Enhanced release of nitric oxide causes increased cytotoxicity of S-nitroso-N-acetyl-dl-penicillamine and sodium nitroprusside under hypoxic conditions

S-Nitroso-N-acetyl-dl-penicillamine (SNAP) and sodium nitroprusside (SNP), both of which are known to release nitric oxide (•NO), exhibited cytotoxicity against cultivated endothelial cells. Under hypoxic conditions 5 mM SNAP and 20 mM SNP induced a loss in cell viability of about 90% and 80% respectively, after an 8 h incubation. Under normoxic conditions, cell death was only 45% and 42% respectively within the same time period. Concentrations of •NO liberated from SNAP and SNP were measured by the oxyhaemoglobin method and by two of the recently developed nitric oxide cheletropic traps (NOCTs). The •NO concentrations from SNAP and SNP increased from 74 µM and 28 µM to 136 µM and 66 µM respectively within 15 min of hypoxic incubation, and then decreased to 36 µM and 28 µM. In the respective normoxic incubations the •NO levels from SNAP and SNP remained in the region of about 30 µM and 20 µM respectively. In contrast, spermine/NO adduct (spermineNONOate) was shown to be more toxic under normoxic than under hypoxic conditions. Under either of these conditions, the concentration of •NO liberated from 2 mM spermineNONOate was about 20 µM. The results demonstrate that the cytotoxicity of SNAP and SNP, but not of spermineNONOate, is significantly enhanced under hypoxic compared with normoxic incubations. Studies on the •NO-releasing behaviour of these compounds indicate that the increased toxicity of SNAP and SNP under hypoxic conditions is related to the influence of O2 on the chemical processes by which •NO is produced from the precursors, rather than to an increased sensitivity of the hypoxic cells towards •NO.

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Effect of nitric oxide on Sertoli cell microtubule of piglets

Chinese Journal of Agricultural Biotechnology ◽

10.1017/s1479236209990064 ◽

2009 ◽

Vol 6 (3) ◽

pp. 257-263 ◽

Cited By ~ 1

Author(s):

Yang Li ◽

Wang Xian-zhong ◽

Yang Meng-bo ◽

Zhang Jia-hua

Keyword(s):

Nitric Oxide ◽

Enzyme Activity ◽

Protein Kinase ◽

Cell Viability ◽

Sodium Nitroprusside ◽

Sertoli Cell ◽

Sertoli Cells ◽

High Concentration ◽

Treatment Results ◽

Anti Oxidant

AbstractTo illustrate the effect of nitric oxide (NO) on the microtubules of Sertoli cells (SC), SCs of piglets were treated with sodium nitroprusside (SNP). Changes in cell viability, anti-oxidant activity, enzyme activity and p38 mutagen-activated protein kinase (p38MAPK) activation were detected. The results were as follows. A low concentration of NO can keep SC microtubule and cell viability normal, and a high concentration of NO could increase p38MAPK activation, decrease anti-oxidant activity and transferrin secretion, and destroy the structure and distribution of the microtubules. The results suggest that SNP treatment results in an increase in NO in SCs and decreased cell anti-oxidant activity. The high concentration of NO destroys cell microtubules by activating p38MAPK.

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Amidative peptide processing and vascular function

AJP Cell Physiology ◽

10.1152/ajpcell.1997.273.6.c1908 ◽

1997 ◽

Vol 273 (6) ◽

pp. C1908-C1914 ◽

Cited By ~ 44

Author(s):

Charlie D. Oldham ◽

Cuizhen Li ◽

Jun Feng ◽

Robert O. Scott ◽

Wen Z. Wang ◽

...

Keyword(s):

Nitric Oxide ◽

Endothelial Cells ◽

Cell Death ◽

Vascular Function ◽

Direct Action ◽

Sensory Nerves ◽

Peptide Processing ◽

Marked Inhibition ◽

Amidated Peptide ◽

Oxide Synthase

Substance P (SP), an amidated peptide present in many sensory nerves, is known to affect cardiovascular function, and exogenously supplied SP has been shown to activate nitric oxide synthase (NOS) in endothelial cells. We now report that SP-Gly, the glycine-extended biosynthetic precursor of SP (which is enzymatically processed to the mature amidated SP), causes relaxation of rat aortic strips with an efficacy and potency comparable to that of SP itself. Pretreatment of the aortic strips with 4-phenyl-3-butenoic acid (PBA), an irreversible amidating enzyme inactivator, results in marked inhibition of the vasodilation activity induced by SP-Gly but not of that induced by SP itself. Isolated endothelial cell basal NOS activity is also decreased by pretreatment with PBA, with no evidence of cell death or direct action of PBA on NOS activity. Both bifunctional and monofunctional forms of amidating enzymes are present in endothelial cells, as evidenced by affinity chromatography and Western blot analysis. These results provide evidence for a link between amidative peptide processing, NOS activation in endothelial cells, and vasodilation and suggest that a product of amidative processing provides intrinsic basal activation of NOS in endothelial cells.

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Nitric Oxide Inhibits Caspase Activation and Apoptotic Morphology but Does Not Rescue Neuronal Death

Journal of Cerebral Blood Flow & Metabolism ◽

10.1038/sj.jcbfm.9600036 ◽

2005 ◽

Vol 25 (3) ◽

pp. 348-357 ◽

Cited By ~ 29

Author(s):

Ping Zhou ◽

Liping Qian ◽

Costantino Iadecola

Keyword(s):

Nitric Oxide ◽

Cell Death ◽

Cell Viability ◽

Cell Survival ◽

Caspase 3 ◽

Apoptotic Cell ◽

Cysteine Residue ◽

Caspase Activity ◽

No Donors ◽

Apoptotic Morphology

Nitric oxide (NO) has been shown to inhibit apoptotic cell death by S-nitrosylation of the catalytic-site cysteine residue of caspases. However, it is not clear whether in neurons NO-mediated caspase inactivation leads to improved cell survival. To address this issue, we studied the effect of NO donors on caspase activity and cell survival in cortical neuronal culture treated with the apoptosis inducer staurosporine (STS) and camptothecin. In parallel, cell viability was assessed by the MTS assay and MAP2 staining. We found that NO donors ((±)- S-nitroso- N-acetylpenicillamine, S-nitrosoglutathione, and NONOates) dose-dependently inhibited caspase-3 and -9 activity induced by STS and camptothecin. The reduction in caspase-3 activity was, in large part, because of the blockage of the proteolytic conversion of pro-caspase-3 to active caspase-3. NO donors also inhibited the appearance of the classical apoptotic nuclear morphology. However, inhibition of both caspase activity and apoptotic morphology was not associated with enhancement of cell viability. Thus, inhibition of caspase and apoptotic morphology by NO donors does not improve neuronal survival. The data suggest that inhibition of caspase by NO unmasks a caspase-independent form of cell death. A better understanding of this form of cell death may provide new strategies for neuroprotection in neuropathologies, such as ischemic brain injury, associated with apoptosis.

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Puerarin protects endothelial cells from oxidized low density lipoprotein induced injuries via the suppression of LOX-1 and induction of eNOS

Canadian Journal of Physiology and Pharmacology ◽

10.1139/cjpp-2013-0322 ◽

2014 ◽

Vol 92 (4) ◽

pp. 299-306 ◽

Cited By ~ 8

Author(s):

Mei-hua Bao ◽

Yi-wen Zhang ◽

Xiao-ya Lou ◽

Yan Xiao ◽

Yu Cheng ◽

...

Keyword(s):

Nitric Oxide ◽

Endothelial Cells ◽

Cell Viability ◽

Nuclear Translocation ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

No Production ◽

Low Density ◽

Oxidized Low Density Lipoprotein ◽

Cox 2

Oxidized low density lipoprotein (oxLDL) induced injury of endothelial cells is considered to be the first step in the pathogenesis of atherosclerosis. This study aimed to investigate some of the effects and mechanisms of puerarin on oxLDL-induced endothelial injuries. We measured cell viability, and the release of lactate dehydrogenase (LDH), nitric oxide (NO), and interleukin-8 (IL-8) to evaluate the protective effects of puerarin. Intracellular reactive oxygen species (ROS) were detected using 2′,7′-dichlorofluorescein diacetate (DCFH-DA). The expression of lectin-like low-density lipoprotein receptor-1 (LOX-1), endothelial nitric oxide synthase (eNOS), cyclooxygenase 2 (COX-2), p38MAPK, and protein kinase B (PKB) phosphorylation, nuclear factor-κB (NF-κB) nuclear translocation, and inhibitor of κB (IκB) degradation were detected using quantitative real-time PCR or Western blot. The results showed that oxLDL significantly decreased cell viability, increased LDH and IL-8 release, inhibited NO production, and induced COX-2 expression. Pretreatment with puerarin led to a strong inhibition of these effects. OxLDL stimulated the expression of LOX-1, the overproduction of ROS, the phosphorylation of p38MAPK, the dephosphorylation of PKB, activation of NF-κB, and the degradation of IκB. These oxLDL-induced effects were suppressed after puerarin pretreatment. These results suggest that puerarin inhibits oxLDL-induced endothelial cell injuries, at least in part, via inhibition of the LOX-1-mediated p38MAPK–NF-κB inflammatory and the PKB–eNOS signaling pathways.

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Nitric oxide releasing iron oxide magnetic nanoparticles for biomedical applications: cell viability, apoptosis and cell death evaluations

Journal of Physics Conference Series ◽

10.1088/1742-6596/429/1/012034 ◽

2013 ◽

Vol 429 ◽

pp. 012034 ◽

Cited By ~ 10

Author(s):

R de Lima ◽

J L de Oliveira ◽

A Ludescher ◽

M M Molina ◽

R Itri ◽

...

Keyword(s):

Nitric Oxide ◽

Cell Death ◽

Iron Oxide ◽

Magnetic Nanoparticles ◽

Cell Viability ◽

Biomedical Applications ◽

Iron Oxide Magnetic Nanoparticles ◽

Nitric Oxide Releasing

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Paraquat Modulates Immunological Function in Bone Marrow-Derived Macrophages

10.21203/rs.3.rs-407029/v1 ◽

2021 ◽

Author(s):

Piyarat Srinont ◽

Jaroon Wandee ◽

Worapol Angwanich

Keyword(s):

Nitric Oxide ◽

Reactive Oxygen Species ◽

Bone Marrow ◽

Cell Death ◽

Cell Viability ◽

Inducible Nitric Oxide Synthase ◽

Ros Production ◽

Oxygen Species ◽

Oxide Synthase ◽

Inducible Nitric Oxide

Abstract Paraquat (PQ) is an herbicide commonly used worldwide. This herbicide is known to alter the human and animal immune systems. Many reports indicated that PQ impacts immune cell viability and functions. However, the underlying mechanism critical is still unknown. Therefore, the aim of this study was to evaluate effects of PQ on free radical production, oxidative stress, cell death, and pro-inflammatory gene expression of murine bone marrow-derived macrophages (BMDMs) from female C57BL/6NJcl mice in vitro. BMDMs were incubated with PQ at 0, 200, 400 µM for 24 h. Intracellular reactive oxygen species (ROS) production, apoptosis, cell viability, nitric oxide, inducible nitric oxide synthase (iNOS), and IL-6 expression of murine BMDMs were measured. The results revealed that PQ treatments led to decrease the cell viability and induced apoptotic cell death in a dose-dependent manner. Additionally, PQ induced reactive oxygen species (ROS) generation. The mRNA expression level of pro-inflammatory mediator gene IL-6 and inducible nitric oxide synthase (iNOS) were elevated, while the level of lipid peroxides (MDA) production was unaltered by PQ treatment. Interestingly, PQ led to a decrease in nitric oxide production depends on its concentration. These phenomena indicated that PQ increased cellular ROS production which induced apoptosis, and the herbicide triggers production of iNOS and IL-6 in murine BMDMs.

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C1q/TNF–Related Protein-3 attenuates endothelial dysfunction in diabetic retinopathy via the Nitric Oxide signaling pathway

10.21203/rs.3.rs-26602/v1 ◽

2020 ◽

Author(s):

Zheyi Yan ◽

Xiaoming Cao ◽

Chunfang Wang ◽

Sha Liu ◽

Lu Gan ◽

...

Keyword(s):

Nitric Oxide ◽

Endothelial Cells ◽

Cell Death ◽

Diabetic Retinopathy ◽

Endothelial Dysfunction ◽

Signaling Pathway ◽

Molecular Mechanisms ◽

Apoptotic Cell ◽

Related Protein

Abstract Background Diabetic retinopathy (DR) is a severe microvasculature complication of diabetes. Restoration of dysfunctional endothelial cells represents a promising approach to treatment of DR. It has been demonstrated that a number of CTRP (C1q/tumor necrosis factor-related protein) members improves vascular endothelial function of the aortic vasculature. However, the role of CTRPs in the treatment of DR remains largely unresolved. Therefore, the aim of this study was to determine whether members of the CTRP family improve diabetes-induced endothelial dysfunction of retinal vasculature, thus exhibiting a protective effect against diabetic injury of retina. Methods The vasoactivity of currently identified murine CTRP family members was assessed in vascular rings and the underlying molecular mechanisms elucidated in human retinal microvascular endothelial cells. We then mimicked diabetic retinopathy both in vitro and in vivo, after which they were treated with CTRP3, and the vasoactivity, apoptotic cell death and vascular leakage in the retina were evaluated. Discovery-drive approaches followed by cause-effect analysis were used to uncover the molecular mechanisms of CTRP3. Results Our results demonstrate that CTRP3, CTRP5, and CTRP9 exert vasorelaxant effects on macro- and micro-vessels, with CTRP3 being the most potent in micro-vessels. The effects of CTRP3 were found to be endothelium-dependent via the AdipoR1/AMPK/eNOS/Nitric Oxide (NO) pathway. In in vitro microvascular reactivity studies, CTRP3 successfully improved high glucose/high lipid-induced impairment of endothelium-dependent vasodilatation. Blockade of either AMPK or eNOS completely abolished the previously observed effects of CTRP3. In addition, in the murine diabetic retinopathy model, CTRP3 treatment increased endothelium-dependent relaxation and NO levels in microvessels, and inhibited apoptotic cell death and vascular leakage in the retina. Finally,blockade of NO synthesis completely abolished the effects of CTRP3 that had been measured previously. Conclusion Taken together, our findings reveal that the AdipoR1/AMPK/eNOS/NO signaling pathway, through which CTRP3 reverses endothelial dysfunction of the microvasculature by normalization of impaired vasodilatation, represents a novel intervention effective against diabetic injury of retina.

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Differential responses of pancreatic β-cells to ROS and RNS

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00424.2012 ◽

2013 ◽

Vol 304 (6) ◽

pp. E614-E622 ◽

Cited By ~ 22

Author(s):

Gordon P. Meares ◽

Dominique Fontanilla ◽

Katarzyna A. Broniowska ◽

Teresa Andreone ◽

Jack R. Lancaster ◽

...

Keyword(s):

Nitric Oxide ◽

Dna Damage ◽

Cell Death ◽

Cell Viability ◽

Cell Fate ◽

Β Cells ◽

Pancreatic Β Cells ◽

Unfolded Protein ◽

Heat Shock Responses ◽

Reduce Cell Viability

Reactive oxygen species (ROS) and reactive nitrogen species (RNS) direct the activation of distinct signaling pathways that determine cell fate. In this study, the pathways activated and the mechanisms by which ROS and RNS control the viability of pancreatic β-cells were examined. Although both nitric oxide and hydrogen peroxide (H2O2) induce DNA damage, reduce cell viability, and activate AMPK, the mechanisms of AMPK activation and cell death induction differ between each reactive species. Nitric oxide activates the unfolded protein and heat shock responses and MAPK kinase signaling, whereas H2O2 stimulates p53 stabilization and poly(ADP-ribose) polymerase (PARP) activation but fails to induce the unfolded protein or heat shock responses or MAPK activation. The control of cell fate decisions is selective for the form of stress. H2O2-mediated reduction in β-cell viability is controlled by PARP, whereas cell death in response to nitric oxide is PARP independent but associated with the nuclear localization of GAPDH. These findings show that both ROS and RNS activate AMPK, induce DNA damage, and reduce cell viability; however, the pathways controlling the responses of β-cells are selective for the type of reactive species.

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