OBJECTIVE: To identify the underlying mechanism by which quercetin (Que) alleviates sepsis-related acute respiratory distress syndrome (ARDS). METHODS: In vivo, C57BL/6 mice were assigned to sham, cecal ligation and puncture (CLP), and CLP+Que (50 mg/kg) groups (n=15 per group) by using a random number table. The sepsisrelated ARDS mouse model was established using the CLP method. In vitro, the murine alveolar macrophages (MH-S) cells were classified into control, lipopolysaccharide (LPS), LPS+Que (10 μmol/L), and LPS+Que+acetylcysteine (NAC, 5 mmol/L) groups. The effect of Que on oxidative stress, inflammation, and apoptosis in mice lungs and MH-S cells was determined, and the mechanism with reactive oxygen species (ROS)/p38 mitogen-activated protein kinase (MAPK) pathway was also explored both in vivo and in vitro. RESULTS: Que alleviated lung injury in mice, as reflected by a reversal of pulmonary histopathologic changes as well as a reduction in lung wet/dry weight ratio and neutrophil infiltration (P<0.05 or P<0.01). Additionally, Que improved the survival rate and relieved gas exchange impairment in mice (P<0.01). Que treatment also remarkedly reduced malondialdehyde formation, superoxide dismutase and catalase depletion, and cell apoptosis both in vivo and in vitro (P<0.05 or P<0.01). Moreover, Que treatment diminished the release of inflammatory factors interleukin (IL)-1β, tumor necrosis factor-α, and IL-6 both in vivo and in vitro (P<0.05 or P<0.01). Mechanistic investigation clarifified that Que administration led to a decline in the phosphorylation of p38 MAPK in addition to the suppression of ROS expression (P<0.01). Furthermore, in LPS-induced MH-S cells, ROS inhibitor NAC further inhibited ROS/p38 MAPK pathway, as well as oxidative stress, inflammation, and cell apoptosis on the basis of Que treatment (P<0.05 or P<0.01). CONCLUSION Que was found to exert anti-oxidative, anti-inflammatory, and anti-apoptotic effects by suppressing the ROS/p38 MAPK pathway, thereby conferring protection for mice against sepsis-related ARDS.
Animals ; Sepsis/drug therapy* ; Quercetin/therapeutic use* ; Respiratory Distress Syndrome/enzymology* ; p38 Mitogen-Activated Protein Kinases/metabolism* ; Mice, Inbred C57BL ; Reactive Oxygen Species/metabolism* ; Apoptosis/drug effects* ; Male ; Oxidative Stress/drug effects* ; MAP Kinase Signaling System/drug effects* ; Lung/drug effects* ; Mice ; Lipopolysaccharides ; Macrophages, Alveolar/pathology* ; Inflammation/pathology* ; Protective Agents/therapeutic use*

OBJECTIVETo evaluate the differences in the autophagy activity of alveolar macrophages between patients with different stages of coal workers' pneumoconiosis (CWP).

METHODSA total of 116 coal workers were investigated in the field. Their lung lavage fluid was collected and purified to obtain alveolar macrophages. The morphological characteristics of autophagy were observed by transmission electron microscopy. The expression of autophagy marker (LC3) and autophagy regulators (Beclin1, mTOR, and p-mTOR) was measured by Western blot. The autophagy activity of alveolar macrophages was compared between dust-exposed subjects and patients with stage I, II, and III CWP.

RESULTSThe autophagy activity of alveolar macrophages differed between patients with different stages of CWP, according to transmission electron microscopy. Patients with stage II CWP had significantly higher protein expression of LC3 II/I and Beclin1 in pulmonary macrophages than those with stage ICWP (P < 0.05); patients with stage III CWP had significantly lower protein expression of LC3 II/I and Beclin1 in pulmonary macrophages than those with stage II CWP (P < 0.05), but had significantly higher protein expression of LC3 II/I and Beclin1 than those with stage I CWP (P < 0.05); patients with stage II CWP had a significantly higher protein expression of Beclin1 than the dust-exposed subjects (P < 0.05). Patients with stage II CWP had significantly lower expression of mTOR and p-mTOR in pulmonary macrophages than the dust-exposed subjects and those with stage I CWP (P < 0.05), while patients with stage III CWP had significantly higher expression of mTOR and p-mTOR than those with stage II CWP (P < 0.05).

CONCLUSIONThe autophagy activity of alveolar macrophages varies between patients with different stages of CWP.

Anthracosis ; pathology ; Apoptosis Regulatory Proteins ; metabolism ; Autophagy ; Beclin-1 ; Biomarkers ; Bronchoalveolar Lavage Fluid ; Coal ; Coal Mining ; Dust ; Humans ; Macrophages, Alveolar ; pathology ; Membrane Proteins ; metabolism ; Microtubule-Associated Proteins ; metabolism ; Occupational Exposure ; Pneumoconiosis ; pathology

OBJECTIVETo analyze the clinical features and investigate the clinical diagnostic methods of hard metal lung disease (HMLD), then provide reference for the diagnostic criteria of occupational HMLD.

METHODSRetrieved the open published case reports associated with HMLD from January, 2000 to June, 2014. Regarding the ages, sex, types and years of work, clinical features and laboratory results for analyzing.

RESULTSCollected 21 clinical cases of HMLD belonged to 6 internal reports and 15 oversea reports. Among them 15 male and 6 female, ages were from 22 to 58, length of service between 1 year and 43 years. Clinical presentations included cough (20 cases), dyspnea on progressive (10 cases), and pulmonary function testing showed a restrictive abnormality. The imaging features presented as bilateral areas of ground-glass attenuation, diffuse small nodules, extensive reticular opacities and traction bronchiectasis. The finding of giant cell interstitial pneumonia (GIP) was almost pathognomonic for hard metal pneumoconiosis. The main pathological findings contained a different levels of lymphocyte, acidophilic cell infiltration, hyperplasia of fibrous tissue and numerous large multinucleated histiocytes which ingested inflammatory cells were admixed with macrophages. 16 cases of the 21 reports showed GIP.

CONCLUSIONSClinical presentations include cough and dyspnea on progressive, and pulmonary function testing show a restrictive abnormality. The imaging features present as bilateral areas of ground-glass attenuation, areas of consolidation, diffuse small nodules, extensive reticular opacities and traction bronchiectasis. The prime pathological findings contain interstitial pneumonia with intra-alveolar macrophages and a large amount of multinucleated histiocytes.

Adult ; Alloys ; Cobalt ; Female ; Humans ; Lung ; physiopathology ; Lung Diseases, Interstitial ; pathology ; Macrophages, Alveolar ; Male ; Middle Aged ; Occupational Diseases ; pathology ; Pneumoconiosis ; pathology ; Tungsten ; Young Adult

OBJECTIVETo investigate the role of oxidative stress in the endoplasmic reticulum stress-induced apoptosis of alveolar macrophages triggered by quartz dust.

METHODSSeventy-two healthy adult Wistar rats were randomly divided into control group, quartz dust group, quartz dust plus N-acetyl cysteine (NAC) group, and NAC group, with 18 rats in each group. One milliliter of sterile saline (for the control and NAC groups) or 1 ml of saline with 5%ultrafine quartz dust (for dust group and dust plus NAC group) was given to each rat by non-exposed endotracheal infusion. From the second day after dust infusion, rats in dust plus NAC group and NAC group received intragastric administration of NAC (100 mg/kg). In each week, the treatment with NAC lasted for 5 consecutive days, followed by 2 days' interval. For each group, 6 rats were randomly selected on the 14th, 28th, or 56th day after dust exposure; they were sacrificed by bloodletting from the femoral artery, and the lungs were collected. Bronchoalveolar lavage fluid was collected to separate macrophages. The protein expression of caspase-12 in alveolar macrophages, the apoptosis rate and reactive oxygen species (ROS) content of alveolar macrophages, and the protein carbonyl content of alveolar macrophages were determined by Western blot, flow cytometry, and colorimetry, respectively.

RESULTSIncreased protein expression of caspase-12, apoptosis rate, and content of ROS and protein carbonyl were discovered on the 14th day in the dust group, in comparison with the control group (P < 0.05), and the increase lasted till the 28th and 56th days. (P < 0.05). Compared with the dust group, the dust plus NAC group showed significant decreases in the content of ROS on the 14th, 28th, and 56th days (P < 0.05), significant decreases in the content of protein carbonyl on the 28th and 56th days (P < 0.05), and significant decreases in the protein expression of caspase-12 and apoptosis rate (P < 0.05).

CONCLUSIONOxidative stress is potentially involved in the endoplasmic reticulum stress-induced apoptosis of alveolar macrophages triggered by quartz dust. Oxidative damage of protein in the endoplasmic reticulum may play an important role in the process.

Animals ; Caspase 12 ; metabolism ; Dust ; Endoplasmic Reticulum Stress ; drug effects ; Macrophages, Alveolar ; drug effects ; pathology ; Male ; Oxidative Stress ; drug effects ; Protein Carbonylation ; Quartz ; toxicity ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; metabolism

OBJECTIVETo investigate the anti-inflammatory and immunoregulatory effects of Yupingfeng (, YPF) Powder and its components in rats.

METHODSA rat chronic bronchitis (CB) model was developed using lipopolysaccharide (LPS) combined with bacillus Calmette Guerin (BCG). YPF, simple recipe Astragalus membranaceus (Fisch.) Bge (AM) and Astragalus membranaceus (Fisch.) Bge plus rhizome of Atractylodes macrocephala Koidz (AM+RA) decoction were administered (intragastric administration, once a day for 21 days) to rats, to prevent and treat CB. Immunoregulatory and anti-inflammatory effects of YPF, AM and AM+RA were tested by serum pharmacology in vitro on splenic lymphocytes of normal rats and alveolar macrophages of CB rats.

RESULTSInflammation in the pulmonary tissue and the bronchus of CB rats was significantly reduced in the YPF-treatment groups, AM and AM+RA groups demonstrating the efficacy of YPF. Serum samples collected at different times from rats after administration of YPF, AM and AM+RA demonstrated increased proliferation of splenic lymphocytes with area under the effect curve (AUE) of 552.6%, 336.3% and 452.0%, respectively. Treatment of alveolar macrophages with serum samples in YPF, AM or AM+RA group inhibited interleukin-8 (IL-8) in the cell culture media, and the effect was much better in the YPF group compared with AM or AM+RA group, with a higher maximal effect (Emax, P<0.05) and larger AUE (P <0.01 and P<0.05). Moreover, serum from rats treated with AM or AM+RA had similar efficacy, while the efficiency was lower than that treated with YPF.

CONCLUSIONYPF demonstrated anti-inflammatory and immunoregulatory effects in a rat model of CB, and timedependent relationships were demonstrated in vitro.

Animals ; Anti-Inflammatory Agents ; pharmacology ; therapeutic use ; Body Weight ; drug effects ; Bronchitis, Chronic ; drug therapy ; pathology ; Cell Proliferation ; drug effects ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Immunologic Factors ; pharmacology ; therapeutic use ; Interleukin-8 ; metabolism ; Lung ; drug effects ; pathology ; ultrastructure ; Lymphocytes ; drug effects ; Macrophages, Alveolar ; drug effects ; metabolism ; Powders ; Rats ; Rats, Sprague-Dawley ; Spleen ; pathology ; Time Factors

Humans ; Lung ; cytology ; Macrophages, Alveolar ; Pulmonary Fibrosis ; pathology ; Silicosis ; pathology

BACKGROUNDAcute lung injury (ALI) is a serious and common condition for which there are currently no specific strategies for treatment. Recent studies have suggested that bone marrow-derived multipotent mesenchymal stem cells (MSCs) may have therapeutic applications in multiple clinical disorders. We explored the biological effects of MSCs during endotoxin-induced ALI and the mechanisms involved.

METHODSMSCs were isolated from male rat bone marrow and the ALI model was induced by intravenous endotoxin injection. Female rats were sacrificed at 6 hours, 24 hours, 4 days, 1 week and 3 weeks post-injection of MSCs or saline and the lung tissue, bronchoalveolar lavage fluid, and serum were harvested for analysis. We further evaluated the survival of the rats and examined the effects of endotoxin-induced injury on the interaction between alveolar macrophages (AMs) and MSCs in ex vivo.

RESULTSThere was a significant decrease in numbers of neutrophils in bronchoalveolar lavage fluid (P < 0.05), and myeloperoxidase activity in the lung (P < 0.01), and of TNF-α and IL-1β in serum (P < 0.05) in the MSC treated rats at 4 days. Furthermore, MSC treated rats exhibited improved survival, lower lung injury score, higher concentration of IL-10 in the serum and a reduced hydroxyproline content, but these differences were not statistically significant. Moreover, co-cultures of MSCs and AMs had significantly reduced levels of TNF-α, IL-1β and macrophage inflammatory protein (MIP)-1α and significantly increased levels of IL-10 (P < 0.05) in the culture supernatants.

CONCLUSIONSTreatment with intravenous injection of bone marrow-derived MSCs have beneficial effects on endotoxin-induced ALI in rats. The beneficial effect might be achieved through the engraftment of differentiated MSCs in the lungs and appears derive more from their capacity to secrete soluble factors that modulate immune responses.

Acute Lung Injury ; chemically induced ; metabolism ; therapy ; Animals ; Bone Marrow Cells ; cytology ; Cells, Cultured ; Coculture Techniques ; Endotoxins ; toxicity ; Female ; Lung ; metabolism ; pathology ; Macrophages, Alveolar ; cytology ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; physiology ; Peroxidase ; metabolism ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo explore the pathological changes of pulmonary fibrosis induced by SiO2 in rats and pigs.

METHODSThe silicosis models in rats and pigs were established by non-exposure method. The pathologic changes in lung tissues of rats and pigs were observed with HE staining under a light microscopy and under a transmission electron microscopy (TEM), the expression of cytokines was detected by immunohistochemistry.

RESULTS(1) The main pathologic changes of silicosis models in rats and pigs included: in 7 ∼ 15 days after treatment, silica dusts, dust cells, a lot of macrophages, lung epithelial cells, a few neutrophils, macrophage alveolar inflammation and nodules of stage I were found in alveolar space; in 30 ∼ 90 days after treatment, many nodules of stage I-III or IV with lymphocytes infiltration were observed in respiratory bronchioles, alveoli, interlobular septa, the subpleural and around blood vessels and bronchi. (2) The expression levels of CK protein, SP-A protein, CD68, b-FGF, TNF-α, IL-6, TGF-β1, NFKappa/P50, Kappa/P65 and VEGF reduced with exposure time, but still were higher than those of the control. (3) The shed alveolar type I cells, proliferation of alveolar type II cells or macrophages and activated cellular function induced by silica were observed under TEM.

CONCLUSIONThe development of pulmonary fibrosis in silicosis models corresponded with the process from macrophages alveolar inflammation to pulmonary fibrosis.

Animals ; Cytokines ; metabolism ; Disease Models, Animal ; Epithelial Cells ; metabolism ; Female ; Lung ; cytology ; pathology ; Macrophages, Alveolar ; metabolism ; Male ; Neutrophils ; metabolism ; Rats ; Rats, Sprague-Dawley ; Silicosis ; pathology ; Swine

OBJECTIVETo study the roles of macrophage apoptosis, IL-1, and IL-8 in the pathogenesis of rat pulmonary fibrosis induced by silica.

METHODSForty eight male Wistar rats were divided into the 4 control groups (24 rats) and 4 experimental groups (24 rats). Rats in the control groups were treated with 1 ml normal saline by trachea instillation, whereas the rats in experimental groups were exposed 1 ml silica suspension (100 mg/ml) by trachea instillation for 1, 7, 14 and 28 days, respectively. Six rats of each group were sacrificed, then the bronchoalveolar lavage fluid and lung tissues were collected, respectively. Pulmonary inflammation, fibrosis and other pathological changes were detected with H.E. staining. Morphological changes of the early stage apoptosis in macrophages were detected with transmission electron microscope (TEM). The early apoptosis rates of macrophages in BALF were also assessed using Annexin V-FITC/PI kit. The IL-1 and IL-8 levels of serum were measured with the ELISA.

RESULTSThe apoptotic rates (11.48% +/- 0.24%, 16.03% +/- 0.68%, 15.53% +/- 1.07%, 18.92% +/- 2.70%, respectively) of macrophage in the experimental groups increased obviously with time, as compared to the controls (5.47% +/- 2.06%, 6.39% +/- 0.215, 9.07% +/- 0.61% and 8.54% +/- 0.16%, Respectively) (P < 0.05). The IL-1 levels of serum in the experimental groups were 23.64 +/- 0.84, 23.38 +/- 1.10, 22.21 +/- 0.86 and 24.29 +/- 1.31 pg/ml, respectively, which were significantly higher than those (18.52 +/- 1.23, 18.40 +/- 1.6, 17.92 +/- 2.21 and 18.53 +/- 2.64 pg/ml, respectively) in the control groups (P < 0.05) without time-effect relationship. The serum IL-8 levels on the 1st, 7th and 14th days in the experimental groups were 21.32 +/- 1.44, 21.90 +/- 2.08 and 22.00 +/- 2.80 pg/ml, respectively, which were significantly higher than those (17.69 +/- 1.09, 16.98 +/- 2.09 and 17.54 +/- 1.62 pg/ml, respectively) in the control groups (P < 0.05).

CONCLUSIONThe early macrophage apoptosis and changes of IL-1 and IL-8 may in lungs may play an important role in the development of pulmonary fibrosis induced by silica.

Animals ; Apoptosis ; drug effects ; Disease Models, Animal ; Interleukin-1 ; blood ; Interleukin-8 ; blood ; Macrophages, Alveolar ; cytology ; drug effects ; Male ; Pulmonary Fibrosis ; Rats ; Rats, Wistar ; Silicon Dioxide ; toxicity ; Silicosis ; blood ; pathology

OBJECTIVETo investigate in vitro cytotoxicity and oxidative stress response induced by multiwalled carbon nanotubes (MWCNTs).

METHODSCultured macrophages (murine RAW264.7 cells) and alveolar epithelium cells type II (human A549 lung cells) were exposed to the blank control, DNA salt control, and the MWCNTs suspensions at 2.5, 10, 25, and 100 μg/mL for 24 h. Each treatment was evaluated by cell viability, cytotoxicity and oxidative stress.

RESULTSOverall, both cell lines had similar patterns in response to the cytotoxicity and oxidative stress of MWCNTs. DNA salt treatment showed no change compared to the blank control. In both cell lines, significant changes at the doses of 25 and 100 μg/mL treatments were found in cell viabilities, cytotoxicity, and oxidative stress indexes. The reactive oxygen species (ROS) generation was also found to be significantly higher at the dose of 10 μg/mL treatment, whereas no change was seen in most of the indexes. The ROS generation in both cell lines went up in minutes, reached the climax within an hour and faded down after several hours.

CONCLUSIONExposure to MWCNTs resulted in a dose-dependent cytotoxicity in cultured RAW264.7 cells and A549 cells, that was closely correlated to the increased oxidative stress.

Animals ; Cell Culture Techniques ; Cell Line ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Lung ; drug effects ; enzymology ; metabolism ; pathology ; Macrophages, Alveolar ; drug effects ; enzymology ; metabolism ; pathology ; Mice ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Microscopy, Fluorescence ; Nanotubes, Carbon ; chemistry ; toxicity ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Surface Properties