Aspiration-induced Lung Injury Research Articles

Restoring discarded porcine lungs by ex vivo removal of neutrophil extracellular traps

BackgroundBy causing inflammation and tissue damage, neutrophil extracellular traps (NETs) constitute an underlying mechanism of aspiration-induced lung injury, a major factor of the low utilization of donor lungs in lung transplantation (LTx). MethodTo determine whether NET removal during ex vivo lung perfusion (EVLP) can restore lung function and morphology in aspiration-damaged lungs, gastric aspiration lung injury was induced in 12 pigs. After confirmation of acute respiratory distress syndrome, the lungs were explanted and assigned to NET removal connected to EVLP (treated) (n=6) or EVLP only (non-treated) (n=6). Hemodynamic measurements were taken, and blood and tissue samples were collected to assess lung function, morphology, levels of cell-free DNA, extracellular histones, and nucleosomes as markers of NETs as well as cytokine levels. ResultsAfter EVLP and NET removal in porcine lungs, PaO2/FiO2 ratios increased significantly compared to those undergoing EVLP alone (p=0.0411). Treated lungs had lower cell-free DNA (p=0.0260) and lower levels of extracellular histones in EVLP perfusate than non-treated lungs (p=0.0260). According to histopathology, treated lungs showed less immune cell infiltration and less oedema compared with non-treated lungs which was reflected in decreased levels of pro-inflammatory cytokines in EVLP perfusate and BALF. ConclusionTo conclude, removing NETs during EVLP improved lung function and morphology in aspiration-damaged donor lungs. The ability to remove NETs during EVLP could represent a new therapeutic approach for LTx and potentially expand the donor pool for transplantation.

Haploinsufficiency of Cnot3 Aggravates Acid-Induced Acute Lung Injury Likely Through Transcriptional and Post-Transcriptional Upregulation of Pro-Inflammatory Genes.

Acute lung injury (ALI) is caused by a variety of illnesses, including aspiration pneumonia and sepsis. The CCR4-NOT complex is a large multimeric protein complex that degrades mRNA through poly(A) tail shortening, whereas it also contributes to regulation of transcription and translation. Cnot3 is a scaffold component of the CCR4-NOT complex and is essential for the integrity of the complex; loss of Cnot3 leads to depletion of whole complex. While the significance of cytokine mRNA degradation in limiting inflammation has been established, the roles of CCR4-NOT complex-mediated in ALI remain elusive. The effects of Cnot3 haploinsufficiency in the pathology and cytokine expression were analyzed in the mouse lungs of acid aspiration-induced acute lung injury. The decay rate and transcription activity of cytokine mRNAs under Cnot3 heterozygous deletion were analyzed in lipopolysaccharide (LPS) -stimulated mouse embryonic fibroblasts (MEFs). Tamoxifen-induced heterozygous deletion of Cnot3 in adult mice (Cnot3 Hetz) did not show body weight loss or any apparent abnormality. Under acid aspiration-induced acute lung injury, Cnot3 Hetz mice exhibited increased pulmonary edema, worse lung pathologies and more severe inflammation compared with wild type mice. mRNA expression of pro-inflammatory genes Il1b and Nos2 were significantly upregulated in the lungs of Cnot3 Hetz mice. Consistently, mRNA expression of Il1b and Nos2 was upregulated in LPS-stimulated Cnot3 Hetz MEFs. Mechanistically, while heterozygous depletion of Cnot3 stabilized both Il1b and Nos2 mRNAs, the nascent pre-mRNA level of Il1b was upregulated in Cnot3 Hetz MEFs, implicating Cnot3-mediated transcriptional repression of Il1b expression in addition to destabilization of Il1b and Nos2 mRNAs. PU.1 (Spi1) was identified as a causative transcription factor to promote Il1b expression under Cnot3 haploinsufficient conditions. CNOT3 plays a protective role in ALI by suppressing expression of pro-inflammatory genes Il1b and Nos2 through both post-transcriptional and transcriptional mechanisms, including mRNA stability control of Spi1.

MicroRNA‑200c‑3p regulates seawater‑induced acute lung injury via ANGII and ACE2/ANG1‑7 pathways.

Apoptosis is a main characteristic of seawater aspiration-induced acute lung injury (ALI). The local angiotensin (ANG) system angiotensin converting enzyme (ACE)-2/ANG1-7/Mas axis and ANGII/angiotensin II receptor type 1 (AT1) play an important role in apoptosis. MicroRNA (miR)-200c-3p is involved in the regulation of the ACE-2 pathway, but its role and mechanism in seawater-induced ALI remain to be elucidated. In the present study, seawater-ALI lung tissue and cell model was established and apoptosis-related proteins, ACE2, ANGII, ANG1-7 were detected by western blotting following downregulation of miR-200c-3p. In addition, miR-200c-3p was detected by reverse transcription-quantitative PCR. The target relationship between miR-200c-3p and ACE2 was confirmed by dual-luciferase reporter assay. Seawater stimulation increased the expression of miR-200c-3p, ANGII and decreased ACE-2/ANG1-7 expression and induced changes of apoptosis-related protein expression. Apoptosis can be inhibited by AT1 blocker and abrogated by addition of ANG1-7 following seawater stimulation. In addition, inhibition of miR-200c-3p suppressed apoptosis and decreased the expression of ANGII, but increased the ACE-2/ANG1-7 expression. These results suggested that increased expression of miR-200c-3p was an important cause in seawater-induced ALI and this phenomenon was through inhibition of ACE2/ANG1-7 pathway.

Efficacy of surfactant therapy of ARDS induced by hydrochloric acid aspiration followed by ventilator-induced lung injury - an animal study.

The development of acute respiratory distress syndrome (ARDS) is known to be independently attributable to aspiration-induced lung injury. Mechanical ventilation as a high pressure/volume support to maintain sufficient oxygenation of a patient could initiate ventilator-induced lung injury (VILI) and thus contribute to lung damage. Although these phenomena are rare in the clinic, they could serve as the severe experimental model of alveolar-capillary membrane deterioration. Lung collapse, diffuse inflammation, alveolar epithelial and endothelial damage, leakage of fluid into the alveoli, and subsequent inactivation of pulmonary surfactant, leading to respiratory failure. Therefore, exogenous surfactant could be considered as a therapy to restore lung function in experimental ARDS. This study aimed to investigate the effect of modified porcine surfactant in animal model of severe ARDS (P/F ratio </=13.3 kPa) induced by intratracheal instillation of hydrochloric acid (HCl, 3 ml/kg, pH 1.25) followed by VILI (V(T) 20 ml/kg). Adult rabbits were divided into three groups: untreated ARDS, model treated with a bolus of poractant alfa (Curosurf®, 2.5 ml/kg, 80 mg phospholipids/ml), and healthy ventilated animals (saline), which were oxygen-ventilated for an additional 4 h. The lung function parameters, histological appearance, degree of lung edema and levels of inflammatory and oxidative markers in plasma were evaluated. Whereas surfactant therapy with poractant alfa improved lung function, attenuated inflammation and lung edema, and partially regenerated significant changes in lung architecture compared to untreated controls. This study indicates a potential of exogenous surfactant preparation in the treatment of experimental ARDS.

893: EXOSOMES FROM MESENCHYMAL STEM CELLS ALLEVIATE ASPIRATION-INDUCED LUNG INJURY IN MICE

Introduction: With acidic fluid, the particulate component of the aspirate and pepsin, and bacteria from the oral cavity, gastric or oropharyngeal contents aspiration readily induces lung injury. Effective therapy remains lacking to date. The exaggerated inflammatory response has been identified as one crucial mechanism that mediates the development of lung injury in this regard. Exosomes from mesenchymal stem cells (MSCs) possess a potent anti-inflammation capacity. Based on these data, we thus conjectured that exosomes from MSCs may mitigate aspiration-induced acute lung injury. To elucidate further on this issue, we thus conducted this murine study. Methods: Adult male C57BL/6 mice were randomly allocated to the Sham group (oropharyngeal aspiration of normal saline), the AP group (oropharyngeal aspiration of mimic gastric content) or the AP-EXO group (AP plus exosomes) (n=6 in each group). The mimic gastric content consisted an acidic (pH 1.6) mixture of 12mg/mL ThickenUp®, 25μL pepsin (2mg/mL) and 20μL lipopolysaccharide (2.5mg/mL). In the AP-EXO group, mice received exosomes (1x10^8 particles/mouse, intratracheal, isolated from human placenta-derived MSCs) at 2 and 24 hours after aspiration. All mice were closely monitored for 48 hours, and each group’s lung injury level was measured. Results: Aspiration induces significant alterations in respiratory function, as our data revealed significantly higher airway resistance and lower forced expiratory volume in 0.1 seconds (FEV0.1) and dynamic compliance in mice of the AP group compared with the Sham group (all p< 0.001). Aspiration also induces significant airway inflammation, lung tissue edema and lung injury, as our data revealed significantly higher cell numbers in bronchoalveolar fluid, wet/dry weight ratio of lung tissue and lung injury score in mice of the AP group compared with the Sham group (all p< 0.001). These adverse effects of aspiration were mitigated by exosomes from MSCs, as our data demonstrated significantly lower airway resistance and, in contrast, higher FEV0.1 and dynamic compliance. Also, cell number in BALF, wet/dry weight ratio of lung tissues and lung injury score decreased in mice of the AP-EXO group compared with the AP group (all p< 0.001). Conclusions: Exosomes from MSCs mitigate aspiration-induced acute lung injury in mice.

Change over time and agreement between clinical markers of disease resolution in dogs with aspiration-induced lung injury.

There is a lack of consensus regarding which markers of disease resolution to prioritise when assessing treatment response in client-owned dogs with aspiration-induced lung injury. This study describes the change over time and the agreement between the clinical markers used to determine disease resolution. Physical examination (PE), owner-reported clinical signs (CS-O), thoracic radiographs (TXR) scores and C-reactive protein (CRP) concentrations were determined at enrolment, after 24, 48 and 72 hours, and after 7, 14 and 28 days. PE scores were significantly improved at 48 hours, while CRP initially increased (24 hours) and then decreased (48 hours). PE, CS-O and CRP significantly improved earlier (7 days) than TXR (14 days). The median number of days to marker normalisation was 7, 9 and 14 for PE, CRP and CS-O, respectively. Marker agreement was excellent/very good at enrolment and fair/poor during disease recovery. Analysis did not control for differences aetiology of aspiration or the lack of standardisation in treatment approach. PE was the earliest and most consistent marker indicating disease resolution. Serial CRP monitoring (72 hours) may provide an objective marker of early treatment response. Alongside PE normalisation, improvement in CS-O, CRP and TRX may assist in determining disease resolution and guide treatments, including limiting antibiotic exposure in dogs with aspiration-induced lung injury.

ACE2-like enzyme B38-CAP suppresses abdominal sepsis and severe acute lung injury.

Angiotensin-converting enzyme 2 (ACE2) is the carboxypeptidase to degrade angiotensin II (Ang II) to angiotensin 1–7 (Ang 1–7) and improves the pathologies of cardiovascular disease and acute respiratory distress syndrome (ARDS)/acute lung injury. B38-CAP is a bacteria-derived ACE2-like carboxypeptidase as potent as human ACE2 and ameliorates hypertension, heart failure and SARS-CoV-2-induced lung injury in mice. Recombinant B38-CAP is prepared with E. coli protein expression system more efficiently than recombinant soluble human ACE2. Here we show therapeutic effects of B38-CAP on abdominal sepsis- or acid aspiration-induced acute lung injury. ACE2 expression was downregulated in the lungs of mice with cecal ligation puncture (CLP)-induced sepsis or acid-induced lung injury thereby leading to upregulation of Ang II levels. Intraperitoneal injection of B38-CAP significantly decreased Ang II levels while upregulated angiotensin 1–7 levels. B38-CAP improved survival rate of the mice under sepsis. B38-CAP suppressed the pathologies of lung inflammation, improved lung dysfunction and downregulated elevated cytokine mRNA levels in the mice with acute lung injury. Thus, systemic treatment with an ACE2-like enzyme might be a potential therapeutic strategy for the patients with severe sepsis or ARDS.

Use of Early Bronchoscopy in Mechanically Ventilated Patients with Aspiration Pneumonitis

Background and objectivesAspiration-induced lung injury accounts for a significant proportion of acute pulmonary dysfunction. Few studies were conducted to study the use of early bronchoscopy in mechanically ventilated patients with aspiration pneumonitis. This study aimed at assessing the clinical impact of early bronchoscopy for removal of gastric fluid and solid particles in the first 24 hours of mechanical ventilation (MV) on the progression of aspiration, MV days, intensive care unit (ICU) stay, development of pneumonia, and ICU mortality.Materials and methodsThe study was an open-label randomized control trial and included 76 adult subjects mechanically ventilated due to aspiration pneumonitis, half the subjects received early bronchoscopy in the first 24 hours after aspiration for removal of aspirated material and bronchoalveolar lavage sampling, the other half received standard treatment.ResultsThe intervention group had a significant reduction in the rate of development of pneumonia at 60.5 vs 81.6%, p = 0.043 through the first week of admission, the intervention group has a significantly better hypoxic index (HI), white blood count, clinical pulmonary infection score, lung injury score, and sepsis-related organ failure assessment (SOFA) score compared to the control group. Although there was a reduction in mechanical ventilation days and ICU mortality in the intervention group vs control group that difference did not reach statistical significance.ConclusionsEarly bronchoscopy in mechanically ventilated patients with aspiration pneumonitis can be beneficial in improving respiratory functions and decreasing the incidence of development of aspiration pneumonia and may guide the de-escalation of antibiotic therapy.How to cite this articleMegahed MM, El-Menshawy AM, Ibrahim AM. Use of Early Bronchoscopy in Mechanically Ventilated Patients with Aspiration Pneumonitis. Indian J Crit Care Med 2021;25(2):146–152.

Hypoxia-Inducible Factor (HIF)-1α Promotes Inflammation and Injury Following Aspiration-Induced Lung Injury in Mice.

Acid aspiration-induced lung injury is a common disease in the intensive care unit (ICU) and acute respiratory distress syndrome (ARDS). Hypoxia-inducible factor (HIF)-1α is a major transcription factor responsible for regulating the cellular response to changes in oxygen tension. A clear understanding of the function of HIF-1α in lung inflammatory response is currently lacking. Here, we sought to determine the role of HIF-1α in type 2 alveolar epithelial cells (AEC) in the generation of the acute inflammatory response following gastric aspiration (GA). GA led to profound hypoxia at very early time points following GA. This correlated to a robust increase in HIF-1α, tissue albumin and pro-inflammatory mediators following GA in AECs. The extent of lung injury and the release of pro/anti-inflammatory cytokines were significantly reduced in HIF-1α (-/-) mice. Finally, we report that HIF-1α upregulation of the acute inflammatory response is dependent on NF-κB following GA.

Immediate Prone Positioning After Massive Gastric Aspiration Reduces Lung Injury Possibly by Attenuating Interleukin-6-Mediated Lung-Tissue Inflammation in Pigs.

Gastric aspiration, which can cause acute, diffuse, inflammatory lung injury, is of particular concern in critically ill patients. This study aimed to determine the effects of immediate prone positioning on the degree of lung injury and inflammatory response induced by gastric aspiration. Following induction of gastric aspiration by injection of gastric fluid, 16 healthy pigs were randomized to one of two groups: supine position (SP) or prone position (PP). After ventilation and monitoring for 6 hr, all pigs were euthanized. The ratio of the partial pressure of arterial oxygen/fraction of inspired oxygen (PaO2/FIO2) and the partial pressure of arterial carbon dioxide (PaCO2) were recorded during the 6-hr study period. Serum levels of interleukin (IL)-6 were measured every 2 hr, and the mean optical density (MOD) of IL-6 in lung tissues and lung-injury scores were measured at the end of the experiment. The PP group showed a significantly higher PaO2/FIO2 ratio, lower serum IL-6 concentration (p = .015), lower lung-injury scores (p = .012), and lower IL-6 concentration and MOD of IL-6 in lung tissue, especially in dorsal (p = .001, p = .021, respectively) and nondependent regions (p = .005, p = .035, respectively) than the SP group. There were no statistically significant differences in PaCO2 between the groups. Lung-injury severity was positively correlated with the IL-6 concentration and MOD of IL-6 in lung tissues (p < .05). These results suggest that immediate prone positioning alleviated the degree of aspiration-induced lung injury, possibly through mitigating IL-6-mediated lung inflammation.

Mechanical Ventilation Strategies Targeting Different Magnitudes of Collapse and Tidal Recruitment in Porcine Acid Aspiration-Induced Lung Injury.

Reducing ventilator-associated lung injury by individualized mechanical ventilation (MV) in patients with Acute Respiratory Distress Syndrome (ARDS) remains a matter of research. We randomly assigned 27 pigs with acid aspiration-induced ARDS to three different MV protocols for 24 h, targeting different magnitudes of collapse and tidal recruitment (collapse&TR): the ARDS-network (ARDSnet) group with low positive end-expiratory pressure (PEEP) protocol (permissive collapse&TR); the Open Lung Concept (OLC) group, PaO2/FiO2 >400 mmHg, indicating collapse&TR <10%; and the minimized collapse&TR monitored by Electrical Impedance Tomography (EIT) group, standard deviation of regional ventilation delay, SDRVD. We analyzed cardiorespiratory parameters, computed tomography (CT), EIT, and post-mortem histology. Mean PEEP over post-randomization measurements was significantly lower in the ARDSnet group at 6.8 ± 1.0 cmH2O compared to the EIT (21.1 ± 2.6 cmH2O) and OLC (18.7 ± 3.2 cmH2O) groups (general linear model (GLM) p < 0.001). Collapse&TR and SDRVD, averaged over all post-randomization measurements, were significantly lower in the EIT and OLC groups than in the ARDSnet group (collapse p < 0.001, TR p = 0.006, SDRVD p < 0.004). Global histological diffuse alveolar damage (DAD) scores in the ARDSnet group (10.1 ± 4.3) exceeded those in the EIT (8.4 ± 3.7) and OLC groups (6.3 ± 3.3) (p = 0.16). Sub-scores for edema and inflammation differed significantly (ANOVA p < 0.05). In a clinically realistic model of early ARDS with recruitable and nonrecruitable collapse, mechanical ventilation involving recruitment and high-PEEP reduced collapse&TR and resulted in improved hemodynamic and physiological conditions with a tendency to reduced histologic lung damage.

Heme oxygenase-1 attenuates seawater drowning-induced acute lung injury through a reduction in inflammation and oxidative stress

ObjectiveHeme oxygenase-1 (HO-1) plays a critical protective role in various insults-induced acute lung injury (ALI) through its strong anti-inflammatory, anti-oxidant, and anti-apoptotic properties, but its protective role and mechanism on seawater aspiration-induced acute lung injury remains unclear. This study aimed to explore the therapeutic potential and mechanism of HO-1 to attenuate seawater aspiration-induced ALI in vivo and in vitro. MethodsThe viability and invasion of A549 cell were analyzed through cell counting kit-8 and lactate dehydrogenase release assay; the transcriptional level of inflammatory cytokines (TNF-α, IL-6, IL-8 and MCP-1) and cell proliferation-related cytokines (FoxM1, Ccnb1 and Cdc25C) in seawater-treated A549 cell were tested by qPCR; apoptotic cells were analyzed by flow cytometryd; HO-1mRNA and protein were determined by qPCR and western blotting; the fluorescent indicators (DCFH-DA, dihydroethidium, MitoSox Red and Fluo-4) were used to monitor generation of ROS and mitochondrial function. The lung wet/dry weight radio and lactate dehydrogenase activity, Sirius red staining, TUNEL assay and immunohistochemical staining with anti-pan Cytokeratin antibody were analyzed in seawater-drowning mice. The role of HO-1 on seawater-drowning pulmonary injury was explored via HO-1 activity inhibitors (Zinc protoporphyrin) in vitro and in vivo. ResultsSeawater exposure decreased the cellular viability, increased the production of pro-inflammatory cytokines (IL-6, IL-8 and TNF-α), induced cellular apoptosis and inhibited the expression of cell proliferation-related cytokines (FoxM1, Ccnb1 and Cdc25C). Moreover, seawater exposure led to mitochondrial dysfunction in A549 cells. Supplement of HO-1 sepcific inducer (heme) or its catalytic product (biliverdin) significantly attenuated seawater-induced A549 damage and promoted cell proliferation. However, Zinc protoporphyrin abolished the beneficial effects of HO-1 on seawater drowning-induced pulmonary tissue injury. ConclusionHO-1 attenuates seawater drowning-induced lung injury by its anti-inflammatory, anti-oxidative, and anti-apoptosis function.

ACE-2/ANG1-7 ameliorates ER stress-induced apoptosis in seawater aspiration-induced acute lung injury.

Previous studies have shown that apoptosis of alveolar cells can be regulated by autocrine of angiotensin (ANG)II and its counter regulatory ACE-2/ANG1-7 axis. Our earlier study has shown that endoplasmic reticulum (ER) stress in response to seawater aspiration eventually led to apoptosis in lung tissue. In this study, we examined the hypothesis that ER stress-induced apoptosis in seawater aspiration-induced acute lung injury (ALI) might also be regulated by the ANGII/ANG1-7 system. ER stress was induced by seawater stimulation and proteasome inhibitor MG132 (an ER stress inductor). Moreover, ER stress in seawater-stimulated lung tissues and rat pulmonary microvascular endothelial cells (RPMVECs) promoted ANGII expression and decreased ACE-2/ANG1-7 expression. ER stress induced by seawater stimulation also led to apoptosis. Apoptosis induced by seawater stimulation and MG132 were inhibited by ANGII receptor blocker and abrogated by the addition of ANG1-7. These results suggest that apoptosis induced by ER stress in seawater aspiration-induced ALI is regulated by ANG II/ANG1-7 in lung tissues and RPMVECs. In addition, the active form of X-box binding protein 1 (XBP1), spliced XBP1 (XBP1s), a transcription factor that regulates ER-associated degradation genes during ER stress was significantly activated in seawater stimulated cells. Based on this phenomenon we designed a tandem gene, Wfs1 promoter (a target gene promoter of XBP1s)- ACE2 and ANG1-7 and transfected this tandem gene into seawater-stimulated cells. ACE-2/ANG1-7 expression were significantly promoted and apoptosis was inhibited in cells transfected with the tandem gene. These results suggest that stimulation of ACE-2/ANG1-7 may be a therapeutic target of ER stress-induced apoptosis in seawater aspiration-induced ALI.

Instillation of hyaluronan reverses acid instillation injury to the mammalian blood gas barrier.

Acid (HCl) aspiration during anesthesia may lead to acute lung injury. There is no effective therapy. We hypothesized that HCl instilled intratracheally in C57BL/6 mice results in the formation of low-molecular weight hyaluronan (L-HA), which activates RhoA and Rho kinase (ROCK), causing airway hyperresponsiveness (AHR) and increased permeability. Furthermore, instillation of high-molecular weight hyaluronan (H-HA; Yabro) will reverse lung injury. We instilled HCl in C57BL/6 wild-type (WT), myeloperoxidase gene-deficient (MPO-/-) mice, and CD44 gene-deficient (CD44-/-) mice. WT mice were also instilled intranasally with H-HA (Yabro) at 1 and 23 h post-HCl. All measurements were performed at 1, 5, or 24 h post-HCl. Instillation of HCl in WT but not in CD44-/- resulted in increased inflammation, AHR, lung injury, and L-HA in the bronchoalveolar lavage fluid (BALF) 24 h post-HCl; L-HA levels and lung injury were significantly lower in HCl-instilled MPO-/- mice. Isolated perfused lungs of HCl instilled WT but not of CD44-/- mice had elevated values of the filtration coefficient ( Kf). Addition of L-HA on the apical surface of human primary bronchial epithelial cell monolayer decreased barrier resistance ( RT). H-HA significantly mitigated inflammation, AHR, and pulmonary vascular leakage at 24 h after HCl instillation and mitigated the increase of Kf and RT, as well as ROCK2 phosphorylation. Increased H- and L-HA levels were found in the BALF of mechanically ventilated patients but not in healthy volunteers. HCl instillation-induced lung injury is mediated by the L-HA-CD44-RhoA-ROCK2 signaling pathway, and H-HA is a potential novel therapeutic agent for acid aspiration-induced lung injury.

Beneficial effects of Caesalpinia digyna extract against acid aspiration-Induced acute lung injury in mice

Objective: Caesalpinia digyna belongs to the genus Caesalpinia, which is known since ancient times for its medicinal properties. The present work was designed to evaluate the beneficial potential of hydroalcoholic extract of the roots of C. dignea against hydrochloric acid (HCl)-induced acute lung injury in mice. Materials and Methods: Ethanolic extract of C. dignea roots at a dose of 50, 100, or 200 mg/kg boy weight was given once orally 90 min before HCl administration. Mice were then analyzed for infiltration of inflammatory cells in bronchoalveolar lavage fluid (BALF) and oxidative stress markers in the lung tissue. Further, the effects of the extract were compared with bergenin isolated from the extract. Results: Our results showed that an oral administration of the extract 90 min before HCl instillation reduced the infiltration of neutrophils in the lungs in a dose-dependent manner. Reduction in lung inflammation was associated with decline in pulmonary edema as the total protein content in the BALF was found to be decreased substantially. The drug also restored the redox balance in the lungs toward normal on HCl treatment as assessed by measuring the levels of reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), and catalase activity. Bergenin, isolated from the plant, was able to suppress the neutrophils but increased the macrophage number in BALF when administered before HCl instillation, suggesting immunoregulatory properties of the key constituent of the extract. Conclusion: Our data suggest that hydroalcoholic extract of Caesalpinia digyna roots constitute the phytochemicals that can protect against HCl-induced acute lung injury in mice. Abbreviations Used: ALI: Acute lung injury, BALF: Bronchoalveolar lavage fluid, MDA: Malondialdehyde, ROS: Reactive oxygen species, DCF-DA: 2' 7'-dichlorofluorescein diacetate

Myeloid but not epithelial tissue factor exerts protective anti-inflammatory properties in acid aspiration-induced acute lung injury

Myeloid but not epithelial tissue factor exerts protective anti-inflammatory properties in acid aspiration-induced acute lung injury

Endothelial semaphorin 7A promotes seawater aspiration-induced acute lung injury through plexin C1 and β1 integrin.

Inflammation and edema are two main characteristics in seawater aspiration‑induced acute lung injury (ALI). In a previous study of the authors, it was demonstrated that endothelial semaphorin 7A (SEMA7A) serves an important role in the development of seawater‑induced inflammation and edema. However, the mechanism of endothelial SEMA7A‑mediated ALI remains unclear. Therefore, the authors explored the effect of SEMA7A in rat pulmonary microvascular endothelial cells (RPMVECs) and the interaction between endothelial SEMA7A and alveolar macrophages during seawater aspiration‑induced ALI. The role of SEMA7A in endothelial permeability was detected using plexin C1 blocking antibody or SEMA7A small interfering (si)RNA. In addition, RPMVECs were co‑cultured with rat alveolar macrophage cell line‑NR8383 cells and pro‑inflammatory cytokine production was detected. Interaction between the β1 integrin and SEMA7A was detected using the β1 integrin blocking antibody or SEMA7A siRNA. Seawater stimulation induced endothelial cytoskeleton remodeling, endothelial permeability, phosphorylation of cofilin, and increased the vascular endothelial growth factor (VEGF) expression in RPMVECs. Moreover, seawater stimulation led to expression of proinflammatory cytokines and activated the nuclear factor‑κB pathway in co‑cultured cells. However, blockage with the plexin C1 antibody inhibited endothelial cytoskeleton remodeling, endothelial permeability, phosphorylation of cofilin, and treatment with SEMA7A siRNA inhibited expression of VEGF in RPMVECs. In addition, blockage with β1 integrin antibody reduced expression of proinflammatory cytokines and inhibited activation of NF‑κB in co‑culture cells. These results suggest that SEMA7A promotes seawater induced lung edema via plexin C1 and stimulates seawater induced lung inflammation via β1 integrin.

1\u03b1,25-Dihydroxyvitamin D3 Ameliorates Seawater Aspiration-Induced Lung Injury By Inhibiting The Translocation Of NF-\u03baB and RhoA

Our previous study have reported that 1α,25-Dihydroxyvitamin D3 (calcitriol) suppresses seawater aspiration-induced ALI in vitro and in vivo. We also have confirmed that treatment with calcitriol ameliorates seawater aspiration-induced inflammation and pulmonary edema via the inhibition of NF-κB and RhoA/Rho kinase pathway activation. In our further work, we investigated the effect of calcitriol on nuclear translocation of NF-κB and membrane translocation of RhoA in vitro. A549 cells and rat pulmonary microvascular endothelial cells (RPMVECs) were cultured with calcitriol or not for 48 h and then stimulated with 25% seawater for 40 min. After these treatments, cells were collected and performed with immunofluorescent staining to observe the translocation of NF-κB and RhoA and the cytoskeleton remodeling. In vitro, seawater stimulation activates nuclear translocation of NF-κB and membrane translocation of RhoA in A549 cells. In addition, seawater administration also induced cytoskeleton remodeling in A549 cells and RPMVECs. However, pretreatment with calcitriol significantly inhibited the activation of NF-κB and RhoA/Rho kinase pathways, as demonstrated by the reduced nuclear translocation of NF-κB and membrane translocation of RhoA in A549 cells. Meanwhile, treatment of calcitriol also regulated the cytoskeleton remodeling in both A549 cells and RPMVECs. These results demonstrated that treatment with calcitriol ameliorates seawater aspiration-induced ALI via inhibition of nuclear translocation of NF-κB and membrane translocation of RhoA and protection of alveolar epithelial and pulmonary microvascular endothelial barrier.

Assessment of Aspiration-Induced lung Injuries among Acute Drug Poisoning Patients; Loghman Hakim Hospital, Poisoning center

Background: Following pulmonary aspiration of gastric contents, a variety of conditions occurs ranging from the acute inflammatory response (aspiration pneumonitis) to an infectious process (aspiration pneumonia). They cause due to the failure of protecting mechanism of airway patency following poisoning toxicity. Methods: Patients with acute drug poisoning for assessment of aspiration-induced lung injury evaluated during the twelfth months. Results: 410 participants followed the study. Of those, 249 (61 %) were male and 161 (39%) female. They ranged between 17-55 years. The mode was 25-39 years with frequency of 38%. The mean±SD of age was 32.3±14.5 (Female 34% and Male 27% respectively). The mean age ±SD of females was lower than males (27.3±12.4 vs. 34.4±14.4 years old). An episode of vomiting was recorded positive in 62% (254) of patients. They were recorded by self-reported or witnessed by someone else within the first hours of poisoning. The chest radiograph abnormality was observed mostly in both lungs (45%, 185). They consisted of multifocal patchy infiltrates in lungs 44% (178), diffuse bilateral infiltration 35% (144), and bilateral perihilar consolidation 18% (72) and lobular consolidation 4% (16) mainly in both lung fields (45%, 185). Aspiration related pulmonary complications attributed to pneumonitis 43.5% (178) including ARDS 35% (144), pneumonia 18% (72), and lung abscess 4% (16). There was a significant relationship with vomiting (P=0. 04). Conclusion: Aspiration pulmonary abnormalities were the commonest lung manifestation of drug toxicities. Chest x-ray abnormalities associated with vomiting presentation are clues to finding pulmonary complications of acute drug toxicity.

Myeloid but not epithelial tissue factor exerts protective anti-inflammatory effects in acid aspiration-induced acute lung injury

Myeloid but not epithelial tissue factor exerts protective anti-inflammatory effects in acid aspiration-induced acute lung injury