In vitro prooxidant effect of caffeine

Caffeine is a methylxanthine compound that acts as a stimulant in humans. It is the most widely consumed behaviourally active substance in the western world and it affects calcium influx and release in living cells. Caffeine is present in various substances such as tea, coffee and some medications. This article is focused on the impact of caffeine on oxidation. Caffeine promoted the peroxidation of linoleic acid emulsions used by 32.5, 48.9, and 54.3%, respectively, at 15, 30 and 45 μg/mL concentrations. Standard antioxidants such as α-tocopherol and trolox, a water-soluble analogue of tocopherol, inhibited 76.2 and 93.2% peroxidation of linoleic acid emulsion at 45 μg/mL concentration. Also, PC₅₀ value (caffeine concentration that produced a 50% increase in linoleic acid peroxidation) of caffeine was found to be 185 μM.     

Antioxidant and radical scavenging properties of curcumin

Curcumin (diferuoyl methane) is a phenolic compound and a major component of Curcuma longa L. In the present paper, we determined the antioxidant activity of curcumin by employing various in vitro antioxidant assays such as 1,1-diphenyl-2-picryl-hydrazyl free radical (DPPH*) scavenging, 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging activity, N,N-dimethyl-p-phenylenediamine dihydrochloride (DMPD) radical scavenging activity, total antioxidant activity determination by ferric thiocyanate, total reducing ability determination by the Fe(3+)-Fe(2+) transformation method, superoxide anion radical scavenging by the riboflavin/methionine/illuminate system, hydrogen peroxide scavenging and ferrous ions (Fe(2+)) chelating activities. Curcumin inhibited 97.3% lipid peroxidation of linoleic acid emulsion at 15 microg/mL concentration (20 mM). On the other hand, butylated hydroxyanisole (BHA, 123 mM), butylated hydroxytoluene (BHT, 102 mM), alpha-tocopherol (51 mM) and trolox (90 mM) as standard antioxidants indicated inhibition of 95.4, 99.7, 84.6 and 95.6% on peroxidation of linoleic acid emulsion at 45 microg/mL concentration, respectively. In addition, curcumin had an effective DPPH* scavenging, ABTS*(+) scavenging, DMPD*(+) scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, ferric ions (Fe(3+)) reducing power and ferrous ions (Fe(2+)) chelating activities. Also, BHA, BHT, alpha-tocopherol and trolox, were used as the reference antioxidant and radical scavenger compounds. According to the present study, curcumin can be used in the pharmacological and food industry because of these properties.     

Antioxidant and antiradical activities of L-carnitine

L-carnitine plays an important regulatory role in the mitochondrial transport of long-chain free fatty acids. In this study, the antioxidant activity of L-carnitine was investigated as in vitro. The antioxidant properties of the L-carnitine were evaluated by using different antioxidant assays such as 1, 1-diphenyl-2-picryl-hydrazyl free radical (DPPH.) scavenging, total antioxidant activity, reducing power, superoxide anion radical scavenging, hydrogen peroxide scavenging and metal chelating activities. Total antioxidant activity was measured according to ferric thiocyanate method. alpha-tocopherol and trolox, a water-soluble analogue of tocopherol, were used as the reference antioxidant compounds. At the concentrations of 15, 30 and 45 microg/mL, l-carnitine showed 94.6%, 95.4% and 97.1% inhibition on lipid peroxidation of linoleic acid emulsion, respectively. On the other hand, 45 microg/mL of standard antioxidant such as alpha-tocopherol and trolox indicated an inhibition of 88.8% and 86.2% on peroxidation of linoleic acid emulsion, respectively. In addition, L-carnitine had an effective DPPH. scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, total reducing power and metal chelating on ferrous ions activities. Also, those various antioxidant activities were compared to alpha-tocopherol and trolox as references antioxidants.     

Turnera ulmifolia L. (Turneraceae): preliminary study of its antioxidant activity

Turnera ulmifolia L. is used in Brazilian folk medicine as an anti-inflammatory. Since this activity may be correlated with the presence of antioxidant compounds, a leaf extract was evaluated for its radical scavenging capacity (RSC). The in vitro RSC of a 50% hydroethanolic (HE) extract was evaluated by beta-carotene/linoleic acid coupled oxidation system for the inhibition of oxidation and the lipid peroxidation inhibition in rat brain homogenates, using thiobarbituric acid reactive substances (TBARS) and chemiluminescence (CL). Results indicated, through peroxidation suppression, that this extract exhibited greater antioxidative activity (77.4% +/- 10%) than alpha-tocopherol (58.4% +/- 3.7%). TBARS and CL inhibition was concentration-dependent and Q(1/2) values were 8.2 and 6.0 microg/mL for TBARS and CL, respectively. For alpha-tocopherol these values were 7.1 microg/mL (TBARS) and 9.8 microg/mL (CL). Phenolic compounds may be responsible for this antioxidant capacity.     

Effects of epidermal growth factor and linoleic acid on lipid contents in human intestinal C2BBe1 cells

Epidermal growth factor (EGF) was reported to regulate triacyl glycerol synthesis in various cells. Linoleic acid and its metabolites were thought to modulate the signal transduction of growth factors. This study determined whether linoleic acid regulated the effect of EGF on lipid contents in human intestinal C2BBe1 cells. Confluent cells were incubated with serum-free medium (control), EGF (45 ng/mL), linoleic acid (42 microg/mL), or combined EGF (45 ng/mL) and linoleic acid (42 microg/mL) for 48 h. The results showed EGF and linoleic acid significantly increased intracellular cholesterol and triglyceride levels compared with the control and combined groups. EGF was a more potent stimulator for triacyl glycerol synthesis in C2BBe1 cells than linoleic acid. However, intracellular cholesterol and triglyceride levels did not differ between the control and combined groups. The secretion of cholesterol and triglyceride into the medium by C2BBe1 cells did not differ among four groups. Both EGF and linoleic acid strongly stimulated the expression of EGF receptor mRNA in C2BBe1 cells at 48 h compared with the control and combined groups. Therefore, EGF and linoleic acid increased triacyl glycerol synthesis in C2BBe1 cells through stimulating the expression of EGF receptor mRNA. The effect of EGF and linoleic acid on this lipogenesis was reversed in the presence of both EGF and linoleic acid by downregulating the expression of EGF receptor mRNA.     

Antioxidant activity of L-adrenaline: a structure-activity insight

L-adrenaline belongs to a group of the compounds known as catecholamines, which play an important role in the regulation of physiological process in living organisms. The antioxidant activity and antioxidant mechanism of L-adrenaline was clarified using various in vitro antioxidant assays including 1,1-diphenyl-2-picryl-hydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), N,N-dimethyl-p-phenylenediamine (DMPD(+)), and superoxide anion radicals (O(2)(-)) scavenging activity, hydrogen peroxide (H(2)O(2)), total antioxidant activity, ferric ions (Fe(3+)) and cupric ions (Cu(2+)) reducing ability, ferrous ions (Fe(2+)) chelating activity. L-adrenaline inhibited 74.2% lipid peroxidation of a linoleic acid emulsion at 30 microg/mL concentration. On the other hand, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), alpha-tocopherol and trolox displayed 83.3, 82.1, 68.1 and 81.3% inhibition on the peroxidation of linoleic acid emulsion at the same concentration, respectively. BHA, BHT, alpha-tocopherol and trolox were used as reference antioxidants and radical scavenger compounds. Moreover, this study will bring an innovation for further studies related to antioxidant properties of L-adrenaline. According to present study, L-adrenaline had effective in vitro antioxidant and radical scavenging activity.     

Antioxidant activity of carotenoid lutein in vitro and in vivo

Carotenoid lutein was evaluated for its antioxidant potential both in vitro and in vivo. Lutein was found to scavenge superoxide radicals, hydroxyl radicals and inhibited in vitro lipid peroxidation. Concentrations needed for 50% inhibition (IC50) were 21, 1.75 and 2.2 microg/mL respectively. It scavenged 2,2-diphenyl-1-picryl hydrazyl (IC50 35 microg/mL) and nitric oxide radicals (IC50 3.8 microg/mL) while 2,2-azobis-3-ethylbenzthiozoline-6-sulfonic acid radicals were inhibited at higher concentration. Ferric reducing power (50%) of lutein was found to be equal 0.3 micromols/mL of FeSO4.7H2O. Its oral administration inhibited superoxide generation in macrophages in vivo by 34.18, 64.32 and 70.22% at doses of 50, 100 and 250 mg/kg body weight. The oral administration of lutein in mice for 1 month significantly increased the activity of catalase, superoxide dismutase, glutathione reductase and glutathione in blood and liver while the activity of glutathione peroxidase and glutathione-S-transferase were found to be increased in the liver tissue. Implication of these results in terms of its role in reducing degenerative diseases is discussed.     

Comparison of in vitro antioxidant and antiradical activities of L-tyrosine and L-Dopa

Summary. Phenolic compounds are interesting because of their antioxidant properties. In the present study, the antioxidant properties of L-tyrosine as a monophenolic and L-Dopa as a diphenolic amino acid were investigated by using different antioxidant assays: (i) 1,1-diphenyl-2-picryl-hydrazyl free radical (DPPH^•) scavenging; (ii) 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical cation decolorization assay; (iii) total antioxidant activity by ferric thiocyanate method; (iv) ferric ions (Fe³⁺) reducing power; (v) superoxide anion radical (O₂ ^•−) scavenging; (vi) hydrogen peroxide (H₂O₂) scavenging, and (vii) ferrous ions (Fe²⁺) chelating activities. Butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), α-tocopherol and trolox, a water-soluble analogue of tocopherol, were used as the reference antioxidant compounds. At the same concentration (20 μg/mL), L-tyrosine and L-Dopa showed 30.6 and 67.9% inhibition of lipid peroxidation of linoleic acid emulsion, respectively. On the other hand, BHA, BHT, α-tocopherol and trolox indicated inhibitions of 74.4, 71.2, 54.7 and 20.1% on the peroxidation of linoleic acid emulsion, respectively, at the above-mentioned concentration. In addition, L-tyrosine and L-Dopa had an effect on DPPH radical scavenging, ABTS radical scavenging, superoxide anion radical scavenging, H₂O₂ scavenging, total ferric ions reducing power and metal chelating on ferrous ions activities.     

In vitro antioxidant activity of silymarin

Silymarin, a known standardized extract obtained from seeds of Silybum marianum is widely used in treatment of several diseases of varying origin. In the present paper, we clarified the antioxidant activity of silymarin by employing various in vitro antioxidant assay such as 1,1-diphenyl-2-picryl-hydrazyl free radical (DPPH(.)) scavenging, 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging activity, total antioxidant activity determination by ferric thiocyanate, total reducing ability determination by Fe3+ - Fe2+ transformation method and Cuprac assay, superoxide anion radical scavenging by riboflavin/methionine/illuminate system, hydrogen peroxide scavenging and ferrous ions (Fe2+) chelating activities. Silymarin inhibited 82.7% lipid peroxidation of linoleic acid emulsion at 30 microg/mL concentration; butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), alpha-tocopherol and trolox indicated inhibition of 83.3, 82.1, 68.1 and 81.3% on peroxidation of linoleic acid emulsion at the same concentration, respectively. In addition, silymarin had an effective DPPH(.) scavenging, ABTS(.)+ scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, ferric ions (Fe3+) reducing power by Fe3+-Fe2+ transformation, cupric ions (Cu2+) reducing ability by Cuprac method, and ferrous ions (Fe2+) chelating activities. Also, BHA, BHT, alpha-tocopherol and trolox, were used as the reference antioxidant and radical scavenger compounds. Moreover, this study, which clarifies antioxidant mechanism of silymarin, brings new information on the antioxidant properties of silymarin. According to the present study, silymarin had effective in vitro antioxidant and radical scavenging activity. It could be used in the pharmacological and food industry because of its antioxidant properties.     

Effect of caffeine on meiotic maturation of porcine oocytes

This study was conducted to evaluate the effect of caffeine on the meiotic maturation of porcine oocytes. Oocyte-cumulus complexes were collected from slaughterhouse-derived ovaries and cultured for 24, 32 or 48 h in medium 199 supplemented with 10% fetal calf serum, 10 microg/ml FSH, 50 microg/ml sodium pyruvate and 50 microg/ml gentamicin in the presence or absence of 2.5 mM caffeine. Caffeine inhibited the meiotic resumption of pig oocytes effectively after 24 h of culture, and 95.5% of oocytes were arrested at the germinal vesicle (GV) stage (control 17.8%, p < 0.05). Prolonged culture with caffeine up to 32 h or 48 h, however, resulted in a significant decrease in the inhibitory effect (GV: 13.8% and 8.2%). The number of oocytes at metaphase II after 48 h of culture in the presence of caffeine was significantly lower than that in the control medium (65.3% vs 94.7%, p < 0.05). The withdrawal of caffeine after 24 h of culture resulted in the resumption of meiotic maturation, and the oocytes reached metaphase II after 48 h. However, the ability of caffeine-treated oocytes to develop to blastocysts after artificial activation was lower than that of the control (5.5% vs 9.1%, p < 0.05). Caffeine treatment significantly increased cAMP levels in the oocytes after 24 h of culture, while both Cdc2 kinase and MAP kinase activation were inhibited in the oocytes. These results suggest that caffeine, similarly to other purine derivatives, prolongs the meiotic arrest of porcine oocytes at the GV stage, perhaps by its action of increasing the cAMP level and by the suppression of Cdc2 kinase and MAP kinase activities in the oocytes.     

Association of different zinc concentrations combined with a fixed caffeine dose on plasma and tissue caffeine and zinc levels in the rat

Because caffeine and tissue levels of Zn are closely related, the objectives of this study were to determine the changes in plasma caffeine levels over a period of 5 h when different concentrations of Zn combined with a fixed concentration of caffeine were injected into the femoral vein of rats and to determine the relationship between tissue levels of caffeine and Zn at 5 h postinjection. Rats were divided into three groups: group 1, 220 μg caffeine; group 2,220 μg caffeine + 8 μg Zn/g body weight (BW); group 3, 220 μg caffeine +16 μg Zn/g BW. Blood from groups 1 and 3 was collected at 3 min, 30 min, 1h, 3h, and 5h to determine the pharmacokinetics of caffeine. All groups were killed at 5 h. Caffeine and Zn concentrations of the brain, kidney, heart, and liver of all groups were determined. The plasma-caffeine curve in group 3 showed a lower concentration at 3 min and a slower caffeine-elimination rate during the first 3 h. Brain and kidney caffeine levels remained constant in all groups, whereas caffeine levels were increased in the heart in group 2 and in the liver in group 3. Zn concentrations in the brain and kidney were lower in group 2 compared with groups 1 and 3 and higher in group 3 compared to groups 1 and 2. Zn concentration in the heart was the same among the three groups but was increased in the liver in group 3 compared to groups 1 and 2. Therefore, we concluded that caffeine combined with Zn affects caffeine pharmacokinetics. With caffeine intake, levels of Zn (16 μg/g BW) that are slightly higher than the daily requirements (12 μg/g BW) may prevent a reduction of Zn in tissue. In addition, caffeine’s effects on Zn concentration among organs are different.     

薤白总皂苷的抗氧化活性

以薤白叶片和鳞茎为材料,测定了一年生、二年生和三年生总皂苷不同浓度下（50、100、200、400、800μg.mL-1）的还原力和抗亚油酸脂质体氧化能力,以及对DPPH（二苯代苦味酰基）、O2-、·OH的清除力,综合评价薤白总皂苷的体外抗氧化活性。结果表明,薤白叶片和鳞茎中的总皂苷随着浓度的增加,其抗氧化能力呈逐渐增强的趋势,对DPPH、O2-和·OH的清除作用分别达77．30％、95．95％和91．96％,相同浓度下对嘎的清除能力要强于抗坏血酸,但还原能力低于抗坏血酸,在浓度为800μg.mL-1 72h时,叶片和鳞茎总皂苷的抗亚油酸脂质体氧化能力均达到最大;薤白叶片的总皂苷在一定浓度下抗氯化能力要强于鳞茎。薤白叶片和鳞茎可作为天然抗氧化剂原料来源。     

Antioxidant activity of 50% aqueous--ethanol extract from Acantholippia deserticola

The antioxidant properties oí Acantholippia deserticola, a herb used in traditional northern Chilean medicine was studied using free radical-generating systems. The 50% aqueous-ethanol extract oí Acantholippia deserticola protected against non-enzymatic lipid peroxidation in microsomal membranes of rat, induced by an Fe++-ascorbate system and measured spectrophotometrically by the TBARS test, and had strong free radical scavenging capacities on stable ABTS and DPPH radicals. The results shows that the IC50 value of the 50% aqueous-ethanolic extract of A.deserticola is 18 +/- 0.5 microg/mL in DPPH radical-scavenging, 15 +/- 0.8 microg/mL in lipid peroxidation , Total Antioxidant Activity (TAA) is 0.95 mM of Trolox per mg/mL of extract. The total phenolics content of extract is 725 +/- 12 mg of gallic acid equivalent per g of dried extract. The results indicate that the 50% aqueous-ethanol extract of Acantholippia deserticola clearly has antioxidant properties.     

Influence of caffeine on sparing effect of dose fractionation in housefly larvae

Summary Caffeine, given during interfraction interval, abolishes the sparing effect of dose fractionation observed for delay of pupariation inMusca domesti ca larvae. When given as postirradiation treatment after single exposure, caffeine increases the delay in a synergistic manner. Pretreatment of the larvae with ascorbic acid for 3 h protects from radiation-induced delay whereas pretreat ment with caffeine does not have any effect. Combination of ascorbic acid and caffeine pretreatment protects the larvae only at low concentration (0.05%) and not at high concentration (0.1%).     

Antioxidant activity of bisbenzylisoquinoline alkaloids from Stephania rotunda: cepharanthine and fangchinoline

In the present study, we determined the antioxidant activity of cepharanthine and fangchinoline from Stephania rotunda by performing different in vitro antioxidant assays, including 1,1-diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging, 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging, N,N- dimethyl-p-phenylenediamine dihydrochloride (DMPD) radical scavenging, superoxide anion (O₂^?–) radical scavenging, hydrogen peroxide scavenging, total antioxidant activity, reducing power, and ferrous ion (Fe²⁺) chelating activities. Cepharanthine and fangchinoline showed 94.6 and 93.3% inhibition on lipid peroxidation of linoleic acid emulsion at 30 μg/mL concentration, respectively. On the other hand, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), α-tocopherol, and trolox indicated inhibitions of 83.3, 92.2, 72.4, and 81.3% on peroxidation of linoleic acid emulsion at the same concentration (30 μg/mL), respectively. According to the results, cepharanthine and fangchinoline have effective antioxidant and radical scavenging activity.     

Allyl mercaptan, a major metabolite of garlic compounds, reduces cellular cholesterol synthesis and its secretion in Hep-G2 cells

The cytotoxicity, cellular cholesterol synthesis, and secretion of allyl mercaptan, a major metabolite of garlic compounds, were studied in Hep-G2 cells. The cells were grown in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum and treated with 5, 25, 50, 100, 125, 250, and 500 microg of allyl mercaptan/mL for 4, 12, and 24 hours. At concentrations up to 125 microg, no significant cytotoxic effect was noted during those incubation periods. However, at a concentration of 250 microg, cell viability decreased approximately 50% compared with the control (P < 0.05) in all three incubation times. At a concentration of 500 microg, allyl mercaptan was highly toxic, causing extensive cell death. The treatment of cells with 5, 10, 25, 50, or 100 microg (noncytotoxic concentration) of allyl mercaptan resulted in a marked inhibition of (3)H-acetate incorporation into cholesterol. At concentrations between 5 and 100 microg, the cholesterol synthesis was inhibited 20 to 80% in cells and the cholesterol secretion into the medium decreased 20 to 50% compared with the control (P < 0.05). The concentration of allyl mercaptan required to suppress cholesterol synthesis by 50% was approximately 25 microg/mL. Allyl mercaptan treatment of cells incubated with 1 mM of oleic acid also resulted in a significant decrease in the cholesterol synthesis compared with the cells incubated with oleic acid alone (19.5 +/- 1.2 x 10(3) dpm/mg protein/4 h vs. 30.0 +/- 2.6 x 10(3) dpm/mg protein/4 h; P < 0.05). The present study demonstrates that allyl mercaptan is effective in inhibiting cholesterol synthesis at concentrations as low as 5 microg, and its inhibition is concentration dependent.     

Inhibition of lipoxygenase-dependent lipid peroxidation by quercetin: Mechanism of antioxidative function

Quercetin inhibited soybean lipoxygenase-1-dependent linoleic acid peroxidation. Two to three μM quercetin was required for 50% inhibition. During the inhibition, quercetin was oxidized. The oxidation was observed as an absorbance decrease at about 380 nm and an absorbance increase at about 335 nm. Inhibition of linoleic acid peroxidation by quercetin seems to be due to reduction by the reagent of the linoleic acid radical formed as an intermediate during lipoxygenation. Quercetin oxidation was suppressed by ascorbate under conditions when ascorbate did not affect lipoxygenase-dependent linoleic acid peroxidation. The results suggest that ascorbate can reduce the quercetin oxidized by the linoleic acid radical back to quercetin. Based on the results, the significance of a redox reaction between oxidized quercetin and ascorbate is discussed.     

Caffeine and theobromine in epicuticular wax of Ilex paraguariensis A. St.-Hil

Caffeine and theobromine were identified and quantified in leaf epicuticular waxes of Ilex paraguariensis A. St.-Hil. (Aquifoliaceae). The total epicuticular leaf wax content was ca. 0.5% on average of dry leaf weight. Epicuticular caffeine and theobromine contents varied from 0.16 to 127.6 microg/mg and from 0 to 9.5 microg/mg of wax, respectively. For some selected samples, the intracellular methylxanthine concentration was also determined. A positive correlation was found between inner and epicuticular caffeine contents.     

Caffeine reduces TNFα up-regulation in human adipose tissue primary culture

Adipose tissue secretions play an important role in the development of obesity-related pathologies such as diabetes. Through inflammatory cytokines production, adipose tissue stromavascular fraction cells (SVF), and essentially macrophages, promote adipocyte insulin resistance by a paracrine way. Since xanthine family compounds such as caffeine were shown to decrease inflammatory production by human blood cells, we investigated the possible effect of caffeine on Tumor Necrosis Factor alpha (TNFalpha) and Interleukin-6 (IL-6) expression by human adipose tissue primary culture. For that purpose, human subcutaneous adipose tissue obtained from healthy non-obese women (BMI: 26.7 +/- 2.2 kg/m2) after abdominal dermolipectomy, was split into explants and cultured for 6 hours with or without caffeine. Three different concentrations of caffeine were tested (0.5 microg/mL, 5 microg/mL and 50 microg/mL). After 6 hours of treatment, explants were subjected to collagenase digestion in order to isolate adipocytes and SVF cells. Then, TNFalpha and IL-6 mRNA were analysed by real-time PCR alternatively in adipocytes and SVF cells. In parallel, we checked gene expression of markers involved in adipocyte differenciation and in SVF cells inflammation and proliferation. Our findings show a strong and dose dependent down-regulation of TNF-alpha gene expression in both adipocyte and SVF cells whereas IL-6 was only down regulated in SVF cells. No effect of caffeine was noticed on the other genes studied. Thus, caffeine, by decreasing TNFalpha expression, could improve adipose tissue inflammation during obesity.     

Acute Caffeine Treatment Increases Extracellular Nucleotide Hydrolysis from Rat Striatal and Hippocampal Synaptosomes

The psychostimulant caffeine promotes behavioral effects such as hyperlocomotion, anxiety, and disruption of sleep by blockade of adenosine receptors. The availability of extracellular adenosine depends on its release by transporters or by the extracellular ATP catabolism performed by the ecto-nucleotidase pathway. This study verified the effect of caffeine on NTP-Dase 1 (ATP diphosphohydrolase) and 5-nucleotidase of synaptosomes from hippocampus and striatum of rats. Caffeine and theophylline tested in vitro were unable to modify nucleotide hydrolysis. Caffeine chronically administered in the drinking water at 0.3 g/L or 1 g/L for 14 days failed to affect nucleotide hydrolysis. However, acute administration of caffeine (30 mg/kg, ip) produced an enhancement of ATP (50%) and ADP (32%) hydrolysis in synaptosomes of hippocampus and striatum, respectively. This activation of ATP and ADP hydrolysis after acute treatment suggests a compensatory effect to increase adenosine levels and counteract the antagonist action of caffeine.