Injection Of L-arginine Research Articles

Effect of nitric oxide pathway regulation on water/sodium balance and renal function in a rodent model of acute liver and renal failure.

BackgroundThe pathomechanism of acute hepatorenal syndrome (HRS), a particular form of acute renal failure that occurs in the course of acute liver injury, is still poorly understood. The aim of our study was to estimate the influence of the activation and inhibition of the nitric oxide pathway on the water/sodium balance and development of acute renal failure in the course of HRS.Material/MethodsWe used male Sprague-Dawley rats in the acute galactosamine (Ga1N) model of HRS. The nitric oxide synthase (NOS) inhibitors L-NAME and L-arginine were administered intraperitoneally before and after liver damage.ResultsHRS developed in all tested groups. L-NAME increased osmotic clearance and urine volume more effectively before liver injury. Furthermore, administration of L-NAME increased creatinine clearance both before and after Ga1N injection. A double dose of L-NAME did not yield further improvement before Ga1N injection, but improved creatinine clearance after Ga1N intoxication. Injection of L-arginine increased sodium excretion and urine volume, but only after liver injury. Moreover, L-arginine injected after Ga1N caused significant improvement of the creatinine clearance in a dose-dependent manner.ConclusionsOur study shows that inhibition of the nitric oxide pathway improves parameters of water and sodium balance and prevents development of acute renal failure in the course of acute liver injury and liver failure. Activation of the nitric oxide system also has a favorable influence on water/sodium balance and renal failure, but only after liver injury.

Intracellular Hmgb1 Inhibits Inflammatory Nucleosome Release and Limits Acute Pancreatitis in Mice

High mobility group box 1 (HMGB1) is an abundant protein that regulates chromosome architecture and also functions as a damage-associated molecular pattern molecule. Little is known about its intracellular roles in response to tissue injury or during subsequent local and systemic inflammatory responses. We investigated the function of Hmgb1 in mice after induction of acute pancreatitis. We utilized a Cre/LoxP system to create mice with pancreas-specific disruption in Hmbg1 (Pdx1-Cre; HMGB1(flox/flox) mice). Acute pancreatitis was induced in these mice (HMGB1(flox/flox) mice served as controls) after injection of l-arginine or cerulein. Pancreatic tissues and acinar cells were collected and analyzed by histologic, immunoblot, and immunohistochemical analyses. After injection of l-arginine or cerulein, Pdx1-Cre; HMGB1(flox/flox) mice developed acute pancreatitis more rapidly than controls, with increased mortality. Pancreatic tissues of these mice also had higher levels of serum amylase, acinar cell death, leukocyte infiltration, and interstitial edema than controls. Pancreatic tissues and acinar cells collected from the Pdx1-Cre; HMGB1(flox/flox) mice after l-arginine or cerulein injection demonstrated nuclear catastrophe with greater nucleosome release when compared with controls, along with increased phosphorylation/activation of RELA nuclear factor κB, degradation of inhibitor of κB, and phosphorylation of mitogen-activated protein kinase. Inhibitors of reactive oxygen species (N-acetyl-l-cysteine) blocked l-arginine-induced DNA damage, necrosis, apoptosis, release of nucleosomes, and activation of nuclear factor κB in pancreatic tissues and acinar cells from Pdx1-Cre; HMGB1(flox/flox) and control mice. Exogenous genomic DNA and recombinant histone H3 proteins significantly induced release of HMGB1 from mouse macrophages; administration of antibodies against H3 to mice reduced serum levels of HMGB1 and increased survival after l-arginine injection. In 2 mouse models of acute pancreatitis, intracellular HMGB1 appeared to prevent nuclear catastrophe and release of inflammatory nucleosomes to block inflammation. These findings indicate a role for the innate immune response in tissue damage.

The influence of L-arginine on circadian rhythm and circadian period genes

To study the effect of L-arginine on circadian system, ICR mice and AR4-2J cells were treated with L-arginine. The locomotor activity of the mice intraperitoneally injected with a high dose of L-arginine was observed. We also detected effects of arginine on Per1 and Per2 gene expressions in AR4-2J cells. Double-plotted actograms were applied to analyze the locomotor activity of the mice, and reverse transcription-polymerase chain reaction (RT-PCR) was used in analysis of the expression of the major circadian genes Per1 and Per2 in AR4-2J cells. The results presented here indicate that the intraperitoneal injection of L-arginine would strongly disrupt the circadian rhythm of locomotor activity in mice. While the co-culture of L-arginine would also disturb the circadian rhythm of Per1 and Per2 genes which were induced by PMA in AR4-2J cells, the expression levels of both genes greatly decreased.

Evaluation of functional relationship between mouse hippocampal cholinergic and nitrergic systems in anxiogenic-like behavior

Although a body of evidence shows the crucial role of hippocampal nitrergic and cholinergic systems in the modulation of anxiety, little is known about their functional relationship with regard to anxiety. The present study investigated the relationship between intra-CA1 administration of a nicotinic acetylcholine receptor antagonist (mecamylamine) and a nitric oxide synthase inhibitor [Nω-nitro-L-arginine methyl ester (L-NAME)] or its precursor (L-arginine) in anxiety-related behaviors. Mice received bilateral intra-CA1 injections of either L-NAME or L-arginine in the presence of mecamylamine and were subsequently tested in the elevated plus maze. A dose of 0.5 μg/0.5 μl mecamylamine bilaterally administered into CA1 did not change the percentage of open arm time (%OAT) or the percentage of open arm entries (%OAE) in the elevated plus maze task and thus was considered as a subeffective dose. Intra-CA1 administration of either L-arginine (1 and 1.5 μg/0.5 μl, bilaterally) or L-NAME (at 60 ng/0.5 μl, bilaterally) decreased %OAT, which represents an anxiogenic-like effect. Coadministration of the subeffective dose of mecamylamine together with the lower doses of L-NAME (10 and 30 ng/0.5 μl, bilaterally) or L-arginine (0.5 μg/0.5 μl, bilaterally) led to a decrease in %OAT and %OAE. Thus, both L-NAME and L-arginine showed anxiogenic-like effects, but the effects of mecamylamine were too small to support a functional relationship between the hippocampal cholinergic and nitrergic systems.

Involvement of opioidergic and nitrergic systems in memory acquisition and exploratory behaviors in cholestatic mice

Bile duct ligation (BDL) is an animal model used in cholestatic disease research. Both opioidergic and nitrergic systems are known to be involved in cholestasis. The aim of this study was to investigate the possible interaction between these two systems in BDL-induced memory formation and exploratory behaviors in mice. Male mice weighing 25-30 g were divided into nonoperated controls, sham-operated, and BDL groups. One-trial step-down and hole-board paradigms were used to assess memory acquisition and exploratory behaviors, respectively. Cholestasis did not alter memory acquisition while increasing exploratory behaviors 7 days after BDL. A pretraining intraperitoneal injection of L-arginine (50, 100, and 200 mg/kg), L-NG-nitroarginine methyl ester (L-NAME) (5, 10, 20, and 40 mg/kg), or naloxone (0.125, 0.25, and 0.5 mg/kg) did not alter memory acquisition or exploratory behaviors, whereas morphine (5 and 7.5 mg/kg) decreased memory acquisition in sham-operated animals. Moreover, although injection of L-NAME and naloxone exerted no effect on memory acquisition in the 7 days post-BDL mice, L-arginine (100 and 200 mg/kg) and morphine (2.5, 5, and 7.5 mg/kg) injection reduced it. In contrast, L-NAME and naloxone, but not morphine or L-arginine, reduced the BDL-induced exploratory behaviors. Coadministration of subthreshold doses of morphine (1.25 mg/kg) and L-arginine (50 mg/kg) caused a memory deficit in 7 days post-BDL mice. However, the memory deficit induced by the effective doses of morphine (2.5 mg/kg) or L-arginine (200 mg/kg) in these mice was restored by the administration of either naloxone (0.5 mg/kg) or L-NAME (40 mg/kg). In addition, naloxone and L-NAME reduced the exploratory behaviors in L-arginine-pretreated mice but not in morphine-pretreated mice. We conclude that there appears to be a synergistic effect between opioidergic and nitrergic systems on memory acquisition and exploratory behaviors in cholestatic mice.

Effects of a mitochondria-targeted antioxidant (MitoQ) in murine experimental acute pancreatitis

Effects of a mitochondria-targeted antioxidant (MitoQ) in murine experimental acute pancreatitis

Effects of intraperitoneal injection of marrow mesenchymal stem cells on intestinal barrier in acute pancreatitis

To explore the effects of intraperitoneal injection of mesenchymal stem cells (MSC) on intestinal barrier in serve acute pancreatitis (SAP) rats. MSC were harvested and cultured from femurs of one male SD rat. And 30 female SD rats were divided into 3 groups: control group (n = 6), SAP group (n = 12) and MSC transplantation group (n = 12). SAP was induced by intraperitoneal injection of L-arginine (2 g/kg) twice in SAP and MSC groups. In MSC group, the third-generation MSC (5×10(6)) were injected intraperitoneally once daily for 3 days. All rats were sacrificed after 72 h. The histomorphologic alternations of small intestine were measured to evaluate the therapeutic effect of MSC transplantation. Reverse transcription (RT)-PCR were used to identify the expression of TNF-α mRNA and IL-1β mRNA in small intestine and pancreas. Small intestine and pancreatic samples were examined for the engraftment of donor-derived MSC by Y chromosome in situ hybridization analysis. Compared with SAP group, histomorphologic alternations of small intestine significantly lower in MSC group (4.17 ± 0.28 vs 3.00 ± 0.33, P < 0.05). The relative expression quantity of TNF-α mRNA and IL-1β mRNA in pancreas were both significant higher in SAP and MSC groups than those in control group (3.10 ± 0.73 and 1.92 ± 0.37 vs 0.51 ± 0.24, 4.60 ± 0.59 and 2.43 ± 0.39 vs 1.15 ± 0.18, all P < 0.05). Compared with SAP group, the expression quantity of TNF-α mRNA and IL-1β mRNA in pancreas significantly lower in MSC group (both P < 0.05). The relative expression quantity of TNF-α mRNA and IL-1β mRNA in small intestine were both significant higher in SAP and MSC groups than those in control group (2.73 ± 0.91 and 1.55 ± 0.48 vs 0.62 ± 0.20, 5.20 ± 0.94 and 2.10 ± 0.34 vs 0.99 ± 0.10, all P < 0.05). The expressions of TNF-α mRNA and IL-1β mRNA in MSC group were lower than those in SAP group (both P < 0.05). Sry gene was not detected in pancreatic and intestinal tissue of MSC-treated rats. Syngraft MSC exert protective effects on pancreas and small intestine injury. And their beneficial effects are primarily mediated via indirect actions but not by their differentiation into target cells.

Involvement of nitrergic system of CA1in harmane induced learning and memory deficits

Harmane (HA) is a β-carboline alkaloid derived from the Peganum harmala plant which induces memory impairment. On the other hand some of the investigations showed that β-carboline alkaloids inhibit NO production. Thus, the aim of the present study was to investigate the role of nitrergic system of the dorsal hippocampus (CA1) in HA-induced amnesia in male adult mice. One-trial step-down passive avoidance and hole-board apparatuses were used for the assessment of memory retrieval and exploratory behaviors respectively. The data indicated that pre-training intraperitoneal (i.p.) administration of HA (12 and 16mg/kg) decreased memory acquisition. Sole pre-training or pre-testing administration of L-NAME, a nitric oxide synthesis inhibitor (5, 10 and 15μg/mice, intra-CA1) did not alter memory retrieval. On the other hand, pre-training (10 and 15μg/mice, intra-CA1) and pre-testing (5, 10μg/mice, intra-CA1) injections of L-NAME restored HA-induced amnesia (16mg/kg, i.p.). Furthermore, neither sole pre-training nor pre-testing administration of l-arginine, a NO precursor (3, 6 and 9μg/mice, intra-CA1), altered memory retrieval. In addition, pre-testing (6 and 9μg/mice, intra-CA1), but not pre-training, injection of l-arginine increased HA-induced amnesia (16mg/kg, i.p.). These results suggest that the nitrergic system of CA1 is involved in HA-induced amnesia.

Netrin-1 protects against L-Arginine-induced acute pancreatitis in mice.

Acute pancreatitis (AP) is a common inflammatory disease mediated by damage to acinar cells and subsequent pancreatic inflammation with infiltration of leukocytes. The neuronal guidance protein, netrin-1, has been shown to control leukocyte trafficking and modulate inflammatory responses in several inflammation-based diseases. The present study was aimed toward investigating the effects of netrin-1 in an in vivo model of AP in mice. AP was induced in C57BL/6 mice by administration of two intraperitoneal injections of L-Arginine (4 g/kg). Mice were treated with recombinant mouse netrin-1 at a dose of 1 µg/mouse or vehicle (0.1% BSA) intravenously through the tail vein immediately after the second injection of L-Arginine, and every 24 h thereafter. Mice were sacrificed at several time intervals from 0 to 96 h after the induction of pancreatitis. Blood and tissue samples of pancreas and lung were collected and processed to determine the severity of pancreatitis biochemically and histologically. Immunohistochemical staining demonstrated that netrin-1 was mainly expressed in the islet cells of the normal pancreas and the AP model pancreas, and the pancreatic expression of netrin-1 was down-regulated at both the mRNA and protein levels during the course of AP. Exogenous netrin-1 administration significantly reduced plasma amylase levels, myeloperoxidase activity, pro-inflammatory cytokine production, and pancreas and lung tissue damages. Furthermore, netrin-1 administration did not cause significant inhibition of nuclear factor-kappa B activation in the pancreas of L-Arginine-induced AP. In conclusion, our novel data suggest that netrin-1 is capable of improving damage of pancreas and lung, and exerting anti-inflammatory effects in mice with severe acute pancreatitis. Thus, our results indicate that netrin-1 may constitute a novel target in the management of AP.

NO/cGMP production is important for the endogenous peripheral control of hyperalgesia during inflammation

Various studies have demonstrated the role of the nitric oxide (NO)/cGMP pathway in pain processing. Our group has also shown that this system participates in opioid-induced antinociception during peripheral inflammation. We have previously observed that inflammation mobilizes an endogenous opioidergic system to control hyperalgesia. Here, we investigated whether the NO/cGMP pathway underlies peripheral endogenous nociception control during inflammation. In this study, a pharmacological approach was used in conjunction with the rat paw pressure test to assess the effects of intraplantar NO synthase inhibitor NG-Nitro-l-arginine (NOArg), guanylyl cyclase inhibitor methylene blue (MB), phosphodiesterase-5 inhibitor zaprinast (ZP), or NO precursor l-arginine injection on carrageenan-induced hyperalgesia, which mimics an inflammatory process, or by prostaglandin E2 (PGE2), which directly sensitizes nociceptors. Intraplantar carrageenan (62.5, 125, 250 or 500μg) or PGE2 (0.1, 0.5 or 2μg) administration produced hyperalgesia, which manifested as a reduction in the rat nociceptive threshold to mechanical stimuli. NOArg (25, 50 or 100μg/paw) and MB (125, 250 or 500μg/paw) induced significant and dose-dependent reductions in the nociceptive threshold of carrageenan-induced (125μg/paw) hyperalgesia, but not PGE2-induced (0.5μg/paw) hyperalgesia. This was a local effect because it did not produce any modifications in the contralateral paw. Both Zaprinast (100, 200 or 400μg/paw) and l-arginine (100, 200 or 400μg/paw) significantly counteracted carrageenan-induced hyperalgesia (250μg/paw), yielding an increase in the nociceptive threshold compared with the control. Zaprinast (200μg/paw) or l-arginine (400μg/paw) did not produce an antinociceptive effect in the contralateral paw, indicating local action. In addition, at the same dose that was able to modify carrageenan-induced hyperalgesia, neither zaprinast nor l-arginine modified PGE2 (2μg) injection-induced hyperalgesia of the rat paw. Taken together, these results indicate that the l-arginine/NO/cGMP pathway functions as an endogenous modulator of peripheral inflammatory hyperalgesia.

The effects of nicotine on nitric oxide induced anxiogenic-like behaviors in the dorsal hippocampus

Nicotine, an active tobacco derived alkaloid, regulates the activity of the neuronal nitric oxide synthase (nNOS) as well as the release of nitric oxide (NO) in the nervous system. Nicotinic acetylcholine receptors and nNOS are abundantly co-expressed in the hippocampal neurons and are found to alter anxiety-like behaviors in rodents. Dorsal hippocampus may be a site for modulation of anxiety. Therefore, in this study, we investigated the possible interactions between nicotine and NO systems of the dorsal hippocampus and the resultant effect on anxiety-like behaviors. The elevated plus-maze (EPM) test has been used to test the anxiety. Intraperitoneal administration of nicotine (0.5mg/kg) decreased the open arm time percentage (%OAT) and open arm entries percentage (%OAE) but not the locomotor activity, indicating an anxiogenic-like response. Intra-CA1 injection of l-arginine (a NO precursor) or l-NAME (a NOS inhibitor) also caused anxiogenic-like effects. On the other hand, injection of the low dose nicotine before different doses of l-arginine or l-NAME blocked the anxiogenic-like response induced by the drugs. Our results suggested that, both NO and nicotinic cholinergic systems not only play a part in the modulation of the anxiety in mice dorsal hippocampus, but also demonstrate a complex interaction in this respect.

L-arginine in the ischemic phase protects against liver ischemia-reperfusion injury

To investigate the effects of intravenous L-arginine (LG) infusion on liver morphology, function and proinflammatory response of cytokines during the early phase of ischemia-reperfusion injury (IRI). Thirty rabbits were subjected to 60 minutes of hepatic ischemia and 120 minutes of reperfusion. An intravenous injection of saline or L-arginine was administered five minutes before the ischemia and five minutes before initiating the reperfusion and at the 55th and 115th minutes after the ischemia. Samples were collected for histological analysis of the liver and measurements of the serum AST, ALT and LDH and the cytokines IL-6 and TNF-alpha. It was observed a significant reduction of sinusoidal congestion, cytoplasmic vacuolization, infiltration of polymorphonuclear leukocyte, nuclear pyknosis, necrosis and steatosis in liver tissue, as well as AST, ALT and LDH after injection of LG in the ischemia (p <0.001). Lower levels of IL-6 and TNF-alpha were associated with LG infusion during ischemia. Higher levels these proteins were observed in animals receiving LG during reperfusion. L-arginine protects the liver against ischemia/reperfusion injury, mainly when is administered during the ischemic phase.

Efficacy of Different Nitric Oxide-Based Strategies in Preventing Experimental Cerebral Malaria by Plasmodium berghei ANKA

BackgroundLow nitric oxide (NO) bioavailability plays a role in the pathogenesis of human as well as of experimental cerebral malaria (ECM) caused by Plasmodium berghei ANKA (PbA). ECM is partially prevented by administration of the NO-donor dipropylenetriamine NONOate (DPTA-NO) at high concentration (1 mg/mouse), which also induces major side effects such as a sharp drop in blood pressure. We asked whether alternative strategies to improve NO bioavailability with minor side effects would also be effective in preventing ECM.Methodology/Principal FindingsMice were infected with PbA and prophylactically treated twice a day with bolus injections of L-arginine, Nω-hydroxy-nor-Arginine (nor-NOHA), tetrahydrobiopterin (BH4), separately or combined, sodium nitrite, sildenafil or sildenafil plus DPTA-NO starting on day 0 of infection. L-arginine and BH4 supplementation, with or without arginase inhibition by nor-NOHA, increased plasma nitrite levels but failed to protect against ECM development. Accordingly, prophylactic treatment with continuous delivery of L-arginine using osmotic pumps also did not improve survival. Similar outcomes were observed with sodium nitrite sildenafil (aimed at inhibiting phosphodiesterase-5) or with DPTA-NO. However, sildenafil (0.1 mg/mouse) in combination with a lower dose (0.1 mg/mouse) of DPTA-NO decreased ECM incidence (82±7.4% mortality in the saline group and 38±10.6% in the treated group; p<0.05). The combined prophylactic therapy did not aggravate anemia, had delayed effects in systolic, diastolic and mean arterial blood pressure and induced lower effects in pulse pressure when compared to DPTA-NO 1 mg/mouse.Conclusions/SignificanceThese data show that sildenafil lowers the amount of NO-donor needed to prevent ECM, resulting also in lesser side effects. Prophylactic L-arginine when given in bolus or continuous delivery and bolus BH4 supplementation, with or without arginase inhibition, were able to increase NO bioavailability in PbA-infected mice but failed to decrease ECM incidence in the doses and protocol used.

P038 Enhancement of cytoprotective mechanisms in case of blockage of COX-2 and introduction of L-arginine in experimental colitis

Background: Ulcerative colitis is one of the most common diseases of large intestine in human. It is indicative of increased expression of NO-synthases, predominantly of iNOS, and COX-2. L-arginine is the substrate for nitric oxide, and also for L-ornithine which is the precursor for the synthesis of collagen and polyamines. Purpose of the research was to study the effect of blockage of COX-2 activity with celecoxib combined with injection of L-arginine on the cytoprotective and metabolic processes under experimental ulcerative colitis. Methods: UC was modeled by 1ml of 4% acetic acid injected into rats’ large intestine. In the MMLI were determined: area and degree of destructive changes (histologically), parameters of NO-synthase system, content of TBA products, activity of SOD, catalase and concentration of L-arginine in the plasma of blood Results: The development of UC was accompanied by the destructive changes in the MMLI, enhanced processes of LP, increased content of NO, and considerable activation of iNOS. Histological investigations revealed disorders of mucous barrier of the MMLI, destructive changes of its components, presence of edema, deep erosions, infiltration of lymphocytes. Blocking COX-2 by celecoxib in case of experimental colitis caused reduction of the area of ulcerogenic lesions to 54.9±14.1mm2. Activity of general NOS reduced by 57% (p < 0.01), of iNOS by 58% (p < 0.01), NO content decreased by 29%, and content of TBA products decreased by 20%. Introduction of of L-arginine in case of experimental colitis caused reduction of the area of ulcerogenic lesions to 41.5±28.7mm2. Activity of general NOS reduced by 57% (p < 0.01), of iNOS by 54% (p < 0.01), NO content decreased by 29%, and content of MDA decreased by 20%. In combined action of introduction of L-arginine with the blockage of COX-2 by celecoxib, area of MMLI lesions considerably decreased and was 37.5±25.5mm2 (p < 0.01). Activity of general NOS diminished to the norm and activity of iNOS by 74% (p < 0.01). Content of MDA and catalase was at the level of intact animals. Conclusions: Combined blockage of COX-2 and introduction of L-arginine as determined in our investigations, strengthened their effects and tended to reduce iNOS activity, content of nitric oxide decreased, lipoperoxiation processes and activity of SOD in the MMLI diminished. This provides substantiation for new approaches to the treatment of ulcerative colitis and for development of the drugs capable of combined effect on iNOS and COX-2

P039 Serum MMPs 7 9 and their inhibitors during glucocorticoid and anti-TNF-alpha therapy in IBD

Background: Ulcerative colitis is one of the most common diseases of large intestine in human. It is indicative of increased expression of NO-synthases, predominantly of iNOS, and COX-2. L-arginine is the substrate for nitric oxide, and also for L-ornithine which is the precursor for the synthesis of collagen and polyamines. Purpose of the research was to study the effect of blockage of COX-2 activity with celecoxib combined with injection of L-arginine on the cytoprotective and metabolic processes under experimental ulcerative colitis. Methods: UC was modeled by 1ml of 4% acetic acid injected into rats’ large intestine. In the MMLI were determined: area and degree of destructive changes (histologically), parameters of NO-synthase system, content of TBA products, activity of SOD, catalase and concentration of L-arginine in the plasma of blood Results: The development of UC was accompanied by the destructive changes in the MMLI, enhanced processes of LP, increased content of NO, and considerable activation of iNOS. Histological investigations revealed disorders of mucous barrier of the MMLI, destructive changes of its components, presence of edema, deep erosions, infiltration of lymphocytes. Blocking COX-2 by celecoxib in case of experimental colitis caused reduction of the area of ulcerogenic lesions to 54.9±14.1mm2. Activity of general NOS reduced by 57% (p < 0.01), of iNOS by 58% (p < 0.01), NO content decreased by 29%, and content of TBA products decreased by 20%. Introduction of of L-arginine in case of experimental colitis caused reduction of the area of ulcerogenic lesions to 41.5±28.7mm2. Activity of general NOS reduced by 57% (p < 0.01), of iNOS by 54% (p < 0.01), NO content decreased by 29%, and content of MDA decreased by 20%. In combined action of introduction of L-arginine with the blockage of COX-2 by celecoxib, area of MMLI lesions considerably decreased and was 37.5±25.5mm2 (p < 0.01). Activity of general NOS diminished to the norm and activity of iNOS by 74% (p < 0.01). Content of MDA and catalase was at the level of intact animals. Conclusions: Combined blockage of COX-2 and introduction of L-arginine as determined in our investigations, strengthened their effects and tended to reduce iNOS activity, content of nitric oxide decreased, lipoperoxiation processes and activity of SOD in the MMLI diminished. This provides substantiation for new approaches to the treatment of ulcerative colitis and for development of the drugs capable of combined effect on iNOS and COX-2

Experimental research Strain of experimental animals and modulation of nitric oxide pathway: their influence on development of renal failure in an experimental model of hepatorenal syndrome

IntroductionPathomechanism of HRS is still poorly understood. The aim of our study was: (1) to test whether different strains of rats could develop typical HRS, and (2) to estimate the influence of activation and inhibition of nitric oxide for development of renal failure in course of HRS.Material and methodsFirst, we used 16 of Wistar and 16 of Sprague-Dawley rats in galactosamine model of HRS. Next, we used 48 of SDR rats, which received saline, N-nitro-L-arginine or L-arginine before and after liver damage. Twenty four hours urine and blood samples were collected 48 h after saline or Ga1N injection. Biochemical parameters were determined in serum or urine and then creatinine clearance and osmolality clearance were calculated. Liver and kidney tissues were collected for histopathological examination.ResultsLiver failure developed in all tested groups with significant increase of bilirubin (p < 0.001), ALT (p < 0.001) and ammonia (p < 0.001). Nevertheless we did not achieve any evidence of renal failure in Wistar, but we found typical renal failure in Sprague-Dawley group with significant decrease in creatinine clearance (p < 0.0012) and increase in concentration of creatinine and urea (p < 0.001) and (p < 0.001) respectively. Inhibition of NOS prevented development of renal failure with significant improvement of GFR both before (p < 0.0017) and after (p < 0.003) Ga1N injection. Injection of L-arginine after Ga1N injection did not caused significant improvement of GFR.ConclusionsOur study showed, that genetic factors might be responsible for development of renal failure in course of HRS and nitric oxide play important role in acute model of this syndrome.

Influence of age on pain sensitivity in response to paw pressure and formalin injection in rats: A role of nitric oxide

The effect of age on pain response to paw pressure and intraplantar formalin injection in rats is elucidated. Pain responses evoked by mechanical pressure on hind paw and intraplantar injection of formaldehyde (5%) into the hind paw were evaluated in groups of adult, young and aged male Sprague Dawley rats, after intraperitoneal (i.p.) or intracerebroventricular (i.c.v.) injection of L-arginine or NG-nitro-L-arginine methyl ester (L-NAME). Nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase staining was done in the two groups. The results show that pain response was reduced in the aged rats and enhanced pain response to paw pressure in aged rats only. L-arginine (i.c.v.) had no effect on pain response to paw pressure in the two groups but enhanced biphasic pain response to formalin. L-NAME (i.p. and i.c.v.) suppressed pain response to paw pressure in the two groups. L-NAME (i.c.v.) suppressed pain response to formalin during the acute phase and enhanced it during the late phase. NADPH-diaphorase activity was significantly greater in young rats. In conclusion, pain response is blunted in the aged rats. NO might be involved in mechanical nociception in aged rats and in formalin-induced nociception in both groups. NO blockade has an antinociceptive effect on pain response. Central NO has dual role in pain response evoked by formalin.

Nitric oxide in the hippocampal cortical area interacts with naloxone in inducing pain

Objective:Role of nitric oxide (NO) in reversing morphine anti-nociception has been shown. However, the interaction between NO and naloxone-induced pain in the hippocampus is unknown. The present study aimed to investigate the involvement of molecule NO in naloxone-induced pain and its possible interaction with naloxone into cortical area 1 (CA1) of hippocampus.Materials and Methods:Male Wistar rats (250–350 g), provided by Pasteur Institute of Iran, were housed two per cage with food and water ad libitum. The animals’ skulls were cannulated bilaterally at coordinates adjusted for CA1 of hippocampus (AP: -3.8; L: ±1.8– 2.2: V: 3) by using stereotaxic apparatus. Each experimental group included 6–8 rats. To induce inflammation pain, the rats received subcutaneous (s.c.) injections of formalin (50 μL at 2.5%) once prior to testing. To evaluate the nociceptive effect of naloxone, the main narcotic antagonist of morphine (0.1–0.4 mg/kg) was injected intraperitoneally (i.p.) 10 min before injection of formalin. Injections of L-arginine, a precursor of NO, and NG-Nitro-L-arginine Methyl Ester (L-NAME), an inhibitor of NO synthase (NOS), intra-CA1, were conducted orderly prior to the administration of naloxone. The pain induction was analyzed by analysis of variance (ANOVA).Results:Naloxone at the lower doses caused a significant (P<0.01) pain in the naloxone-treated animals. However, pre-administration (1–2 min) of L-arginine (0.04, 0.08, 0.15, 0.3, 1.0, and 3.0 μg/rat, intra-CA1) reversed the response to naloxone. But, the response to L-arginine was blocked by pre-microinjection (1–2 min) of L-NAME (0.15, 0.3, 1.0, and 3.0 μg/rat), whilst, L-arginine or L-NAME alone did not induce pain behavior.Conclusion:NO in the rat hippocampal CA1 area is involved in naloxone-induced nociception.

Nitric Oxide Inhibits Nociceptive Transmission by Differentially Regulating Glutamate and Glycine Release to Spinal Dorsal Horn Neurons

Nitric oxide (NO) is involved in many physiological functions, but its role in pain signaling remains uncertain. Surprisingly, little is known about how endogenous NO affects excitatory and inhibitory synaptic transmission at the spinal level. Here we determined how NO affects excitatory and inhibitory synaptic inputs to dorsal horn neurons using whole-cell recordings in rat spinal cord slices. The NO precursor L-arginine or the NO donor SNAP significantly increased the frequency of glycinergic spontaneous and miniature inhibitory postsynaptic currents (IPSCs) of lamina II neurons. However, neither L-arginine nor SNAP had any effect on GABAergic IPSCs. L-arginine and SNAP significantly reduced the amplitude of monosynaptic excitatory postsynaptic currents (EPSCs) evoked from the dorsal root with an increase in paired-pulse ratio. Inhibition of the soluble guanylyl cyclase abolished the effect of L-arginine on glycinergic IPSCs but not on evoked monosynaptic EPSCs. Also, inhibition of protein kinase G blocked the increase in glycinergic sIPSCs by the cGMP analog 8-bromo-cGMP. The inhibitory effects of L-arginine on evoked EPSCs and high voltage-activated Ca(2+) channels expressed in HEK293 cells and dorsal root ganglion neurons were abolished by blocking the S-nitrosylation reaction with N-ethylmaleimide. Intrathecal injection of L-arginine and SNAP significantly increased mechanical nociceptive thresholds. Our findings suggest that spinal endogenous NO enhances inhibitory glycinergic input to dorsal horn neurons through sGC-cGMP-protein kinase G. Furthermore, NO reduces glutamate release from primary afferent terminals through S-nitrosylation of voltage-activated Ca(2+) channels. Both of these actions probably contribute to inhibition of nociceptive transmission by NO at the spinal level.

Oxymatrine Ameliorates l-Arginine-Induced Acute Pancreatitis in Rats

The aim of this study was to determine whether oxymatrine has a protective effect against acute pancreatitis (AP) in a rat model of L-arginine-induced AP. AP was induced by two intraperitoneal injections of L-arginine (250 mg/100 g) at a 1-h interval. Oxymatrine (50 mg/kg) was administered every 6 h after the induction of AP. Oxymatrine significantly reduced the plasma amylase, D-lactic acid and tumor necrosis factor alpha concentration, serum diamine oxidase and lipase activity, and pancreatic myeloperoxidase activity, which were increased in AP rats (P < 0.05). In addition, the pancreatic CD45 expression and the expression of claudin-1, but not zonula occludens-1 (ZO-1) and occludin, in the intestinal tissues were significantly reduced after the induction of AP. However, oxymatrine increased the expression of claudin-1 and CD45, but did not alter the expression of ZO-1 and occludin. In conclusion, our results demonstrated that oxymatrine is potentially capably of protecting against L-arginine-induced AP and attenuating AP-associated intestinal barrier injury by up-regulation of claudin-1.