Application Of L-NAME Research Articles

Nitric oxide in the mechanisms of inhibitory effects of sodium butyrate on colon contractions in a mouse model of irritable bowel syndrome.

Irritable bowel syndrome (IBS) is a multifactorial disorder, with altered intestinal motility, visceral hypersensitivity, and dysfunction of the gut-brain axis. The aim of our study was to analyze the role of nitric oxide (NO) in the inhibitory effects of sodium butyrate on spontaneous contractility of proximal colon in a mouse model of IBS. IBS was induced by intracolonic infusion of acetic acid in the early postnatal period. Spontaneous contractions of proximal colon segments were studied in isometric conditions. The amplitude and frequency of colon contractions were higher in the IBS group. Sodium butyrate exerted inhibitory effects on colon contractions, which were less pronounced in IBS group. NO donors decreased spontaneous colon contractility and prevented the inhibitory effects of sodium butyrate in control and IBS groups. Nitric oxide synthase (NOS) inhibition by L-NAME increased contractile activity more effective in the control group and decreased the inhibitory action of sodium butyrate. In IBS group, preliminary application of L-NAME did not prevent sodium butyrate action. Our data indicate that butyrate exerts its inhibitory effects on colon motility at least partially through activation of NO synthesis. In the IBS model group, the NO-dependent mechanisms were less effective probably due to downregulation of NOS.

Influence of Remote Ischemic Conditioning and Nitrogen Monoxide on Angiogenesis and Microcirculation in a Mouse Ear Burn Model

IntroductionRemote ischemic conditioning (RIC) has been shown to improve tissue resilience against ischemia. The aim of this study was to investigate the influence of RIC and its key factor, nitrogen monoxide (NO), on angiogenesis in a burn wound. Materials and methodsA unilateral burn injury on the ear of hairless mice (n = 48) was generated via a hot air jet in a contact-free manner. In four randomized groups, including the control (NoRIC group), RIC alone (RIConly group), RIC plus NO donor (ISDN group), and RIC plus NO synthase inhibitor (L-NAME group), the impact on angiogenesis, vessel diameter, blood flow, edema formation, and leukocyte-endothelial-cell interaction was evaluated over a 12-d period using intravital fluorescence microscopy. ResultsTissue perfusion was significantly improved by RIC (Day 3: ISDN group showed 182% and RIConly group showed 200% of baseline [BL], P < 0.001), while angiogenesis was not improved by RIC (nonperfused area on Day 12: mean 52% of BL in all groups, P >0.05). The application of ISDN did not further enhance the positive effect of RIC, whereas the application of L-NAME neutralized the effect of RIC. The most pronounced edema formation was observed in the RIConly group (mean 145% of BL, P ≤0.001), while the NoRIC group showed the least edema formation (Day 12: 117% of BL). ConclusionsRIC led to increased tissue perfusion, which did not result in improved angiogenesis, which may have been due to increased edema formation after RIC performance. The results of the present study do not support the establishment of a causal therapy strategy for burn wounds, including RIC.

Transdermal iontophoretic application of l-NAME is available in sweating research induced by heat stress in young healthy adults

Iontophoretic transdermal administration of NG-nitro-l-arginine methyl ester hydrochloride [l-NAME, a nitric oxide synthase (NOS) inhibitor] has been used as a non-invasive evaluation of NOS-dependent mechanisms in human skin. However, the availability has yet to be investigated in sweating research. Prior observations using invasive techniques (e.g., intradermal microdialysis technique) to administer l-NAME have implicated that NOS reduces sweating induced by heat stress but rarely influences the response induced by the administration of cholinergic muscarinic receptor agonists. Therefore, we investigated whether the transdermal iontophoretic administration of l-NAME modulates sweating similar to those prior observations. Twenty young healthy adults (10 males, 10 females) participated in two experimental protocols on separate days. Before each protocol, saline (control) and 1% l-NAME were bilaterally administered to the forearm skin via transdermal iontophoresis. In protocol 1, 0.001% and 1% pilocarpine were iontophoretically administered at l-NAME-treated and untreated sites. In protocol 2, passive heating was applied by immersing the lower limbs in hot water (43 °C) until the rectal temperature increased by 0.8 °C above baseline. The sweat rate was continuously measured throughout both protocols. Pilocarpine-induced sweat rate was not significantly different between the control and l-NAME-treated sites in both pilocarpine concentrations (P ≥ 0.316 for the treatment effect and interaction of treatment and pilocarpine concentration). The sweat rate during passive heating was attenuated at the l-NAME-treated site relative to the control (treatment effect, P = 0.020). Notably, these observations are consistent with prior sweating studies administrating l-NAME into human skin using intradermal microdialysis techniques. Based on the similarity of our results with already known observations, we conclude that transdermal iontophoresis of l-NAME is a valid non-invasive technique for the investigation of the mechanisms of sweating related to NOS during heat stress.

Nitric Oxide Induced by Ammonium/Nitrate Ratio Ameliorates Low-Light Stress in Brassica pekinesis: Regulation of Photosynthesis and Root Architecture.

Low-light intensity affects plant growth and development and, finally, causes a decrease in yield and quality. There is a need for improved cropping strategies to solve the problem. We previously demonstrated that moderate ammonium:nitrate ratio (NH4+:NO3-) mitigated the adverse effect caused by low-light stress, although the mechanism behind this alleviation is unclear. The hypothesis that the synthesis of nitric oxide (NO) induced by moderate NH4+:NO3- (10:90) involved in regulating photosynthesis and root architecture of Brassica pekinesis subjected to low-light intensity was proposed. To prove the hypothesis, a number of hydroponic experiments were conducted. The results showed that in plants exposed to low-light intensity, the exogenous donors NO (SNP) and NH4+:NO3- (N, 10:90) treatments significantly increased leaf area, growth range, and root fresh weight compared with nitrate treatment. However, the application of hemoglobin (Hb, NO scavenger), N-nitro-l-arginine methyl ester (L-NAME, NOS inhibitor), and sodium azide (NaN3, NR inhibitor) in N solution remarkably decreased the leaf area, canopy spread, the biomass of shoot and root, the surface area, and volume and tips of the root. The application of N solution and exogenous SNP significantly enhanced Pn (Net photosynthetic rate) and rETR (relative electron transport rates) compared with solo nitrate. While all these effects of N and SNP on photosynthesis, such as Pn, Fv/Fm (maximum quantum yield of PSII), Y(II) (actual photosynthetic efficiency), qP (photochemical quenching), and rETR were reversed when the application of Hb, L-NAME, and NaN3 in N solution. The results also showed that the N and SNP treatments were more conducive to maintaining cell morphology, chloroplast structure, and a higher degree of grana stacking of low-light treated plants. Moreover, the application of N significantly increased the NOS and NR activities, and the NO levels in the leaves and roots of mini Chinese cabbage seedlings treated with N were significantly higher than those in nitrate-treated plants. In conclusion, the results of this study showed that NO synthesis induced by the appropriate ammonia-nitrate ratio (NH4+:NO3- = 10:90) was involved in the regulation of photosynthesis and root structure of Brassica pekinesis under low-light stress, effectively alleviating low-light stress and contributing to the growth of mini Chinese cabbage under low-light stress.

Nitric oxide synthase (NOS)-dependent nitric oxide generation is involved in putrescine-induced chilling tolerance of Anthurium andraeanum

Previous studies have demonstrated the positive effects of exogenous putrescine (Put) on the chilling tolerance of Anthurium andraeanum. In the present study, anthurium seedlings were pretreated with Put and Put in combination with NO scavenger cPTIO, nitric oxide synthase (NOS) inhibitor L-NAME, or nitrate reductase (NR) inhibitor Na2WO4 solutions for four weeks and then subjected to chilling stress (6 ºC) for three days, followed by recovery (25 ºC) for one day. NO fluorescence, endogenous NO content, and activities of NOS, NR, and copper amine oxidase (CuAO), as well as physiological indexes, were assessed during the experiment. The results were further confirmed at the transcriptional level via transcriptome sequencing and qRT-PCR. Results indicated that the application of Put prompted the accumulation of endogenous NO, and the NOS-dependent pathway probably played a vital role in the endogenous NO generation. The enhancement of chilling tolerance induced by exogenous Put could be notably reversed by the combined application of L-NAME. Moreover, transcriptome sequencing and gene expression analyses confirmed the contribution of the NOS-dependent pathway to the accumulation of endogenous NO and the effects of endogenous NO on the mediation of chilling tolerance. In summary, the NOS-dependent pathway was the primary source of endogenous NO in the Put-pretreated anthurium. Furthermore, endogenous NO was involved in the chilling response of anthurium, playing a synergistic role in the mediation of chilling tolerance with exogenous Put.

Distinct effects of orexin A on spontaneous and evoked synaptic currents in the rat nucleus tractus solitarius

Orexins are produced in hypothalamic areas and orexin-containing neurons are distributed in widespread areas of the central nervous system. Orexins regulate several physiological functions such as arousal, food intake and autonomic control. The presence of orexin-containing neuron terminals and orexin receptors has been confirmed in the nucleus tractus solitarius (NTS), which receives primary afferent fibers from peripheral organs including baroreceptors. However, the neuronal effects of orexin-1 receptor (OX1R) activation were not examined. Here, we aimed to determine the effects of OX1R activation on excitatory synaptic transmission. OX1R activation increased the frequency of spontaneous excitatory synaptic currents (sEPSCs). This effect was blocked by the prior application of L-NAME. In contrast, the amplitude of evoked excitatory postsynaptic currents (eEPSCs) was suppressed by OX1R activation, and this effect was prevented by a cannabinoid receptor 1 blocker, AM251, but not by the pretreatment with L-NAME. Altogether, these results suggest that OX1R activation increases sEPSCs frequency by stimulating NO production, whereas it inhibits eEPSCs by releasing endocannabinoids in the NTS. Thus, OX1R activation had distinct effects on spontaneous and evoked excitatory synaptic transmissions in the NTS.

The positive effects of exogenous sodium nitroprusside on the plant growth, photosystem II efficiency and Calvin cycle of tomato seedlings under salt stress

Although some studies have shown that exogenous sodium nitroprusside (SNP, an NO donor) participates in the tolerance of plants to stress, the protective role of the level of SNP-mediated endogenous nitrous oxide (NO) on the salt-induced inhibition of photosynthesis in salt-stressed tomato seedlings merits understanding. Thus, in this study, with the exception of tomato seedlings treated with 0 or 100 mmol· L−1 NaCl alone or together with 0.1 mmol· L−1 SNP, 100 μmol· L−1 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (PTIO, an NO scavenger) and 100 μmol· L−1 N-nitro-L-arginine methyl ester (L-NAME, an inhibitor of NO synthase) was applied jointly to saline-stressed tomato seedlings by a foliage spray of 0.1 mmol· L−1 SNP, respectively. The application of SNP counteracted the saline-induced growth inhibition and the reduction in net photosynthetic rate of tomato seedlings by enhancing their stomatal conductance, intercellular CO2 concentration, apparent photosynthetic quantum efficiency, PSII maximum photochemical efficiency, actual photochemical efficiency of PSII, non-photochemical quenching coefficient, and a decrease in the dark respiration rate, which is coupled with an increase in the endogenous levels of NO. In addition, treatment with SNP increases the maximum carboxylation rate of ribulose-1,5-bisphosphate carboxylase (Rubisco), the maximum regeneration rate of ribulose-1,5-bisphosphate (RuBP) and the upregulated total and initial activity of Rubisco; the activity in Rubisco activase and fructose-1,6-bisphosphate phosphatase; and the transcriptions of genes that encode Rubisco activase, the Rubisco large subunit, and the Rubisco small subunit, and sedoheptulose-1,7-bisphosphatase, 3-phosphogiyceric acid kinase, transketolase and ribulose-5-phosphate kinase, which are the key enzymes involved in the Calvin-Benson cycle. However, the positive effect of SNP was impaired by the application of PTIO and L-NAME to eliminate the generation of endogenous NO, respectively. Our findings conclusively revealed that SNP triggers the upregulation of endogenous NO signal, which, in turn, conferred tolerance to salinity in tomato seedlings and positively regulated photosynthesis by overcoming stomatal limitations, enhancing the efficiency of photosystem II and maintaining the activation states of the Calvin-Benson cycle through the regulation of gene expressions and activities of key CO2 assimilation enzymes.

Caveolin-1 Involves in the Rearrangement of F-actin through NF-κB Pathway in Endothelial Cells Induced by Lipopolysaccharide

Objective: To explore the role and possible mechanism of Caveolin-1 in the rearrangement of F-actin in endothelial cell induced by lipopolysaccharides (LPS).Methods: EA.hy926 cells were divided into 4 Groups: Group C (control Group), Group L (LPS 10μg/mL Group), Group S (cav-1-siRNA548 + LPS 10μg/mL Group) and Group N (cav-1-siRNA548 and L-NAME 10μM +LPS 10μg/mL Group). Real-Time PCR analysis was performed to measure the mRNA expression. Immunofluorescence staining was carried out to observe the changes of F-actin in different group.Results: We found that treatment of siRNA548 reduced the mRNA expression of caveolin-1; the mRNA expression of peNOS-s1177 in Group S significantly increased when compared with Group C and Group L, respectively. Six hours after LPS treatment, the mRNA expression of peNOS-s1177 is higher than any other time point in Group L, while the mRNA expression of peNOS-s1177 is highest after LPS treatment for 3 hours in Group S. We also found that the mRNA expression of NF-κB in Group L, S and N remarkably upregulated when compared with Group C, respectively. Moreover, the mRNA expression of NF-κB in Group L and N significantly upregulated compared with Group S. The immunofluorescence staining showed that EA.hy926 cell strain manifested F-actin depolymerization and reconstitution after LPS treatment for 6 hours, and the phenomenon had a time-dependent manner. Application of cav-1-siRNA548 could delay the depolymerization and reconstruction of F-actin, and with a lesser degree, when application of L-NAME could reverse the response.Conclusions: These data indicate that Caveolin-1 may have an important role in the modulation of LPS- induced endothelial F-actin depolymerization. And Caveolin-1 may modulate F-actin depolymerization and reconstitution through eNOS-NF-κB pathway.

Oxidative stress in the skin: Impact and related protection.

Skin, our first interface to the external environment, is subjected to oxidative stress caused by a variety of factors such as solar ultraviolet, infrared and visible light, environmental pollution, including ozone and particulate matters, and psychological stress. Excessive reactive species, including reactive oxygen species and reactive nitrogen species, exacerbate skin pigmentation and aging, which further lead to skin tone unevenness, pigmentary disorder, skin roughness and wrinkles. Besides these, skin microbiota are also a very important factor ensuring the proper functions of skin. While environmental factors such as UV and pollutants impact skin microbiota compositions, skin dysbiosis results in various skin conditions. In this review, we summarize the generation of oxidative stress from exogenous and endogenous sources. We further introduce current knowledge on the possible roles of oxidative stress in skin pigmentation and aging, specifically with emphasis on oxidative stress and skin pigmentation. Meanwhile, we summarize the science and rationale of using three well-known antioxidants, namely vitamin C, resveratrol and ferulic acid, in the treatment of hyperpigmentation. Finally, we discuss the strategy for preventing oxidative stress-induced skin pigmentation and aging.

BPC 157 as a Therapy for Retinal Ischemia Induced by Retrobulbar Application of L-NAME in Rats.

Providing NO-system importance, we suggest that one single application of the NOS-blocker L-NAME may induce retinal ischemia in rats, and that the stable pentadecapeptide BPC 157 may be the therapy, since it may interact with the NO-system and may counteract various adverse effects of L-NAME application. A rat retinal ischemia study was conducted throughout 4 weeks, including fundoscopy, behavior presentation, tonometry, and histology assessment. Retrobulbar L-NAME application (5 mg/kg; 0.5 mg/0.1 ml saline/each eye) in rats immediately produced moderate generalized irregularity in the diameter of blood vessels with moderate atrophy of the optic disc and faint presentation of the choroidal blood vessels, and these lesions rapidly progressed to the severe stage. The specific L-NAME–induced vascular failure points to normal intraocular pressure (except to very transitory increase upon drug retrobulbar administration). When BPC 157 (10 μg; 10 ng/kg, as retrobulbar application, 1 μg; 1 ng/0.1 ml saline/each eye) is given at either 20 min after L-NAME or, lately, at 48 h after L-NAME, the regular retrobulbar L-NAME injection findings disappear. Instead, fundoscopy demonstrated only discrete generalized vessel caliber irregularity with mild atrophy of the optic disc, and then, quite rapidly, normal eye background and choroidal blood vessels, which remain in all of the subsequent periods. Also, histology assessment at 1, 2, and 4 weeks shows that BPC 157 counteracted the damaged inner plexiform layer and inner nuclear layer, and revealed normal retinal thickness. The poor behavioral presentation was also rescued. Thus, while further studies will be done, BPC 157 counteracted L-NAME–induced rat retinal ischemia.

Inhibition of neuroinflammatory nitric oxide signaling suppresses glycation and prevents neuronal dysfunction in mouse prion disease

Several neurodegenerative diseases associated with protein misfolding (Alzheimer's and Parkinson's disease) exhibit oxidative and nitrergic stress following initiation of neuroinflammatory pathways. Associated nitric oxide (NO)-mediated posttranslational modifications impact upon protein functions that can exacerbate pathology. Nonenzymatic and irreversible glycation signaling has been implicated as an underlying pathway that promotes protein misfolding, but the direct interactions between both pathways are poorly understood. Here we investigated the therapeutic potential of pharmacologically suppressing neuroinflammatory NO signaling during early disease progression of prion-infected mice. Mice were injected daily with an NO synthase (NOS) inhibitor at early disease stages, hippocampal gene and protein expression levels of oxidative and nitrergic stress markers were analyzed, and electrophysiological characterization of pyramidal CA1 neurons was performed. Increased neuroinflammatory signaling was observed in mice between 6 and 10 wk postinoculation (w.p.i.) with scrapie prion protein. Their hippocampi were characterized by enhanced nitrergic stress associated with a decline in neuronal function by 9 w.p.i. Daily in vivo administration of the NOS inhibitor L-NAME between 6 and 9 w.p.i. at 20 mg/kg prevented the functional degeneration of hippocampal neurons in prion-diseased mice. We further found that this intervention in diseased mice reduced 3-nitrotyrosination of triose-phosphate isomerase, an enzyme involved in the formation of disease-associated glycation. Furthermore, L-NAME application led to a reduced expression of the receptor for advanced glycation end-products and the diminished accumulation of hippocampal prion misfolding. Our data suggest that suppressing neuroinflammatory NO signaling slows functional neurodegeneration and reduces nitrergic and glycation-associated cellular stress.

The Effect of Mesenchymal Stem Cells on the Reactivity of Smooth Muscle Cells of Pial Arteries in Nephrectomized Rats

The aim of this work is to study the deterioration of the contractile function of smooth muscle cells in pial arteries after subtotal nephrectomy, as well as the possibility of restoring this function by transplantation of human mesenchymal stem cells. A self-designed visualizing device for studying microcirculation (160× magnification) was used to study the reactivity of sensorimotor cortex pial vessels to hydrogen sulfide (H2S), L-NAME nonselective nitric oxide synthase inhibitor, and the combined application of H2S and L-NAME in sham-operated rats, nephrectomized rats, and nephrectomized animals that received human mesenchymal stem cells (hMSCs) intravenously. In parallel, the myogenic tone of cerebral vessels was assessed with a LAKK-M laser doppler. The results showed that, 4 months after nephrectomy in rats, the reactivity of pial arteries to H2S and L-NAME seriously deteriorated (the number of dilated arteries decreased by 1.2–1.7 times, and the extent of constriction was 6–17% smaller). The myogenic tone of cerebral vessels after nephrectomy was 1.5 times higher than in control rats. Intravenous hMSC transplantation allowed the myogenic tone and the reactivity of smooth muscle cells (SMCs) to be maintained at the level of control animals.

The Role of Hydrogen Sulfide in the Dilatation of Mesenteric Lymphatic Vessels in Bulls.

We studied the effects and mechanisms of action of NaHS, an H2S donor, on bovine isolated mesenteric efferent lymphatic vessels pre-contracted with phenylephrine. NaHS induced concentration-dependent relaxation of lymphatic vessels. Removal of the endothelium reduced, but did not completely abolish the relaxing effect of NaHS. Application of NO synthase inhibitor L-NAME, soluble guanylyl cyclase inhibitor ODQ, blocker ATP-sensitive K+ channels glibenclamide, and a combination of blockers of Ca-activated K+ channels of small and intermediate conductance charybdotoxin and apamin attenuated relaxation of lymphatic vessels. Thus, H2S produces a pleiotropic effect on lymphatic vessels; vasorelaxant effect is achieved by several parallel mechanisms. H2S induces relaxation of lymphatic vessels and modulates the rate of lymph transport, thereby affecting the development of immune processes in the body.

H2S induces NO in the regulation of AsA-GSH cycle in wheat seedlings by water stress.

In current study, we investigated the relationship between hydrogen sulfide (H2S) and nitric oxide (NO) in the regulation of ascorbate-glutathione (AsA-GSH) cycle in wheat seedlings by water stress. Findings showed that water stress significantly stimulated the production of H2S and NO, the transcript levels and activities of enzymes in AsA-GSH cycle, as well as malondialdehyde (MDA) production and electrolyte leakage, but significantly decreased AsA/DHA and GSH/GSSG. Meanwhile, water stress significantly decreased plant height and dry biomass. Except MDA and electrolyte leakage, above changes induced by water stress were reversed by H2S synthesis inhibitor aminooxyacetic acid (AOA) and NO synthesis inhibitor NG-nitro-L-arginine methyl ester (L-NAME). However, AOA and L-NAME significantly enhanced MDA production and electrolyte leakage, which further decreased plant height and dry biomass of wheat seedlings under water stress. Application of exogenous H2S donor sodium hydrosulfide (NaHS) to AOA-treated plants and application of exogenous NO donor sodium nitroprusside (SNP) to L-NAME-treated plants reversed above effects of AOA and L-NAME, respectively. Application of L-NAME plus water stress significantly decreased NO production induced by water stress. However, application of L-NAME plus water stress had no obvious influence on H2S production induced by water stress, while application of AOA plus water stress significantly reduced the production of H2S and NO induced by water stress. Current findings suggested that H2S acted upstream of NO in the regulation of AsA-GSH cycle in wheat seedlings by water stress.

Nitrate reductase rather than nitric oxide synthase activity is involved in 24-epibrassinolide-induced nitric oxide synthesis to improve tolerance to iron deficiency in strawberry (Fragaria × annassa) by up-regulating the ascorbate-glutathione cycle

Involvement of nitrate reductase (NR) and nitric oxide synthase (NOS)-like enzyme in 24-epibrassinolide (EB)-triggered nitric oxide (NO) synthesis to improve iron deficiency (ID) tolerance in strawberry plants was studied. EB was sprayed to strawberry plants every two days for two weeks. Then, the EB-treated plants were pre-treated with inhibitors of NR, tungstate, or NOS, L-NAME for 3 h. During the first three weeks, Fe was supplied as 100 μM EDTA-Fe or FeSO4 to Fe-sufficient or Fe-deficient plants, respectively. Thereafter, plants were subjected for further three weeks to control (100 μM EDTA-Fe) and Fe deficiency (ID; without Fe). ID reduced biomass, chlorophyll, and chlorophyll fluorescence, while increased oxidative stress parameters, ascorbate (AsA), glutathione (GSH), endogenous NO, and the activities of NR, NOS, and antioxidant enzymes. Pre-treatments with EB and EB + SNP improved ID tolerance of strawberry by improving leaf Fe2+, plant growth, and antioxidant enzyme activities, and causing a further elevation in AsA, GSH, NO, NR and NOS. L-NAME application reversed NOS activity, but it did not eliminate NO, however, tungstate application reversed both NR activity and NO synthesis in plants exposed to ID + EB, suggesting that NR is the main contributor of EB-induced NO synthesis to improve ID tolerance in strawberry plants.

Exogenous hydrogen sulfide alleviates surgery-induced neuroinflammatory cognitive impairment in adult mice by inhibiting NO signaling

BackgroundTo investigate the effect and mechanisms of exogenous hydrogen sulfide in surgery-induced neuroinflammatory cognitive dysfunction.MethodsC57BL/6 J male mice (n = 140) were used and randomly divided into seven groups: the sham group, surgery group, GYY4137 group, L-NAME group, surgery+GYY4137 group, surgery +L-NAME group, and surgery+GYY4137 + L-NAME group. After the interventions, open field tests (OFT) and the Morris water maze (MWM) test were conducted to evaluate learning and memory abilities in the mice. ELISAs, nitrate reductase assays, and Western blots (WB) were conducted to evaluate interleukin-1 beta (IL-1β), tumor necrosis factor-alpha (TNF-α), nitric oxide (NO), inducible nitric oxide synthase (iNOS), malondialdehyde (MDA), and antioxidant enzyme superoxide dismutase (SOD) levels. Furthermore, the expression level of microglial marker ionized calcium binding adaptor molecule 1 (IBA) in the hippocampal CA1 and CA3 areas was detected by an immunohistochemical (IHC) assay and apoptotic cells were observed using terminal deoxynucleotidyl transferase dUTP end-labeling (TUNEL) staining kits.ResultsWe found that surgery induced neuroinflammatory cognitive dysfunction, oxidative stress, microglial activation, and cell apoptosis in the hippocampus. Moreover, following surgery, NO and iNOS levels were elevated in the hippocampus. Notably, all the effects caused by surgery were reversed by the H2S donor GYY4137 or the iNOS inhibitor N(gamma)-nitro-L-arginine methyl ester (L-NAME). However, the combined application of GYY4137 and L-NAME was not superior to treatment with either agent alone and the effect of GYY4137 was similar to that of L-NAME.ConclusionThe long-acting hydrogen sulfide donor GYY4137 had an ability to reversed the cognitive deficits and inflammation caused by carotid artery exposure surgery. This implies that NO signaling pathways might participate in this process. These results indicate that exogenous H2S may be a promising therapy for POCD.

Antihypertensive effect of methanol leaf extract of Azadirachta indica is mediated through suppression of renal caspase 3 expressions on Nω-Nitro-l-arginine methyl ester induced hypertension

Background: Azadirachta indica (AI) Adr Juss (Meliaceae), known as neem, has been used traditionally for the treatment of various disease conditions including obesity and hypertension. Objective: The antihypertensive effect and mechanism of action of modulatory effect of AI were investigated after the induction of hypertension using Nω-nitro-L-arginine methyl ester (L-NAME). Materials and Methods: Five groups of ten rats divided as follows: Control; L-NAME (40 mg/kg); L-NAME + 100 mg/kg AI; L-NAME and 200 mg/kg ?AI; and L-NAME and Enalapril (25 mg/kg) were used. Results: following the application of L-NAME, hypertension (elevated systolic, diastolic, mean arterial blood pressures) and increased levels of oxidative stress markers were observed in rats. Immunohistochemistry showed increased caspase-3 expressions in hypertensive rats compared to normotensive rats. Conversely, AI treatment resulted in restoration of physiological antioxidant status and normotension, comparable to the standard antihypertensive agent enalapril. Conclusion: AI leaf is a good candidate for the management of high blood pressure.

Pentadecapeptide BPC 157 shortens duration of tetracaine- and oxybuprocaine-induced corneal anesthesia in rats.

SUMMARYWe focused on the relationship of 0.5% tetracaine- and 0.4% oxybuprocaine-induced corneal anesthesia in rats, and pentadecapeptide BPC 157 (0.4 µg/eye), along with nitric oxide synthase (NOS) inhibitor N(gamma)-nitro-L-arginine methyl ester (L-NAME) (0.1 mg/eye) and/or NOS substrate L-arginine (2 mg/eye), applied in the form of eye drops. We assessed corneal sensitivity recovery (Cochet-Bonnet esthesiometer), corneal lesion elimination (staining with 10% fluorescein) and decrease in tear volume (Schirmer test). BPC 157 administration had a full counteracting effect. Recovery also occurred in the presence of NOS blockade and NOS substrate application. L-arginine eventually shortened duration of corneal insensitivity and exerted corneal lesion counteraction (and counteraction of tetracaine-induced decrease of tear volume) only in earlier but not in later period. L-NAME application led to longer duration of corneal insensitivity, increase in corneal lesions and decrease in tear volume. When L-NAME and L-arginine were applied together, they antagonized each other’s effect. These distinctions may indicate particular NOS involvement (corneal insensitivity vs. corneal lesion along with tear production), distinctively affected by the administration of NO agents. However, additional BPC 157 co-administration would re-establish counteraction over topical ophthalmic anesthetic-induced effect, be it in its early or late course. We suggest BPC 157 as an antidote to topical ophthalmic anesthetics.

Age-Dependent Effects of NO on Rhythmic Activity of Postganglionic Sympathetic Fibers.

We studied the effects of exogenous NO donor (sodium nitroprusside) and NO synthesis blocker (100 μM L-NAME) on baseline electrical activity of postganglionic fibers in the sympathetic superior cervical in rats during postnatal ontogeny. Starting the age of 20 days, sodium nitroprusside increased the mean discharge amplitude and the spectrum power in the respiratory (0.7-1.5 Hz) and cardiac (4-7 Hz) frequency bands. In contrast, application of L-NAME for 1 h decreased the spectrum power in these bands. Both agents produced no significant effect on the rhythmic sympathetic discharges at frequencies >8 Hz.

Acute spatial spread of NO-mediated potentiation during hindpaw ischaemia in mice.

Key points Neuropathic pain spreads spatially beyond the injured sites, and the mechanism underlying the spread has been attributed to inflammation occurring in the spinal cord.However, the spatial spread of spinal/cortical potentiation induced by conduction block of the peripheral nerves can be observed prior to inflammation.In the present study, we found that spreading potentiation and hypersensitivity acutely induced by unilateral hindpaw ischaemia are nitric oxide (NO)‐dependent and that NO is produced by ischaemia and quickly diffuses within the spinal cord.We also found that NO production induced by ischaemia is not observed in the presence of an antagonist for group II metabotropic glutamate receptors (mGluRs) and that neuronal NO synthase‐positive dorsal horn neurons express group II mGluRs.These results suggest strongly that NO‐mediated spreading potentiation in the spinal cord is one of the trigger mechanisms for neuropathic pain. Cortical/spinal responses to hindpaw stimulation are bilaterally potentiated by unilateral hindpaw ischaemia in mice. We tested the hypothesis that hindpaw ischaemia produces nitric oxide (NO), which diffuses in the spinal cord to induce spatially spreading potentiation. Using flavoprotein fluorescence imaging, we confirmed that the spreading potentiation in hindpaw responses was induced during ischaemia in the non‐stimulated hindpaw. This spreading potentiation was blocked by spinal application of l‐NAME, an inhibitor of NO synthase (NOS). Furthermore, no spreading potentiation was observed in neural NOS (nNOS) knockout mice. Spinal application of an NO donor was enough to induce cortical potentiation and mechanical hypersensitivity. The spatial distribution of NO during unilateral hindpaw ischaemia was visualized using 4‐amino‐5‐methylamino‐2′,7′‐difluorofluorescein (DAF‐FM). An increase in fluorescence derived from the complex of DAF‐FM with NO was observed on the ischaemic side of the spinal cord. A similar but smaller increase was also observed on the contralateral side. Somatosensory potentiation after hindpaw ischaemia is known to be inhibited by spinal application of LY354740, an agonist of group II metabotropic glutamate receptors (mGluRs). We confirmed that the spinal DAF‐FM fluorescence increases during hindpaw ischaemia were not observed in the presence of LY354740. We also confirmed that approximately half of the nNOS‐positive neurons in the superficial laminae of the dorsal horn expressed mGluR2 mRNA. These results suggest that disinhibition of mGluR2 produces NO which in turn induces a spreading potentiation in a wide area of the spinal cord. Such spreading, along with the consequent non‐specific potentiation in the spinal cord, may trigger neuropathic pain.