Irisin attenuates fine particulate matter induced acute lung injury by regulating Nod2/NF-κB signaling pathway.

Air pollution consisting of fine particulate matter (PM2.5) can induce or aggravate pulmonary inflammatory injury. Irisin has been shown to inhibit inflammation and help to protect against acute kidney, lung or brain injury. However, the role of irisin in lung inflammation after exposure to PM2.5 remains unclear. The aim of this study was to investigate the effect and molecular mechanism of irisin supplementation on in vitro and in vivo models of PM2.5-induced acute lung injury（ALI). C57BL/6 mice and alveolar macrophage cell line (MH-S) were treated with PM2.5. Histopathological examination and FNDC5/ irisin immunofluorescence staining was performed on lung tissue sections. MH-S cell viability was determined by CCK-8 assay. The levels of Nod2, NF-κB p65 and NLRP3 were detected by qRT-PCR and western blotting. The levels of cytokines (IL-1ß, IL-18 and TNF-α) were detected by ELISA. PM2.5 exposure induced increased secretion of pro-inflammatory factors and activation of Nod2, NF-κB p65 and NLRP3 as well as endogenous levels of irisin. In vivo and in vitro inflammation was alleviated by irisin supplementation. Irisin significantly decreased IL-1ß, IL-18, and TNF-α production at both mRNA and protein level. Expression levels of Nod2, NF-κB p65, and NLRP3 were all significantly affected by irisin. In vivo the degree of pulmonary injury and inflammatory infiltration was weakened after irisin administration. In vitro, irisin could inhibit the activation of the NLRP3 inflammasome for a sustained period of 24 h, and its inhibitory ability was gradually enhanced. In conclusion, our findings indicate that irisin can modulate the inflammatory injury of lung tissue caused by PM2.5 through the Nod2/NF-κB signaling pathway, suggesting that irisin can be a candidate for the therapeutic or preventive intervention in acute lung inflammation.

Irisin attenuates acute lung injury by suppressing the pyroptosis of alveolar macrophages.

Irisin is a hormone­like myokine that regulates cell signaling pathways and exerts anti­inflammatory effects. However, the specific molecular mechanisms involved in this process are currently unknown. The present study explored the role and mechanisms underlying the functions of irisin in alleviating acute lung injury (ALI). The present study used MH­S, an established murine alveolar macrophage­derived cell line, and a mouse model of lipopolysaccharide (LPS)­induced­ALI to examine the efficacy of irisin against ALI in vitro and in vivo, respectively. Fibronectin type III repeat­containing protein/irisin was expressed in the inflamed lung tissue, but not in normal lung tissue. Exogenous irisin reduced alveolar inflammatory cell infiltration and pro­inflammatory factor secretion in mice following LPS stimulation. It also inhibited the polarization of M1­type macrophages and promoted the repolarization of M2­type macrophages, thus reducing the LPS­induced production and secretion of interleukin (IL)­1ß, IL­18 and tumor necrosis factor­α. In addition, irisin reduced the release of the molecular chaperone heat shock protein 90 (HSP90), inhibited the formation of nucleotide­binding and oligomerization domain­like receptor protein 3 (NLRP3) inflammasome complexes, and decreased the expression of caspase­1 and the cleavage of gasdermin D (GSDMD), leading to reduced pyroptosis and the accompanying inflammation. On the whole, the findings of the present study demonstrate that irisin attenuates ALI by inhibiting the HSP90/NLRP3/caspase­1/GSDMD signaling pathway, reversing macrophage polarization and reducing the pyroptosis of macrophages. These findings provide a theoretical basis for understanding the role of irisin in the treatment of ALI and acute respiratory distress syndrome.

Idiopathic mesenteric phlebosclerosis associated with long-term oral intake of geniposide.

BACKGROUND: Idiopathic mesenteric phlebosclerosis (IMP) is a rare disease, and its etiology and risk factors remain uncertain. AIM: To investigate the possible influence of Chinese herbal liquid containing geniposide on IMP. METHODS: The detailed formula of herbal liquid prescriptions of all patients was studied, and the herbal ingredients were compared to identify the toxic agent as a possible etiological factor. Abdominal computed tomography (CT) and colonoscopy images were reviewed to determine the extent and severity of mesenteric phlebosclerosis and the presence of findings regarding colitis. The disease CT score was determined by the distribution of mesenteric vein calcification and colon wall thickening on CT images. The drinking index of medicinal liquor was calculated from the daily quantity and drinking years of Chinese medicinal liquor. Subsequently, Spearman's correlation analysis was conducted to evaluate the correlation between the drinking index and the CT disease score. RESULTS: The mean age of the 8 enrolled patients was 75.7 years and male predominance was found (all 8 patients were men). The patients had histories of 5-40 years of oral Chinese herbal liquids containing geniposide and exhibited typical imaging characteristics (e.g., threadlike calcifications along the colonic and mesenteric vessels or associated with a thickened colonic wall in CT images). Calcifications were confined to the right-side mesenteric vein in 6 of the 8 patients (75%) and involved the left-side mesenteric vein of 2 cases (25%) and the calcifications extended to the mesorectum in 1 of them. The thickening of colon wall mainly occurred in the right colon and the transverse colon. The median disease CT score was 4.88 (n = 7) and the median drinking index was 5680 (n = 7). After Spearman's correlation analysis, the median CT score of the disease showed a significant positive correlation with the median drinking index (r = 0.842, P < 0.05). CONCLUSION: Long-term oral intake of Chinese herbal liquid containing geniposide may play a role in the pathogenesis of IMP.

Role of CCL20/CCR6 and the ERK signaling pathway in lung adenocarcinoma.

Previous studies have revealed that carcinoma-associated fibroblasts communicate microenvironment-derived signals through chemokine/chemokine receptor interaction, resulting in carcinogenesis. C-C motif chemokine ligand 20 (CCL20)/C-C motif chemokine receptor 6 (CCR6) interactions are involved in the pathogenesis of colonic malignancies. The present study aimed to characterize the roles of CCL20/CCR6 and the extracellular signal-regulated kinase (ERK) signaling pathway in lung adenocarcinoma growth. Lung adenocarcinoma samples obtained at surgery were assessed for the expression, tissue localization and production of CCL20/CCR6. In addition, colony formation, ERK signaling and chemokine production were measured to assess the responsiveness of the A549 cell line to CCL20 stimulation. CCL20 and CCR6 were found to be highly expressed in the majority of samples in the recurrence group (76 and 66%, respectively). The staining indexes of CCL20 and CCR6 in the recurrence group were 149.3 and 134.4, respectively, which were significantly higher than those in the non-recurrence group (57.2 and 58.0, respectively); the protein and mRNA expression levels determined by western blot and reverse transcription-quantitative polymerase chain reaction were also found to be high in the recurrence group For A549 cells, the colony-forming capacity was increased by CCL20 stimulation, and this effect was dependent in part on ERK phosphorylation. Collectively, the findings suggest that CCR6 and CCL20 may serve a role in lung adenocarcinoma, leading to proliferation and migration via autocrine or paracrine mechanisms. The disruption of CCL20/CCR6 interactions may be a promising strategy for the treatment of cancer.

In vitro modeling of COPD inflammation and limitation of p38 inhibitor - SB203580.

BACKGROUND: Systemic inflammation and steroid resistance are the hallmarks of COPD. We examined the impact of p38 inhibitor (SB203580) in in vitro assays of systemic inflammation using pulmonary cells and patients' sera. OBJECTIVE AND METHODS: Data from 66 COPD patients and 15 age-/sex-matched healthy controls were compared. Interleukin-10 (IL-10), tumor necrosis factor-α (TNF-α), and CCL5 were measured in serum samples and culture media from peripheral blood mononuclear cells. The impact of sera on IL-10 and CCL5 expression in alveolar macrophage cell line (MH-S) was examined. The in vitro effects of SB203580 on lipopolysaccharide-induced inflammation were investigated. RESULTS: Peripheral blood mononuclear cells from Global initiative for chronic Obstructive Lung Disease (GOLD) D patients produced more CCL5 and TNF-α, and less IL-10 compared to GOLD A-C patients. SB203580 treatment suppressed CCL5 and TNF-α and stimulated IL-10 production; however, the effect of SB203580 on IL-10 was lower in the COPD group. Culture of MH-S cells with COPD serum showed a significant increase in CCL5 and a significant decrease in IL-10 compared to healthy serum. This effect was not suppressed with SB203580 treatment. CONCLUSION: COPD serum has a potent proinflammatory effect on pulmonary cells. Inhibition of p38 phoshorylation had a limited effect in restoring impaired lymphocyte function and suppressing inflammation induced by COPD serum, implying important p38-independent inflammatory mechanisms in COPD.

Additive Suppression of LPS-Induced IL-10 and TNF-α by Pre-treatment of Dexamethasone and SB203580 in a Murine Alveolar Macrophage Cell Line (MH-S).

P38 inhibitors are potent anti-inflammatory agents with distinctive mechanism of action from corticosteroid; the potential combined use of both anti-inflammatory agents could be an effective treatment for inflammatory lung disease; however, the impact of such combination on the homeostasis of immune response was poorly understood. To investigate the combined effect of dexamethasone (DEX) and/or SB203580 on tumor necrosis factor (TNF)-α (pro-inflammatory) and interleukin (IL)-10 (anti-inflammatory) secretion in mouse alveolar macrophage cell line. Secreted TNF-α and IL-10 were measured by ELISA. Phosphorylated STAT3 were investigated using Western blotting and immunocytochemistry. Pre-treatment of DEX or SB203580 inhibited lipopolysaccharide (LPS)-stimulated IL-10, TNF-α secretion, and STAT3 phosphorylation. Combined use of both agents showed stronger inhibitory effect. Combining DEX and SB203580 showed strong inhibition on the LPS-induced IL-10 secretion and STAT3 phosphorylation, which might reflect a very important drawback from the combined use of both anti-inflammatory agents.

Role of p38 MAPK and STAT3 in lipopolysaccharide-stimulated mouse alveolar macrophages.

Excessive production of inflammatory mediators is an important feature of inflammatory lung disease. In macrophages, mitogen-activated protein kinase (MAPK) and signal transducer and activator of transcription-3 (STAT3) are crucial mediators for the production of proinflammatory cytokines. In the present study, the role of MAPK and STAT3 on tumor necrosis factor (TNF)-α and interleukin (IL)-10 production was investigated in mouse alveolar macrophages. The levels of TNF-α and IL-10 in lipopolysaccharide (LPS; 100 ng/ml)-stimulated MH-S cell lines were measured by an enzyme-linked immunosorbent assay, with or without p38 inhibitor (SB203580; 5, 10 or 15 µM) intervention. Phosphorylated STAT3 (p-STAT3) expression was examined by western blot analysis and immunocytochemistry following LPS stimulation for 15 or 30 min. Antibodies against STAT3 were used to verify comparable sample loading. Cells stimulated with LPS showed significantly increased levels of p-STAT3 protein (P<0.05) when compared with the baseline levels. TNF-α and IL-10 protein levels also increased following LPS stimulation (P<0.05). By contrast, treatment with the p38 inhibitor, SB203580, decreased the levels of p-STAT3, TNF-α and IL-10 (P<0.05) following LPS stimulation. SB203580 was shown to inhibit LPS-stimulated TNF-α expression (P<0.05) in a concentration-dependent manner, reaching significance at a concentration of 10 µM. However, the inhibition of IL-10 expression was not concentration-dependent. Therefore, LPS-stimulated overproduction of TNF-α and IL-10 is mediated at least partially by the MAPK pathway. Inhibition of p38 prevented LPS-induced STAT3 phosphorylation, indicating an interaction between the STAT3 and MAPK signaling pathways.

High serum macrophage inflammatory protein-3α is associated with the early recurrence or metastasis of non-small cell lung cancer following primary pulmonary resection.

The present study sought to characterize the role of macrophage inflammatory protein-3α (MIP-3α) in non-small cell lung cancer (NSCLC) patients with early recurrence or metastasis after primary pulmonary resection. Follow-up examinations were conducted for 203 NSCLC patients with primary pulmonary resection for two years post-operatively, and data was also collected for 20 healthy subjects. Serum MIP-3α levels were determined prior to surgery and at post-operative days (PODs) 30, 90 and 180, and the relevant clinical and operative variables were collected. Serum MIP-3α was measured using a commercially available enzyme-linked immunosorbent assay. There were no significant differences in age, gender and histological type among all groups (P>0.05). Serum MIP-3α levels on POD 180 were significantly higher in the recurrence group than in the non-recurrence group and healthy subjects (P=0.001). There was no significant difference in the serum MIP-3α level at PODs 90 and 180 in the patients with or without adjuvant chemotherapy (P>0.05). The recurrence rate in the high serum MIP-3α level group was 41.67%, much higher than the 23.53% observed in the low level group (P=0.006). The patients with high serum levels of MIP-3α had a significantly shorter overall recurrence-free time compared with those with low levels (P=0.004). Multivariate Cox's regression analyses showed that only serum MIP-3α level was significant, with a hazard ratio of 1.061, a 95% confidence interval of 1.044-1.078 and a P-value of 0.001. The serum MIP-3α level in the patients with liver and bone metastases were remarkably higher than those with recurrence at other sites. The high post-operative serum MIP-3α levels were associated with an increased risk of post-operative early recurrence or metastasis in the lung cancer patients, specifically in those with bone or liver metastases.

Classification and comparison of municipal solid waste based on thermochemical characteristics.

UNLABELLED: Municipal solid waste (MSW) has been normally sorted into six categories, namely, food residue, wood waste, paper textiles, plastics, and rubber In each category, materials could be classified further into subgroups. Based on proximate and ultimate analysis and heating value, statistical methods such as analysis of variance (ANOVA) and cluster analysis were applied to analyze the characteristics of MSW in every subgroup and to try to distinguish their relative properties. The chemical characteristics analysis of MSW showed that polyethylene (PE), polypropylene (PP), and polystyrene (PS) had the highest volatile matter content, with almost no ash and fixed carbon, while polyethylene terephthalate (PET) had high carbon content but low hydrogen content. Bones and vegetables had the highest ash content, while nutshells and rubber had the highest fixed carbon content. Paper and starch food had the highest oxygen content, and wool and bones had the highest nitrogen and sulfur content. Polyvinyl chloride (PVC) had the highest chlorine content at about 55%. PE, PP and PS had the highest heating value, followed by chemical products such as rubber and chemical fiber. Conversely, paper, vegetables and bones had the lowest heating value. The results of cluster analysis of MSW components showed that fruit peel, weeds, wood, bamboo, leaves and nutshells could be classified as the lignocellulose category; starch food, cotton, toilet paper, printing paper and cardboard could be classified as the glucose monomer category; wood and chemical fiber could be classified as the high nitrogen and sulfur category; and PE, PP, and PS could be cluster as the polyolefin category. IMPLICATIONS: The yield of municipal solid waste (MSW) is constantly increasing and waste to energy (WTE) has been used extensively all over the world. During the processes of incineration, pyrolysis, or gasification, the impact of physical and chemical properties of MSW is of great significance. However, the traditional classification of MSW is too general to provide more detailed information in many investigations. It is necessary to perform the investigation of characteristics of combustible MSW to distinguish different categories of MSW and find out their subclassification.

Investigation of heavy metal partitioning influenced by flue gas moisture and chlorine content during waste incineration.

The impact of moisture on the partitioning of the heavy metals including Pb, Zn, Cu and Cd in municipal solid waste (MSW) was studied in a laboratory tubular furnace. A thermodynamic investigation using CHEMKIN software was performed to compare the experimental results. Simulated waste, representative of typical MSW with and without chlorine compounds, was burned at the background temperature of 700 and 950 degrees C, respectively. In the absence of chlorine, the moisture content has no evident effect on the volatility of Pb, Zn and Cu at either 700 or 950 degrees C, however, as flue gas moisture increasing the Cd distribution in the bottom ash increased at 700 degrees C and reduced at 950 degrees C, respectively. In the presence of chlorine, the flue gas moisture reduced the volatility of Pb, Zn and Cu due to the transformation of the more volatile metal chlorides into less volatile metal oxides, and the reduction became significant as chlorine content increase. For Cd, the chlorine promotes its volatility through the formation of more volatile CdCl2. As a result, the increased moisture content increases the Pb, Zn, Cu and Cd concentrations in the bottom ash, which limits the utilization of the bottom ash as a construction material. Therefore, in order to accumulate heavy metals into the fly ash, MSW should be dried before incineration.

Experimental and thermodynamic investigation on transfer of cadmium influenced by sulfur and chlorine during municipal solid waste (MSW) incineration.

We used two approaches to investigate the impact of sulfur and chlorine compounds on transfer of a semivolatile heavy metal, cadmium, during municipal solid waste (MSW) incineration: experiments using a tubular furnace reactor and thermodynamic equilibrium calculations. Artificial wastes representative of typical MSW in China with and without the presence of sulfur and chlorine compounds were combusted at 850 degrees C, and the partitioning of Cd among bottom ash, fly ash and flue gas was quantified. The results indicate that sulfur compounds in the elemental form and reduced state could stabilize Cd in the form of CdS due to local reducing environment, while sulfur in the oxidized forms slightly increased Cd volatilization during incineration. In contrast, the presence of chlorine compounds significantly increased the partitioning of Cd on fly ash. Chemical equilibrium calculations show that sulfur binds with Cd and alters Cd speciation at low temperatures (<700 degrees C), while chlorine significantly increases the volatilization of Cd through formation of volatile CdCl(2) and thus its partitioning on the fly ash between 400 and 1000 degrees C. The equilibrium calculation results also suggest that SiO(2)- and Al(2)O(3)-containing minerals could function as sorbents stabilizing Cd as condensed phase solids (CdSiO(3) and CdAl(2)O(4)). These findings provide useful information on understanding the partitioning behavior of Cd and can help development of strategies to control volatilization of Cd during MSW incineration.

CCK-8 inhibits expression of TNF-alpha in the spleen of endotoxic shock rats and signal transduction mechanism of p38 MAPK.

AIM: To study the effect of sulfated cholecystokinin-octapeptide (CCK-8) on systemic hypotension, gene and protein expression of TNF-alpha in the spleen of lipopolysaccharide (LPS)-induced endotoxic shock (ES) rats, and further investigate the signal transduction mechanism of p38 mitogen-activated protein kinase (MAPK). METHODS: The changes of blood pressure were observed using physiological record instrument in the four groups of rats: LPS (8 mg x kg(-1), iv), CCK-8 (40 microg x kg(-1), iv) pretreatment 10 min before LPS (8 mg x kg(-1)), CCK-8 (40 microg x kg(-1), iv) or normal saline (control) group. The content of TNF-alpha in the spleen was assayed 2 h after LPS administration using ELISA kit and the expression of TNF-alpha mRNA was examined 30 min, 2 h and 6 h after LPS administration by reverse transcribed polymerase chain reaction (RT-PCR). Activation of p38 MAPK was detected with Western blot 30 min after LPS administration. RESULTS: CCK-8 reversed LPS-induced decrease of mean arterial pressure (MAP) in rats. The content of TNF-alpha in the spleen was (282+/-30) ng x L(-1) in control group, while it increased to (941+/-149) ng x L(-1) in LPS group, P<0.01. CCK-8 significantly inhibited the LPS-induced increase of TNF-alpha content in spleen. It decreased to (462 +/-87) ng x L(-1) in CCK-8+LPS group, P<0.01. The expression of TNF-alpha mRNA 30 min and 2 h after treatment was stronger in LPS group, while it was lowered after CCK-8 pretreatment. The p38 MAPK expression increased significantly in LPS group (5.84 times of control) and CCK-8 increased the activation of p38 MAPK in ES rats (10.74 times of control). CONCLUSION: CCK-8 reverses the decrease of MAP in ES rats and has inhibitory effect on the gene and protein expression of TNF-alpha in spleen, and p38 MAPK may be involved in its signal transduction mechanisms.

Graft expression of p38 and tumor necrosis factor-alpha in heart transplantation in rats.

OBJECTIVES: To investigate the expression of p38 mitogen-activated protein kinase and its relationship with myocardial apoptosis and tumor necrosis factor-alpha during acute cardiac allograft rejection and to study the effects of tacrolimus on the expression of the kinase. METHODS: Rats were divided into 3 groups: isograft (Lewis heart to Lewis rat; control group), allograft (Brown Norway heart to Lewis rat), and tacrolimus-treated allograft (Brown Norway heart to tacrolimus-treated Lewis rat). Grafts were collected 1, 3, 5, and 7 days after transplantation for determination of histopathological features, apoptosis of cardiac cells (by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick labeling), number of cells positive for both p38 and CD8 (by laser scanning confocal imaging), and expression of the kinase (by Western immunoblotting) and tumor necrosis factor-alpha (by reverse-transcriptase polymerase chain reaction). RESULTS: Compared with isografts from the control group, grafts from the untreated allograft group had significantly more apoptotic cells, greater expression of tumor necrosis factor-alpha and p38 mitogen-activated protein kinase, and more CD8-p38 double-positive cells at 5 and 7 days (P < .05). The increases were prevented by treatment with tacrolimus. CONCLUSIONS: The findings that the number of apoptotic cells, the number of CD8-p38 double-positive cells, the expression of tumor necrosis factor-alpha and p38 mitogen-activated protein kinase all increased during the same period in the allografts in nonimmunosuppressed recipients suggests that intragraft expression of p38 would be associated with the rejection in acute cardiac allograft rejection. Tacrolimus may alleviate rejection partly by inhibiting p38 mitogen-activated protein kinase.