Rutin restores adenosine deaminase activity in serum and the liver and improves biochemical parameters in streptozotocin-induced diabetic rats / Rutina restaura a atividade da Adenosina desaminase no soro e no fígado e melhora os parâmetros bioquímicos em ratos diabéticos induzidos por estreptozotocina

ABSTRACT denosine deaminase (ADA) is a critical control point in the regulation of adenosine levels. This study aimed to investigate the effects of a polyphenolic flavonoid, rutin, on the activity of ADA in serum, the cerebral cortex, liver, kidney, and biochemical parameters in diabetic rats. The animals were divided into four groups (n=6) for the following treatments: control; diabetic (streptozotocin 55 mg/kg); diabetic with rutin (100 mg/kg/day); diabetic with glibenclamide (10 mg/kg/day). After 30 days, ADA activity and biochemical parameters were analyzed. The ADA activity in the serum was significantly elevated in the diabetic group compared to the control group (p<0.01). The treatment with rutin prevented the increase in ADA activity in the STZ-induced rats when compared to control group. Our data showed that rutin reduced glucose, LDL levels, and hepatic enzymes in comparison with the control group. These results demonstrate that the increase of ADA activity observed in diabetic rats may be an important indicator of the immunopathogenesis of hyperglycemic disorders and suggest that rutin is important for regulating the enzymatic activities associated with immune, hyperglycemic, and inflammatory response in diabetes mellitus.

RESUMO A Adenosina desaminase (ADA) representa um ponto de controle crítico na regulação dos níveis de adenosina. A rutina, um flavonóide polifenólico presente em muitas plantas, foi testado para verificar a sua influência na atividade da ADA no soro, córtex cerebral, fígado rim e parâmetros bioquímicos em ratos diabéticos. Os animais foram divididos em quatro grupos cada grupo com 6 animais), tal como: controle; diabética (estreptozotocina 55 mg/kg); diabética + rutina (100 mg/kg/dia); diabético + glibenclamida (10 mg/kg/dia). Após 30 dias foram analisadas a atividade da ADA sérica e tecidual e parâmetros bioquímicos. A atividade de ADA no soro foi significativamente elevada no grupo diabético quando comparado ao grupo controle (p<0,01). O tratamento com Rutina preveniu o aumento na atividade da ADA nos ratos diabéticos, quando comparado com o grupo controle. Os resultados mostraram que a rutina reduziu a glicose, os níveis de LDL e as enzimas hepáticas, em comparação com o grupo controle. Estes resultados mostram que o aumento da atividade da ADA observado em ratos diabéticos pode ser um indicador importante da imuno-patogênese de perturbações hiperglicêmicas e sugerem que a Rutina é importante na regulação das atividades enzimáticas associadas com a resposta imunitária, hiperglicêmica e inflamatória no Diabetes mellitus.

Anti-inflammatory evaluation of Coronopus didymus in the pleurisy and paw oedema models in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Mastrunço (Coronopus didymus--CD) is currently considered as a medicinal specie often used in Brazil, especially in southeast region, for the treatment of several diseases in which pain and inflammation are common. Treatment with the plant can be done by infusion, decoction, or through food. The aim of this study was: to investigate the anti-inflammatory effect of hydroalcoholic extract obtained from the leaves of CD following the traditional procedure. MATERIALS AND METHODS: The anti-inflammatory activity was determined using mouse of pleurisy and paw oedema models, both process being induced by different flogistic agents such as: carrageenan (Cg), bradykinin (BK), histamine (HIS), substance P (SP), dextran (DEX) or prostaglandin E(2) (PGE(2)). We evaluated the effect of CD (200-600 mg/kg) administered by oral route (p.o.) upon leukocytes migration, myeloperoxidase (MPO), and adenosine-deaminase (ADA) activities and nitric oxide (NO) levels. RESULTS: CD (200-600 mg/kg) inhibited the leukocytes by 60.0+/-1.42%, neutrophils by 82.75+/-1.29%, MPO by 42.30+/-4.23%, and ADA activities by 57.89+/-1.94%, as well as NO levels by 64.28+/-2.15% in Cg induced pleurisy. CD also inhibited total and differential leukocytes in the pleurisy induced by BK (1.30+/-0.11/0.29+/-0.02), HIS (1.20+/-0.09/0.42+/-0.05) and SP (0.74+/-0.06/0.14+/-0.01). In addition, CD was effective in reducing paw oedema induced by Cg by 72.79+/-1.13%, SP by 68.26.+/-0.78%, BK by 66.66.+/-0.77%, PGE(2) by 53.346.+/-1.18 and DEX by 65.14+/-2.35%. CONCLUSION: Several mechanisms, including the inhibition of enzymes (MPO and ADA) and mediators (BK, HIS, SP, NO and PGE(2)) release and/or action, appear to account for the anti-inflammatory effect of Coronopus didymus.

Adenosine deaminase enzyme therapy prevents and reverses the heightened cavernosal relaxation in priapism.

INTRODUCTION: Priapism featured with painful prolonged penile erection is dangerous and commonly seen in sickle cell disease (SCD). The preventive approaches or effective treatment options for the disorder are limited because of poor understanding of its pathogenesis. Recent studies have revealed a novel role of excess adenosine in priapism caused by heightened cavernosal relaxation, and therefore present an intriguing mechanism-based therapeutic possibility. AIM: The aim of this study was to determine the therapeutic effects of adenosine deaminase (ADA) enzyme therapy to lower adenosine in priapism. METHODS: Both ADA-deficient mice and SCD transgenic (Tg) mice display priapism caused by excessive adenosine. Thus, we used these two distinct lines of mouse models of priapism as our investigative tools. Specifically, we treated both of these mice with different dosages of polyethylene glycol-modified ADA (PEG-ADA) to reduce adenosine levels in vivo. At the end points of the experiments, we evaluated the therapeutic effects of PEG-ADA treatment by measuring adenosine levels and monitoring the cavernosal relaxation. MAIN OUTCOME MEASURES: Adenosine levels in penile tissues were measured by high-performance liquid chromatography, and cavernosal relaxation was quantified by electrical field stimulation (EFS)-induced corporal cavernosal strip (CCS) assays. RESULTS: We found that lowering adenosine levels in penile tissues by PEG-ADA treatment from birth in ADA-deficient mice prevented the increased EFS-induced CCS relaxation associated with priapism. Intriguingly, in both ADA-deficient mice and SCD Tg mice with established priapism, we found that normalization of adenosine levels in penile tissues by PEG-ADA treatment relieved the heightened EFS-induced cavernosal relaxation in priapism. CONCLUSIONS: Our studies have identified that PEG-ADA is a novel, safe, and mechanism-based drug to prevent and correct excess adenosine-mediated increased cavernosal relaxation seen in two independent priapic animal models, and suggested its therapeutic possibility in men suffering from priapism.

Aqueous extract of Urtica dioica makes significant inhibition on adenosine deaminase activity in prostate tissue from patients with prostate cancer.

AIM: Investigation of possible effects of aqueous extract of Urtica dioica leaves on adenosine deaminase activity in prostate tissue from patients with prostate cancer. METHODS: Ten prostate tissues from patients with pathologically proven localized prostate cancer (Gleason scores 4 to 7) were used in the study. In the tissues, ADA activities with and without preincubation with different amounts of Urtica dioica extracts were performed. RESULTS: Aqueous extract of Urtica dioica results in significant inhibition on adenosine deaminase (ADA) activity of prostate tissue. CONCLUSION: ADA inhibition by Urtica dioica extract might be one of the mechanisms in the observed beneficial effect of Urtica dioica in prostate cancer.

Central inhibitory effect of adenosine deaminase on carotid blood flow increase at high pressure.

Carotid blood flow in rats was measured by implanted transit-time ultrasonic flowprobes throughout hyperbaric experiments conducted up to 70 bar (7 MPa) with a helium-oxygen hyperoxic (PO2 = 400 mbar) mixture. Before the hyperbaric experiment, an intracerebroventricular (ICV) injection of phosphate saline-buffered solution (PBS) or adenosine deaminase (ADA, 100 U.ml-1) in PBS was performed. Throughout the hyperbaric experiment carotid blood flow increased with ambiant pressure in PBS-treated rats. Conversely, the increase in carotid blood flow was attenuated by ADA treatment. These results suggest that the increase in carotid blood flow at high ambiant pressure could result from an increase of adenosine concentration in the rat brain.

PEG-ADA: an alternative to haploidentical bone marrow transplantation and an adjunct to gene therapy for adenosine deaminase deficiency.

PEG-ADA is a long-circulating form of adenosine deaminase (ADA) that has been in use for > 8 years as replacement therapy for severe combined immunodeficiency disease due to ADA deficiency. Treatment with PEG-ADA almost completely corrects metabolic abnormalities, allowing the recovery of a variable degree of immune function. Although not normal, the level of function achieved has in most cases been sufficient to protect against opportunistic and life-threatening infections. PEG-ADA has been used as an alternative for patients who lack an HLA-identical bone marrow donor, but are judged to be at too high a risk for undergoing HLA-haploidentical marrow transplantation. To date, mortality and morbidity with PEG-ADA have been less than for the latter procedure. PEG-ADA has also been an important adjunct to attempts to develop somatic cell gene therapy for ADA deficiency, although its continued use poses a problem for evaluation of the benefit of gene therapy. As a true "orphan drug" developed to treat a very small patient population, the cost per patient of PEG-ADA is very high.

Effects of short term ischemia and reperfusion on coronary vascular reactivity and myocardial function.

Ischemia and reperfusion have been shown to cause damage to the endothelium as well as to the cardiac myocyte. Although the vasodilator response has been shown to be impaired following ischemia and reperfusion, the effect of a short period of global ischemia on the contractile response of the coronary vasculature is not clear. In the present study, coronary vasoconstriction in response to U46619, PGF2 alpha, 5-HT, and KCl was found to be depressed for at least 15 min following 15 min of in vitro global ischemia in rats hearts. Vasodilator blockers or inactivators were used in an effort to restore this depressed coronary response. Indomethacin (5 microM) was used to block production of vasodilator prostaglandins, L-NAME (30 microM) to block production of nitric oxide (NO), and adenosine deaminase (2.4 units/ml of coronary flow) to inactivate adenosine. None of these agents restored the normal coronary constrictor response following ischemia. When superoxide dismutase and catalase (both 20 micrograms/ml of coronary flow) were infused for 5 min before and after ischemia, the coronary response recovered more than 100% of its preischemic value by 15 min of reperfusion, but still remained depressed at 5 min reperfusion. These data suggest that free radicals produced during ischemia and/or reperfusion may be at least partly responsible for this temporary "stunning" of the coronary vasculature. Since the impaired contractile response was still present at 5 min reperfusion when the buffer was supplemented with oxygen radical scavengers, another mechanism must also be involved in this "stunning" process.

Non-eicosanoid functions of essential fatty acids: regulation of adenosine-related functions in cultured neuroblastoma cells.

Studies have demonstrated that augmenting the omega 6 polyunsaturated-fatty-acid (PUFA) content of N1E-115 neuroblastoma cells by media supplementation with linoleic acid results in greater than or equal to 2-fold increases in basal levels of intracellular cyclic AMP (cAMP). Data suggested some involvement of increased production of adenosine from endogenous metabolites; however, increases in adenosine were not related to increased activity of 5'-nucleotidase or decreased uptake of extracellular adenosine. PUFA-dependent elevations in basal cAMP were evident within 1 min of exposure to a phosphodiesterase inhibitor; this phenomenon did not appear to be due to PUFA-dependent changes in Ca2+ uptake or to increases in sensitivity of adenylate cyclase to Ca2+. Forskolin-stimulated cAMP formation was 3-fold higher in PUFA-enriched cells than in control cells, which suggested a direct effect on the functioning of the catalytic unit. Linoleic acid supplementation resulted in a 2-fold increase in the maximum amounts of cAMP produced in response to the stable adenosine analogue, 5'-N'ethylcarboxy-amidoadenosine (NECA). The altered stimulatory response did not involve eicosanoid formation, but may have been related to an increase in the number of stimulatory adenosine receptors, as judged by binding of [3H]NECA. These studies indicate that membrane PUFA modulate adenosine-related functions in neuroblastoma cells, and suggest that a complex series of mechanisms is involved in this regulation.

Duodenal rapeseed oil infusion in early and midlactation cows. 4. In vivo and in vitro adipose tissue lipolytic responses.

In vitro glycerol and FFA releases from adipose tissue were studied in early (wk 3, trial 1) and midlactation (wk 19 to 26, trial 2) multiparous Holstein Friesian cows receiving a duodenal rapeseed oil infusion (1.0 to 1.1 kg/d). In trial 2, in vitro basal FFA release, basal FFA: glycerol ratio, and isoproterenol-stimulated FFA and glycerol releases were higher in perirenal adipose tissue from oil-infused cows. Plasma FFA concentration also was higher in oil-infused cows before and after intravenous isoproterenol injection. In trail 1, basal and stimulated glycerol and FFA releases from perirenal (but not subcutaneous) adipose tissue tended to be lower in oil-infused cows. This was probably linked to a lower milk production potential of oil-infused than of control cows which introduced a bias in energy balance. The basal FFA:glycerol ratio tended to be higher in oil-infused cows in both adipose tissues, suggesting a lower rate of reesterification inducted by oil, as was the case in trial 2. The alpha 2-agonist clonidine decreased perirenal adipose tissue glycerol release in cows and treatments in which the responses to 4 x 10(-7) M isoproterenol were higher. In vivo and in vitro lipolytic responses were lower in trial 2 than in trial 1, except the in vitro maximally stimulated lipolytic rate, which probably reflected a long lasting teleophoretic adaptation to ensure energy needs of lactation. This study indicated that oil infusion affected both beta- and alpha 2-adrenergic responses and that postpartum lipid mobilization did not seem to be reduced by the exogenous unsaturated fatty acid supply.

Adenosine formation and energy metabolism: a 31P-NMR study in isolated rat heart.

Temporal and quantitative relations between cytosolic energy metabolism, adenosine efflux, and coronary flow were examined during 10 min of isoproterenol (ISO) infusion (60 nM) or hypoxia (5% O2) in isolated isovolumic rat heart. Myocardial metabolism was monitored using 31P-nuclear magnetic resonance spectroscopy, and venous effluent was collected and assayed for adenosine. During ISO infusion, coronary flow increased to approximately 170%, and [ATP]/[ADP] [Pi] (cytosolic phosphorylation potential) declined to less than 25% of preinfusion levels, respectively (P less than 0.001). During hypoxia, coronary flow increased to 190%, and [ATP]/[ADP] [Pi] declined to less than 25% of normoxic levels (P less than 0.001). Release of adenosine into the coronary venous effluent increased greater than 10-fold and displayed significant inverse linear correlations with log[ATP]/[ADP] [Pi] and positive linear correlations with free cytosolic [AMP] and coronary flow during ISO infusion and hypoxia. Adenosine deaminase (ADA) treatment reduced coronary vasodilation by approximately 30% during ISO infusion and 40% during hypoxia (P less than 0.001) and augmented chronotropic and inotropic responses to ISO infusion (P less than 0.01). Infusion of ADA potentiated changes in [ATP]/[ADP] [Pi] and [AMP] observed during ISO infusion and hypoxia (P less than 0.05). These results indicate that 1) endogenous adenosine mediates metabolic vasodilation in the heart, 2) adenosine modulates the response of isolated myocardium to catecholamines, 3) myocardial adenosine formation appears to be linked to cytosolic metabolism via changes in [ATP]/[ADP] [Pi] and [AMP], and 4) endogenous adenosine provides a significant, metabolically beneficial action in isolated hearts during hypoxia and inotropic stimulation.

Regulation of thymocyte proliferation by endogenous adenosine and adenosine deaminase.

Spontaneous proliferation of thymocytes after 20-25 h of culture was significantly increased by the presence of adenosine deaminase (ADA) or theophylline. The effect of ADA was counteracted by the ADA inhibitor EHNA. When given alone, EHNA inhibited proliferation. This effect was not blocked by inhibition of adenosine uptake with dipyridamol. These results suggest that proliferation in culture is regulated by a balance between endogenous adenosine and ADA, controlling the influence of adenosine on the intracellular cyclic AMP level via an adenosine receptor on the surface of thymocytes. According to the hypothesis, ADA would stimulate proliferation by decreasing extracellular adenosine levels and theophylline by blocking adenosine receptors on thymocytes. EHNA would inhibit proliferation by increasing extracellular adenosine levels. In accordance with this interpretation, the adenosine analogue phenylisopropyl adenosine (PIA) inhibited proliferation and the effect could be inhibited by theophylline. The postulated effect of endogenous adenosine could not be mimicked by a single administration of exogenous adenosine. Whereas most doses of adenosine were without effect, a high dose of adenosine (0.1 mM) in combination with EHNA unexpectedly stimulated proliferation. Since the effect was blocked by dipyridamol, an intracellular site of action for adenosine is suggested in this case.