Levels Of TNF-α In Bronchoalveolar Lavage Fluid Research Articles

Integrating Network Pharmacology, Transcriptome and Artificial Intelligence for Investigating Into the Effect and Mechanism of Ning Fei Ping Xue Decoction Against the Acute Respiratory Distress Syndrome.

Acute respiratory distress syndrome (ARDS) is a high-mortality disease and lacks effective pharmacotherapy. A traditional Chinese medicine (TCM) formula, Ning Fei Ping Xue (NFPX) decoction, was demonstrated to play a critical role in alleviating inflammatory responses of the lung. However, its therapeutic effectiveness in ARDS and active compounds, targets, and molecular mechanisms remain to be elucidated. The present study investigates the effects of NFPX decoction on ARDS mice induced by lipopolysaccharides (LPS). The results revealed that NFPX alleviated lung edema evaluated by lung ultrasound, decreased lung wet/Dry ratio, the total cell numbers of bronchoalveolar lavage fluid (BALF), and IL-1β, IL-6, and TNF-α levels in BALF and serum, and ameliorated lung pathology in a dose-dependent manner. Subsequently, UPLC-HRMS was performed to establish the compounds of NFPX. A total of 150 compounds in NFPX were characterized. Moreover, integrating network pharmacology approach and transcriptional profiling of lung tissues were performed to predict the underlying mechanism. 37 active components and 77 targets were screened out, and a herbs-compounds-targets network was constructed. Differentially expressed genes (DEGs) were identified from LPS-treated mice compared with LPS combined with NFPX mice. GO, KEGG, and artificial intelligence analysis indicated that NFPX might act on various drug targets. At last, potential targets, HRAS, SMAD4, and AMPK, were validated by qRT-PCR in ARDS murine model. In conclusion, we prove the efficacy of NFPX decoction in the treatment of ARDS. Furthermore, integrating network pharmacology, transcriptome, and artificial intelligence analysis contributes to illustrating the molecular mechanism of NFPX decoction on ARDS.

Prognostic Value of Angiopoietin-like 4 in Patients with Acute Respiratory Distress Syndrome.

Angiopoietin-like 4 (ANGPTL4) is a secreted glycoprotein that plays an important role in endothelial injury and the inflammatory response. Experimental models have implicated ANGPTL4 in acute respiratory distress syndrome (ARDS), but its impact on the progression of ARDS is unclear. Paired bronchoalveolar lavage fluid (BALF) and serum samples were obtained from patients with ARDS (n = 56) within 24 h of diagnosis and from control subjects (n = 32). ANGPTL4, angiopoietin-2, interleukin (IL)-6, and TNF-α levels were measured by magnetic Luminex assay. BALF albumin (BA) and serum albumin (SA) were evaluated by enzyme-linked immunosorbent assay. BALF and serum ANGPTL4 concentrations were higher in patients with ARDS than in controls and were even higher in non-survivors than in survivors. The serum ANGPTL4 level was higher in indirect (extrapulmonary) ARDS than in direct (pulmonary) ARDS. Furthermore, BALF and serum ANGPTL4 levels correlated well with angiopoietin-2, IL-6, and TNF-α levels in BALF and serum. BALF ANGPTL4 was positively correlated with the BA/SA ratio (an indicator of pulmonary vascular permeability), and serum ANGPTL4 was associated with the severity of multiple organ dysfunction syndrome based on SOFA and APACHE II scores. Moreover, serum ANGPTL4 was better able to predict 28-day ARDS-related mortality (AUC 0.746, P < 0.01) than the APACHE II score or PaO2/FiO2 ratio. Serum ANGPTL4 was identified as an independent risk factor for mortality in a univariate Cox regression model (P < 0.001). ANGPTL4 levels were elevated in patients with ARDS and significantly correlated with disease severity and mortality. ANGPTL4 may be a novel prognostic biomarker in ARDS.

IL-33 Treatment Attenuates the Systemic Inflammation Reaction in Acinetobacter baumannii Pneumonia by Suppressing TLR4/NF-κB Signaling.

Interleukin (IL)-33 treatment has been reported to reduce mortality in a rat model of sepsis, and the present study aimed to determine whether this effect of IL-33 is achieved through a reduction in the systemic inflammatory response in Acinetobacter baumannii pneumonia. After induction of pneumonia, rats were treated with normal saline or IL-33, and mortality over 5days was recorded. Inflammation within lung tissues was evaluated by hematoxylin and eosin staining as well as measurement of the concentrations of IL-8 and tumor necrosis factor alpha (TNF-α) in the bronchoalveolar lavage fluid (BALF) and plasma by enzyme-linked immunosorbent assay. In addition, the expression of Toll-like receptor 4 (TLR4), ST2, and nuclear factor kappa B (NF-κB) in rat lung tissues was assessed by western blotting. The result showed that the mortality rate and systemic inflammation were significantly increased in rats upon infection with A. baumannii, as evidenced by significant increases in the IL-8 and TNF-α levels in BALF and plasma as well as increased NF-κB activity and TLR4 expression in rat lung tissues. Importantly, IL-33 (1μg/kg) treatment significantly decreased mortality and pulmonary inflammation in A. baumannii-infected rats. Moreover, IL-33 treatment suppressed the elevation of IL-8 and TNF-α levels and inhibited TLR4 expression and NF-κB activation. Overall, these results suggest that IL-33 may decrease the mortality and inhibit the systematic inflammatory response associated with A. baumannii pneumonia by suppressing TLR4/NF-κB signaling.

Effects of p38 mitogen-activated protein kinase in rats with oleic acid-induced acute lung injury

To study the effects of p38 mitogen-activated protein kinase (p38MAPK) signal transduction pathway inhibitor SB203580 on the inflammatory reaction and lung water clearance, and to explore the role of p38MAPK in acute lung injury, to provide new way for p38MAPK inhibitor -SB203580 intervene fat embolism syndrome induced lung injury. Twenty-four adult male SD rats were randomly assigned to normal control group (OA group) (n=8), oleic acid-induced lung injury group (OA group, n=8)and SB203580 pretreatment group (n=8). OA-group was administered oleic acid (0.20 ml/kg) via right jugular vein; In SB203580-group, SB203580(5 mg/kg) was injected via jugular vein, followed 30 min before by OA infusion; At the 4 hours animals were sacrificed. Arterial blood gas, the wet/dry weight(W/D)of the right lower lung were examined, lung index(LI), pulmonary permeability index(PPI) and levels of tumor necrosis factor α(TNF-α) in bronchoalveolar lavage fluid(BALF) were examined. The expressions of p38MAPK and phospho-p38MAPK (p-p38MAPK) were determined by Western blot and immunohistochemical method. Pathological changes of the lung tissue were examined with light microscrope. Compared to control group, arterial oxygen partial pressure (PaO2) and PaO2/FiO2 were decreased in the animals of OA-group, while right lower lung wet/dry ratio, lung index, PPI, levels of TNF-α in BALF and the protein expression of p-p38MAPK were increased significantly (P<0.01). The pathological changes were observed significantly in injured lung tissue. Compared to OA-group, those indexes were improved in SB203580 pretreated group. p38MAPK signal transaction path mediated inflammatory response process and played an important role in acute lung injury. SB203580 could inhibit the expression of inflammatory cytokines, reduce lung edema, protect lung tissue of rats from OA-induced lung injury obviously. Therefore, inhibition of p38MAPK activity provides a new way for the clinical treatment of fat embolism syndrome induced lung injury.

Effect of aminophylline and simvastatin on airway inflammation and mucus hypersecretion in rats with chronic obstructive pulmonary disease

To observe the role of aminophylline and simvastatin in preventing and curing chronic obstructive pulmonary disease (COPD), and to explore the underlying mechanisms based on airway inflammation and mucus hypersecretion. The rat model of COPD was established by combination of cigarette smoking with intratracheal lipopolysaccharide (LPS) injection. Male SD rats were randomly divided into 4 groups (n=10 per group): a control group, a COPD group, an aminophylline group and a simvastatin group. The rats in the control group and the COPD group were treated with normal saline once a day via intragastric administration, while the rats in the aminophylline group and the simvastatin group were treated with aminophylline (5 g/L) and simvastatin (0.5 g/L) 1 mL/100 g once a day via intragastric administration, respectively. Pulmonary function and pathological changes in bronchus and lung were observed. The levels of IL-8, IL-17, and TNF-α in bronchoalveolar lavage fluid (BALF) were measured by enzyme-linked immunosorbent assay (ELISA). The mRNA and protein expressions of TLR4 and mucin 5AC (MUC5AC) in bronchi and lung tissues were detected by real-time PCR and Western blot, respectively. Pulmonary function and the pathophysiologic changes in bronchi and lung tissues in the COPD rats were consistent with typical phenotype of COPD. Compared with the control group, lung function indexes were significantly attenuated in the COPD group, while the levels of IL-8, IL-17, and TNF-α in BALF as well as the mRNA and protein levels of MUC5AC and TLR4 were significantly increased. Compared with the COPD group, lung function indexes were significantly increased in the aminophylline group and simvastatin group (P<0.01), while pulmonary pathological damages, the levels of IL-8, IL-17, and TNF-α in BALF as well as the mRNA and protein levels of MUC5AC and TLR4 were significantly decreased (P<0.01). Compared with the aminophylline group, the peak expiratory flow as well as the levels of IL-8, IL-17, and TNF-α in the simvastatin group were elevated (P<0.05). There are no significant difference in the mRNA and protein levels of MUC5AC and TLR4 between the 2 groups (P﹥0.05). Aminophylline and simvastatin can decrease IL-8, IL-17, and TNF-α levels in BALF and inhibit the expression of MUC5AC and TLR4 in airway and lung tissues in COPD rats, suggesting that they may have a preventive and therapeutic effect on COPD through reducing the airway inflammation and mucus hypersecretion.

Febuxostat protects rats against lipopolysaccharide-induced lung inflammation in a dose-dependent manner.

The aim of the present work was to investigate possible protective effects of febuxostat, a highly potent xanthine oxidase inhibitor, against acute lung injury (ALI) induced by lipopolysaccharide (LPS) in rats. Male Sprague Dawley rats were randomly divided into six groups, as follows: (i) vehicle control group; (ii) and (iii) febuxostat 10 and febuxostat 15 groups, drug-treated controls; (iv) LPS group, receiving an intraperitoneal injection of LPS (7.5 mg/kg); (v) and (vi) febuxostat 10-LPS and febuxostat 15-LPS groups, receiving oral treatment of febuxostat (10 and 15 mg/kg/day, respectively) for 7 days before LPS. After 18 h administration of LPS, blood was collected for C-reactive protein (CRP) measurement. Bronchoalveolar lavage fluid (BALF) was examined for leukocyte infiltration, lactate dehydrogenase (LDH) activity, protein content, and total nitrate/nitrite. Lung weight gain was determined, and lung tissue homogenate was prepared and evaluated for oxidative stress. Tumor necrosis factor-α (TNF-α) was assessed in BALF and lung homogenate. Moreover, histological changes of lung tissues were evaluated. LPS elicited lung injury characterized by increased lung water content (by 1.2 fold), leukocyte infiltration (by 13 fold), inflammation and oxidative stress (indicated by increased malondialdehyde (MDA), by 3.4 fold), and reduced superoxide dismutase (SOD) activity (by 34 %). Febuxostat dose-dependently decreased LPS-induced lung edema and elevations in BALF protein content, infiltration of leukocytes, and LDH activity. Moreover, the elevated levels of TNF-α in BALF and lung tissue of LPS-treated rats were attenuated by febuxostat pretreatment. Febuxostat also displayed a potent antioxidant activity by decreasing lung tissue levels of MDA and enhancing SOD activity. Histological analysis of lung tissue further demonstrated that febuxostat dose-dependently reversed LPS-induced histopathological changes. These findings demonstrate a significant dose-dependent protection by febuxostat against LPS-induced lung inflammation in rats.

The protective role of mesenchymal stem cells in ventilator-induced lung injury in rats

Objective To investigate the protective role of mesenchymal stem cells(MSCs)in ventilator- induced lung injury(VILI)in rats and its mechanisms. Methods Rat VILI model was established by high tidal volume ventilation in SD rats and randomly divided into 3 groups:sham operated group, control group and MSCs treatment group(n= 10 per groups).The lung tissue injury, lung polymorphonuclear(PMN) cells counts and pro- inflammatory cytokines[interleukin(IL)- 1β, IL- 6 and tumor necrosis factor-α(TNF-α)]levels in bronchoalveolar lavage fluid(BALF) were measured 4 h after mechanical ventilation. Results 4 h after mechanical ventilation, the lung tissue injury score of MSCs group was 4.23±1.64, significantly lower than that of control group(7.31±2.15, t= 6.427, P 0.05). The lung PMN counts of three groups were 9.27±4.55,26.37±9.44 and 15.33±6.62, respectively.The PMN counts of MSCs group was significantly lower than that of saline group(t= 3.011, P 0.05). The IL- 1β, IL- 6 and TNF-αlevels in BALF of MSCs group was(70.40±31.40),(41.37±14.34)and(47.60±10.23)ng/L, respectively,significantly lower than that of control group[(170.20±58.90) ng/L, t= 5.04, P 0.05;(29.20±9.11) ng/L, t= 2.33, P> 0.05;(39.44± 12.14) ng/L,t= 0.91, P> 0.05]. Conclusion MSCs has a protective effect on VILI by attenuating the degree of lung tissue injury, PMN lung infiltration and production of local pro- inflammatory cytokines. Key words: Mesenchymal stem cells; Ventilator-induced lung injury; Polymorphonuclear; Inflammatory

Effects of combining tumor necrosis factor-α inhibitor with adenosine A2b receptor antagonist on asthmatic lung inflammation in mice

Objective To explore the effects of combining tumor necrosis factor-α(TNF-α) inhibitor with adenosine A2b receptor antagonist CVT-6883 on asthmatic lung inflammation in mice. Methods A total of 40 female Balb/c mice were evenly randomized into 5 groups, including normal control group, asthma group, CVT-6883 group, CVT-6883 + etanercept group, and etanercept group.The pathological changes in the lungs were determined and the number of white blood cells(WBC) and eosinophil(EOS) in the bronchoalveolar lavage fluid(BALF) was counted by cell count in each group.The levels of TNF-α in BALF were evaluated by enzyme-linked immunosorbent assay (ELISA). The expression of adenosine A2b receptor mRNA in the lung tissues were measured by reverse transcription-polymerase chain reaction(RT-PCR). Results 1.The lung tissue in asthma group, dyed by HE, was found to have a large number of airway inflammatory cell infiltration, thickening of the bronchial mucosa, the alveolar septa widened and fracture.In the CVT-6883, CVT-6883 + etanercept and etanercept group, the pathological changes were relieved.2.The WBC and EOS counts in BALF of the asthma group[(413.8±5.8)/L, (139.3±1.4)/L] were higher than those of the normal control group[(24.0±1.3)/L, (1.8±0.1)/L, P<0.05]. The WBC and EOS counts of the CVT-6883 group[(111.5±3.8)/L, (3.3±0.1)/L], the etanercept group + the CVT-6883 group[(173.8±3.9)/L, (10.4±0.2)/L], and the etanercept group[(138.4±3.0)/L, (4.1±0.1)/L] were lower than those of the asthma group (P<0.05).3. Compared with the control group(100.4±5.7) ng/L, the TNF-α concentration of the asthma group(145.2±8.8) ng/L was significantly higher (P<0.05); the TNF-α concentration of CVT-6883 group(130.9±5.9) ng/L, CVT-6883 + etanercept group(115.7±8.2) ng/L and the etanercept group(122.0±8.7) ng/L, were significantly decreased compared with asthma group (P<0.05). 4. In asthma group(8.9±1.1)compared with the control group(0.6±0.2), the A2bAR(adenosine A2b receptor) mRNA expression was upregulated (P<0.05); CVT-6883 group(1.6±0.3), CVT-6883 + etanercept group(2.5±0.6)and the etanercept group(5.3±0.4), the A2bAR(adenosine A2b receptor) mRNA expression was significantly decreased compared with asthma group (all P<0.05). Conclusion Combination of TNF-α inhibitor with adenosine A2b receptor antagonist can reduce asthmatic lung inflammation. Key words: Tumor necrosis factor-α; Adenosine A2b receptor; Asthma; Etanercept

Effects of Schisandra chinensis Baillon (Schizandraceae) on lipopolysaccharide induced lung inflammation in mice

Ethnopharmacological relevanceSchisandra chinensis Baillon (Sc), an anti-inflammatory herb that has been used in traditional Chinese medicine for thousands of years, is frequently used to treat upper respiratory tract infections. Aim of the studyThis study was conducted to evaluate the ability of a water extract of Sc to prevent airway inflammation both in vitro and in vivo. Materials and methodsHuman lung alveolar epithelial-derived A549 cells were stimulated with to interleukin-1β, tumor necrosis factor-α, and interferon-γ (IL-1β, TNF-α, and INF-γ; cytokine mixture; CM) and treated with Sc extracts. They were then evaluated using nitric oxide (NO), IL-8 and monocyte chemotactic protein-1 (MCP-1) secretions. In the in vivo study, BALB/c mice were challenged with lipopolysaccharide (LPS) to induce acute airway inflammation. After this challenge, the mice were treated with Sc extracts (10, 50 and 100mg/kg) by oral administration, and inflammatory cells in the bronchoalveolar lavage (BAL) fluid were counted. IL-6 and TNF-α secretions were measured using an enzyme-linked immunosorbent assay. Lung tissues of the LPS treated mice were prepared and stained with hematoxylin and eosin (HE) for histological examination. ResultsIn the A549 cells, Sc extracts dose-dependently and significantly inhibited CM-induced NO production and reduced IL-8 and MCP-1 secretions. Sc extracts efficiently suppressed neutrophil and macrophage infiltrations of lung tissues and increased IL-6 and TNF-α levels in BAL fluid in LPS-instilled BALB/c mice. In addition, Sc extracts treatment inhibited pathologic progress in the lung tissues, as confirmed by H&E staining. These findings indicate that Sc extracts could be potentially useful for the treatment of acute lung inflammation and acute lung injury.

Diagnostic Accuracy of Cytokine Levels (TNF-α, IL-2 and IFN-γ) in Bronchoalveolar Lavage Fluid of Smear-Negative Pulmonary Tuberculosis Patients

Background: The determination of cytokine concentrations in serum and bronchoalveolar lavage fluid (BALF) may contribute to the diagnosis of tuberculosis (TB) since cytokines have been ascribed an important role in TB pathogenesis. Objective: To assess the diagnostic accuracy of TNF-α, IFN-γ and IL-2 levels in serum and BALF of smear-negative pulmonary TB patients. Method: BALF was obtained from the affected lobe in patients with smear-negative TB or other pulmonary diseases (OPD), and from the right middle lobe in healthy controls. ELISA and a nephelometric method were used to detect cytokine and albumin levels. Results: TNF-α levels in BALF were significantly elevated in the TB group (n = 15) compared with the OPD patients (n = 40) and controls (n = 17; p < 0.001). Although these three cytokines correlated well with each other in BALF (p < 0.0001, and r ≧ 0.7, respectively), BALF IL-2 and IFN-γ levels were not significantly different among the groups (p > 0.05). BALF TNF-α or IFN-γ levels were significantly higher in patients with cavitary disease (n = 11) versus those without (n = 61; p < 0.05). However, no significant difference was found between cavitary (n = 7) and non-cavitary TB in cytokine levels (p > 0.05). Neither gender nor smoking status showed any statistical differences in cytokines in the groups (p > 0.05). Sensitivity and specificity of BALF TNF-α were found to be 73 and 76%, respectively. The positive and negative predictive values for BALF TNF-α were 44 and 91%, respectively. Conclusion: In cases of smear-negative TB, BALF TNF-α can be a useful tool to identify healthy subjects rather than smear-negative TB patients.

Murine TLR4 Is Implicated in Cigarette Smoke-Induced Pulmonary Inflammation

Background: Chronic obstructive pulmonary disease (COPD) is associated with an abnormal inflammatory response of the lungs to noxious particles or gases. We investigated whether Toll-like receptor 4 (TLR4) is implicated in cigarette smoke (CS)-induced pulmonary inflammation in a murine model of COPD. Methods: C3H/HeOuJ (Tlr4^WT) and C3H/HeJ (Tlr4^defective) mice were exposed to air or CS for 5 weeks (subacute) and 26 weeks (chronic), and pulmonary inflammation was evaluated. Results: In Tlr4^WT mice, subacute and chronic CS exposure induced a substantial pulmonary infiltration of macrophages, neutrophils, lymphocytes and dendritic cells (DCs), that was absent in air-exposed mice. CS exposure increased the costimulatory marker expression on DCs, the levels of monocyte chemoattractant protein-1 (MCP-1) and tumor necrosis factor-α (TNF-α) in bronchoalveolar lavage (BAL) fluid and induced the pulmonary expression of matrix metalloproteinase-12 (MMP-12), TLR4 and TLR2. In contrast, after subacute CS exposure, Tlr4^defective mice showed a limited (5-fold lower) increase of DCs and lymphocytes in BAL fluid, lower costimulatory marker expression on DCs and lower MCP-1 and TNF-α levels in BAL fluid compared to Tlr4^WT animals. After chronic CS exposure, however, the difference in pulmonary inflammation between Tlr4^WT and Tlr4^defective mice was less pronounced and both strains showed similar MCP-1 and TNF-α levels in BAL and similar pulmonary MMP-12, TLR4 and TLR2 expression. Conclusions: We demonstrated that the TLR4 mutation in C3H/HeJ mice is protective against CS-induced pulmonary influx of neutrophils, DCs and lymphocytes upon subacute CS exposure. However, TLR4 is only of minor importance in chronic CS-induced inflammation in mice.

Pneumocystis cariniiinfection sensitizes lung to radiation-induced injury after syngeneic marrow transplantation: role of CD4+T cells

A murine model of bone marrow transplant (BMT)-related lung injury was developed to study how infection sensitizes lung to the damaging effects of total body irradiation (TBI) at infectious and TBI doses that individually do not cause injury. Mice infected with Pneumocystis carinii exhibited an asymptomatic, rapid, and transient influx of eosinophils and T cells in bronchoalveolar lavage fluid (BALF). In contrast, mice infected with P. carinii 7 days before receiving TBI and syngeneic BMT (P. carinii/TBI mice) exhibited severe pulmonary dysfunction, surfactant aggregate depletion, and surfactant activity reductions at 17 days post-BMT. BALF from P. carinii/TBI mice contained a disproportionate initial influx of CD4(+) T cells (CD4(+):CD8(+) ratio of 2.7) that correlated with progressive lung injury (from 8 to 17 days post-BMT). Levels of TNF-alpha in BALF were significantly increased in P. carinii/TBI mice compared with mice given either insult alone, with peak values found at 11 days post-BMT. In vivo depletion of CD4(+) T cells in P. carinii/TBI mice abrogated pulmonary dysfunction and reduced TNF-alpha levels in BALF, whereas depletion of CD8(+) T cells did not affect lung compliance or TNF-alpha. Lung injury was not attributable to direct P. carinii damage, since CD4-depleted P. carinii/TBI mice that exhibited no injury had higher average lung P. carinii burdens than either mice given P. carinii alone or undepleted P. carinii/TBI mice. Together, these results indicate that P. carinii infection can sensitize the lung to subsequent TBI-mediated lung injury via a process dependent on non-alloreactive CD4(+) T cells.