Alveolar Wall Thickness Research Articles

P48 Hydrogen sulfide mediated Atf3 activation is involved in protection against ventilator induced lung injury

Rational Our group has recently shown that inhalation of hydrogen sulfide (H2S) during mechanical ventilation protects against VILI [1] . Here we explored the gene expression profile of this model in order to identify underlying molecular mechanisms. Methods C57/BL6 mice were randomized into four groups (n = 5/group). Two groups of control animals were breathing spontaneously air with or without H2S (80 ppm). Two groups were ventilated with synthetic air with or without H2S as described previously [1] . Lung injury was assessed by an acute lung injury scoring system in hematoxylin and eosin (H& E) stained tissue sections. Bronchoalveolar lavage fluid (BALF) was analysed for inflammatory infiltration. Micro array analysis was performed with RNA from lung tissue samples. Genes with different expression (ANOVA p ⩽ 0.05; fold induction or suppression ⩾2.2) were considered for further analysis. Gene expression of Atf3 was validated by sqPCR and Western Blot. The functional relevance of Atf3 up-regulation was validated in Atf3-Morpfolino knockout mice ventilated in presence of H2S. Results Mechanical ventilation led to lung injury, i.e., H& E-stained samples revealed clear thickening of alveolar walls and increased amounts of inflammatory cell infiltrates in lung sections and BALF from animals ventilated with synthetic air. In contrast, the presence of H2S prevented lung injury as well as neutrophil and macrophage infiltration. Spontaneous breathing control animals regardless of the presence or absence of H2S showed no significant effects on lung injury or inflammation. H2S application led to differential expression of 137 genes. Among these, Atf3 was chosen for further analysis. In contrast to the protective effects of H2S in ventilated mice, Atf3-Morpholino mice developed despite the presence of H2S sings of lung injury, i.e., enlarged alveolar walls, increased inflammatory cell infiltrates, and occurrence of sporadic blood vessel leakages. Further analysis revealed significant increase of the VILI score in the Atf3-Morpholino group as compared to the air control and control-Morpholino groups. However, histology demonstrated that down-regulated Atf3 protein synthesis failed to completely abolish H2S mediated protection in VILI. Thus, Atf3 significantly contributes to the protective effect of H2S, but seems not to be the only player. Likewise and with respect to the inflammatory response, inhibition Atf3 protein synthesis in H2S ventilated mice resulted in significant increase of neutrophils in BALF as compared to H2S ventilation alone, but neutrophil counts were still lower as compared to injurious ventilation without any treatment. Conclusions (1) VILI can be prevented by application of H2S. (2) 137 genes were differentially expressed in the presence of H2S during mechanical ventilation. (3) Our data suggest that the Atf3 signaling pathway significantly contributes to the lung protective and anti-inflammatory effects of H2S.

The pulmonary toxicity study of nano-silica particles on rats through dynamic inhalation

To investigate the pulmonary toxicity of different concentrations of nano-silica (nano-SiO2) under continuous dynamic inhalation conditions in the rat. 48 male Sprague-Dawley rats were randomly divided into four groups, including the dispersant control group (saline) and nano-SiO2 low-dose group (0.3%, w/v), the middle-dose group (1%) and the high-dose group (3%). Animals were sacrificed at 28 d after exposure under continuous dynamic inhalation conditions, and bronchoalveolar lavage fluid (BALF) and lung tissue were collected. And following items were observed: body coefficient, BALF related items (leukocytes and classification, total protein content, LDH activity), lung tissue pathological changes (HE staining), and pulmonary fibrosis forming (collagen fiber VG staining). Compared to the dispersant control group, there was no significant change on lung organ coefficient in Nano-SiO2 group (P < 0.05). The BALF total WBC count in 1% and 3% in nano-SiO2 groups showed higher value than the dispersant control group (P < 0.05). No obvious changes were found on total protein content and LDH activity in nano-SiO2 groups compared to the dispersant control group (P > 0.05). For differential WBC counts, lymphocyte count in BALF in nano-SiO2 groups was significantly decreased (P < 0.05), monocyte and macrophage counts were significantly increased (P < 0.05), but there was no difference on the proportion of neutrophils (P > 0.05). HE staining results showed that the obvious thickening of alveolar wall in nano-SiO2 groups, inflammatory cell infiltration also increased around the bronchial and vascular wall. Lung fibrosis VG staining showed no significant change of collagen fiber distribution. Under our experimental conditions, the continuous dynamic inhalation of nano-SiO2 only caused the significant inflammation in rat lungs, pulmonary fibrosis phenomenon could not be observed significantly.

Amelioration of carbon tetrachloride-induced pulmonary toxicity with Oxalis corniculata

This research work was planned to investigate the antioxidant potential of methanolic crude extract of Oxalis corniculata (OCME) against lung injuries initiated by carbon tetrachloride (CCl4) in rats at histological and biochemical level. A total of 42 female Sprague Dawley rats were randomly distributed in to seven groups and each group comprised of six rats. Experiment was completed in 22 days (10 doses at alternate days). Group I was not treated (control rats), while group II was administered with vehicles (olive oil and dimethyl sulfoxide), groups III, IV, and V were treated with 1 ml kg(-1) body weight (b.w.) of CCl4 (20% in olive oil). Group III received only CCl4, whereas groups IV and V were administered with 100 and 200 mg kg(-1) b.w. of OCME, respectively. Group VI was administered with OCME (200 mg kg(-1) b.w.) alone. Group VII was treated with sylimarin (50 mg kg(-1) b.w.). CCl4 enhanced the lipid peroxidation while reduced the glutathione in lung samples. Activities of antioxidant enzymes, catalase, peroxidase, superoxide dismutase, and glutathione-S-transferase decreased in lung homogenates with CCl4. Treatment of CCl4 induced deleterious changes in the microanatomy of lungs by rupturing the alveolar septa, thickening of alveolar walls, and damaging the cells with subsequent collapse of blood vessels due to the accumulation of degenerated blood cells. OCME, dose dependently, prevented the alterations in these parameters. These results suggest that OCME protected the lungs due to its intrinsic properties by scavenging of free radicals generated by CCl4.

Abstract 264: Edaravone Injected at the Start of Reperfusion Suppresses Lung Damage Caused by Myonephropathic Metabolic Syndrome in Rats

Introduction &amp; Hypothesis Free radicals have been implicated in myonephropathic metabolic syndrome (MNMS), which damages not only muscles but also lungs. At ATVB 2012, we previously reported that radical scavenger, edaravone (Radicut®, Mitsubishi Tanabe Pharma Co., Japan) injected at the start of reperfusion suppresses muscle injury following leg ischemia in rats. In this study, we evaluated whether edaravone can suppress MNMS-induced lung damage following leg ischemia, if injected at the start of reperfusion. Methods The MNMS models (Male Lewis rats, 525 ± 78 g, n = 10) were induced, by clamping the bilateral common femoral arteries for 5 hours and then de-clamping. The rats were intraperitoneally injected with either 9.0 mg / kg of edaravone (edaravone group; n = 5), or saline (MNMS group; n = 5) at the same time as de-clamping, corresponding to the start of reperfusion. Five hours after de-clamping, the both lungs were harvested and stained with hematoxylin &amp; eosin (HE). Normal lungs without reperfusion were also harvested as control (n = 3). Lung damage was expressed as the percentage area of the alveolar wall thickness, determined by computerized densitometry (NIH Image Program J, Ver. 1.37). Results The lungs of MNMS group had extensive swelling. But the lungs of the edaravone group were filled with air, similar to that of normal lungs (Figure attached, original 200x). The mean percentage area in the edaravone group was significantly lower than that in MNMS group (17.2 ± 5.2 vs. 63.6 ± 5.6%, p &lt; 0.001). Conclusions Our results suggest that edaravone injected at the start of reperfusion can suppress MNMS-induced lung damage following leg ischemia in rats.

Pulmonary inflammation after ethanol exposure and burn injury is attenuated in the absence of IL-6

Alcohol consumption leads to an exaggerated inflammatory response after burn injury. Elevated levels of interleukin-6 (IL-6) in patients are associated with increased morbidity and mortality after injury, and high systemic and pulmonary levels of IL-6 have been observed after the combined insult of ethanol exposure and burn injury. To further investigate the role of IL-6 in the pulmonary inflammatory response, we examined leukocyte infiltration and cytokine and chemokine production in the lungs of wild-type and IL-6 knockout mice given vehicle or ethanol (1.11 g/kg) and subjected to a sham or 15% total body surface area burn injury. Levels of neutrophil infiltration and neutrophil chemoattractants were increased to a similar extent in wild-type and IL-6 knockout mice 24 h after burn injury. When ethanol exposure preceded the burn injury, however, a further increase of these inflammatory markers was seen only in the wild-type mice. Additionally, signal transducer and activator of transcription-3 (STAT3) phosphorylation did not increase in response to ethanol exposure in the IL-6 knockout mice, in contrast to their wild-type counterparts. Visual and imaging analysis of alveolar wall thickness supported these findings and similar results were obtained by blocking IL-6 with antibody. Taken together, our data suggest a causal relationship between IL-6 and the excessive pulmonary inflammation observed after the combined insult of ethanol and burn injury.

The effect of grand maternal nicotine exposure during gestation and lactation on lung integrity of the F2 generation

Maternal nicotine exposure during gestation and lactation adversely affects lung development in the offspring. It has been suggested that the "program" that control long-term maintenance of the structural integrity of the lung may be compromised. The aim of the study was to establish whether the effect of grand-maternal nicotine exposure during gestation and lactation can be transferred to the F2 generation. After mating, rats were randomly divided into two groups (F0). One group received nicotine (1 mg/kg body weight/day). The controls receive saline. Body weight (BW), lung volume (Lv), linear intercept (Lm), alveolar wall thickness (Tsept), senescent and proliferating cell numbers were used to evaluate changes in the lung structure of the offspring (F1). The F1 generation was divided into four groups, namely, (1) control (F1 males mated with F1 females, (2) NmCf (F1 nicotine exposed male mated with F1 control female), (3) NfCm (F1 nicotine exposed female mated with F1 control male), and (4) NmNf (F1 male exposed to nicotine mated with F1 female also exposed to nicotine). The F1 nicotine exposed males and females were exposed to nicotine via the placenta and mother's milk (F0 generation) only. The F2 progeny was never exposed to nicotine. Grand-maternal nicotine (F0) resulted in parenchymal deterioration and emphysema in the F2 progeny due to increased numbers of premature senescent cells together with a slower cell proliferation. The transfer of premature aging characteristics from the F1 progeny to the F2 progeny is via the male and female germ cell line. Grand-maternal nicotine exposure induces structural changes in the lungs of the F2 generation that resembled premature aging.

The influence of intermittent hypoxia and hyperoxia on the respiratory part of the lungs

Summary. An influence of normobaric hypoxic (10 % of oxygen in nitrogen) and hyperoxic (40 % of oxygen in nitrogen) breathing sessions on the respiratory part of the lungs was investigated in rats of 3 month of age. After 28 daily hypoxic sessions, 30 min each, we detected lung hyperinflation, an enlargement of the total alveolar area and reduction in connecting tissue elements in the lungs. After 28 daily hyperoxic sessions, 60 min each, we found reduction in mean diameter, depth, cross-sectional area and the entrance size of alveoli, increased the amount of collagen fibers, the alveolar wall thickness and increased oxyproline concentration in the lungs. These findings could indicate the connecting tissue growth in the respiratory part of the lungs and worsening of oxygen diffusion through the blood-air barrier.

CCK1-Receptor Stimulation Protects Against Gut Mediator-Induced Lung Damage During Endotoxemia

Background/Aims: Cholecystokinin 1-receptor (CCK1-R) activation by long chain fatty acid (LCFA) absorption stimulates vago-vagal reflex pathways in the brain stem. The present study determines whether this reflex also activates the cholinergic anti-inflammatory pathway, a pathway known to modulate cytokine release during endotoxemia. Methods:Mesenteric lymph was obtained from wild type (WT) and CCK1-R knockout (CCK1-R^-/-) mice intraperitoneally challenged with Lipopolysaccharid (LPS) (endotoxemic lymph, EL) and intestinally infused with vehicle or LCFA-enriched solution. The lymph was analyzed for TNFα, IL-6 and IL-10 concentration and administered to healthy recipient mice via jugular infusion. Alveolar wall thickness, myeloperoxidase (MPO) and TUNEL positive cells were determined in lung tissue of recipient mice. Results: LCFA infusion in WT mice reduced TNFα concentration in EL by 49% compared to vehicle infusion, but had no effect in CCK1-R^-/- mice. EL significantly increased the alveolar wall thickness, the number of MPO-positive and TUNEL-positive cells compared to control lymph administration. LCFA infusion in WT, but not in CCK1R^-/- mice, significantly reduced these pathological effects of EL. Conclusion: During endotoxemia enteral LCFA absorption reduces TNFα release into mesenteric lymph and attenuates histomorphologic parameters of lung dysfunction. Failure to elicit this effect in CCK1R^-/- mice demonstrates that anti-inflammatory properties of LCFAs are mediated through CCK1-Rs.

Rho-kinase–dependent F-actin rearrangement is involved in the release of endothelial microparticles during IFN-α–induced endothelial cell apoptosis

Activation of cytoskeleton regulator Rho-kinase during inflammatory stimulations plays a major role in cellular dysfunction and apoptosis. Because endothelial dysfunction may be influenced by increased circulating membrane microparticles (MPs), we hypothesized that inhibition of Rho-kinase can prevent interferon-α (IFN-α)-induced endothelial cell (EC) apoptosis and that protective effects of Rho-kinase inhibition are facilitated by prevention of F-actin rearrangement. In this study, Lewis rats were subjected to an intraperitoneal injection of IFN-α or IFN-α + Y-27632. FCM was performed to analyze circulating endothelial MPs (EMPs) from the blood samples of these animals by detecting the expression of CD144, CD62E, CD31, CD51, and CD54 on EMPs. IFN-α-induced pulmonary injury was assessed by measurement of lung wet-to-dry weight ratios and measurement of alveolar wall thickness. Human pulmonary microvascular ECs (HPMECs) were cultured with IFN-α or EMPs to elucidate the probable mechanisms of the release of EMPs. Injection of IFN-α resulted in much higher levels of CD144 EMPs, CD62E EMPs, CD31 EMPs, CD51 EMPs, and CD54 EMPs. Pulmonary injury was also observed after injection of IFN-α. Furthermore, IFN-α induced F-actin rearrangement and apoptosis of HPMECs in vitro, and the Toll-like receptor 4/MyD88 and nuclear factor-κB pathways and EMPs per se played important roles in this process. The results demonstrate that increased Rho-kinase activity causes the release of EMPs and cellular apoptosis. Moreover, HPMEC apoptosis that resulted from EMP stimulation indicates that EMPs can be considered as a potential target to regulate the rearrangement of cytoskeleton during endothelial cell apoptosis.

Acute mechanical forces cause deterioration in lung structure and function in elastase-induced emphysema.

The relation between the progression of chronic obstructive pulmonary disease (COPD) and exacerbations is unclear. Currently, no animal model of acute exacerbation of COPD (AECOPD) exists. The objectives of this study were to evaluate the effects of mechanical forces induced by deep inspirations (DIs) on short-term deterioration of lung structure and function to mimic AECOPD. At 2, 7, or 21 days after treatment with elastase, mice were ventilated with or without DIs (35 cmH(2)O airway pressure for 3 s, 2 times/min) for 1 h. Functional residual capacity (FRC) was measured with body plethysmography, and respiratory compliance, resistance, and hysteresivity were obtained via forced oscillations. From hematoxylin and eosin-stained sections, equivalent airspace diameters (D), alveolar wall thickness (W(t)), number of septal ruptures (N(sr)), and attachment density (A(d)) around airways were determined. FRC, compliance, and hysteresivity statistically significantly increased with time, and both increased due to DIs. Interestingly, DIs also had an effect on FRC, compliance, resistance, and hysteresivity in control mice. The development of emphysema statistically significantly increased D and W(t) in time, and the DIs caused subtle differences in D. At 21 days, the application of DIs changed the distribution of D, increased W(t) and N(sr), and decreased A(d). These results suggest that once a critical remodeling of the parenchyma has been reached, acute mechanical forces lead to irreversible changes in structure and function, mimicking COPD exacerbations. Thus, the acute application of DIs in mice with emphysema may serve as a useful model of AECOPD.

Exploratory study using proton induced X-ray emission analysis and histopathological techniques to determine the toxic burden of environmental pollutants

The aim of this novel research was to determine the toxic burden of increased elements in water resources on the inhabitant wild animals (squirrels, turtles, bats), using particle induced x-ray emission (PIXE) and histopathological approaches. PIXE analysis of skin, muscle, lung, liver and kidney revealed significant increase in Al, Cl, Fe, Mg, Mn, Si and V. Moreover, data clearly reflect a significant (P < 0.001) deposition of toxic elements (Al, Cl, Fe and K) in the lung producing interstitial/proliferative pneumonitis, intra-alveolar hemorrhages, and thickening of alveolar capillary walls. The results obtained from the liver samples emphasized that majority of the animals were intoxicated with Cl, Mg, S, Si and V, which have produced profound deterioration and swelling of the hepatocytes. Likewise, histopathology of the kidney sections spotlighted severe nephritis and degenerative changes, which could be associated with the elevated amount of Al, Cl and Mg. This data undoubtedly provide relevant information on the heavy burden of toxic elements and their pathological outcomes in wild animals and highlight their potential risks for human exposure. Thus, the information provided is critical for developing effective strategies in dealing with health hazards associated with elemental exposures.

Kaempferol regulates MAPKs and NF-κB signaling pathways to attenuate LPS-induced acute lung injury in mice

Recent studies show that mitogen-activated protein kinases (MAPKs) and nuclear factor-kappa B (NF-κB) signaling pathways are two pivotal roles contributing to the development of lipopolysaccharide (LPS)-induced acute lung injury (ALI). The present study aimed to investigate the protective effect of kaempferol (Kae), a naturally occurring flavonoid compound, on ALI and explore its possible mechanisms. Male BALB/c mice with ALI, induced by intranasal instillation of LPS, were treated or not with Kae (100mg/kg, intragastrically) 1h prior to LPS exposure. Kae treatment attenuated pulmonary edema of mice with ALI after LPS challenge, as it markedly decreased the lung W/D ratio of lung samples, protein concentration and the amounts of inflammatory cells in BALF. Similarly, LPS mediated overproduction of proinflammatory cytokines in BALF, including TNF-α, IL-1β and IL-6, was strongly reduced by Kae. Histological studies demonstrated that Kae substantially inhibited LPS-induced alveolar wall thickness, alveolar hemorrhage and leukocytes infiltration in lung tissue with evidence of reduced myeloperoxidase (MPO) activity. Kae also efficiently increased superoxide dismutase (SOD) activity of lung sample when compared with LPS group, which was obviously reduced by LPS administration. In addition, Western blot analysis indicated that the activation of MAPKs and NF-κB signaling pathways stimulated by LPS was significantly blocked by Kae. Taken together, our results suggest that Kae exhibits a protective effect on LPS-induced ALI via suppression of MAPKs and NF-κB signaling pathways, which may involve the inhibition of tissue oxidative injury and pulmonary inflammatory process.

Kinetic effects of carbon monoxide inhalation on tissue protection in ventilator-induced lung injury

Mechanical ventilation causes ventilator-induced lung injury (VILI), and contributes to acute lung injury/acute respiratory distress syndrome (ALI/ARDS), a disease with high morbidity and mortality among critically ill patients. Carbon monoxide (CO) can confer lung protective effects during mechanical ventilation. This study investigates the time dependency of CO therapy with respect to lung protection in animals subjected to mechanical ventilation. For this purpose, mice were ventilated with a tidal volume of 12 ml/kg body weight for 6 h with air in the absence or presence of CO (250 parts per million). Histological analysis of lung tissue sections was used to determine alveolar wall thickening and the degree of lung damage by VILI score. Bronchoalveolar lavage fluid was analyzed for total cellular influx, neutrophil accumulation, and interleukin-1β release. As the main results, mechanical ventilation induced pulmonary edema, cytokine release, and neutrophil recruitment. In contrast, application of CO for 6 h prevented VILI. Although CO application for 3 h followed by 3-h air ventilation failed to prevent lung injury, a further reduction of CO application time to 1 h in this setting provided sufficient protection. Pre-treatment of animals with inhaled CO for 1 h before ventilation showed no beneficial effect. Delayed application of CO beginning at 3 or 5 h after initiation of ventilation, reduced lung damage, total cell influx, and neutrophil accumulation. In conclusion, administration of CO for 6 h protected against VILI. Identical protective effects were achieved by limiting the administration of CO to the first hour of ventilation. Pre-treatment with CO had no impact on VILI. In contrast, delayed application of CO led to anti-inflammatory effects with time-dependent reduction in tissue protection.

Mast Cell Infiltration Discriminates between Histopathological Phenotypes of Chronic Obstructive Pulmonary Disease

COPD is a complex disease with heterogeneous manifestations. Attempts have been made to define different phenotypes that could guide toward better disease understanding. We described before that smokers can develop either panlobular (PLE) or centrilobular emphysema (CLE). The latter has worse small airways remodeling and narrowing, which account for the airflow obstruction similar to asthma. Because of the small airways involvement in CLE similar to asthma, we hypothesized a role for mast cells in CLE but not in PLE. Hence, we investigated mast cell infiltration, along with overall inflammation, and their relation with hyperreactivity and emphysema type in COPD. We studied lung function, emphysema type, mast cells, and overall inflammation in small airways and alveolar walls, along with alveolar wall thickening in 67 subjects undergoing lung resection (59 smokers, 8 nonsmokers). Twenty-seven smokers had CLE, 24 had PLE, and 8 had no emphysema. Mast cells were significantly increased in CLE compared with PLE and control subjects. Especially relevant was the mast cell increase in airway smooth muscle in CLE, which related significantly to airway hyperreactivity. CD4(+)T cells, neutrophils, and macrophages, but not eosinophils and CD8(+)T cells, were significantly higher in CLE than PLE. Alveolar wall thickness was increased in all smokers, but significantly more in CLE. The pathological phenotypes of COPD CLE and PLE show important differences in their overall inflammation with a protagonism of mast cells, which are related to airway reactivity. These findings highlight the distinctness of these COPD phenotypes and the role of mast cells in the pathophysiology of COPD.

Significance of celecoxib on enhanced metastatic potential due to hepatic artery occlusion in hepatocellular carcinoma

Objective To investigate the local or systemic inflammatory reaction after hepatic artery ligation ( HAL),and to explore the therapeutic effect of celecoxib on HAL-enhanced metastatic potential of hepatocellular carcinoma (HCC).Methods The spontaneous metastatic model in nude mice was established with MHCC97 cells via orthotopic implantation and further to HAL.The proinflammatory cytokines such as tumor necrosis factor (TNF)-α,interleukin (IL)-6,and hepatocyte growth factor ( HGF ) in serum in HAL group were compared with those in sham-operation group by enzyme linked immunosorbent assay (ELISA).The signs of inflammation in primary tumor or the lung from two groups were observed by immunohistochemistry and the expression of cyclooxgenase 2 (COX-2) in xenografts was evaluated.The influence of celecoxib at different doses on tumor growth,pulmonary metastatic ratio,and COX-2 expression in tumor tissues due to HAL was investigated.Results The serum levels of proinflammatory cytokines (TNF-α,IL-6 and HGF) were elevated after HAL (3.78 times; 2.91 times; 1.82 times,P ＜ 0.05 ).Also the necrotic area in the xenograft as scored with hepatic replacement area (HRA) was increased by HAL [ HAL group:(5 1.27 ± 19.39 ) ％ vs.sham-operation group:( 19.69 ± 5.49 ) ％ ; P ＜ 0.01 ].Especially,the lungs of nude mice in the HAL group showed more inflammatory damage,characterized by intense inflammatory infiltration,thickening of alveolar walls,and severe vascular margination.As compared with normal saline (NS) treatment following HAL,HAL in combination with high dose of celecoxib (50 mg/kg every day) resulted in significantly reduced tumor size [( 1091.81 ± 870.14) mm3 vs. (2735.06 ±474.97 ) mm3,P ＜0.01 ],decreased pulmonary metastasis (0/6 vs.8/10,P ＜0.01 ),and prolonged survival [ ( 81.67 ± 8.51 ) days vs.( 60.17 ± 5.42 ) days,P ＜ 0.05 ].HAL followed by celecoxib at a dose of 12.5 mg/kg every day neither slowed down residual tumors growth nor prolonged survival,but inhibited the formation of pulmonary metastatic nodules though the difference (compared with sham operation) was not statistically significant (2/6 vs.8/10,P ＞0.05).Interestingly,as shown in supplementary study,we did not detect the conspicuous changes of COX-2 expression in tumor tissues-derived HAL with low dose celecoxib-trcated mice,while the striking decrease of COX-2 expression was observed in treatments by HAL combined with moderate or high dose celecoxib.Conclusion HLA leads to local or systemic inflammatory reaction,which is inhibited by celecoxib as accompanied with arrest of HAL-induced metastatic potential in residual HCC cells. Key words: Hepatic artery blocking; Hepatocellular carcinoma; Inflammation; Celecoxib

Grape seed extract attenuates lung parenchyma pathology in ovalbumin-induced mouse asthma model: An ultrastructural study

Due to the growing incidence of asthma and because of the non-specificity and side effects of the conventional drugs, the development of novel agents for the treatment of asthma has become considerably important. Natural plant products offer promising alternatives for the development of effective and safe treatments. Grape seed extract (GSE) is one such phytochemical supplement that has been shown to have potent antioxidant and anti-inflammatory effects. Thus, the present study aimed to investigate the effect of GSE to suppress lung parenchyma pathology and inflammation in ovalbumin-induced murine asthma model. Ovalbumin exposure was associated with many pathological and morphometric alterations in the lungs of asthmatic mice. The alterations involved alveolar size reduction, alveolar wall thickening, cellular infiltration and blood capillary congestion, as well as significant increase in the number of type II pneumocytes and lamellar bodies. However, GSE significantly ameliorated of the pathological changes of ovalbumin-induced asthma. The results support the possibility of GSE as an effective, safe anti-inflammatory dietary supplement to attenuate the pathogenicity of asthma. While these preliminary results appear promising, further studies are required to elucidate the precise mechanism of the modulatory effect of GSE on asthma remodeling.

Licochalcone A Inhibits Lipopolysaccharide-Induced Inflammatory Response in Vitro and in Vivo

Licochalcone A (Lico A), a flavonoid found in licorice root (Glycyrrhiza glabra), is known for its antimicrobial activity and its reported ability to inhibit cancer cell proliferation. In the present study, we found that Lico A exerted potent anti-inflammatory effects in in vitro and in vivo models induced by lipopolysaccharide (LPS). The concentrations of TNF-α, interleukin (IL)-6, and IL-1β in the culture supernatants of RAW 264.7 cells were determined at different time points following LPS administration. LPS (0.5 mg/kg) was instilled intranasally (i.n.) in phosphate-buffered saline to induce acute lung injury, and 24 h after LPS was given, bronchoalveolar lavage fluid was obtained to measure pro-inflammatory mediator and total cell counts. The phosphorylation of mitogen-activated protein kinases (MAPKs) and nuclear factor-κB (NF-κB) p65 protein was analyzed by Western blotting. Our results showed that Lico A significantly reduced the amount of inflammatory cells, the lung wet-to-dry weight (W/D) ratio, protein leakage, and myeloperoxidase activity and enhances oxidase dimutase activity in mice with LPS-induced acute lung injury (ALI). Enzyme-linked immunosorbent assay results indicated that Lico A can significantly down-regulate TNF-α, IL-6, and IL-1β levels in vitro and in vivo, and treatment with Lico A significantly attenuated alveolar wall thickening, alveolar hemorrhage, interstitial edema, and inflammatory cells infiltration in mice with ALI. In addition, we further demonstrated that Lico A exerts an anti-inflammation effect in an in vivo model of acute lung injury through suppression of NF-κB activation and p38/ERK MAPK signaling in a dose-dependent manner.

Patterns of Interstitial Lung Disease During Everolimus Treatment in Patients with Metastatic Renal Cell Carcinoma

To elucidate the patterns of interstitial lung disease during everolimus treatment in patients with metastatic renal cell carcinoma, we reviewed seven cases of everolimus-induced interstitial lung disease. Seven patients with metastatic renal cell carcinoma, which continued to progress despite treatment with sunitinib or sorafenib, developed interstitial lung disease after treatment with everolimus. Chest X-ray demonstrated diffuse infiltrates in lung fields, and chest computed tomography showed bilateral reticular and ground-glass opacities. Serum levels of lactate dehydrogenase (7/7), C-reactive protein (6/7), pulmonary surfactant associated protein D (1/7) and Krebs von den Lungen 6 (5/7) were elevated. The bronchoalveolar lavage fluid obtained from four patients with Grade 3 interstitial lung disease showed lymphocytosis. The transbronchial lung biopsy specimens showed interstitial lymphocytic infiltration and septal thickening of alveolar walls. In two cases with mild interstitial lung disease, the everolimus therapy was successfully continued. In four cases with Grade 3 interstitial lung disease, the drug was discontinued and steroid therapy was initiated. Pulmonary symptoms and radiological abnormalities resolved within 2 months. Serum Krebs von den Lungen 6 was elevated compared with baseline in all cases with interstitial lung disease. Some patients who developed mild interstitial lung disease during everolimus treatment could continue to receive the treatment. Even when severe interstitial lung disease developed, withdrawal of the drug and short-term use of high-dose steroids resulted in rapid recovery. Prompt recognition of interstitial lung disease exacerbation as well as exclusion of progressive disease or infection is of primary importance.

The Volatile Anesthetic Isoflurane Prevents Ventilator-Induced Lung Injury via Phosphoinositide 3-Kinase/Akt Signaling in Mice

Mechanical ventilation leads to ventilator-induced lung injury in animals, and can contribute to acute lung injury/acute respiratory distress syndrome in humans. Acute lung injury/acute respiratory distress syndrome currently causes an unacceptably high rate of morbidity and mortality among critically ill patients. Volatile anesthetics have been shown to exert anti-inflammatory and organ-protective effects in vivo. We investigated the effects of the volatile anesthetic isoflurane on lung injury during mechanical ventilation. C57BL/6N mice were ventilated with a tidal volume of 12 mL/kg body weight for 6 hours in the absence or presence of isoflurane, and, in a second series, with or without the specific phosphoinositide 3-kinase/Akt inhibitor LY294002. Lung injury was determined by comparative histology, and by the isolation of bronchoalveolar lavage for differential cell counting and analysis of cytokine levels using enzyme-linked immunosorbent assays. Lung homogenates were analyzed for protein expression by Western blotting. Mechanical ventilation caused increases in alveolar wall thickening, cellular infiltration, and an elevated ventilator-induced lung injury score. Neutrophil influx and cytokine (i.e., interleukin-1β, and macrophage inflammatory protein-2) release were enhanced in the bronchoalveolar lavage of ventilated mice. The expression levels of the stress proteins hemeoxygenase-1 and heat shock protein-70 were elevated in lung tissue homogenates. Isoflurane ventilation significantly reduced lung damage, inflammation, and stress protein expression. In contrast, phosphorylation of Akt protein was substantially increased during mechanical ventilation with isoflurane. Inhibition of phosphoinositide 3-kinase/Akt signaling before mechanical ventilation completely reversed the lung-protective effects of isoflurane treatment in vivo. Inhalation of isoflurane during mechanical ventilation protects against lung injury by preventing proinflammatory responses. This protection is mediated via phosphoinositide 3-kinase/Akt signaling.

Effects of Postnatal Dexamethasone or Hydrocortisone in a Rat Model of Antenatal Lipopolysaccharide and Neonatal Hyperoxia Exposure

The aim of our study was to investigate the differential effects of dexamethasone (DXM) and hydrocortisone (HCS) on somatic growth and postnatal lung development in a rat model of bronchopulmonary dysplasia (BPD). A rat model of BPD was induced by administering intra-amniotic lipopolysaccharide (LPS) and postnatal hyperoxia. The rats were treated with a 6-day (D1-D6) tapering course of DXM (starting dose 0.5 mg/kg/day), HCS (starting dose 2 mg/kg/day), or an equivalent volume of normal saline. DXM treatment in a rat model of BPD induced by LPS and hyperoxia was also associated with a more profound weight loss compared to control and LPS + O2 groups not exposed to corticosteroid, whereas HCS treatment affected body weight only slightly. Examination of lung morphology showed worse mean cord length in both LPS + O2 + DXM and LPS + O2 + HCS groups as compared to the LPS + O2 alone group, and the LPS + O2 + DXM group had thicker alveolar walls than the LPS + O2 group at day 14. The HCS treatment was not significantly associated with aberrant alveolar wall thickening and retarded somatic growth. The use of postnatal DXM or HCS in a rat model of BPD induced by intra-amniotic LPS and postnatal hyperoxia appeared detrimental to lung growth, but there was less effect in the case of HCS. These findings suggest that effect of HCS on somatic growth and pulmonary outcome may be better tolerated in neonates for preventing and/or treating BPD.