Effects of NO/cGMP inhibitors in a rat model of anaphylactoid shock

Anaphylactic shock can be defined as an acute syndrome, and it is the most severe clinical manifestation of allergic diseases. Anaphylactoid reactions are similar to anaphylactic events but differ in the pathophysiological mechanism. Nitric oxide (NO) inhibitors during anaphylaxis suggest that NO might decrease the signs and symptoms of anaphylaxis but exacerbate associated vasodilation. Therefore, blocking the effects of NO on vascular smooth muscle by inhibiting the guanylate cyclase (GC) would be a reasonable strategy. This study aimed to investigate the effects of NO/cGMP pathway inhibitors methylene blue (MB), Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME), and indigo carmine (IC) in shock induced by compound 48/80 (C48/80) in rats. The effect was assessed by invasive blood pressure measurement. Shock was initiated by C48/80 intravenous bolus injection 5 min before (prophylactic) or after (treatment) the administration of the inhibitors MB (3 mg/kg), L-NAME (1 mg/kg), and IC (3 mg/kg). Of the groups that received drugs as prophylaxis for shock, only the IC group did not present the final systolic blood pressure (SBP) better than the C48/80 group. Regarding shock treatment with the drugs tested, all groups had the final SBP similar to the C48/80group. Altogether, our results suggested that inhibition of GC and NO synthase in NO production pathway was not sufficient to revert hypotension or significantly improve survival.

Effects of nitric oxide inhibitors in mice with bladder outlet obstruction

ABSTRACT Purpose To investigate the lower urinary tract changes in mice treated with L-NAME, a non-selective competitive inhibitor of nitric oxide synthase (NOS), or aminoguanidine, a competitive inhibitor of inducible nitric oxide synthase (iNOS), after 5 weeks of partial bladder outlet obstruction (BOO), in order to evaluate the role of constitutive and non-constitutive NOS in the pathogenesis of this experimental condition. Materials and Methods C57BL6 male mice were partially obstructed and randomly allocated into 6 groups: Sham, Sham + L-NAME, Sham + aminoguanidine, BOO, BOO + L-NAME and BOO + aminoguanidine. After 5 weeks, bladder weight was obtained and cystometry and tissue bath contractile studies were performed. Results BOO animals showed increase of non-voiding contractions (NVC) and bladder capacity, and also less contractile response to Carbachol and Electric Field Stimulation. Inhibition of NOS isoforms improved bladder capacity and compliance in BOO animals. L-NAME caused more NVC, prevented bladder weight gain and leaded to augmented contractile responses at muscarinic and electric stimulation. Aminoguanidine diminished NVC, but did not avoid bladder weight gain in BOO animals and did not improve contractile responses. Conclusion It can be hypothesized that chronic inhibition of three NOS isoforms in BOO animals leaded to worsening of bladder function, while selective inhibition of iNOS did not improve responses, what suggests that, in BOO animals, alterations are related to constitutive NOS.

Cytotoxicity, nitric oxide and acetylcholinesterase inhibitory activity of three limonoids isolated from Trichilia welwitschii (Meliaceae)

BACKGROUND: Limonoids are highly oxygenated compounds with a prototypical structure. Their occurrence in the plant kingdom is mainly confined to plant families of Meliaceae and Rutaceae. Owing to their wide range of pharmacological and therapeutic properties, this study was aimed at investigating the potential nitric oxide (NO) and acetylcholinesterase (AChE) inhibitory activity and the cytotoxicity of three limonoids: trichilia lactone D5 (1), rohituka 3 (2) and dregeanin DM4 (3), isolated from Trichilia welwitschii C.DC. RESULTS: Results indicated that the three limonoids had low cytotoxicity towards Vero cells with LC50 values ranging from 89.17 to 75.82 µg/mL. Compounds (2) and (3) had lower cytotoxicity compared to puromycin and doxorubicin used as reference cytotoxic compounds. Compound (1) (LC50 of 23.55 µg/mL) had good antiproliferative activity against RAW 264.7 cancer cells. At the lowest concentration tested (0.5 µg/mL), compound (2) and (3) released the lowest amount of nitric oxide (2.97 and 2.93 µM, respectively). The three limonoids had anti-AChE activity with IC50 values ranged of 19.13 µg/mL for (1), 34.15 µg/mL for (2) and 45.66 µg/mL for (3), compared to galantamine (IC50 of 8.22 µg/mL) used as positive control. CONCLUSION: The limonoid compounds studied in this work inhibited nitric oxide production in LPS-stimulated macrophages and had anti-AChE activity. Trichilia lactone D5 had potential antiproliferative activity against RAW 264.7 cancer cells. The limonoids had low cytotoxicity towards Vero cells lines. This study provided further examples of the importance of limonoids compounds as potential AChE inhibitors and anti-inflammatory agents targeting the inhibition of NO production.

Anti-inflammatory effects of Zingiber officinale roscoe involve suppression of nitric oxide and prostaglandin E2 production

Inflammation is a physiological response to injury and infection. However, chronic inflammation causes tissue damage and is a feature of most chronic diseases. Despite significant progress in developing therapies to target chronic inflammation over the years, almost all current therapies have serious side effects. The current investigation is to identify naturally-existing anti-inflammatory therapies with fewer side effects. The anti-inflammatory effects and mechanisms of action of extracts and fractions obtained using vacuum liquid chromatography [VLC] from ginger [Zingiber officinale] on the production of nitric oxide [NO] and prostaglandin E[2] [PGE[2]] were investigated. NO and PGE[2] production were induced by stimulating the mouse RAW264.7 monocyte/macrophage cell line with lipopolysaccharide [LPS]. Levels of NO and PGE[2] were deter-mined using the Griess method and enzyme linked sorbent assay [ELISA], respectively. Extracts of two Zingiber officinale species obtained with chloroform showed po-tent inhibitory effects on NO and PGE[2] production. The extracts had a higher potency than N[G]-nitro-L-arginine methyl ester [L-NAME], a known specific inducible nitric oxide synthase [iNOS] inhibitor and were comparable in their effects on PGE[2] with Indomethacin, a specific PGE[2] inhibitor. Further, we identified a fraction [F6] that had most potent inhibi-tory effects. The study shows that extract of Zingiber officinale have strong inhibitory effects on key pro-inflammatory mediators involved in chronic inflammation. Both the extracts and F6 had better inhibitory effects than established pharmaceutical inhibitors of NO and PGE[2]

El sistema renina angiotensina y el estrés oxidativo en un modelo experimental de preeclampsia en ratas / The renin angiotensin system and the oxidative stress in a animal model of preeclampsia in rats

La preeclampsia es un síndrome exclusivo de la gestación humana y responsable de una alta morbimortalidad perinatal, cuyas manifestaciones incluyen: hipertensión arterial, proteinuria y edema. Un mecanismo postulado en la fisiopatología de la preeclampsia, es la reducción de la perfusión placentaria y el desarrollo del síndrome clínico materno ocasionado por la liberación de factores placentarios que afectan la regulación de la presión arterial y la función renal. Uno de los factores que ocasiona el trastorno endotelial son las especies reactivas de oxígeno, el incremento de elementos vasoactivos, así como la disminución de agentes vasorelajantes como el óxido nítrico. Todas estas alteraciones vasculares conducen no sólo a la hipertensión sino también a la disfunción renal. Debido a la importancia del papel del óxido nítrico y su desregulación en la preeclampsia, en el presente trabajo se caracterizó un modelo experimental de preeclampsia que resulta de la inhibición de la síntesis de óxido nítrico mediante la administración de L-NAME a ratas preñadas y no preñadas. En el mismo se evaluó el estatus oxidativo y, el papel del sistema renina angiotensina en la contribución de la disfunción renal. Los resultados demuestran el papel primordial del óxido nítrico y su desregulación en este modelo de preeclampsia experimental. En efecto, se demostró que el tratamiento durante siete días con L-NAME incrementó la presión arterial media, aumentó la sensibilidad vascular, inhibió la actividad de la sintasa del óxido nítrico renal y redujo el guanilil monofosfato cíclico urinario. La disfunción endotelial renal en este modelo experimental se manifiesto por proteinuria, incremento de la creatinina plasmática, disminución de la excreción urinaria de sodio, potasio y creatinina, así como, evidencia morfológica de endoteliosis glomerular. Al caracterizar el papel de las enzimas antioxidantes renales se encontró una reducción significativa de la actividad de las mismas, y un incremento de la peroxidación lipídica asociada a una elevada concentración de agentes pro-oxidantes. Nuestro modelo experimental constituye una buena aproximación a la preeclampsia humana y no un efecto inespecífico del L-NAME, ya que aún cuando la inhibición crónica de la síntesis de óxido nítrico en ratas no preñadas induce un incremento de la presión arterial media, proteinuria, reducción de la actividad de la sintasa del óxido nítrico renal y de la excreción urinaria de guanilil monofosfato cíclico similar a las ratas preñadas, los efectos sobre la proteinuria, las acciones morfológicas renales, la excreción urinaria de sodio, potasio y creatinina, y sobre el sistema renina angiotensina son específicos de la preeclampsia experimental en ratas. Así, se demostró que en las ratas preñadas tratadas con L-NAME la actividad de la enzima convertidora de angiotensina plasmática, los niveles de renina plasmática, y la aldosterona amniótica se encuentran marcadamente disminuidos, cuando se comparan con las ratas preñadas normotensas. Estos hallazgos sugieren que la preeclampsia experimental se caracteriza por la supresión de los componentes circulantes del sistema renina angiotensina, que podrían ser responsables del desbalance entre los sistemas vasoconstrictores y vasodilatadores observados en la preeclampsia, así como de algunos de los signos de la preeclampsia, similar a lo que ocurre en la mujer embarazada hipertensa. Por otra parte, al evaluar la contribución del estrés oxidativo en el daño renal en la preeclampsia experimental, se demostró una disminución de la actividad de las enzimas antioxidantes renales. Asimismo, la disminución de la actividad de la glutatión peroxidasa plasmática y la tendencia a la reducción en la glutatión peroxidasa en el líquido amniótico, con el simultáneo incremento de los valores de las sustancias que reaccionan con el ácido tiobarbitúrico (TBARS) plasmático, sugiriéndose que la desregulación generalizada y renal está asociada a una baja protección oxidativa durante la preeclampsia, que favorece a la insuficiencia renal. El incremento temprano del estrés oxidativo placentario juega un papel fundamental en la disfunción endotelial generalizada y el daño renal en la preeclampsia. Debido a ello, nos planteamos que el tratamiento temprano con antioxidantes o con desacoplantes de la NAD (P) H oxidasa podría interrumpir el proceso de este síndrome. Efectivamente, el tratamiento crónico con un compuesto que mimetiza a la superóxido dismutasa, el tempol, o el desacoplante del ensamblaje de la NAD(P)H oxidasa (apocinina), fueron capaces de reducir significativamente la hipertensión inducida por el L-NAME. Igualmente, ambos compuestos fueron capaces de prevenir la proteinuria y la reducción de la actividad de las enzimas antioxidantes renales estudiadas. En conclusión, la preeclampsia experimental inducida por la inhibición crónica de la síntesis de óxido nítrico en ratas preñadas, reproduce los signos clásicos de la preeclampsia humana, y se acompaña de la desregulación del sistema renina angiotensina y de disfunción renal. Esto nos permite aseverar que este modelo experimental de preeclampsia constituye una buena aproximación a la preeclampsia humana. Se demuestra que el daño renal encontrado en este modelo experimental se asocia a una disminución de los mecanismos antioxidantes renales, que lleva a un incremento del estrés oxidativo, y a una reducción de la protección de la función renal. Estos resultados indican que la sobreproducción de especies reactivas de oxígeno tanto placentaria como renal, son causa fundamental de la disfunción endotelial generalizada y del daño renal. Finalmente, la inhibición del estrés oxidativo mediante el uso de agentes antioxidantes como el tempol o la apocinina, pudiese ser una de las posibles estrategias terapéuticas en el tratamiento de la hipertensión inducida por el embarazo humano y abre nuevos horizontes en el tratamiento de este síndrome.

Rol del sistema kallicreína kinina y su interrelación con sistemas vasoactivos durante la preñez / Role of the kallikrein kinin system and its interrelationship with vasoactive systems in pregnancy

La hiperfiltración glomerular y el aumento de la reabsorción de sodio son factores fundamentales para el desarrollo de la unidad feto placentaria. Dichos factores resultan de adaptaciones hemodinámicas y renales en las que participan sistemas vasoactivos. Se pudo demostrar en ratas que la activación del sistema kallicreína kinina (SKK) precede a la instalación de la hiperfiltración glomerular, dado que su inhibición por aprotinina previene el aumento del filtrado glomerular. Además, la inhibición individual o asociada de los efectores específicos del SKK, las prostaglandinas (PGs) y el óxido nítrico (ON), confirman la dependencia del filtrado glomerular del SKK durante la preñez. Encontramos también que el sistema renina angiotensina (SRA) participa en la generación de la hiperfiltración glomerular dado que ésta es afectada por la administración de bloqueantes del SRA. La inhibición máxima sobre el pico de hiperfiltración se obtuvo con el bloqueo de ambos sistemas (SKK y SRA). Además, estrategias para alterar la hiperfiltración glomerular y la reabsorción de sodio de la preñez evidenciaron alteraciones en el desarrollo de la unidad feto placentaria, menor número de crías, mayor cantidad de reabsorciones intrauterinas y retardo en el crecimiento. El tratamiento combinado de inhibidores del SKK asociados a bloqueantes del SRA o de óxido nítrico mostraron los mayores efectos. En consecuencia, demostramos que el SKK juega un rol central en los fenómenos de adaptación que acompañan la preñez normal. La interrelación del SKK con varios sistemas vasoactivos parecería formar una red que participa en las adaptaciones hemodinámicas para un adecuado desarrollo de la gestación y de la unidad feto-placentaria.

Glomerular hyperfiltration and increased sodium reabsorption are key factors for the development of the fetus and placenta in pregnancy. These adjustments result from hemodynamic and renal factors involving vasoactive systems. It was demonstrated in rats that activation of KKS precedes the installation of glomerular hyperfiltration as aprotinin prevents the increase in glomerular filtration. In addition, individual or associated inhibition of specific kallikrein kinin system effectors, prostaglandins (PGs) and nitric oxide (NO), confirm the glomerular filtration rate dependence of KKS during pregnancy. It was also found that the renin-angiotensin system (RAS) contributes to glomerular hyperfiltration as this is affected by the administration of RAS blockers. The peak of hyperfiltration maximum inhibition was obtained by the blockade of both systems (KKS and RAS). In addition, strategies used to alter the glomerular hyperfiltration and increased sodium reabsorption during pregnancy, showed abnormalities in the development of the fetus and placenta, fewer offspring, more fetus resorptions and intrauterine growth retardation. KKS inhibitors associated with RAS or nitric oxide blockers showed the greatest impact. As a consequence, it was demonstrated that KKS plays a central role in the adaptation phenomenom that accompanies normal pregnancy. The interplay of KKS with several vasoactive systems, seem to arrange a network involved in the hemodynamic adaptations to allow the proper development of pregnancy and the fetus and placenta.

Effect of nitric oxide inhibitor and donor substances on the infammatory process caused by endodontic irrigants

Nitric oxide (NO) has been considered a key molecule in infammation. OBJECTIVE: The aim of this study was to evaluate the effect of treatment with L-NAME and sodium nitroprussiate, substances that inhibit and release NO, respectively, on tissue tolerance to endodontic irrigants. MATERIAL AND METHODS: The vital dye exudation method was used in a rat subcutaneous tissue model. Injections of 2 percent Evans blue were administered intravenously into the dorsal penial vein of 14 male rats (200-300 g). The NO inhibitor and donor substances were injected into the subcutaneous tissue in the dorsal region, forming two groups of animals: G1 was inoculated with L-NAME and G2 with sodium nitroprussiate. Both groups received injections of the test endodontic irrigants: acetic acid, 15 percent citric acid, 17 percent EDTA-T and saline (control). After 30 min, analysis of the extravasated dye was performed by light absorption spectrophotometry (620 nm). RESULTS: There was statistically signifcant difference (p<0.05) between groups 1 and 2 for all irrigants. L-NAME produced a less intense infammatory reaction and nitroprussiate intensifed this process. CONCLUSIONS: Independently of the administration of NO inhibitors and donors, EDTA-T produced the highest irritating potential in vital tissue among the tested irrigating solutions.

[Effects of nitric oxide production inhibitor named, N[G]-nitro-L-arginine methyl ester [L-NAME], on rat mesenchymal stem cells differentiation]

Recently, the findings of some studies have shown that, nitric oxide [NO] probably has an important role in differentiation of mesenchymal stem cells to osteoblasts. The aim of the present investigation was to study the effects of nitric oxide production inhibitor named, N[G]-nitro-L-arginine methyl ester [L-NAME], on rat mesenchymal stem cells differentiation to osteoblasts in vitro. This was an experimental study conducted at Hamedan University of Medical Sciences in 2009, in which rat bone marrow stem cells were isolated in an aseptic condition and cultured in vitro. After third passage, the cells were cultured in osteogenic differentiation medium. To study the effects of L-NAME on osteogenic differentiation, the L-NAME was added to the culture medium at a concentration of 125, 250, and 500 micro M in some culture plates. During the culture procedure, the media were replaced with fresh ones, with a three days interval. After 28 days of culturing; the mineralized matrix was stained using Alizarian red staining method. The gathered data were analyzed by SPSS software version 12 using one way ANOVA. The findings of this study showed that in the presence of L-NAME, differentiation of bone marrow mesenchymal stem cells to osteoblasts was disordered and matrix mineralization significantly decreased in a dose dependent manner. This study revealed that, inhibition of nitric oxide production using L-NAME can prevent the differentiation of rat bone marrow mesenchymal stem cells to osteoblast. The results imply that NO is an important constituent in differentiation of mesenchymal stem cell to osteoblasts

Divergent effects of Nitric oxide on airway epithelial cell activation

Nitric oxide (NO*) is a gaseous mediator synthesized by Nitric oxide sinthases. NO* is involved in the modulation of inflammation, but its role in airway inflammation remains controversial. We investigated the role of NO* in the synthesis of the chemok Nes Interleukin-8 and Monocyte Chemotactic Protein-1, and of Intercellular Adhesion Molecule-1 by human airway epithelial cells. normal human bronchial epithelial cells and the bronchial epithelial cell line BEAS-2B were used. Neterleukin-8 (IL-8) and Monocyte Chemotactic Protein-1 (MCP-1) secretion and Intercellular Adhesion Molecule-1 (ICAM-1) expression were measured by ELISA. mRNA was assessed by semiquantitative RTI-PCR. Neterleukin-8 secretion was significantly reduced after 24h incubation with the NO* donor, sodium nitroprusside. The effect was dose-dependent. Similar results were obta Ned with S-Nitroso-N-D,L-penicillam Ne and S-Nitroso-L-glutathione. Inhibition of endogenous NO* with the Nitric oxide synthase inhibitor N-Nitro-L-arg N Ne-methyl-esther caused an increase in IL-8 secretion by lypopolisaccharide- and cytok Ne-stimulated BEAS-2B cells. Sodium nitroprusside also caused a reduction in Monocyte Chemotactic Protein-1 secretion by both cell types. In contrast, Intercellular Adhesion Molecule-1 expression was upregulated by sodium NItroprusside. RTI-PCR results indícate that the modulation of protein levels was paralleled by modification in mRNA levels. NO* has divergent effects on the synthesis of different inflammatory mediators in human bronchial epithelial cells.

Efeito do óxido nítrico no transporte mucociliar / Effects of nitric oxide in mucociliary transport

As vias aéreas, constituídas por epitélio ciliado e secretor de muco, promovem ao trato respiratório mecanismo de defesa que livra esta superfície das partículas inaladas durante a respiração. É de fundamental importância o entendimento da fisiologia e dos mecanismos envolvidos com a atividade mucociliar. A literatura sugere que o NO, em especial o produzido pela expressão da iNOS, mantém a função mucociliar e a defesa imune da cavidade nasal. Objetivo: Avaliar o envolvimento do NO e das vias enzimáticas da produção do NO no transporte mucociliar, utilizando inibidores da NO sintase constitutiva e indutiva, L-NAME e aminoguanidina, respectivamente. Materiais e métodos: Preparações de palatos de rã foram imersos em soluções de ringer (controle), L-NAME ou aminoguanidina. Os palatos foram imersos nestas soluções por quatro períodos de 15 minutos. Medidas da velocidade do transporte mucociliar foram feitas antes e após cada exposição. Resultos: Palatos controles mantiveram estável a velocidade do transporte. O L-NAME aumentou, enquanto a aminoguanidina reduziu a velocidade de transporte do muco. Conclusão: O bloqueio inespecífico da cNOS com L-NAME e bloqueio relativamente específico da iNOS com aminoguanidina permitiu propor que dependendo da via o NO pode aumentar ou diminuir o transporte mucociliar em palatos de rã.

The airways are made up of ciliated epithelium which secretes mucous, protecting the respiratory tract from particles inhaled during breathing. Its is paramount to understand the physiology and the mechanisms involved in mucociliary activity. Literature suggests that Nitric oxide (NO), especially the one produced by iNOS expression, maintains the mucociliary function and the immune defense of the nasal cavity. AIM: to assess NO participation and the enzymatic pathways in the production of NO and mucociliary transport, using constructive and inductive NO synthetase inhibitors, L-NAME and aminoguanidine, respectively. Materials and methods: frog palates were prepared and immerse in ringer (control), L-NAME or aminoguanidine solutions. The palates were immerse in these solutions for four periods of 15 minutes. Mucociliary transport measures were carried out before and after each exposure. Results: control palates maintained stable their transportation speed. L-NAME increased, while aminoguanidine reduced mucous transportation velocity. Conclusion: unspecific cNOS block with L-NAME and relatively specific iNOS block with aminoguanidine results leads us to propose that depending on the pathway, the NO can increase or reduce mucociliary transport in frog palates.

Oxidative stress and nitric oxide deficiency in inflammation of chronic renal failure. Possible preventive role of L-arginine and multiple antioxidants

To evaluate the effect of L-arginine and multiple antioxidants on the inflammatory cytokines level, renal functions, blood pressure and dyslipidemia in chronic renal failure [CRF] rats. This study was carried out between December 2007 and November 2008 in the Department of Physiology, Faculty of Medicine, King Saud University, Kingdom of Saudi Arabia. Chronic renal failure was induced in 40 rats by renal mass reduction [RMR] and 10 rats were sham operated. Renal mass reduction rats were treated for 12 weeks by L-arginine and/or a mixture of antioxidants [L-carnitine, Catechin, Vitamins E and C] and the effect of the treatments on plasma cytokines, soluble intercellular adhesion molecule-1 [sICAM-1], nitrate [NO2] and nitrites [NO3], lipid profile, blood pressure, and renal function was examined. Chronic renal failure increased plasma Interleukin [IL]-1alpha, IL-1beta, IL-6, tumor necrosis factor-alpha, soluble intercellular adhesion molecule-1 [sICAM-1] levels and decreased anti-inflammatory cytokines IL-4 and 10 levels. In addition, hypertension, and dyslipidemia were found. L-arginine treatment improved kidney functions, decreased systolic blood pressure and decreased inflammatory cytokines levels. Antioxidants administration decreased inflammatory cytokines and sICAM-1 levels and increased IL-4 levels. Combined use of L-arginine and the antioxidants mixture were very effective in their tendency to recover normal values of kidney functions, plasma cytokines, sICAM-1, blood pressure, NO2/NO3, cholesterol and triglycerides concentrations. Restoration of the pro-oxidant/ antioxidants balance with increased NO bio-availability counteracts inflammation, renal impairment and dyslipidemia in CRF. This may open new perspectives for the role of antioxidants and NO precursors in the treatment of uremia and its complications

protective effect of N[G]-nitro-L-arginine methyl ester and insulin on nitric oxide inhibition and pathology in experimental diabetic rat liver

To determine on protective role of NG-nitro-L-arginine methyl ester L-NAME, and insulin on the liver in streptozotocin STZ induced diabetic rats. This study was performed in the Department of Biochemistry, Faculty of Medicine, Eskisehir Osmangazi University, Eskisehir, Turkey in 2007. Forty male Wistar albino rats were divided into 5 groups. These were untreated, diabetic control, STZ+insulin, STZ+L-NAME and STZ+insulin+L-NAME induced groups. The STZ was intraperitonally injected into 3 groups, and includes insulin, L-NAME, and their joint administrations as protective agents. The blood glucose and nitric oxide NO levels were determined. The tissue samples were obtained at the end of the fourth week. The liver tissue distortions were evaluated using hematoxylin and eosin staining. The serum glucose level was significantly higher in diabetic control p = 0.000, than the untreated group. Nitric oxide level was significantly lower in STZ+L-NAME p = 0.000 than the untreated group. The focal pseudo lobular structures without vena centralis increased portal fibrillary necrosis, and bile duct stenosis with coagulation necrosis of the peripheral hepatocytes were more observed in diabetic group than the protective agent groups. In addition, insulin, and L-NAME lead to hepatocyte regeneration; and minimal mononuclear cell infiltration was noted. NG-nitro-L-arginine methyl ester inhibits NO level in STZ+L-NAME induced group. NG-nitro-L-arginine methyl ester either alone, or with insulin combination significantly attenuates the liver morphological disarrangements in STZ induced diabetic rats

Role of nitric oxide and prostaglandin in the maintenance of cortical and renal medullary blood flow

This study was undertaken in anesthetized dogs to evaluate the relative participation of prostaglandins (PGs) and nitric oxide (NO) in the maintenance of total renal blood flow (TRBF), and renal medullary blood flow (RMBF). It was hypothesized that the inhibition of NO should impair cortical and medullary circulation because of the synthesis of this compound in the endothelial cells of these two territories. In contrast, under normal conditions of perfusion pressure PG synthesis is confined to the renal medulla. Hence PG inhibition should predominantly impair the medullary circulation. The initial administration of 25 µM kg-1 min-1 NG-nitro-L-arginine methyl ester produced a significant 26 percent decrease in TRBF and a concomitant 34 percent fall in RMBF, while the subsequent inhibition of PGs with 5 mg/kg meclofenamate further reduced TRBF by 33 percent and RMBF by 89 percent. In contrast, the initial administration of meclofenamate failed to change TRBF, while decreasing RMBF by 49 percent. The subsequent blockade of NO decreased TRBF by 35 percent without further altering RMBF. These results indicate that initial PG synthesis inhibition predominantly alters the medullary circulation, whereas NO inhibition decreases both cortical and medullary flow. This latter change induced by NO renders cortical and RMBF susceptible to a further decrease by PG inhibition. However, the decrease in medullary circulation produced by NO inhibition is not further enhanced by subsequent PG inhibition.

Biological activity predictions, crystallographic comparison and hydrogen bonding analysis of cholane derivatives.

A total of eighteen molecules of cholane derivatives (I-XVIII) (a series of steroids) have been included to predict their pharmacological effects, specific mechanisms of action, known toxicities, drug-likeness, etc, by using the statistics of multilevel neighbourhoods of atoms (MNA) descriptors for active and inactive fragments. The biological activity spectra for substances have been correlated on SAR base (structure-activity relationships data and knowledge base), which provides the different P(a) (possibility of activity) and P(i) (possibility of inactivity). Most of the probable activities have been characterized by P(a) and P(i) values, which depict that all the molecules have high value of teratogen activity. The Lipinski's thumb rule predicts that all the cholane derivatives have stronger preponderance for "cancer-like-drug" molecules and some of their related analogous have entered in the ANCI (American National Cancer Institute) database. Some selected bond distances and bond angles of interest have been taken into account and deviation of bond distances/bond angles, vis-a-vis the substitutional group and X-H...A intra/intermolecular hydrogen bonds has been discussed in detail. X-H...A intra and intermolecular hydrogen bonds in the molecules have been described with the standard distance and angle cut-off criteria. D-theta and d-theta. scatter plots for intra- and intermolecular interactions are presented for better understanding of packing interactions existing among these derivatives. There exists only one C-H...O intramolecular bifurcated hydrogen bond. while high tendency of intermolecular bifurcated hydrogen bonds based on a defined O-H...O has been observed, in which O atom acts as a prototype donor as well as acceptor. The frequency of occurrence of C-H...O hydrogen bonds is predominant (i.e. 85.7%) in intramolecular interactions, whereas in intermolecular interactions, frequency of occurrence for O-H...O interactions is 62.9%. Solvent-solute/solute-solvent interactions have also been investigated to understand more complicated processes that occur for biomolecules in aqueous solutions. The number of hydrogen donors in each derivative is less than 5, except for molecule XVIII and which has 91.3% of drug-likeness, instead of observed range of 96.5-99.39%.

Antagonic effect of the inhibition of inducible nitric oxide on the mortality of mice acutely infected with Escherichia coli and Bacteroides fragilis

Sepsis, the leading cause of death in intensive care units, is associated with overproduction of nitric oxide (NO) due to inducible NO synthase (iNOS), responsible for some of the pathologic changes. Aminoguanidine (AG) is a selective iNOS inhibitor with reported inconsistent actions in sepsis. To investigate the influence of iNOS, we studied models of acute bacterial sepsis using acute challenges with aerobic (Escherichia coli) and anaerobic (Bacteroides fragilis) bacteria in the presence of AG. Six-week-old, 23 g, male and female BALB/c and C57Bl/6j mice, in equal proportions, were inoculated (ip) with bacteria in groups of 4 animals for each dose and each experiment in the absence or presence of AG (50 mg/kg, ip, starting 24 h before challenge and daily until day 6) and serum nitrate was measured by chemiluminescence. Both types of bacteria were lethal to mice, with an LD50 of 6 nephelometric units (U) for E. coli and 8 U for B. fragilis. Nitrate production peaked on the second day after E. coli inoculation with 8 and 6 U (P < 0.05), but was absent after non-lethal lower doses. After challenge with B. fragilis this early peak occurred at all tested doses after 24 h, including non-lethal ones (P < 0.05). AG-treated mice challenged with E. coli presented higher survival (P < 0.05) and increased LD50. AG-treated mice challenged with B. fragilis had lower LD50 and higher mortality. Control AG-treated animals presented no toxic effects. The opposite effect of iNOS blockade by AG in these models could be explained by restriction of oxygen for immune cells or an efficient action of NO in anaerobic localized infections. The antagonic role of NO production observed in our bacterial models could explain the reported discrepancy of NO action in sepsis.

Persistent hypertension and progressive renal injury induced by salt overload after short term nitric oxide inhibition

INTRODUCTION: Administration of the NO inhibitor Nwð-nitro-L-arginine methyl ester (NAME) and a high-salt diet (HS) promotes severe albuminuria and renal injury, which regresses upon discontinuation of treatments. OBJECTIVE: We investigated whether these changes reappear after reinstitution of HS, and whether they are prevented by treatment with the antilymphocyte agent mycophenolate mofetil (MMF) or the AT-1 receptor blocker losartan (L). Adult male Munich-Wistar rats received NAME and HS. A control Group (C) received only HS. After 20 days, rats receiving HS and NAME exhibited severe hypertension and albuminuria. After a 30-day recovery period, hypertension was attenuated and albuminuria had virtually disappeared. MATERIAL AND METHODS: Rats were then distributed among the following groups: HS, receiving HS; NS, receiving a normal salt (NS) diet; HS-MMF, receiving HS and MMF; HS-LOS, receiving HS and L; HS-HDZ, receiving HS and hydralazine (HDZ). Sixty days later, NS rats showed only slight albuminuria and renal damage or inflammation. In contrast, HS rats developed severe hypertension, marked glomerulosclerosis with interstitial expansion and renal infiltration by macrophages and angiotensin II-positive cells. The group treated with losartan had lowered blood pressure and a lack of albuminuria or renal injury. MMF provided similar protection without altering blood pressure, suggesting a nonhemodynamic effect, a hypothesis reinforced by the finding that HDZ lowered blood pressure without preventing renal injury. RESULTS: These results indicate that treatment with HS and NAME predisposes to the development of hypertension and renal injury upon salt overload, characterizing a new model of chronic nephropathy. CONCLUSION: The response to MMF or L, but not HDZ, suggests a key role for inflammatory rather than hemodynamic factors.

INTRODUÇÃO: A administração de Nômega-nitro-L-arginina metiléster (NAME), um inibidor da produção de NO, com dieta rica em sal (HS) promove albuminúria e dano renal graves, reversíveis ao interromperem-se os tratamentos. OBJETIVO: Investigamos se tais alterações recrudescem ao reinstituir-se a HS e se são prevenidas pelo micofenolato mofetil (MMF), um agente antilinfócito, ou losartan, um bloqueador do receptor AT-1. MATERIAL E MÉTODOS: Ratos Münich-Wistar machos adultos receberam NAME e HS. Um grupo controle (C) recebeu apenas HS. Após 20 dias, os ratos que receberam HS e NAME exibiam hipertensão e albuminúria graves. Após recuperação de 30 dias, a hipertensão atenuou-se e a albuminúria praticamente desapareceu. Formaram-se então os grupos: HS, recebendo HS; NS, recebendo dieta normal em sal (NS); HS-MMF, recebendo HS e MMF; HS-LOS, recebendo HS e losartan; HS-HDZ, recebendo HS e hidralazina. Após sessenta dias os ratos NS tinham albuminúria e dano/inflamação renal apenas discretos. Já os ratos HS desenvolveram hipertensão e glomerulosclerose acentuadas, expansão intersticial e infiltração renal por macrófagos e células positivas para angiotensina II. Losartan baixou a pressão arterial e preveniu albuminúria e lesão renal. MMF proporcionou proteção semelhante sem alteração pressórica, sugerindo a ação de mecanismos não hemodinâmicos, hipótese reforçada pelo achado de que a HDZ baixou a pressão arterial sem prevenir a nefropatia. RESULTADOS: Esses resultados indicam que o tratamento com HS e NAME predispõe ao desenvolvimento de hipertensão e lesão renal induzidos por excesso de sal, caracterizando um novo modelo de nefropatia crônica. CONCLUSÃO: A resposta ao MMF ou losartan, mas não à hidralazina, sugere o predomínio de fatores inflamatórios.

Sepsis: emerging role of nitric oxide and selectins

Sepse – um estado de infecção bacteriana sistêmica – frequentemente leva à falência múltipla de órgãos e associa-se a altos índices de mortalidade, apesar de progressos recentes no manejo de pacientes em unidades de terapia intensiva. Muitos dos efeitos maléficos associados à sepse são atribuídos a uma resposta inflamatória patologicamente ampliada que leva a recrutamento neutrofílico e ativação das moléculas de adesão do grupo das selectinas, durante as fases iniciais do processo . O óxido nítrico e sua diversas isoformas também foram implicados nas diversas manifestações vasculares da sepse como participantes diretos da toxicidade celular. Esta revisão descreve o papel das selectinas e do óxido nítrico em situações clínicas e experimentais de sepse, bem como os respectivos efeitos de processos terapêuticos de bloqueio.

Semecarpus anacardium L, nuts inhibit lipopolysaccharide induced NO production in rat macrophages along with its hypolipidemic property.

Traditionally S. anacardium is used for rejuvenation, rheumatoid arthritis, fever and neurological disorders. In the present study it was observed that a fraction of S. anacacrdium at dose of 1 mg/100 g body wt, significantly reduced serum cholesterol from 378.87 mg/dl in the rats fed with atherogenic diet (AD) to 197.99 mg/dl (45-52%) in the rats fed with AD diet and increased serum HDL-cholesterol (33-37%). The same fraction also inhibited LPS induced NO production in the culture activated rat peritoneal macrophages in the dose dependent manner with IC50 value at 50 ng/ml of the culture medium. The drug in the above doses was completely safe and non-toxic, (no change in the enzymes), to liver and kidney functions.

Participação do óxido nítrico no processo inflamatório articular / Nitric oxide participation in the articular inflammatory process

O presente trabalho avalia a participação do óxido nítrico (NO) na fisiopatologia da resposta inflamatória articular. O papel pró-inflamatório deste medidor, tanto no componente vascular quanto no celular foi evidenciado em diferentes modelos experimentais, utilizando animais tratados cronicamente com o inibidor inespecífico de síntese de NO, NG-nitro-L-arginine metil ester (L-NAME, 20mg/Kg/dia/14 dias). Investigamos os mecanismos envolvidos na alteração de parâmetros inflamatórios como: interação leucócito endotélio, expressão de moléculas de adesão, reatividade da microcirculação, parâmetros hemodinâmicos, alterações na permeabilidade vascular, além da interação entre produção de NO e outros mediadores da resposta inflamatória, como eicosanóides, sistema de cininas, citocinas e histamina, bem como influência do tratamento na. hematopoiese e hemograma. Em conjunto, os estudos contribuem para esclarecer o controverso papel do NO no processo inflamatório. /The present study describes the participation of nitric oxide (NO) in the patophisiology of experimental arthritis induced by different antigens through chronicle administration of the unspecific inhibitor of NO synthases NG-nitro-L-arginine metyl ester (L-NAME, 20mg/Kg/day/14 days). The pro-inflammatory effect of NO was observed such as on cellular as on vascular events of the process. The mechanisms involved were assessed by investigations on haematopoeisis and haemogram, leukocyte endothelial interaction, adhesion molecules expression, haemodynamic properties, microvascular reactivity, protein leakage, as well as interaction between NO and eicosanoids, bradykinin, cytokines and histamine production. Together these studies contribute to clarify the controversial role of NO on the inflammatory process...