Intranasal metformin treatment ameliorates cognitive functions via insulin signaling pathway in ICV-STZ-induced mice model of Alzheimer's disease.

AIMS: The relationship between type 2 diabetes and Alzheimer's disease (AD) provides evidence that insulin and insulin sensitizers may be beneficial for the treatment of AD. The present study investigated the effect and mechanism of action of intranasal metformin treatment on impaired cognitive functions in an experimental mice model of AD. MAIN METHODS: Intracerebroventricularly (ICV) streptozotocin (STZ)-injected mice were treated with intranasal or oral metformin for 4 weeks. Learning and memory functions were evaluated using Morris water maze. Metformin and Aß42 concentrations were determined by liquid chromatography tandem mass spectrometry and ELISA respectively. The expressions of insulin receptor, Akt and their phosphorylated forms were determined in the hippocampi and cerebral cortices of mice. KEY FINDINGS: ICV-STZ-induced AD mice displayed impaired learning and memory functions which were improved by metformin treatment. ICV-STZ injection or intranasal/oral metformin treatments had no effect on blood glucose concentrations. Intranasal treatment yielded higher concentration of metformin in the hippocampus and lower in the plasma compared to oral treatment. ICV-STZ injection and metformin treatments did not change amyloid ß-42 concentration in the hippocampus of mice. In hippocampal and cortical tissues of ICV-STZ-induced AD mice, insulin receptor (IR) and Akt expressions were unchanged, while phosphorylated insulin receptor (pIR) and pAkt expressions decreased compared to control. Metformin treatments did not change IR and Akt expressions but increased pIR and pAkt expressions. SIGNIFICANCE: The present study showed for the first time that intranasal metformin treatment improved the impaired cognitive functions through increasing insulin sensitivity in ICV-STZ-induced mice model of AD.

Trimethylamine-N-oxide (TMAO) Selectively Disrupts Endothelium-Dependent Hyperpolarization-Type Relaxations in a Time-Dependent Manner in Rat Superior Mesenteric Artery.

The vascular action of trimethylamine-N-oxide (TMAO)-the gut microbiota-derived metabolite-in contributing cardiovascular disease is a controversial topic. A recent study has shown that acute exposure of TMAO at moderate concentrations inhibits endothelium-dependent hyperpolarization (EDH)-type relaxations selectively in rat isolated femoral arteries, but not in mesenteric arteries. Here we determined the efficacy of higher TMAO concentrations with longer exposure times on vascular reactivity in rat isolated superior mesenteric arteries. Acetylcholine-induced EDH-type relaxations were examined before and after incubation with TMAO (0.1-10 mM) at increasing exposure times (1-24 h). One- and 4-h-incubations with TMAO at 0.1-3 mM did not cause any change in EDH-type relaxations. However, when the incubation time was increased to 24 h, responses to acetylcholine were reduced in arteries incubated with 1-3 mM TMAO. In addition, at higher TMAO concentration (10 mM) the decrease in EDH relaxations could be detected both in 4-h- and 24-h-incubations. The EDH-relaxations were preserved in rings incubated with 10 mM TMAO for 24 h in the presence of SKA-31 (10 µM), the small (SKCa)- and intermediate (IKCa)-conductance calcium-activated potassium channel activator. Contractile responses to phenylephrine increased in arteries exposed to 10 mM TMAO for 24 h. Interestingly, nitric oxide (NO)-mediated relaxations remained unchanged in arteries treated for 24 h at any TMAO concentration. Our study revealed that TMAO selectively disrupted EDH-type relaxations time-dependently without interfering with NO-induced vasodilation in rat isolated mesenteric arteries. Disruption of these relaxations may help explain the causal role of elevated TMAO levels in certain vascular diseases.

The effects of 5-hydroxytryptophan on carrageenan-induced mouse paw oedemas / Efeito de 5-hidroxitriptofano no edema de pata de camundongo induzido por carragenina

ABSTRACT Objective 5-Hydroxytryptophan is the precursor compound of serotonin biosynthesis. The oral absorption of 5-Hydroxytryptophan is close to 100% and, unlike serotonin, it crosses the blood-brain barrier freely. 5-Hydroxytryptophan has been used as a food supplement for many years to treat anxiety and depression. Recent studies have shown that 5-Hydroxytryptophan suppresses the pro-inflammatory mediators and is effective in some inflammatory diseases, such as arthritis and allergic asthma. However, the role of 5-Hydroxytryptophan supplements on acute peripheral inflammation has not been investigated yet. In this study, the in vivo anti-inflammatory activity of 5-Hydroxytryptophan was evaluated with a carrageenan-induced paw oedema test in mice. Methods For the investigation of the acute antiinflammatory activity, single oral doses of 5-Hydroxytryptophan (1.5, 5 and 20mg/kg) were given to mice 1.5 hours prior to the carrageenan test. For chronic activity, the same oral doses were administered daily for two weeks prior to the carrageenan test on the 14th day. To induce inflammation, 0.01mL of 2% carrageenan was injected into the paws of mice. Results Supplementation with 5-Hydroxytryptophan significantly reduced inflammation in a dose-independent manner which was irrespective of the duration of exposure (per cent inhibition in acute experiments was 35.4%, 20.9%, 24.0%, and per cent inhibition in chronic experiments was 29.5%, 35.3%, 40.8% for the doses of 1.5, 5, and 20mg/kg, respectively). Conclusion Our findings demonstrate for the first time that 5-HTP supplements have the potential of suppressing the measures of acute peripheral inflammation. It is suggested that, apart from several diseases where serotonin is believed to play an important role, including depression, patients with inflammatory conditions may also benefit from 5-HTP.

RESUMO Objetivo O 5-hidroxitriptofano (5-HTP) é o composto precursor da biossíntese da serotonina. A absorção oral do 5-HTP é próxima a 100% e, ao contrário da serotonina, atravessa a barreira hematoencefálica livremente. O 5-HTP tem sido usado como suplemento alimentar por muitos anos na ansiedade e na depressão. Estudos recentes demonstraram que o 5-HTP suprime os mediadores pró-inflamatórios e é eficaz em algumas doenças inflamatórias, como artrite e asma alérgica. No entanto, o papel dos suplementos de 5-HTP na inflamação periférica aguda ainda não foi investigado. Neste estudo, a atividade anti-inflamatória in vivo do 5-HTP foi avaliada por meio do teste de edema de pata induzido por carragenina em ratos. Métodos Para a atividade aguda, doses orais únicas de 5 -HTP (1,5, 5 e 20 mg/kg) foram dados aos ratos 1,5 horas antes do teste da carragenina. Para a atividade crônica, as mesmas doses orais foram dadas cada dia durante duas semanas antes do teste da carragenina no 14º dia. 0,01ml da carragenina a 2% foi injetado nas patas dos ratos a fim de induzir a inflamação. Resultados A suplementação com 5-HTP reduziu significativamente a inflamação de uma maneira independente da dose, que foi independente da duração da exposição (por cento de inibição em experimentos agudos; 35,4%, 20,9%, 24,0% e por cento de inibição em experimentos crônicos; 29,5%, 35,3%, 40,8% para as doses de 1.5, 5 e 20 mg/kg respectivamente). Conclusão Nossas conclusões demonstram pela primeira vez que os suplementos de 5-HTP têm potencial para suprimir os sintomas de inflamação periférica aguda. É sugerido que, além de várias doenças em que se acredita que a serotonina tem uma função importante, incluindo a depressão, os pacientes com doenças inflamatórias também podem se beneficiar do 5-HTP.

Insulin deprivation decreases insulin degrading enzyme levels in primary cultured cortical neurons and in the cerebral cortex of rats with streptozotocin-induced diabetes.

BACKGROUND: Many studies have indicated a relationship between diabetes and Alzheimer's disease (AD). However, the molecular mechanism underlying this association has not been clarified. Among several factors, insulin degrading enzyme (IDE), which plays roles in the degradation of both insulin and amyloid ß (Aß), has gained interest as a potential target in efforts to solve this puzzle. This study sought to examine the effects of varying insulin and/or glucose concentrations on IDE expression. METHODS: Experiments were performed on primary cultured rat neurons and cortices of rats with streptozotocin (STZ)-induced diabetes. IDE protein and mRNA expression levels were measured by western blot and RT-PCR, respectively. RESULTS: In primary cultured cortical neurons, removal of insulin for 5days reduced the expression of IDE. A five-day treatment with a high concentration of glucose in insulin-free media reduced IDE levels, while a high concentration of glucose in the presence of insulin had no effect. In groups treated with glucose or insulin intermittently, the reduction in IDE levels was observed only in neurons exposed to high glucose together with no insulin for 5days. Shorter incubation periods (48h), either continuously or intermittently, did not affect IDE levels. IDE expression in the cortex of rats with STZ-induced diabetes was found to be decreased. CONCLUSION: Our data suggest that insulin deprivation, rather than high glucose, is a significant determinant of IDE regulation. As evidence indicates potential roles for IDE in diabetes and AD, understanding the mechanisms regulating IDE expression may be important in developing new treatment strategies.

ß-adrenergic Receptor Blocker ICI 118,551 Selectively Increases Intermediate-Conductance Calcium-Activated Potassium Channel (IKCa )-Mediated Relaxations in Rat Main Mesenteric Artery.

Endothelial IKCa and/or SKCa channels play an important role in the control of vascular tone by participating in endothelium-dependent relaxation. Whether ß-AR antagonists, mainly used in hypertension, affect endothelial KCa channel function is unknown. In this study, we examined the effect of the ß2-AR antagonist and inverse agonist ICI 118,551 on the IKCa /SKCa channel activity by assessing functional relaxation responses to several agonists that stimulate these channels. Mesenteric arterial rings isolated from male Sprague Dawley mounted to organ baths. Acetylcholine elicited IKCa - and SKCa -mediated relaxations that were abolished by TRAM-34 and apamin, respectively. ICI 118,551, which did not dilate the arteries per se, increased the IKCa -mediated relaxations, whereas SKCa -mediated relaxations remained unaltered. Same potentiating effect was also detected on the IKCa -mediated relaxations to carbachol and A23187, but not to NS309. Neither acetylcholine-induced nitric oxide-mediated relaxations nor SNP relaxations changed with ICI 118,551. The PKA inhibitor KT-5720, the selective ß2-AR agonist salbutamol, the selective ß2-AR antagonist butoxamine, the non-selective ß-AR antagonist propranolol, and the inverse agonists carvedilol or nadolol failed to affect the IKCa -mediated relaxations. ICI 118,551-induced increase was not reversed by salbutamol or propranolol as well. Besides, low potassium-induced relaxations in endothelium-removed arteries remained the same in the presence of ICI 118,551. These data demonstrate a previously unrecognized action of ICI 118,551, the ability to potentiate endothelial IKCa channel-mediated vasodilation, through a mechanism independent of ß2-AR antagonistic or inverse agonistic action. Instead, the enhancement of acetylcholine relaxation seems likely to occur by a mechanism secondary to endothelial calcium increase.

Lack of insulin results in reduced seladin-1 expression in primary cultured neurons and in cerebral cortex of STZ-induced diabetic rats.

Several studies demonstrated that Diabetes mellitus (DM) enhances the risk for Alzheimer's disease (AD). Although hyperglycemia and perturbed function of insulin signaling have been proposed to contribute to AD pathogenesis, the molecular mechanisms behind this association is not clear yet. Seladin-1 is an enzyme catalyzing the last step in cholesterol biosynthesis converting desmosterol to cholesterol. The neuroprotective function of seladin-1 has gained interest in AD research recently. Seladin-1 has anti-apoptotic properties and regulates the expression of ß-secretase (BACE-1). Here we measured seladin-1 mRNA and protein expressions in rat primary cultured neurons under diabetic conditions and also in the brains of rats with streptozotocine (STZ)-induced diabetes. We show that constant lack of insulin for 5days decreased seladin-1 levels in cultured rat primary neurons. Similarly, a decrease in seladin-1 was found in the brains of rats with STZ-induced diabetes. However, if the lack of insulin and/or high glucose treatment was intermittent, neuronal seladin-1 levels were not affected in vitro. On the other hand, treatment of neurons with metformin resulted in a significant increase in seladin-1. Constant lack of insulin for 5days, as well as high glucose treatment, increased the neuronal expression of BACE-1 in vitro, but not in the in vivo model. Our study defines insulin as a regulator of seladin-1 expression for the first time. The relevance of these findings for the association of DM with AD is discussed.

Electrical field stimulation (EFS)-induced relaxations turn into contractions upon removal of extracellular calcium in rat mesenteric artery.

In the present study, we aimed to examine the effect of blockade of L-type Ca(2+) channels (LTCC) and in addition the removal of extracellular Ca(2+), on EFS-induced relaxations in rings of rat mesenteric artery. EFS applied to the tissues precontracted with phenylephrine caused relaxations which were markedly inhibited by nifedipine (10(-7)M) and tetraethylammonium (TEA) (1mM). Addition of LTCC opener BAY K 8644 (10(-7)M) failed to enhance the relaxations. Upon removal of Ca(2+), EFS with the same stimulation parameters produced frequency-dependent transient contractions. Tetrodotoxin (10(-6)M), capsaicin (10(-5)M) and removal of endothelium did not alter these contractions suggesting that they were not neural in origin and endothelium-derived contracting factors were unlikely to be involved. However, they were increased by nearly 40% in response to BAY K 8644 (10(-7)M) and were inhibited by nifedipine (10(-7)M), indicating that activation of the LTCCs was essential. Inositol triphosphate (InsP3) receptor antagonist 2-APB (10(-4)M) significantly reduced, and high concentration of caffeine (20mM) almost totally suppressed the contractions. These results suggest that in the absence of extracellular Ca(2+) EFS through membrane depolarization, evokes the opening of the LTCCs which subsequently leads to the release of Ca(2+) from internal stores via InsP3 receptors, a phenomenon known as Ca(2+) channel-induced Ca(2+) release (CCICR), to trigger vasoconstriction. That activation of LTCCs causes arterial relaxation or contraction depending on the Ca(2+) status apparently exemplifies how the same messenger fulfils opposing physiological functions in a given cell.

Preparation and in vitro/in vivo evaluation of microparticle formulations containing meloxicam.

In this study, we have formulated chitosan-coated sodium alginate microparticles containing meloxicam (MLX) and aimed to investigate the correlation between in vitro release and in vivo absorbed percentages of meloxicam. The microparticle formulations were prepared by orifice ionic gelation method with two different sodium alginate concentrations, as 1% and 2% (w/v), in order to provide different release rates. Additionally, an oral solution containing 15 mg of meloxicam was administered as the reference solution for evaluation of in vitro/in vivo correlation (ivivc). Following in vitro characterization, plasma levels of MLX and pharmacokinetic parameters [elimination half-life (t(1/2)), maximum plasma concentration (C(max)), time for C(max) (t(max))] after oral administration to New Zealand rabbits were determined. Area under plasma concentration-time curve (AUC(0-∞)) was calculated by using trapezoidal method. A linear regression was investigated between released% (in vitro) and absorbed% (in vivo) with a model-independent deconvolution approach. As a result, increase in sodium alginate content lengthened in vitro release time and in vivo t(max) value. In addition, for ivivc, linear regression equations with r(2) values of 0.8563 and 0.9402 were obtained for microparticles containing 1% and 2% (w/v) sodium alginate, respectively. Lower prediction error for 2% sodium alginate formulations (7.419 ± 4.068) compared to 1% sodium alginate formulations (9.458 ± 5.106) indicated a more precise ivivc for 2% sodium alginate formulation.

Effects of atorvastatin and L-arginine treatments on electrical field stimulation-mediated relaxations in pulmonary arterial rings of monocrotaline-induced pulmonary hypertensive rats.

This study aimed to examine the effect of monocrotaline (MCT)-induced pulmonary hypertension on electrical field stimulation (EFS)-mediated relaxation in rings of rat main pulmonary artery and to see whether treatment with atorvastatin or L-arginine would prevent the action of MCT. Rats were killed 21 days after MCT injection (60 mg/kg), and the main pulmonary arteries were isolated. EFS (40 V, 0.2 milliseconds, 5 seconds, 10 Hz)-induced relaxations in vessels precontracted with phenylephrine (10(-6) to 3 × 10(-6) M) were abolished in MCT-injected group compared with control group. Treatment of MCT group with atorvastatin (10 mg/kg, orally) completely, whereas treatment with L-arginine (500 mg/kg, intraperitoneally) partially but significantly prevented the inhibition. Similarly, acetylcholine (10(-9) to 3 × 10(-5) M)-evoked relaxations that were markedly inhibited in MCT-group were also protected from inhibition after pretreatment with atorvastatin or L-arginine. Responses to endothelium-independent relaxants sodium nitroprusside (10(-9) to 10(-5) M), pinacidil (10(-10) to 10(-4) M), and papaverine (10(-8) to 10(-4) M) were unaltered in MCT-induced pulmonary hypertensive rats. The present findings suggest that MCT-induced pulmonary hypertension inhibits the EFS-mediated relaxation through suppression of endothelial NO production. Reversal of this inhibition by atorvastatin treatment presumably results from stimulation of endothelial nitric oxide synthase expression. Relatively weak protection elicited by L-arginine might be secondary to impaired endothelial nitric oxide synthase activity caused by MCT-induced pulmonary hypertension.

Does endothelium-derived hyperpolarizing factor play a role in endothelium-dependent component of electrical field stimulation-induced vasorelaxation of rat mesenteric arterial rings?

Electrical field stimulation (EFS)-induced, nonadrenergic, noncholinergic vasodilation was investigated in rat mesenteric arterial rings. Tetrodotoxin (10(-6) M), capsaicin (10(-5) M), or L-NAME (10(-4) M) failed to change the EFS-induced relaxations, whereas they were increased with indomethacin (10(-5) M). Removal of the endothelium caused approximately 20% reduction in the maximum response, whereas precontraction with 40 mM KCI abolished the relaxations at all frequencies. Iberiotoxin (3 x 10(-7) M) attenuated the relaxations in endothelium-intact tissues but blocked completely those in endothelium-denuded arteries. Combination of TRAM-34 (10(-5) M) with apamin (5 x 10(-7) M) and single administrations of NiCI2 (5 x 10(-4) M), ruthenium red (3 x 10(-5) M), and 18[alpha]-glycyrrhetinic acid (10(-4) M) significantly reduced the responses only in endothelium-intact tissues. These data indicate that in rat mesenteric arteries, EFS leads to vasodilation through both endothelium-dependent and endothelium-independent mechanisms. The major component of the relaxation is endothelium independent and seems to occur via BK(Ca) channels, whereas endothelium-dependent component is likely to be mediated by endothelium-derived hyperpolarizing factor rather than nitric oxide, prostacyclin, or a neural substance. We propose that Ca2+ entry into endothelial cells via nonspecific cation channels in response to EFS induces hyperpolarization by activating endothelial IK(Ca) and SK(Ca) channels, which is spread to the smooth muscle via gap junctions to produce relaxation.

Torsion/detorsion of the testis does not modify responses to nitric oxide in rat isolated penile bulb.

Ischaemia-reperfusion damage induced by torsion/detorsion of the testicles may be a causative factor leading to erectile dysfunction through oxidative stress-dependent changes in the responses of the penile bulb, an erectile tissue of the penis. We aimed at investigating the effects of unilateral testicular torsion/detorsion (2 or 24 hr) treatment on relaxations induced by electrical field stimulation and sodium nitroprusside in rat isolated penile bulb. Male Sprague-Dawley rats used in the study were divided into two groups. The treatment group was subjected to unilateral torsion followed by detorsion for 2 or 24 hr, while the control group underwent only sham operation. For in vitro organ bath experiments, penile bulbs were isolated and responses to relaxant agents and electrical field stimulation (70 V, 1 msec., 0.5-8 Hz, 5 sec.) were recorded on a computer-based data acquisition system via a force displacement transducer. In tissues precontracted with phenylephrine (3 x 10(-6 )M), relaxations induced by electrical field stimulation were not significantly different before and after 2 or 24 hr of detorsion. Similarly sodium nitroprusside- (10(-8)-3 x 10(-6 )M) and papaverine-induced (10(-7)-10(-4 )M) relaxations were also found unchanged in the detorsion group compared to control. In conclusion, spermatic cord torsion did not lead to impairment in nitric oxide-mediated relaxant responses of the rat isolated penile bulb.

The effects of superoxide anion generators on responses to exogenous nitric oxide and non-adrenergic, non-cholinergic nerve stimulation in rat isolated penile bulb.

The effects of the superoxide anion generators, pyrogallol and hydroquinone on relaxations induced by electrical field stimulation (70 V, 0.7 msec., 0.5-8 Hz for 5 sec.) and exogenous nitric oxide donor sodium nitroprusside, were investigated in rat penile bulb precontracted with phenylephrine (10(-4) M). Pyrogallol (10(-4) M, 3 x 10(-4) M) and hydroquinone (3 x 10(-4) M) reduced the relaxations induced by sodium nitroprusside, but had no effect on relaxations elicited by nitrergic nerve stimulation. After treatment with diethyldithiocarbamate (3 x 10(-3) M), an inhibitor of Cu/Zn superoxide dismutase, both agents reduced the relaxations induced by electrical field stimulation. Superoxide dismutase, at 300 U/ml, significantly reversed the inhibitory action of pyrogallol and hydroquinone on responses to sodium nitroprusside. This concentration of superoxide dismutase failed to reverse the inhibitory action of pyrogallol on responses to electrical field stimulation observed in the presence of diethyldithiocarbamate, while at 600 U/ml it significantly prevented the reduction in relaxations. However, even at 600 U/ml, superoxide dismutase did not alter the decrease in responses to electrical field stimulation evoked by hydroquinone in tissues pretreated with diethyldithiocarbamate. These results suggest that the nitrergic transmitter in rat penile bulb is protected against superoxide anions by endogenous Cu/Zn superoxide dismutase in a manner similar to gastric fundus and anococcygeus muscles.

Inhibition of acetylcholine-induced EDHF response by elevated glucose in rat mesenteric artery.

The effects of high glucose on endothelium-derived hyperpolarizing factor (EDHF)-mediated relaxations of isolated rat mesenteric artery and the possible involvement of reactive oxygen species in these responses were investigated. After precontraction with phenylephrine (3 x 10(-8)-10(-7) M), acetylcholine (10(-8)-3 x 10(-6) M) and A 23187 (10(-8)-3 x 10(-6) M), a calcium ionophore, induced concentration-dependent relaxations in the presence of N(W)-nitro-l-arginine methyl ester (L-NAME) (10(-4) M) and indomethacin (10(-5) M). These relaxations were abolished in the presence of charybdotoxin (2 x 10(-7) M) plus apamin (10(-7) M) and were assumed to be mediated by EDHF. Effects of elevated glucose were examined by incubating the arterial rings for 6 h in Krebs-Henseleit solution containing 22.2 mM glucose. Under these conditions relaxation to acetylcholine was significantly attenuated but was unchanged when the tissues were incubated for 6 h in solution containing 11.1 mM mannitol used as hyperosmotic control. Addition of superoxide dismutase (SOD) (75 U/ml) and combination of SOD with catalase (200 U/ml) during incubation with high glucose significantly preserved the impairment of EDHF-mediated relaxations to acetylcholine. A 23187-induced endothelium-dependent relaxation was not affected by high glucose. Similarly, relaxations to pinacidil (10(-10)-10(-5) M) and to sodium nitroprusside (SNP) (10(-10)-3 x 10(-7) M) were also unchanged in the rings exposed to high glucose. These results suggest that in rat mesenteric arteries exposed to elevated glucose receptor-dependent EDHF-mediated relaxations (acetylcholine-induced) are impaired whereas receptor-independent ones (A 23187-induced) and responses to smooth muscle relaxants that exert their effects through mechanisms independent of endothelium are unaffected. Our findings lead us to propose that reactive oxygen species like superoxide ((.)O(2)(-)) and hydrogen peroxide (H(2)O(2)) do seem to play a role in the impairment of EDHF-mediated relaxations in the presence of elevated glucose.

Some arylacridine derivatives possessing potassium channel opening activity.

In this study, six new 2,2,7,7-tetramethyl-9-aryl-2,3,4,5,6,7,9,10-octahydro-1,8-acridinedione derivatives (1-6) were synthesised and their functional effects on vascular potassium channels and mechanism of induced relaxations on phenylephrine-induced contractile responses in isolated rat mesenteric arteries were investigated. Pinacidil was used as standard potassium channel opener. Compounds 1, 2, 5, 6 and pinacidil induced concentration-dependent relaxation response of vessel rings previously contracted with phenylephrine.

Two new nitric oxide synthase inhibitors: pyridoxal aminoguanidine and 8-quinolinecarboxylic hydrazide selectively inhibit basal but not agonist-stimulated release of nitric oxide in rat aorta.

Structural modification at one of the guanidine nitrogens of L-arginine has led to the development of a number of compounds N(G)-monomethyl-L-arginine (L-NMMA), N(G)-nitro-L-arginine (L-NOARG), N(G)-nitro-L-arginine methyl ester (L-NAME) that competitively inhibit nitric oxide synthase (NOS). It was reported that another chemically related compound known as a glycation inhibitor, aminoguanidine also inhibits NOS. Recently, two new glycation inhibitors, structurally related to aminoguanidine (AG), pyridoxal aminoguanidine (PLAG) and 8-quinoline carboxylic hydrazide (8Q) were synthesized. In this study, the effects of these two inhibitors on responses mediated by constitutive nitric oxide (NO) were investigated in vitro. For this purpose, in the present study vascular responses to phenylephrine and acetylcholine in isolated aortas were evaluated. Incubation (15 min) with PLAG and 8Q (10(-4)M for each) induced potentiation of phenylephrine-induced contraction in endothelium intact but not in endothelium denuded rings of rat aorta. The ability of PLAG or 8Q to augment phenylephrine-induced tone in endothelium containing rings was completely prevented by preincubation with L-arginine (1mM), but not with D-arginine. Both compounds (PLAG, 8Q) did not affect acetylcholine-induced relaxation. These results suggest that both of the new compounds produced a selective inhibition of basal but not agonist stimulated production of nitric oxide in rat aorta.

Omeprazole-induced relaxation in rat aorta is partly dependent on endothelium.

We investigated the effect of omeprazole (1 x 10(-5)-3 x 10(-4)M), an inhibitor of H(+),K(+)-ATPase, on rat aortic rings pre-contracted with phenylephrine (10(-6)M). Omeprazole relaxed the tissue in a concentration-dependent manner. Either removal of the endothelium or incubation with nitric oxide (NO) synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME, 3 x 10(-5)M) significantly attenuated the relaxations. Pre-treatment with L-arginine (10(-3)M), but not with D-arginine, reversed the inhibitory action of L-NAME. Indomethacin (10(-6)M) and tetraethylammonium (TEA, 10(-2)M) did not affect the relaxant responses to omeprazole indicating the lack of involvement of cyclooxygenase products and K(+) channels, respectively. These results suggest a role of NO in the mechanism of action of omeprazole.