BACKGROUNDSepsis is a major cause of mortality in Intensive Care Units. Anesthetic dose isoflurane and 100% oxygen were proved to be beneficial in sepsis; however, their application in septic patients is limited because long-term hyperoxia may induce oxygen toxicity and anesthetic dose isoflurane has potential adverse consequences. This study was scheduled to find the optimal combination of isoflurane and oxygen in protecting experimental sepsis and its mechanisms.

METHODSThe effects of combined therapy with isoflurane and oxygen on lung injury and sepsis were determined in animal models of sepsis induced by cecal ligation and puncture (CLP) or intraperitoneal injection of lipopolysaccharide (LPS) or zymosan. Mouse RAW264.7 cells or human peripheral blood mononuclear cells (PBMCs) were treated by LPS to probe mechanisms. The nuclear factor kappa B (NF-κB) signaling molecules were examined by Western blot and cellular immunohistochemistry.

RESULTSThe 0.5 minimum alveolar concentration (MAC) isoflurane in 60% oxygen was the best combination of oxygen and isoflurane for reducing mortality in experimental sepsis induced by CLP, intraperitoneal injection of LPS, or zymosan. The 0.5 MAC isoflurane in 60% oxygen inhibited proinflammatory cytokines in peritoneal lavage fluids (tumor necrosis factor-alpha [TNF-β]: 149.3 vs. 229.7 pg/ml, interleukin [IL]-1β: 12.5 vs. 20.6 pg/ml, IL-6: 86.1 vs. 116.1 pg/ml, and high-mobility group protein 1 [HMGB1]: 323.7 vs. 449.3 ng/ml; all P< 0.05) and serum (TNF-β: 302.7 vs. 450.7 pg/ml, IL-1β: 51.7 vs. 96.7 pg/ml, IL-6: 390.4 vs. 722.5 pg/ml, and HMGB1: 592.2 vs. 985.4 ng/ml; all P< 0.05) in septic animals. In vitro experiments showed that the 0.5 MAC isoflurane in 60% oxygen reduced inflammatory responses in mouse RAW264.7 cells, after LPS stimulation (all P< 0.05). Suppressed activation of NF-κB pathway was also observed in mouse RAW264.7 macrophages and human PBMCs after LPS stimulation or plasma from septic patients. The 0.5 MAC isoflurane in 60% oxygen also prevented the increases of phospho-IKKβ/β, phospho-IκBβ, and phospho-p65 expressions in RAW264.7 macrophages after LPS stimulation (all P< 0.05).

CONCLUSIONCombined administration of a sedative dose of isoflurane with 60% oxygen improves survival of septic animals through reducing inflammatory responses.

Adult ; Anesthesia ; methods ; Animals ; Blotting, Western ; Bronchoalveolar Lavage Fluid ; Disease Models, Animal ; Female ; Humans ; Inflammation ; drug therapy ; Isoflurane ; therapeutic use ; Leukocytes, Mononuclear ; metabolism ; Lipopolysaccharide Receptors ; metabolism ; Lipopolysaccharides ; pharmacology ; Lung Injury ; drug therapy ; immunology ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; NF-kappa B ; metabolism ; Oxygen ; therapeutic use ; Peroxidase ; metabolism ; RAW 264.7 Cells ; Rats, Sprague-Dawley ; Sepsis ; drug therapy ; immunology ; Tumor Necrosis Factor-alpha ; metabolism

The present study was designed to synthesize 2-Cyano-3, 12-dioxooleana-1, 9(11)-en-28-oate-13β, 28-olide (1), a lactone derivative of oleanolic acid (OA) and evaluate its anti-inflammatory activity. Compound 1 significantly diminished nitric oxide (NO) production and down-regulated the mRNA expression of iNOS, COX-2, IL-6, IL-1β, and TNF-α in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Further in vivo studies in murine model of LPS-induced acute lung injury (ALI) showed that 1 possessed more potent protective effects than the well-known anti-inflammatory drug dexamethasone by inhibiting myeloperoxidase (MPO) activity, reducing total cells and neutrophils, and suppressing inflammatory cytokines expression, and thus ameliorating the histopathological conditions of the injured lung tissue. In conclusion, compound 1 could be developed as a promising anti-inflammatory agent for intervention of LPS-induced ALI.
Acute Lung Injury ; drug therapy ; genetics ; immunology ; Animals ; Anti-Inflammatory Agents ; administration & dosage ; chemical synthesis ; Bronchoalveolar Lavage Fluid ; immunology ; Cyclooxygenase 2 ; genetics ; immunology ; Female ; Humans ; Interleukin-1beta ; genetics ; immunology ; Interleukin-6 ; genetics ; immunology ; Lipopolysaccharides ; adverse effects ; Lung ; drug effects ; immunology ; Macrophages ; drug effects ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Neutrophils ; drug effects ; immunology ; Oleanolic Acid ; administration & dosage ; analogs & derivatives ; chemical synthesis ; Peroxidase ; genetics ; immunology ; RAW 264.7 Cells ; Tumor Necrosis Factor-alpha ; genetics ; immunology

OBJECTIVETo investigate the Th17 cell and Treg cell levels in patients with sarcoidosis, and their relation to disease activation and glucocorticoids treatment.

METHODSTwenty-three sarcoidosis patients admitted in Yinzhou People's Hospital from January 2009 to December 2013 and 25 healthy subjects (controls) were included in this study. The blood samples and bronchoalveolar lavage fluid (BALF) samples were collected in all patients before and after glucocorticoids treatment. The serum angiotensin converting enzyme (SACE) levels were detected. The percentages of Th17 cells and Treg cells in peripheral blood and BALF were determined by flow cytometry, the concentrations of cytokines in serum and supernatants of BALF were measured by enzyme-linked immunosorbent assay (ELISA). The levels of ROR-γt and Foxp3 mRNA transcripts in peripheral blood mononuclear cells (PBMC) were determined by real-time quantitative PCR. The potential correlation between the percentages of Th17 or Treg cells and SACE levels was evaluated.

RESULTSCompared with healthy controls, significantly higher frequencies of Th17 cells (4.34%±0.89% vs 1.60% ± 0.42%), lower frequencies of Treg cells (1.28% ± 0.37% vs 3.39% ± 0.50%) in peripheral blood were observed. Higher level of ROR-γt mRNA (21.31 ± 3.55 vs 3.63 ± 1.00) and lower level of Foxp3 mRNA (1.60 ± 0.24 vs 3.12 ± 0.76) in peripheral blood were detected in sarcoidosis patients in active stage (before glucocorticoids treatment) (all P<0.01). After the treatment of glucocorticoids, these index in peripheral blood were significantly improved (Th17 cells 2.16% ± 0.68%，Treg cells 2.21% ± 0.42%, ROR-γt mRNA 10.15 ± 1.93, Foxp3 mRNA 2.44 ± 0.38) ( all P<0.05). The changing trends of Th17 and Treg cell cytokines levels in serum were consistent with two type cells. Meanwhile, the changing trends of above index in BALF of patients treated by glucocorticoids were consistent with those in sarcoidosis patients in active stage. The increased ratios of Th17 cells to Treg cells were positively correlated with the level of serum SACE (r= 0.781).

CONCLUSIONThe imbalance of Th17 cells and Treg cells in peripheral blood and airway may be involved in the pathogenesis of sarcoidosis, which was associated with the activity of disease, and the treatment of glucocorticoids may achieve a therapeutic effect by correcting the immune imbalance.

Bronchoalveolar Lavage Fluid ; Case-Control Studies ; Cytokines ; immunology ; Enzyme-Linked Immunosorbent Assay ; Forkhead Transcription Factors ; metabolism ; Humans ; Leukocytes, Mononuclear ; metabolism ; Nuclear Receptor Subfamily 1, Group F, Member 3 ; metabolism ; Sarcoidosis ; immunology ; T-Lymphocytes, Regulatory ; immunology ; Th17 Cells ; immunology

OBJECTIVETo observe the changes in Th17 cell levels in bronchoalveolar lavage fluid in children of sepsis with acute lung injury and the relationship between the Th17 cell levels and prognosis.

METHODSFifty children of sepsis with acute lung injury were enrolled in the study. The percentages of Th17 cells in bronchoalveolar lavage fluid were measured by flow cytometry. The patients of sepsis with acute lung injury were classified into three groups based on the Pediatric Critical Illness Score (PCIS): extremely critical, critical and non-critical. According to the clinical prognosis, the patients were classified into survical and death groups. Th17 cell levels were compared between the two groups. The relationship between Th17 cell levels and the PCIS scores was analyzed.

RESULTSWith the increase in the severity of sepsis, Th17 cell levels in bronchoalveolar lavage fluid were gradually increased (P<0.05). The Th17 cell levels were negatively correlated to the PCIS scores (r=-0.853; P<0.01). The Th17 cell levels were significantly higher in the death group than in the survival group. Moreover, compared with the survival group, the PCIS scores were lower in the death group (P<0.05).

CONCLUSIONSThe increased Th17 cell levels in children of sepsis with acute lung injury are closely related to the severity and prognosis of patients, suggesting that Th17 cell levels can be used as a predictor of the severity and prognosis.

Acute Lung Injury ; immunology ; Bronchoalveolar Lavage Fluid ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Prognosis ; Sepsis ; immunology ; Severity of Illness Index ; Th17 Cells ; physiology

In the present study, we analyzed the role of Ginkgo biloba extract in lipopolysaccharide(LPS)-induced acute lung injury (ALI). ALI was induced in mice by intratracheal instillation of LPS. G. biloba extract (12 and 24 mg·kg(-1)) and dexamethasone (2 mg·kg(-1)), as a positive control, were given by i.p. injection. The cells in the bronchoalveolar lavage fluid (BALF) were counted. The degree of animal lung edema was evaluated by measuring the wet/dry weight ratio. The superoxidase dismutase (SOD) and myeloperoxidase (MPO) activities were assayed by SOD and MPO kits, respectively. The levels of inflammatory mediators, tumor necrosis factor-a, interleukin-1b, and interleukin-6, were assayed by enzyme-linked immunosorbent assay. Pathological changes of lung tissues were observed by H&E staining. The levels of NF-κB p65 and COX-2 expression were detected by Western blotting. Compared to the LPS group, the treatment with the G. biloba extract at 12 and 24 mg·kg(-1) markedly attenuated the inflammatory cell numbers in the BALF, decreased NF-κB p65 and COX-2 expression, and improved SOD activity, and inhibited MPO activity. The histological changes of the lungs were also significantly improved. The results indicated that G. biloba extract has a protective effect on LPS-induced acute lung injury in mice. The protective mechanism of G. biloba extract may be partly attributed to the inhibition of NF-κB p65 and COX-2 activation.
Acute Lung Injury ; chemically induced ; drug therapy ; metabolism ; Animals ; Bronchoalveolar Lavage Fluid ; cytology ; Cell Count ; Cyclooxygenase 2 ; genetics ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; drug effects ; Ginkgo biloba ; chemistry ; Interleukin-1beta ; analysis ; Interleukin-6 ; analysis ; Lipopolysaccharides ; Lung ; immunology ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Peroxidase ; metabolism ; Phytotherapy ; Plant Extracts ; pharmacology ; Pulmonary Edema ; Superoxide Dismutase ; metabolism ; Transcription Factor RelA ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; analysis

Although some studies have explained the immunomodulatory effects of statins, the exact mechanisms and the therapeutic significance of these molecules remain to be elucidated. This study not only evaluated the therapeutic potential and inhibitory mechanism of simvastatin in an ovalbumin (OVA)-specific asthma model in mice but also sought to clarify the future directions indicated by previous studies through a thorough review of the literature. BALB/c mice were sensitized to OVA and then administered three OVA challenges. On each challenge day, 40 mg kg-1 simvastatin was injected before the challenge. The airway responsiveness, inflammatory cell composition, and cytokine levels in bronchoalveolar lavage (BAL) fluid were assessed after the final challenge, and the T cell composition and adhesion molecule expression in lung homogenates were determined. The administration of simvastatin decreased the airway responsiveness, the number of airway inflammatory cells, and the interleukin (IL)-4, IL-5 and IL-13 concentrations in BAL fluid compared with vehicle-treated mice (P<0.05). Histologically, the number of inflammatory cells and mucus-containing goblet cells in lung tissues also decreased in the simvastatin-treated mice. Flow cytometry showed that simvastatin treatment significantly reduced the percentage of pulmonary CD4+ cells and the CD4+/CD8+ T-cell ratio (P<0.05). Simvastatin treatment also decreased the expression of the vascular cell adhesion molecule 1 and intercellular adhesion molecule 1 proteins, as measured in homogenized lung tissues (P<0.05) and human epithelial cells. The reduction in the T cell influx as a result of the decreased expression of cell adhesion molecules is one of the mechanisms by which simvastatin attenuates airway responsiveness and allergic inflammation. Rigorous review of the literature together with our findings suggested that simvastatin should be further developed as a potential therapeutic strategy for allergic asthma.
Animals ; Anti-Inflammatory Agents/*therapeutic use ; Asthma/*drug therapy/immunology ; Bronchoalveolar Lavage Fluid/immunology ; CD4-Positive T-Lymphocytes/drug effects/immunology ; CD8-Positive T-Lymphocytes/drug effects/immunology ; Female ; Humans ; Hydroxymethylglutaryl-CoA Reductase Inhibitors/*therapeutic use ; Inflammation/*drug therapy/immunology ; Interleukins/analysis/immunology ; Lung/*drug effects/immunology ; Mice, Inbred BALB C ; Simvastatin/*therapeutic use

OBJECTIVETo investigate the effect of Mycoplasma pneumoniae (MP) infection on the function of T lymphocytes in the bronchoalveolar lavage fluid (BALF) of asthmatic children in acute and stable periods and the relationship between MP infection and asthma.

METHODSSeventy-one hospitalized children (with bronchitis, pneumonia, and asthma) were divided into non-MP infection control group (group A, pneumonia and bronchitis without MP infection), non-MP infection asthma group (group B), and MP infection asthma group (group C). Flow cytometry was used to determine CD3(+), CD4(+), and CD8(+) T cell counts and CD4(+)/CD8(+) ratio in BALF among all children in acute and stable periods.

RESULTSCompared with group A, groups B and C showed significant differences in CD3(+), CD4(+), and CD8(+) T cell counts and CD4(+)/CD8(+) ratio (P<0.05) in acute and stable periods, had decreased CD3(+) and CD4(+) T cell counts, an increased CD8(+) T cell count, and a significantly decreased CD4(+)/CD8(+) ratio (P<0.05) in the acute period, and had decreased CD3(+) and CD4(+) T cell counts and CD4(+)/CD8(+) ratio and an increased CD8(+) T cell count (P<0.05) in the stable period. Compared with group B, group C had significantly decreased CD3(+) and CD4(+) T cell counts and CD4(+)/CD8(+) ratio (P<0.05) and a significantly increased CD8(+) T cell count (P<0.05) in the acute period and showed no significant differences in CD3(+), CD4(+), and CD8(+) T cell counts (P>0.05) and a significant decrease in CD4(+)/CD8(+) ratio (P<0.05) in the stable period.

CONCLUSIONSThe immunological function of T lymphocytes in the airway declines significantly among asthmatic children with MP infection in acute and stable periods, leading to immue system disorder. MP may be associated with the pathogenesis of asthma.

Asthma ; etiology ; immunology ; Bronchoalveolar Lavage Fluid ; immunology ; CD4-CD8 Ratio ; Child ; Child, Preschool ; Female ; Humans ; Male ; Pneumonia, Mycoplasma ; complications ; immunology ; T-Lymphocytes ; immunology

BACKGROUNDA variety of inflammatory mediators and effector cells participate together in acute lung injury, and lead to secondary injury that is due to an inflammatory cascade and secondary diffuse lung parenchyma injury. Inflammation is associated with an oxidative stress reaction, which is produced in the development of airway inflammation, and which has positive feedback on inflammation itself. Resolvin D1 can reduce the infiltration of neutrophils, regulate cytokine levels and reduce the inflammation reaction, and thereby promote the resolution of inflammation. The purpose of this study is to investigate the effects of resolvin D1 on an inflammatory response and oxidative stress during lipopolysaccharide (LPS)-induced acute lung injury.

METHODSLPS (3 mg/kg) was used to induce the acute lung injury model. Pretreatment resolvin D1 (100 ng/mouse) was given to mice 30 minutes before inducing acute lung injury. Mice were observed at 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days and 7 days after LPS was administrated, then they were humanely sacrificed. We collected bronchoalveolar lavage fluid (BALF) and the lung tissues for further analysis. Paraffin section and HE staining of the lung tissues were made for histopathology observations. Parts of the lung tissues were evaluated for wet-to-dry (W/D) weight ratio. tumor necrosis factor (TNF)-α, inter leukin (IL)-1β, IL-10 and myeloperoxidase (MPO) were detected by enzyme-linked immunosorbent assay (ELISA). A lipid peroxidation malondialdehyde (MDA) assay kit was used to detect MDA. A total superoxide dismutase assay kit with WST-1 was used to analyze superoxide dismutase (SOD). We determined the apoptosis of neutrophils by Flow Cytometry. A real-time quantitative PCR Detecting System detected the expression of mRNA for heme oxygenase (HO)-1.

RESULTSPretreatment with resolvin D1 reduced the pathological damage in the lung, decreased the recruitment of neutrophils and stimulated their apoptosis. It markedly decreased the expressions of TNF-α, IL-1β and increased the expressions of IL-10, and decreased the production of MDA and increased the expressions of SOD. The mRNA expression of HO-1 was also significantly increased.

CONCLUSIONSResolvin D1 displays potent anti-inflammatory actions by regulating cytokines, inhibiting aberrant neutrophil recruitment and stimulating apoptosis of neutrophils. Resolvin D1 can also relieve the injury due to oxidative stress. The mechanisms might be related to increase HO-1 expression.

Acute Lung Injury ; chemically induced ; drug therapy ; immunology ; Animals ; Bronchoalveolar Lavage Fluid ; immunology ; Docosahexaenoic Acids ; therapeutic use ; Interleukin-10 ; metabolism ; Interleukin-1beta ; metabolism ; Lipopolysaccharides ; toxicity ; Male ; Mice ; Mice, Inbred BALB C ; Oxidative Stress ; drug effects ; Peroxidase ; metabolism ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVES: We established a Wistar rat model of asthma caused by toluene diisocyanate (TDI) exposure, and investigated the relationship between TDI exposure concentrations and respiratory hypersensitivity, airway inflammation, and cytokine secretions in animals, to better understand the mechanism of TDI induced occupational asthma. METHODS: Wistar rats were exposed to two different concentrations of TDI vapor four hours a day for five consecutive days. Bronchoalveolar lavage (BAL) was performed, and differential leucocytes from the BAL fluid were analyzed. Lung histopathological examination was carried out to investigate the inflammatory status in the airways. Production of cytokines interleukin (IL)-4 and IL-5 productions in the BAL fluid in vivo was determined with enzyme-linked immunosorbent assay kits. RESULTS: The TDI-exposed rats exhibited greater airway hypersensitivity symptoms than the control rats. The BAL differential cell count and lung histopathological examination demonstrated that inflammation reactions were present in both the central and peripheral airways, characterized with marked infiltration of eosinophils in the TDI-exposed rats. The cytokine assay showed that IL-4 and IL-5 were predominantly produced in the BAL fluid in vivo. CONCLUSIONS: These findings imply that TDI exposure concentrations may greatly affect the occurrence and extent of inflammatory events and that Th2 type cytokines may play an important role in the immunopathogenesis of TDI-induced occupational respiratory hypersensitivity.
Animals ; Bronchoalveolar Lavage Fluid/chemistry/cytology ; Enzyme-Linked Immunosorbent Assay ; Eosinophils/cytology/immunology ; Female ; Gases/chemistry ; Hypersensitivity/pathology ; Interleukin-4/*analysis ; Interleukin-5/*analysis ; Lung/*drug effects/pathology/secretion ; Rats ; Rats, Wistar ; Toluene 2,4-Diisocyanate/*toxicity

AIM: The aim of the study was to investigate the effect and mechanism of action of synthetic salidroside in an ovalbumin (OVA)-induced asthma model in mice. METHOD: BALB/c mice were sensitized with an intraperitoneal injection of ovalbumin (OVA) to induce a mouse model of asthma in paracmasis. The mice were treated with dexamethasone as the positive control. At the end of the study, respiratory reactivity was detected, the numbers of various kinds of white blood cells in the bronchoalveolar lavage fluid (BALF) were counted, and the levels of IL-4 and INF-γ in BALF were determined. Quantitative PCR was used to detect the mRNA contents of IL-4 and INF-γ in lung tissue. Histologic examination was performed to observe inflammatory cellular infiltration. RESULTS: Salidroside treatment virtually eliminated airway hyper-reactivity, markedly reduced the eosinophil percent, obviously reduced the levels of IL-4 and raised INF-γ in the bronchoalveolar lavage fluid (BALF) compared with the sham-treated group. Quantitative PCR on the mRNA content of IL-4 and INF-γ provided confirmation. Lung histologic observations showed that salidroside reduced inflammation and edema. These effects were equivalent to the effects of dexamethasone. CONCLUSION Synthetic salidroside exhibits an anti-asthma effect which is related to the regulation of Th1/Th2 balance. This provides a new possibility for treatment of allergic asthma.
Animals ; Anti-Asthmatic Agents ; administration & dosage ; Asthma ; drug therapy ; genetics ; immunology ; Bronchoalveolar Lavage Fluid ; immunology ; Female ; Glucosides ; administration & dosage ; chemical synthesis ; Humans ; Interferon-gamma ; genetics ; immunology ; Interleukin-4 ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Phenols ; administration & dosage ; chemical synthesis ; Th1 Cells ; drug effects ; immunology ; Th2 Cells ; drug effects ; immunology