with interplay of release, acts as a powerful chronobiotic maintaining
reuptake, metabolism and excretion of synchronicity and preventing desynchrony of
neurotransmitters (Julien, 1996; Eteng et al, 1997; circadian rhythms (Armstrong, 1989; Attenburrow et
Obochi, 2006). Thus, blockade of adenosine receptors al, 1996); improves sleep (Garfinkel et al, 1995) and
by caffeine results in alteration in behavioural pattern sex life (Laudon et al, 1988).
by delaying neuronal tube closure (Eteng et al, 1997); Melatonin stimulates gonadotrophin release
activates phosphorylase and lipase, thereby enhancing hormone, estrogen levels and androgen responses
glycogenolysis and lipolysis, resulting in loss of (Terzollo et al, 1993; Laudon et al, 1996), and may
weight (Julien, 1996). The blockade also inhibits inhibit ovarian function in women and may be
glutamate release, resulting in low protein synthesis, beneficial as oral contraceptive (Voordouw et al,
poor growth, low protein efficiency ratio, poor 1992). It is involved in the regulation of calcium and
nitrogen retention (Eteng et al, 1997), and decreases phosphorous metabolism by stimulating the
bone mineral density, leading to osteoporosis, parathyroid gland and by inhibiting calcitonin release
palpitations, tremor, headache, anxiety, peptic ulcer and prostaglandins synthesis (Sandyk et al, 1992).
and insomnia (Julien, 1996). The blockade of Melatonin stimulates immunoenhancing effects
adenosine receptors by caffeine inhibits androgen (Maestroni, 1993), improves leucopenia and winter
binding protein, resulting in decreased cauda depression (Neri et al, 1994; Webb and Pulg-
epididymis sperm reserve, seminiferous tubular fluid Domingo, 1995). Because of these therapeutic
volume, low sperm production and infertility (Eteng properties of melatonin, it is often bought over the
et al, 1997; Obochi, 2006). Thus, the toxicity of counter in health food shops and is consumed for
caffeine is viewed as a phenomenon of rapid improved sleep, sex life, longevity and lowering of
absorption but slow excretion (Obochi, 2006). blood pressure, leading to self medication. The
Therapeutic properties of caffeine include relief of current study is therefore focused on assessment of
pain, headache and cold (through inhibition of the how melatonin could impact upon caffeine induced
inflammatory action of prostaglandins), depression oxidative stress and sleep disorders in animals
and in appetite-suppressant medications due to its exposed daily to caffeine and melatonin.
diuretic effects (Obochi, 2006). Melatonin (N-acetyl-
5-mehoxytryptamine is a hormone synthesized from
tryptophan in the pineal gland of the brain in humans MATERIALS AND METHODS
and animals (Reiter, 1995); stimulated by the dark
and inhibited by light (Reiter, 1995). It also occurs Fifteen Wistar albino rats weighing between 150g
naturally in plants such as oats, tomatoes, banana and to 250g obtained from the disease free stock of the
barley (Webb and Pulg-Domingo, 1995). The animal house, Department of Biochemistry, College
principal role of melatonin involves regulation of of Medical Sciences, Benue State University,
neuro-endocrine reproductive functions (Reiter et al, Makurdi, Nigeria, were used for the study. The
1995; Webb and Pulg-Domingo, 1995). It also plays a animals were randomly assigned into three study
role in signaling changes in reproductive hormone groups of five animals per group. Each rat in a study
levels (Maestroni, 1993). Melatonin is metabolized to group was individually housed in a stainless steel
6-hydroxy-melatonin in the liver, and the main cage with plastic bottom grid and a wire screen top.
metabolite, 6-sulphatoxy-melatonin is excreted The animal room was adequately ventilated, and kept
(Maestroni, 1993; Cavallo, 1993); lowers body at a room temperature and relative humidity of 29 ±
temperature and blood flow to the brain, (Lamberg, 2oC and 40 – 70% respectively with 12 hour natural
1996; Brzezinski, 1997). light-dark cycle. The animals were fed ad libitum
Melatonin is an effective antioxidant that protects with water and rat chow (Livestock Feeds Ltd,
DNA, cells and tissues from free radical assault Makurdi, Nigeria). Good hygiene was maintained by
(Sharma et al, 1989; Stokkan et al, 1991; Pierpaoli et constant cleaning and removal of faeces and spilled
al, 1993; Tan et al, 1993; Lesnikov et al, 1994; feeds from cages daily. All animal experiments were
Pierpaoli et al, 1994; Reiter, 1995). It stimulates the approved by the Animal Care and Use Committee of
main antioxidant ezyme of the brain, glutathione the Medical College, Benue State University,
peroxidase (Reiter, 1995) and inhibits proliferation Makurdi, Nigeria.
and growth of cancer cells (Blask et al, 1991; Lissoni
et al, 1992; Cos et al, 1994; Neri et al, 1994; Aldeghi
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activation of cAMP-dependent protein kinase, PKA. receptors on the surface of the plasma membrane to
PKA phosphorylates phosphoprotein phophatase generate signals, since these molecules cannot cross
inhibitor – 1 (PP1 – 1) leading to an increase in its the plasma membrane (blood-brain-barrier). The
activity. Phosphoprotein phosphotase inhibitor – 1 amplification and subsequent transmission of such
(PP1 – 1) can inhibit the activity of numerous signals to the cell interiors require the participation of
phosphatases including HMG-CoA reductase a second messenger, usually a cyclic nucleotide,
phosphatase, which remove phosphates from HMGR. cAMP, while the hormones, peptides or receptors
This maintains HMGR in the phosphorylated and serve as the first messengers (Eteng et al, 1997).
inactive state, resulting in decreased activity of The effect of melatonin on protein levels could be
HMGR, and reduced cholesterol synthesis (Reiter, attributed to activation of transcriptional promoter
1995). Also, increases in cAMP lead to activation of and enhancer elements used for the control of gene
cAMP-dependent protein kinase, PKA, which in turn, expression, which promoted the nucleotide at the
inhibits HMGR activity, resulting in decreased initiation stage, thereby increasing protein synthesis
cholesterol synthesis (Reiter, 1995; Chan and Tang, and stimulating the gonadotrophin release, estrogen
1995). The effect of caffeine on cholesterol levels and androgen levels. This could account for use of
could also be attributed to hormonal stimuli, melatonin for improved sex life.
particularly lipolytic hormones such as glucagon, The effect of melatonin on blood urea nitrogen
adrenaline, noradrenaline, adrenocorticotrophin could be attributed to the antioxidant properties, by
hormone (ACTH), growth hormones and stimulating the main antioxidant of the brain,
parathormone, which increase the concentration of glutathione peroxidase, which neutralized free
cAMP, thereby increasing the activity of cAMP- radicals. This property of melatonin could account for
dependent protein kinase, PKA, which in turn, its use for longevity and lowering of blood pressure.
inhibited the activity of HMGR, resulting in The effect of melatonin on cholesterol levels could be
decreased cholesterol synthesis. However, insulin attributed to inhibition of the enzyme, HMGR
decreases the concentration of cAMP, and thereby activity, which catalyzes the conversion of HMG-
activates cholesterol synthesis. Thus, the ability of CoA to mevalonate. This enzyme contains SH group
insulin to stimulate, and glucagon to inhibit, HMGR and requires NADPH as cofactor (supplied by HMP
activity is consistent with the effects of these pathway) in the conversion of SH group to S-S group
hormones on other metabolic pathways. The basic which increases the hydrophobic properties, thereby
function of these two hormones is to control the distorting the lipid components of the membrane
availability and delivery of energy to all cells of the structure and function, resulting in decreased
body (Reiter, 1995; Chan and Tang, 1995). Therefore, cholesterol synthesis (Chan and Tang, 1995). In this
hormones act by influencing a change in study, melatonin acted synergistically with caffeine,
intracelluloar concentration of cAMP through the and decreased cholesterol synthesis, and could be
adenylate cyclase system. Adenylate cyclase used as therapy for hypercholesteromia. Thus, low
catalyzes the convesion of ATP to cAMP, thus, levels of melatonin could be a factor in multiple
increasing the intracellular concentration of cAMP. sclerosis, coronary heart disease, sudden infant death
The action of cAMP is mainly to activate some syndrome, epilepsy and post menopausal osteoporosis
protein kinases (Reiter, 1995). (Sturner et al, 1990; Brugger et al, 1995;
The effects of caffeine on brain tryptophan levels Constantinescu et al, 1995). The menopause is
could be attributed to inhibition of 3-hydroxy associated with a decline in melatonin secretion and
anthranilate oxidase, which catalyzes the rate limiting increased calcification (Sandyk, 1994).
step in the kynurenine-anthranilate pathway, which The effect of melatonin on tryptophan levels could
converts tryptophan (an essential amino acid) into be attributed to activation of the enzymes,
acetyl CoA. 3-hydroxy kynurenine could not be kynureninase and 3-hydroxy anthranilate oxidase,
converted to 3-hydroxy-anthranilic acid, resulting in which catalyze the conversion of 3-hydroxy
accumulation of kynurenine and 3-hydroxy kynurenine to 3-hydroxy anthranilate and 3-hydroxy
kynurenine. These products are converted to anthranilate to 2-acroleyl-3-amino fumarate, a
xanthurenic acid in the extra hepatic tissues, leading precursor for the synthesis of niacin and acetyl-CoA.
to tissue wasting and negative nitrogen balance. Activation of kynureninase and 3-hydroxy
Xanthurenic acid excretion in urine is an index of B6- anthranilate oxidase lead to synthesis of thiamin (B6),
deficiency and could lead to anemia (Chatterjee and a vitamin of B complex, thereby preventing
Shinde, 2000). accumulation of xanthurenic acid and pellagra
Mechanism of action of caffeine, therefore, (Chatterjee and Shinde, 2000).
involves interaction with hormones, peptides and
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In conclusion, this study has shown that melatonin susceptibility to autoimmune demyelinaiton: A
interacted with caffeine to promote synthesis of rationale for light therapy in multiple sclerosis.
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treatment of impotence (androgen depression), melatonin on MCF-7 human breast cancer cell
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