Nitric oxide participates in early events associated  with NNMU-induced acute lung injury in rats

In this study, the biochemical mechanisms by which N-nitroso- N-methylurethane (NNMU) induces acute lung injury are examined. Polymorphonuclear neutrophil infiltration into the lungs first appears in the bronchoalveolar lavage (BAL) fluid 24 h after NNMU injection (10.58 ± 3.00% of total cells; P < 0.05 vs. control animals). However, NNMU-induced elevation of the alveolar-arterial O2 difference requires 72 h to develop. Daily intraperitoneal injections of the inducible nitric oxide (⋅ NO) synthase (iNOS)-selective inhibitor aminoguanidine (AG) initiated 24 h after NNMU administration improve the survival of NNMU-treated animals. However, AG administration initiated 48 or 72 h after NNMU injection does not significantly improve the survival of NNMU-treated animals. These results suggest that ⋅ NO participates in events that occur early in NNMU-induced acute lung injury. BAL cells isolated from rats 24 and 48 h after NNMU injection produce elevated ⋅ NO and express iNOS during a 24-h ex vivo culture. AG attenuates ⋅ NO production but does not affect iNOS expression, whereas actinomycin D prevents iNOS expression and attenuates ⋅ NO production by BAL cells during this ex vivo culture. These results suggest that NNMU-derived BAL cells can stimulate iNOS expression and ⋅ NO production during culture. In 48-h NNMU-exposed rats, iNOS expression is elevated in homogenates of whole lavaged lungs but not in BAL cells derived from the same lung. These findings suggest that the pathogenic mechanism by which NNMU induces acute lung injury involves BAL cell stimulation of iNOS expression and ⋅ NO production in lung tissue.

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Reduction of Acute Lung Injury by Administration of Spironolactone After Intestinal Ischemia and Reperfusion in Rats

Clinical & Investigative Medicine ◽

10.25011/cim.v39i1.26326 ◽

2016 ◽

Vol 39 (1) ◽

pp. 15 ◽

Cited By ~ 6

Author(s):

Figen Barut ◽

V Haktan Ozacmak ◽

Inci Turan ◽

Hale Sayan-Ozacmak ◽

Erol Aktunc

Keyword(s):

Acute Lung Injury ◽

Lung Injury ◽

Intestinal Ischemia ◽

Neutrophil Infiltration ◽

Artery Occlusion ◽

Polymorphonuclear Neutrophils ◽

Polymorphonuclear Neutrophil ◽

Albino Rats ◽

Ischemia And Reperfusion ◽

Neutrophil Sequestration

Purpose: Multiple organ failure, including acute lung injury, is a common complication of intestinal ischemia and reperfusion (I/R) injury and contributes to its high mortality rate. Activated polymorphonuclear neutrophils and reactive oxygen species contribute to the lung injury caused by intestinal I/R. Mineralokortikoid receptor antagonist spironolactone has a protective effect against I/R injury in animal models of retina, kidney, heart, and brain. The aim of the present study is to investigate the effect of aldosteron receptor blocker spironolactone on lung injury induced by intestinal I/R. Methods: Wistar albino rats were divided into four groups: (1) sham control; (2) intestinal I/R (30 min of ischemia by superior mesenteric artery occlusion followed by 3 h of reperfusion); (3) spironolactone pretreatment (20 mg/kg) + I/R; and, (4) spironolactone pretreatment without I/R. Spironolactone was given orally 3 days prior to intestinal I/R. A marker for lipid peroxidation (malondialdehyde; MDA), an indicator or oxidation state (reduced glutathione; GSH), an index of polymorphonuclear neutrophil sequestration (myeloperoxidase; MPO), inducible nitric oxide synthase (iNOS) immunoreactivity, and the histopathology of the lung tissue were analyzed. Results: Spironolactone pretreatment markedly reduced intestinal I/R-induced lung injury as indicated by histology and MDA and MPO levels. Moreover, the pretreatment decreased the iNOS immunoreactivity. Conclusion: The present study strongly suggests that spironolactone pretreatment decreased neutrophil infiltration, iNOS induction, oxidative stress, and histopathological injury in an experimental model of intestinal I/R induced-lung injury of rats.

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Exhalation of gaseous nitric oxide by rats in response to endotoxin and its absorption by the lungs

Journal of Applied Physiology ◽

10.1152/jappl.1997.82.1.305 ◽

1997 ◽

Vol 82 (1) ◽

pp. 305-316 ◽

Cited By ~ 17

Author(s):

John T. Stitt ◽

Arthur B. Dubois ◽

James S. Douglas ◽

Steven G. Shimada

Keyword(s):

Nitric Oxide ◽

Acute Lung Injury ◽

Lung Injury ◽

No Synthase ◽

No Production ◽

Alveolar Air ◽

Lipopolysaccharide Endotoxin ◽

Early Biomarker ◽

Dose Dependent ◽

Expired Air

Stitt, John T., Arthur B. DuBois, James S. Douglas, and Steven G. Shimada. Exhalation of gaseous nitric oxide by rats in response to endotoxin and its absorption by the lungs. J. Appl. Physiol. 82(1): 305–316, 1997.—Rats injected with a lipopolysaccharide endotoxin produce detectable concentrations of nitric oxide gas (NO) in the expired air within 60 min. The concentration of NO reaches a plateau at 3 h. Production of the NO is dose dependent on lipopolysaccharide, and at a dose of 1 mg/kg iv, lipopolysaccharide alveolar concentrations of >260 parts per billion are observed. NO synthase inhibitors suppress this NO production in response to endotoxin. Experiments were conducted to ascertain the site of origin of this NO and to measure the capacity of the lungs to absorb NO from alveolar air. Results indicate that the endotoxin-induced NO originates from within the lungs themselves and that the lungs have the capacity to absorb >60% of NO that is presented to them. Lung tissues absorb ∼44–47% of the NO load, blood carries away between 15 and 19%, while the remainder is exhaled in the expired air. It is proposed that the exhalation of NO might prove useful as an early biomarker for acute lung injury.

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Deficiency of cationic amino acid transporter-2 protects mice from hyperoxia-induced lung injury

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00223.2018 ◽

2019 ◽

Vol 316 (4) ◽

pp. L598-L607 ◽

Cited By ~ 1

Author(s):

Yi Jin ◽

Yusen Liu ◽

Leif D. Nelin

Keyword(s):

Nitric Oxide ◽

Acute Lung Injury ◽

Amino Acid ◽

Lung Injury ◽

Weight Ratio ◽

No Production ◽

Myeloperoxidase Activity ◽

Wild Type ◽

Cationic Amino Acid ◽

Significant Difference

The pathology of acute lung injury (ALI) involves inducible nitric oxide (NO) synthase (iNOS)-derived NO-induced apoptosis of pulmonary endothelial cells. In vitro, iNOS-derived NO production has been shown to depend on the uptake of l-arginine by the cationic amino acid transporters (CAT). To test the hypothesis that mice deficient in CAT-2 ( slc7a2−/− on a C57BL/6 background) would be protected from hyperoxia-induced ALI, mice ( slc7a2−/− or wild-type) were placed in >95% oxygen (hyperoxia) or 21% oxygen (control) for 60 h. In wild-type mice exposed to hyperoxia, the exhaled nitric oxide (exNO) was twofold greater than in wild-type mice exposed to normoxia ( P < 0.005), whereas in slc7a2−/− mice there was no significant difference between exNO in animals exposed to hyperoxia or normoxia ( P = 0.95). Hyperoxia-exposed wild-type mice had greater ( P < 0.05) lung resistance and a lower ( P < 0.05) lung compliance than did hyperoxia-exposed slc7a2−/− mice. The lung wet-to-dry weight ratio was greater ( P < 0.005) in the hyperoxia-exposed wild-type mice than in hyperoxia-exposed slc7a2−/− mice. Neutrophil infiltration was lower ( P < 0.05) in the hyperoxia-exposed slc7a2−/− mice than in the hyperoxia-exposed wild-type mice as measured by myeloperoxidase activity. The protein concentration in bronchoalveolar lavage fluid was lower ( P < 0.001) in the hyperoxia-exposed slc7a2−/− mice than in similarly exposed wild-type mice. The percent of TUNEL-positive cells in the lung following hyperoxia exposure was significantly lower ( P < 0.001) in the slc7a2−/− mice than in the wild-type mice. These results are consistent with our hypothesis that lack of CAT-2 protects mice from acute lung injury.

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Adoptive transfer of acute lung injury

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.2000.279.5.l985 ◽

2000 ◽

Vol 279 (5) ◽

pp. L985-L993 ◽

Cited By ~ 10

Author(s):

Michael A. Moxley ◽

Tracey L. Baird ◽

John A. Corbett

Keyword(s):

Acute Lung Injury ◽

Lung Injury ◽

Adoptive Transfer ◽

Cell Activation ◽

Ex Vivo ◽

Inos Expression ◽

Arterial Oxygen ◽

Blue Dye ◽

Intratracheal Administration ◽

Oxygen Difference

In this study, we describe a novel adoptive transfer protocol to study acute lung injury in the rat. We show that bronchoalveolar lavage (BAL) cells isolated from rats 5 h after intratracheal administration of lipopolysaccharide (LPS) induce a lung injury when transferred to normal control recipient rats. This lung injury is characterized by increased alveolar-arterial oxygen difference and extravasation of Evans blue dye (EBD) into lungs of recipient rats. Recipient rats receiving similar numbers of donor cells isolated from healthy rats do not show adverse changes in the alveolar-arterial oxygen difference or in extravasation of EBD. The adoptive transfer-induced lung injury is associated with increased numbers of neutrophils in the BAL, the levels of which are similar to the numbers observed in BAL cells isolated from rats treated for 5 h with LPS. As an indicator of BAL cell activation, donor BAL cell inducible nitric oxide synthase (iNOS) expression was compared with BAL cell iNOS expression 48 h after adoptive transfer. BAL cells isolated 5 h after LPS administration expressed iNOS immediately after isolation. In contrast, BAL cells isolated 48 h after adoptive transfer did not express iNOS immediately after isolation but expressed iNOS following a 24-h ex vivo culture. These findings indicate that the activation state of donor BAL cells differs from BAL cells isolated 48 h after adoptive transfer, suggesting that donor BAL cells may stimulate migration of new inflammatory cells into the recipient rats lungs.

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Induction of nitric oxide synthase by rotavirus enterotoxin NSP4: implication for rotavirus pathogenicity

Journal of General Virology ◽

10.1099/vir.0.82618-0 ◽

2007 ◽

Vol 88 (7) ◽

pp. 2064-2072 ◽

Cited By ~ 25

Author(s):

Mohamed A. Borghan ◽

Yoshio Mori ◽

Abu-Baker El-Mahmoudy ◽

Naoto Ito ◽

Makoto Sugiyama ◽

...

Keyword(s):

Nitric Oxide ◽

Mrna Expression ◽

Ex Vivo ◽

Inos Expression ◽

Intestinal Cells ◽

Inos Mrna ◽

No Production ◽

Post Inoculation ◽

Structural Protein ◽

Rotavirus Infection

Rotavirus non-structural protein (NSP) 4 can induce aqueous secretion in the gastrointestinal tract of neonatal mice through activation of an age- and Ca2+-dependent plasma membrane anion permeability. Accumulating evidence suggests that nitric oxide (NO) plays a role in the modulation of aqueous secretion and the barrier function of intestinal cells. This study investigated transcriptional changes in inducible NO synthase (iNOS), an enzyme responsible for NO production, after rotavirus infection in mice and after treatment of intestinal cells with NSP4. Diarrhoea was observed in 5-day-old CD-1 mice from days 1 to 3 after inoculation with 107 focus-forming units of different rotavirus strains. Ileal iNOS mRNA expression was induced as early as 6 h post-inoculation, before the onset of clinical diarrhoea in infected mice, and was upregulated during the course of rotavirus-induced diarrhoea. Ex vivo treatment of ilea excised from CD-1 suckling mice with NSP4 resulted in upregulation of ileal iNOS mRNA expression within 4 h. Furthermore, NSP4 was able to induce iNOS expression and NO production in murine peritoneal macrophages and RAW264.7 cells. The specificity of NSP4 inducibility was confirmed by the inhibitory effect of anti-NSP4 serum. Using a series of truncated NSP4s, the domain responsible for iNOS induction in macrophages was mapped to the reported enterotoxin domain, aa 109–135. Thus, rotavirus infection induces ileal iNOS expression in vivo and rotavirus NSP4 also induces iNOS expression in the ileum and macrophages. Together, these findings suggest that NO plays a role in rotavirus-induced diarrhoea.

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H2S Attenuates LPS-Induced Acute Lung Injury by Reducing Oxidative/Nitrative Stress and Inflammation

Cellular Physiology and Biochemistry ◽

10.1159/000453210 ◽

2016 ◽

Vol 40 (6) ◽

pp. 1603-1612 ◽

Cited By ~ 54

Author(s):

Hong-Xia Zhang ◽

Shu-Juan Liu ◽

Xiao-Lu Tang ◽

Guo-Li Duan ◽

Xin Ni ◽

...

Keyword(s):

Nitric Oxide ◽

Acute Lung Injury ◽

Treatment Group ◽

Lung Injury ◽

Control Group ◽

No Production ◽

Sod Activity ◽

Nitrative Stress ◽

Anti Oxidant ◽

Anti Inflammatory

Background: Hydrogen sulfide (H2S), known as the third endogenous gaseous transmitter, has received increasing attention because of its diverse effects, including angiogenesis, vascular relaxation and myocardial protection.We aimed to investigate the role of H2S in oxidative/nitrative stress and inflammation in acute lung injury (ALI) induced by endotoxemia. Methods: Male ICR mice were divided in six groups: (1) Control group; (2) GYY4137treatment group; (3) L-NAME treatment group; (4) lipopolysaccharide (LPS) treatment group; (5) LPS with GYY4137 treatment group; and (6) LPS with L-NAME treatment group. The lungs were analysed by histology, NO production in the mouse lungs determined by modified Griess (Sigma-Aldrich) reaction, cytokine levels utilizing commercialkits, and protein abundance by Western blotting. Results: GYY4137, a slowly-releasing H2S donor, improved the histopathological changes in the lungs of endotoxemic mice. Treatment with NG-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase (NOS) inhibitor, increased anti-oxidant biomarkers such as thetotal antioxidant capacity (T-AOC) and theactivities of catalase (CAT) and superoxide dismutase (SOD) but decreased a marker of peroxynitrite (ONOO-) action and 3-nitrotyrosine (3-NT) in endotoxemic lung. L-NAME administration also suppressed inflammation in endotoxemic lung, as evidenced by the decreased pulmonary levels of interleukin (IL)-6, IL-8, and myeloperoxidase (MPO) and the increased level of anti-inflammatory cytokine IL-10. GYY4137 treatment reversed endotoxin-induced oxidative/nitrative stress, as evidenced by a decrease in malondialdehyde (MDA), hydrogenperoxide (H2O2) and 3-NT and an increase in the antioxidant biomarker ratio of reduced/oxidized glutathione(GSH/GSSG ratio) and T-AOC, CAT and SOD activity. GYY4137 also attenuated endotoxin-induced lung inflammation. Moreover, treatment with GYY4137 inhibited inducible NOS (iNOS) expression and nitric oxide (NO) production in the endotoxemia lung. Conclusions: GYY4137 conferred protection against acute endotoxemia-associated lung injury, which may have beendue to the anti-oxidant, anti-nitrative and anti-inflammatory properties of GYY4137. The present findings warrant further exploration of the clinical applicability of H2S in the prevention and treatment of ALI.

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Immunomodulatory Effect of Chinese Herbal Medicine Formula Sheng-Fei-Yu-Chuan-Tang in Lipopolysaccharide-Induced Acute Lung Injury Mice

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2013/976342 ◽

2013 ◽

Vol 2013 ◽

pp. 1-12 ◽

Cited By ~ 4

Author(s):

Chia-Hung Lin ◽

Ching-Hua Yeh ◽

Li-Jen Lin ◽

Shulhn-Der Wang ◽

Jen-Shu Wang ◽

...

Keyword(s):

Nitric Oxide ◽

Acute Lung Injury ◽

Lung Injury ◽

Inflammatory Cytokines ◽

Interleukin 1 ◽

Reactive Oxygen Species Generation ◽

Interleukin 10 ◽

Interleukin 4 ◽

No Production ◽

Anti Inflammatory

Traditional Chinese medicine formula Sheng-Fei-Yu-Chuan-Tang (SFYCT), consisting of 13 medicinal plants, was used to treat patients with lung diseases. This study investigated the immunoregulatory effect of SFYCT on intratracheal lipopolysaccharides- (LPS-) challenged acute lung injury (ALI) mice. SFYCT attenuated pulmonary edema, macrophages, and neutrophils infiltration in the airways. SFYCT decreased inflammatory cytokines, including tumor necrosis factor-α(TNFα), interleukin-1β, and interleukin-6 and inhibited nitric oxide (NO) production but increased anti-inflammatory cytokines, interleukin-4, and interleukin-10, in the bronchoalveolar lavage fluid of LPS-challenged mice. TNFαand monocyte chemotactic protein-1 mRNA expression in the lung of LPS-challenged mice as well as LPS-stimulated lung epithelial cell and macrophage were decreased by SFYCT treatment. SFYCT treatment also decreased the inducible nitric oxide synthase expression and phosphorylation of nuclear factor-κB (NF-κB) in the lung of mice and macrophage with LPS stimulation. SFYCT treatment dose dependently decreased the LPS-induced NO and reactive oxygen species generation in LPS-stimulated macrophage. In conclusion, SFYCT attenuated lung inflammation during LPS-induced ALI through decreasing inflammatory cytokines production while increasing anti-inflammatory cytokines production. The immunoregulatory effect of SFYCT is related to inhibiting NF-κB phosphorylation.

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Hypoxia compared with normoxia alters the effects of nitric oxide in ischemia-reperfusion lung injury

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1997.273.3.l504 ◽

1997 ◽

Vol 273 (3) ◽

pp. L504-L512 ◽

Cited By ~ 2

Author(s):

Y. C. Huang ◽

P. W. Fisher ◽

E. Nozik-Grayck ◽

C. A. Piantadosi

Keyword(s):

Nitric Oxide ◽

Lung Injury ◽

Average Rate ◽

Ischemia Reperfusion ◽

Oxidant Stress ◽

No Production ◽

Protective Effects ◽

Lung Weight ◽

Edema Formation ◽

No Donors

Because both the biosynthesis of nitric oxide (NO.) and its metabolic fate are related to molecular O2, we hypothesized that hypoxia would alter the effects of NO. during ischemia-reperfusion (IR) in the lung. In this study, buffer-perfused lungs from rabbits underwent either normoxic IR (AI), in which lungs were ventilated with 21% O2 during ischemia and reperfusion, or hypoxic IR (NI), in which lungs were ventilated with 95% N2 during ischemia followed by reoxygenation with 21% O2. Lung weight gain (WG) and pulmonary artery pressure (Ppa) were monitored continuously, and microvascular pressure (Pmv) was measured after reperfusion to calculate pulmonary vascular resistance. We found that both AI and NI produced acute lung injury, as shown by increased WG and Ppa during reperfusion. In AI, where perfusate PO2 was > 100 mmHg, the administration of the NO. synthase inhibitor N-nitro-L-arginine methyl ester (L-NAME) before ischemia worsened WG and Ppa. Pmv also increased, suggesting a hydrostatic mechanism involved in edema formation. The effects of L-NAME could be attenuated by giving L-arginine and exogenous NO. donors before ischemia or before reperfusion. Partial protection was also provided by superoxide dismutase. In contrast, lung injury in NI at perfusate PO2 of 25-30 mmHg was attenuated by L-NAME; this effect could be reversed by L-arginine. Exogenous NO. donors given either before ischemia or before reperfusion, however, did not increase lung injury. NO. production was measured by quantifying the total nitrogen oxides (NOx) accumulating in the perfusate. The average rate of NOx accumulation was greater in AI than in NI. We conclude that hypoxia prevented the protective effects of NO on AI lung injury. The effects of hypoxia may be related to lower NO. production relative to oxidant stress during IR and/or altered metabolic fates of NO.-mediated production of peroxynitrite by hypoxic ischemia.

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