Neutrophil Proteinases in Hydrochloric Acid- and Endotoxin-Induced Acute Lung Injury: Evaluation of Interstitial Protease Activity by in situ Zymography

Adrenomedullin (ADM) is upregulated independently by hypoxia and LPS, two key factors in the pathogenesis of acute lung injury (ALI). This study evaluates the expression of ADM in ALI using experimental models combining both stimuli: an in vivo model of rats treated with LPS and acute normobaric hypoxia (9% O2) and an in vitro model of rat lung cell lines cultured with LPS and exposed to hypoxia (1% O2). ADM expression was analyzed by in situ hybridization, Northern blot, Western blot, and RIA analyses. In the rat lung, combination of hypoxia and LPS treatments overcomes ADM induction occurring after each treatment alone. With in situ techniques, the synergistic effect of both stimuli mainly correlates with ADM expression in inflammatory cells within blood vessels and, to a lesser extent, to cells in the lung parenchyma and bronchiolar epithelial cells. In the in vitro model, hypoxia and hypoxia + LPS treatments caused a similar strong induction of ADM expression and secretion in epithelial and endothelial cell lines. In alveolar macrophages, however, LPS-induced ADM expression and secretion were further increased by the concomitant exposure to hypoxia, thus paralleling the in vivo response. In conclusion, ADM expression is highly induced in a variety of key lung cell types in this rat model of ALI by combination of hypoxia and LPS, suggesting an essential role for this mediator in this syndrome.

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СУБМІКРОСКОПІЧНІ ЗМІНИ КОМПОНЕНТІВ АЕРОГЕМАТИЧНОГО БАР’ЄРУ РЕСПІРАТОРНОГО ВІДДІЛУ ЛЕГЕНЬ ЩУРІВ У РАННІ ТЕРМІНИ ПІСЛЯ ГОСТРОГО УРАЖЕННЯ ХЛОРИДНОЮ КИСЛОТОЮ

Вісник наукових досліджень ◽

10.11603/2415-8798.2018.2.9154 ◽

2018 ◽

Author(s):

V. O. Beskyy ◽

Z. M. Nebesna ◽

M. I. Marushchak ◽

L. A. Hryshchuk

Keyword(s):

Acute Lung Injury ◽

Hydrochloric Acid ◽

Lung Injury ◽

Early Period ◽

Alveolar Epithelium ◽

Male Rats ◽

Control Group ◽

Ultrastructural Organization ◽

Type I ◽

White Rats

Submicroscopic studies of the respiratory part of the lungs after 2 and 6 hours after the experimental acute lung injury with hydrochloric acid established adaptive-compensatory and destructive changes in the components of the air-blood barrier.The aim of the study – to learn submicroscopic changes in the components of the air-blood barrier of the lungs in the early period after acute lung injury.Materials and Methods. The experiments were carried out on 30 white mature non-linear male rats weighing 200–220 g. The animals were divided into 3 groups: 1 – control group, 2 – hydrochloric acid damage after 2 hours, 3 – hydrochloric acid damage after 6 hour.Results and Discussion. In an experiment on mature white rats, a study was made of the submicroscopic state of the components of the air-blood barrier in the early periods after acute lung injury. It has been established that adaptive-compensatory and initial destructive changes of the alveolar epithelium and the walls of the hemocapillary take place at 2 o'clock in the experiment. The cytoplasm of respiratory epitheliocytes during this period of the experiment was focal-edematous and enlightened, organelles were destructively altered. For alveolocytes of type I, there was a significant swelling and clarification of the cytoplasm. During this period of the experiment, an increased number of actively phagocytizing macrophages appeared, which acquired a rounded shape, clearly contoured membranes of the cariolema, their invaginations were determined, and in the karyoplasm euchromatin predominated. In alveolocytes of type II, after 6 hours, the progression of destructive changes was established. For which there were peculiarity hypertrophied nuclei with deep invagination of the cariolema, in which there were few nuclear pores, locally expanded perinuclear space. In the edematous cytoplasm, organelles were found to be destructively altered.Conclusions. Acute damage to the lungs leads to a disruption of the ultrastructural organization of the air-blood barrier. Established adaptive-compensatory processes and signs of destructive changes in the alveolar epithelium and the walls of hemocapillaries, which leads to deterioration of gas-exchange processes in the lungs.

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Skin barrier lipid enzyme activity in Netherton patients is associated with protease activity and ceramide abnormalities

The Journal of Lipid Research ◽

10.1194/jlr.ra120000639 ◽

2020 ◽

Vol 61 (6) ◽

pp. 859-869 ◽

Cited By ~ 1

Author(s):

Jeroen van Smeden ◽

Hanin Al-Khakany ◽

Yichen Wang ◽

Dani Visscher ◽

Nicole Stephens ◽

...

Keyword(s):

Serine Protease ◽

Protease Activity ◽

Skin Barrier ◽

Skin Inflammation ◽

Acid Sphingomyelinase ◽

Netherton Syndrome ◽

Serine Protease Activity ◽

In Situ Zymography ◽

Ichthyosis Linearis Circumflexa

Individuals with Netherton syndrome (NTS) have increased serine protease activity, which strongly impacts the barrier function of the skin epidermis and leads to skin inflammation. Here, we investigated how serine protease activity in NTS correlates with changes in the stratum corneum (SC) ceramides, which are crucial components of the skin barrier. We examined two key enzymes involved in epidermal ceramide biosynthesis, β-glucocerebrosidase (GBA) and acid-sphingomyelinase (ASM). We compared in situ expression levels and activities of GBA and ASM between NTS patients and controls and correlated the expression and activities with i) SC ceramide profiles, ii) in situ serine protease activity, and iii) clinical presentation of patients. Using activity-based probe labeling, we visualized and localized active epidermal GBA, and a newly developed in situ zymography method enabled us to visualize and localize active ASM. Reduction in active GBA in NTS patients coincided with increased ASM activity, particularly in areas with increased serine protease activity. NTS patients with scaly erythroderma exhibited more pronounced anomalies in GBA and ASM activities than patients with ichthyosis linearis circumflexa. They also displayed a stronger increase in SC ceramides processed via ASM. We conclude that changes in the localization of active GBA and ASM correlate with i) altered SC ceramide composition in NTS patients, ii) local serine protease activity, and iii) the clinical manifestation of NTS.

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Investigation of anti-inflammatory effects of oxygen nanobubbles in a rat hydrochloric acid lung injury model

Nanomedicine ◽

10.2217/nnm-2020-0338 ◽

2020 ◽

Vol 15 (27) ◽

pp. 2647-2654

Author(s):

Keisuke Yoshida ◽

Yukihiro Ikegami ◽

Shinju Obara ◽

Keiko Sato ◽

Masahiro Murakawa

Keyword(s):

Acute Lung Injury ◽

Mean Arterial Pressure ◽

Hydrochloric Acid ◽

Lung Injury ◽

Arterial Pressure ◽

Rat Model ◽

Injury Model ◽

Lung Injury Model ◽

Anti Inflammatory ◽

Inflammatory Effect

Aim: To investigate the anti-inflammatory effect of oxygen nanobubbles (ONBs) in an acute lung injury rat model. Materials & methods: In a rat hydrochloric acid lung injury model, ONB fluid was administered intravenously in the ONB group (n = 6) and normal saline was administered in the control group (n = 6). 4 h later, arterial partial pressure of oxygen (PaO2), mean arterial pressure and plasma inflammatory cytokines were measured. Results: There were no significant differences in the PaO2, mean arterial pressure or TNF-α and IL-6 levels between the two groups. Conclusions: No anti-inflammatory effect could be confirmed at the present ONB dose in the rat model of acute lung injury.

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Changes in central respiratory patterns and activity following acute lung injury in the in situ rat preparation

The FASEB Journal ◽

10.1096/fasebj.26.1_supplement.1148.6 ◽

2012 ◽

Vol 26 (S1) ◽

Author(s):

Abigail Rose Zaylor ◽

David Nethery ◽

Yee-Hsee Hsieh ◽

Thomas E Dick ◽

Frank J Jacono

Keyword(s):

Acute Lung Injury ◽

Lung Injury

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Intravenous Lidocaine Attenuates Acute Lung Injury Induced by Hydrochloric Acid Aspiration in Rabbits

Anesthesiology ◽

10.1097/00000542-199805000-00022 ◽

1998 ◽

Vol 88 (5) ◽

pp. 1300-1309 ◽

Cited By ~ 59

Author(s):

Kahoru Nishina ◽

Katsuya Mikawa ◽

Yumiko Takao ◽

Makoto Shiga ◽

Nobuhiro Maekawa ◽

...

Keyword(s):

Acute Lung Injury ◽

Hydrochloric Acid ◽

Lung Injury ◽

Weight Ratio ◽

The Other ◽

Lung Damage ◽

Lung Mechanics ◽

Intravenous Lidocaine ◽

Superoxide Anion Production ◽

Acid Aspiration

Background Neutrophils play a crucial role in the pathogenesis of acid-induced acute lung injury. Lidocaine inhibits the function of neutrophils. This study aimed to determine whether lidocaine attenuates acute lung injury induced by hydrochloric acid (HCl) instillation. Methods In study 1, rabbits were divided into four groups (n = 7 each). Lung injury was induced by intratracheal HCl (0.1 N, 3 ml/kg) in two groups. The other two groups received saline intratracheally. Lidocaine given intravenously (2 mg/ g bolus + 2 mg x kg(-1) x h(-1) infusion) was started 10 min before intratracheal instillation in one HCl and one saline group, and saline was given intravenously in the other two groups. In study 2, rabbits (four groups of seven animals each) received HCl (0.1 N, 3 ml/kg) intratracheally. Treatment with intravenous lidocaine was started 10 min before, 10 min after, or 30 min after acid instillation, or saline was given intravenously 10 min before instillation. Results In study 1, HCl caused deterioration of the partial pressure of oxygen (PaO2), lung leukosequestration, decreased lung compliance, and increased the lung wet-to-dry weight ratio and albumin, interleukin-6 (IL-6), and IL-8 levels in bronchoalveolar lavage fluid. Lidocaine pretreatment attenuated these changes. Hydrochloric acid increased superoxide anion production by neutrophils and caused morphologic lung damage, both of which were lessened by lidocaine. In study 2, lidocaine given 10 min after acid instillation was as effective as pretreatment in PaO2, lung mechanics, and histologic examination. However, PaO2 changes in lidocaine 30 min after injury were similar to those in saline given intravenously. Conclusions Intravenous lidocaine started before and immediately after acid instillation attenuated the acute lung injury, in part by inhibiting the sequestration and activation of neutrophils.

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Acute lung injury isolated to an in situ lung preparation causes sustained reflex cardiovascular depression in dogs

Journal of Applied Physiology ◽

10.1152/jappl.1994.77.4.1850 ◽

1994 ◽

Vol 77 (4) ◽

pp. 1850-1857 ◽

Cited By ~ 1

Author(s):

D. A. Allen ◽

E. R. Schertel ◽

S. E. Weisbrode ◽

P. D. Myerowitz

Keyword(s):

Acute Lung Injury ◽

Lung Injury ◽

Left Lung ◽

Left Ventricular ◽

Rate Of Change ◽

Efferent Nerve ◽

Isolated Lung ◽

Cardiovascular Depression ◽

Lung Preparation

We tested the hypothesis that acute lung injury (ALI) isolated to a perfused in situ left lung preparation results in sustained reflex cardiovascular depression. Phorbol myristate acetate (PMA), an agent that activates neutrophils, administered into the isolated lung preparation of chloralose-anesthetized dogs resulted in ALI, as assessed by wet-to-dry weight ratios and histopathology, and significant decreases in heart rate (43%), mean arterial pressure (27%), aortic blood flow (29%) and maximum rate of change in left ventricular pressure (30%). Significant reflex effects occurred by 20 min after PMA administration and were sustained for 40 min (n = 7). Hemodynamic variables recovered when the left lung was denervated 60 min after PMA administration. Indomethacin administered into the isolated circulation before PMA (n = 5) did not significantly influence the ALI or reflex effects. Systemic atropinization (n = 6) prevented only the bradycardia. Left lung denervation before ALI (n = 3) prevented all reflex effects. We conclude that PMA administration into an isolated in situ lung preparation results in ALI and sustained reflex cardiovascular depression that is most likely elicited by pulmonary C-fiber stimulation and mediated by withdrawal of sympathetic efferent nerve activity.

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Peculiarities of dynamics indicators of endogenous intoxication in acute lung injury induced by hydrochloric acid

Medical and Clinical Chemistry ◽

10.11603/mcch.2410-681x.2016.v0.i3.6946 ◽

2016 ◽

Author(s):

O. O. Kostina ◽

A. A. Hudyma

Keyword(s):

Molecular Weight ◽

Acute Lung Injury ◽

Hydrochloric Acid ◽

Lung Injury ◽

Average Molecular Weight ◽

Experimental Studies ◽

Injury Rates ◽

Endogenous Intoxication ◽

Serum Fractions

In experimental studies on rats we found out that in acute lung injury rates there is increasing an endogenous intoxication – content serum fractions of average molecular weight within 48 hours of observation and erythrocytic index of intoxication for 24 hours followed by 72 hours of decrease in the experiment.

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