Pulmonary Inflammation In Mice Research Articles

Cynaropicrin attenuates inflammatory cytokines in LPS‐induced RAW264.7 cells and ovalbumin‐induced asthmatic mice

AbstractThis study examines the anti‐inflammatory activity of cynaropicrin against lipopolysaccharide (LPS) in vitro and ovalbumin (OVA)‐challenged asthma in mice. Cynaropicrin's antimicrobial effects were tested on Escherichia coli (E. coli) and Streptococcus pyogenes (S. pyogenes) using the disc diffusion technique. Cytotoxicity was assessed with an (3‐(4, 5‐dimethylthiazolyl‐2)‐2, 5‐diphenyltetrazolium bromide) assay. The anti‐inflammatory property was evaluated in LPS‐induced RAW264.7 cells, while OVA‐challenged asthmatic mice were treated with 10 mg/kg of cynaropicrin. Key inflammatory and antioxidant markers were quantified, and lung histology was examined to confirm therapeutic roles. The antimicrobial studies proved that cynaropicrin effectively inhibited the growth of E. coli and S. pyogenes. Cynaropicrin displayed no cytotoxicity on RAW264.7 cells. Furthermore, it significantly inhibited inflammatory cytokine synthesis upon LPS induction. Cynaropicrin treatment decreased the inflammatory cell counts and also suppressed specific allergic markers in OVA‐challenged mice. It also decreased nitric oxide and myeloperoxidase levels and reduced pulmonary edema. Cynaropicrin increased antioxidant levels and decreased proinflammatory cytokines in the asthmatic mice. Lung histological examination confirms the ameliorative potency of cynaropicrin against OVA‐induced asthmatic pulmonary inflammation in mice. Our findings suggest cynaropicrin possesses significant ameliorative potency against allergen‐induced pulmonary inflammation.

The Effect of Green Tea Extract on Pulmonary Inflammation in Nanoparticles-Exposed Mice.

Titanium dioxide nanoparticles (TiO2 NPs) are air pollutants that exacerbate chronic respiratory diseases such as asthma and Chronic Obstructive Pulmonary Disease (COPD) However, little is known about the mechanism underlying the antipollutant effects of green tea extract (GTE). This study evaluates the efficacy and mechanism of GTE on lung inflammation and fibrosis in mice exposed to TiO2 NPs. The TiO2 NPs model is induced by having mice inhale TiO2 NPs, while controls receive an equivalent volume of saline. Treatment with oral GTE is initiated after TiO2 NPs inhalation and is given once daily for 4 weeks. Airway resistance and pulmonary inflammation are increased in mice exposed to TiO2 NPs. GTE treatment reduces the airway inflammation and airway resistance, and attenuates the pathological changes including lung fibrosis compared to the mice exposed to TiO2 NPs. With GTE, there are no significant increases in cytokines and immunoglobulin E (IgE) in mice exposed to TiO2 NPs. GTE inhibits matrix metalloproteinases (MMPs) and apoptotic factors induced by TiO2 NPs exposure, and these protective effects of GTE are closely related to the mitogen-activated protein kinase (MAPK) signaling pathway. GTE modulates pulmonary inflammation in mice exposed to air pollutants, suggesting that GTE may be beneficial in respiratory diseases exacerbated by such pollutants.

CPT1A-IL-10-mediated macrophage metabolic and phenotypic alterations ameliorate acute lung injury.

Acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) is a common acute respiratory failure due to diffuse pulmonary inflammation and oedema. Elaborate regulation of macrophage activation is essential for managing this inflammatory process and maintaining tissue homeostasis. In the past decades, metabolic reprogramming of macrophages has emerged as a predominant role in modulating their biology and function. Here, we observed reduced expression of carnitine palmitoyltransferase 1A (CPT1A), a key rate-limiting enzyme of fatty acid oxidation (FAO), in macrophages of lipopolysaccharide (LPS)-induced ALI mouse model. We assume that CPT1A and its regulated FAO is involved in the regulation of macrophage polarization, which could be positive regulated by interleukin-10 (IL-10). After nasal inhalation rIL-10 and/or LPS, wild type (WT), IL-10-/-, Cre-CPT1Afl/fl and Cre+CPT1Afl/fl mice were sacrificed to harvest bronchoalveolar lavage fluid, blood serum and lungs to examine cell infiltration, cytokine production, lung injury severity and IHC. Bone marrow-derived macrophages (BMDMs) were extracted from mice and stimulated by exogenous rIL-10 and/or LPS. The qRT-PCR, Seahorse XFe96 and FAO metabolite related kits were used to test the glycolysis and FAO level in BMDMs. Immunoblotting assay, confocal microscopy and fluorescence microplate were used to test macrophage polarization as well as mitochondrial structure and function damage. In in vivo experiments, we found that mice lacking CPT1A or IL-10 produced an aggravate inflammatory response to LPS stimulation. However, the addition of rIL-10 could alleviate the pulmonary inflammation in mice effectively. IHC results showed that IL-10 expression in lung macrophage decreased dramatically in Cre+CPT1Afl/fl mice. The in vitro experiments showed Cre+CPT1Afl/fl and IL-10-/- BMDMs became more "glycolytic", but less "FAO" when subjected to external attacks. However, the supplementation of rIL-10 into macrophages showed reverse effect. CPT1A and IL-10 can drive the polarization of BMDM from M1 phenotype to M2 phenotype, and CPT1A-IL-10 axis is also involved in the process of maintaining mitochondrial homeostasis. CPT1A modulated metabolic reprogramming and polarisation of macrophage under LPS stimulation. The protective effects of CPT1A may be partly attributed to the induction of IL-10/IL-10 receptor expression.

Enzymatic Modulation of the Pulmonary Glycocalyx Enhances Susceptibility to Streptococcus pneumoniae.

The pulmonary epithelial glycocalyx is rich in glycosaminoglycans such as hyaluronan and heparan sulfate. Despite their presence, the importance of these glycosaminoglycans in bacterial lung infections remains elusive. To address this, we intranasally inoculated mice with Streptococcus pneumoniae in the presence or absence of enzymes targeting pulmonary hyaluronan and heparan sulfate, followed by characterization of subsequent disease pathology, pulmonary inflammation, and lung barrier dysfunction. Enzymatic degradation of hyaluronan and heparan sulfate exacerbated pneumonia in mice, as evidenced by increased disease scores and alveolar neutrophil recruitment. However, targeting epithelial hyaluronan in combination with Streptococcus pneumoniae infection further exacerbated systemic disease, indicated by elevated splenic bacterial load and plasma levels of pro-inflammatory cytokines. In contrast, enzymatic cleavage of heparan sulfate resulted in increased bronchoalveolar bacterial burden, lung damage and pulmonary inflammation in mice infected with Streptococcus pneumoniae. Accordingly, heparinase-treated mice also exhibited disrupted lung barrier integrity as evidenced by higher alveolar edema scores and vascular protein leakage into the airways. This finding was corroborated in a human alveolus-on-a-chip platform, confirming that heparinase treatment also disrupts the human lung barrier during Streptococcus pneumoniae infection. Notably, enzymatic pre-treatment with either hyaluronidase or heparinase also rendered human epithelial cells more sensitive to pneumococcal-induced barrier disruption, as determined by transepithelial electrical resistance measurements, consistent with our findings in murine pneumonia. Taken together, these findings demonstrate the importance of intact hyaluronan and heparan sulfate in limiting pneumococci-induced damage, pulmonary inflammation, and epithelial barrier function and integrity.

Ultrashort wave diathermy inhibits pulmonary inflammation in mice with acute lung injury in a HSP70 independent way: a pilot study.

Acute lung injury (ALI) is a clinical syndrome characterized by pulmonary inflammation. Ultrashort wave diathermy (USWD) has been shown to be effective at in inhibiting ALI inflammation, although the underlying mechanism remains unclear. Previous studies have demonstrated that USWD generates a therapeutic thermal environment that aligns with the temperature required for heat shock protein 70 (HSP70), an endogenous protective substance. In this study, we examined the correlation between HSP70 and USWD in alleviating lung inflammation in ALI. Forty-eight male C57BL/6 mice were randomly divided into control, model, USWD intervention (LU) 1, 2, and 3, and USWD preintervention (UL) 1, 2, and 3 groups (n = 6 in each group). The mice were pretreated with LPS to induce ALI. The UL1, 2, and 3 groups received USWD treatment before LPS infusion, while the LU1, 2, and 3 groups received USWD treatment after LPS infusion. Lung function and structure, inflammatory factor levels and HSP70 protein expression levels were detected. USWD effectively improved lung structure and function, and significantly reduced IL-1β, IL-10, TGF-β1, and TNF-α levels in both the USWD preintervention and intervention groups. However, HSP70 expression did not significantly differ across the experimental groups although the expression of TLR4 was significantly decreased, suggesting that USWD may have anti-inflammatory effects through multiple signaling pathways or that the experimental conditions should be restricted. Both USWD intervention and preintervention effectively reduced the inflammatory response, alleviated lung injury symptoms, and played a protective role in LPS-pretreated ALI mice. HSP70 was potentially regulated by USWD in this process, but further studies are urgently needed to elucidate the correlation and mechanism.

Gypsum alleviates pneumonia via the gut–lung axis by mediating ILC2 compartmental migration

IntroductionBacterial pneumonia is a common lower respiratory tract infectious disease. Gypsum, a type of calcium sulfate mineral, primarily consists of calcium sulfate and contains trace amounts of other metallic elements. In traditional Chinese medicine, gypsum has been widely applied, possessing effects such as heat-clearing, detoxification, cooling blood, arresting bleeding, reducing swelling, and alleviating pain. The purpose of this study was to investigate the mechanism of gypsum inhibition of Streptococcus pneumoniae -induced pneumonia by mediating interregional migration of ILC2 through the gut-lung axis. MethodsNormal pneumonia model was induced by Streptococcus pneumoniae, and pseudo-sterile mouse model was established using broad-spectrum antibiotics. The influence of gypsum on the intestinal flora was analyzed using 16S rDNA. Flow cytometry was used to analyze the typing and content of ILC2 in mouse mesenteric lymph nodes and lung tissues. The expression levels of inflammatory cytokines and specific targets associated with immune migration were assessed using ELISA, RT-qPCR, and western blotting methods. And the effects of gypsum on mucosal barrier using IHC. ResultsGypsum effectively alleviates pulmonary inflammation in mice with pneumonia. Gypsum restores the gut microbiota in mice with Streptococcus pneumoniae and gypsum can regulate the expression of miR-155, miR-146a, IL-25, and S1P, promoting the migration of intestinal ILC2s to the lungs. Finally promotes type 2 immunity, alleviating pneumonia and restore lung mucosal barrier. DiscussionOur study contributes to the understanding of gypsum's function in treating infectious pulmonary conditions. Its mechanism involves remedying gut dysbiosis and initiating ILC2 migration, culminating in decreased lung inflammation and enhanced mucosal immunity in the lungs.

Repositioning baloxavir marboxil as VISTA agonist that ameliorates experimental asthma

V-type immunoglobulin domain–containing suppressor of T-cell activation (VISTA), a novel negative checkpoint regulator, plays an essential role in allergic pulmonary inflammation in mice. Treatment with a VISTA agonistic antibody could significantly improve asthma symptoms. Thus, for allergic asthma treatment, VISTA targeting may be a compelling approach. In this study, we examined the functional mechanism of VISTA in allergic pulmonary inflammation and screened the FDA-approved drugs for VISTA agonists. By using mass cytometry (CyTOF), we found that VISTA deficiency primarily increased lung macrophage infiltration in the OVA-induced asthma model, accompanied by an increased proportion of M1 macrophages (CD11b+F4/80+CD86+) and a decreased proportion of M2 macrophages (CD11b+F4/80+CD206+). Further in vitro studies showed that VISTA deficiency promoted M1 polarization and inhibited M2 polarization of bone marrow–derived macrophages (BMDMs). Importantly, we discovered baloxavir marboxil (BXM) as a VISTA agonist by virtual screening of FDA-approved drugs. The surface plasmon resonance (SPR) assays revealed that BXM (KD = 1.07 µM) as well as its active form, baloxavir acid (BXA) (KD = 0.21 µM), could directly bind to VISTA with high affinity. Notably, treatment with BXM significantly ameliorated asthma symptoms, including less lung inflammation, mucus secretion, and the generation of Th2 cytokines (IL-5, IL-13, and IL-4), which were dramatically attenuated by anti-VISTA monoclonal antibody treatment. BXM administration also reduced the pulmonary infiltration of M1 macrophages and raised M2 macrophages. Collectively, our study indicates that VISTA regulates pulmonary inflammation in allergic asthma by regulating macrophage polarization and baloxavir marboxil, and an old drug might be a new treatment for allergic asthma through targeting VISTA.

Effect of E. cava and C. indicum Complex Extract on Phorbol 12-Myristate 13-Acetate (PMA)-Stimulated Inflammatory Response in Human Pulmonary Epithelial Cells and Particulate Matter (PM)2.5-Induced Pulmonary Inflammation in Mice.

This study explores the potential of a natural composite formulation known as ED, consisting of Ecklonia cava (E. cava, family: Lessoniaceae) and Chrysanthemum indicum Linne (C. indicum, family: Asteraceae), in alleviating lung inflammation induced by fine particulate matter (PM2.5). Initial assessments confirmed that neither ED nor one of its components, dieckol, exhibited cytotoxic effects on A549 cells. Subsequently, the impact of ED and dieckol on MUC5AC gene expression in A549 cells stimulated by phorbol 12-myristate 13-acetate (PMA) was investigated, revealing promising results that demonstrated a dose-dependent inhibition of MUC5AC gene expression. The study also delves into the underlying mechanisms, demonstrating that ED and dieckol effectively suppressed the phosphorylation of mitogen-activated protein kinases (MAPKs), including JNK, ERK, and p38, which are known to be involved in the regulation of MUC5AC gene expression. In in vivo experiments using a PM2.5-induced pulmonary inflammation mouse model, the research findings showed that ED mitigated cellular accumulation in the airways, leading to a significant reduction in the total cell count in bronchoalveolar lavage fluid (BALF). Moreover, ED exhibited protective effects against PM2.5-induced pulmonary damage, characterized by reduced inflammatory cell infiltration and decreased mucus secretion in pulmonary tissues. Additionally, ED's anti-inflammatory properties were evident in its ability to decrease the levels of key inflammatory cytokines, TNF-α and IL-6, both in the serum and lung tissue of the PM2.5-induced pulmonary inflammation mouse model. These findings suggest the potential of ED as a therapeutic agent for inflammatory respiratory diseases.

Inhalation of electronic cigarettes slightly affects lung function and inflammation in mice.

Electronic cigarettes have become increasingly popular, but the results of previous studies on electronic cigarette exposure in animals have been equivocal. This study aimed to evaluate the effects of electronic cigarette smoke (ECS) and cigarette smoke (CS) on lung function and pulmonary inflammation in mice to investigate whether electronic cigarettes are safer when compared to cigarettes. 32 specific pathogen-free BALB/c male mice were randomly grouped and exposed to fresh air (control), mint-flavored ECS (ECS1, 6mg/kg), cheese-flavored ECS (ECS2, 6mg/kg), and CS (6mg/kg). After 3weeks exposure to ECS or CS, we measured lung function (PIF and Penh) and blood oxygen saturation. The levels of TNF-α and IL-6 in the bronchoalveolar lavage fluid (BALF) and serum were measured using ELISA. HE staining was performed to observe the pathological changes in the lung tissues. The levels of IL-6 in BALF and serum, and TNF-α in BALF, were elevated similarly in the ECS and CS groups compared to the control group. Significant elevation was observed in serum TNF-α levels in the CS group. The total count of cells in BALF were increased after ECS1 exposure and CS exposure. PIF and oxygen saturation decreased, and Penh increased markedly in the CS group but not in the ECS groups. Compared with the ECS groups, mice in the CS group had widened lung tissue septa and increased inflammatory cell infiltration. However, we did not detect significant differences between mint-flavored and cheese-flavored e-cigarettes in our study. Overall, our findings suggested that both ECS and CS impair lung function and histopathology while promoting inflammation. In contrast, ECS has a less negative impact than CS.

Sphingosine-1-Phosphate Receptor 4 Attenuates Neutrophilic Airway Inflammation in Experimental Asthma via Repressing Proinflammatory Macrophage Activation.

Patients with eosinophilic asthma react well to conventional treatment of asthma while individualized therapy for non-eosinophilic endotypes have yet to be developed. Dysregulated sphingosine metabolites are associated with the pathophysiology of different asthma endotypes with their receptors involved. However, whether the sphingosine-1-phosphate receptor 4 (S1PR4) contributes to disease progression of asthma remains underappreciated. In this study, we demonstrated that sphingosine metabolism was disturbed in asthma while it could not be used to distinguish between different endotypes of asthma. S1PR4, a vital receptor of bioactive sphingosine metabolites and mainly expressed in macrophages, exhibited lower expression both in patients and experimental mice with neutrophilic airway inflammation. Additionally, S1pr4 deficiency aggravated the OVA/LPS-induced pulmonary inflammation in mice along with a significant up-regulation in M1 macrophage activation. Mechanistic studies showed that S1PR4 was strongly connected to bioactive oxylipins concurrent with bounding to formyl peptide receptor 2 to influence the phosphorylation of JNK and contributed to the macrophage M1 program, which in turn secreted amounts of inflammatory cytokines associated to the inflammatory response of neutrophilic asthma. Furthermore, treating mice with S1PR4 agonist CYM50308 was characterized by less pulmonary inflammatory infiltration. Our research indicates S1PR4 a promising therapeutic target for non-eosinophilic phenotypes of asthma.

Effects of Purinergic Receptor Deletion or Pharmacologic Modulation on Pulmonary Inflammation in Mice.

COVID-19 disease is associated with progressive accumulation of SARS-CoV-2-specific mRNA, which is recognized by innate immune receptors, such as TLR3. This in turn leads to dysregulated production of multiple cytokines, including IL-6, IFN-γ, CXCL1, and TNF-α. Excessive production of these cytokines leads to acute lung injury (ALI), which consequently compromises alveolar exchange of O2 and CO2. It is therefore of considerable interest to develop novel therapies that reduce pulmonary inflammation and stem production of pro-inflammatory cytokines, potentially for COVID-19 patients that are at high risk of developing severe disease. Purinergic signaling has a central role in fine-tuning the innate immune system, with P2 (nucleotide) receptor antagonists and adenosine receptor agonists having anti-inflammatory effects. Accordingly, we focused here on the potential role of purinergic receptors in driving neutrophilic inflammation and cytokine production in a mouse model of pulmonary inflammation. To mimic the effects of SARS-CoV-2-specific RNA accumulation in mice, we administered progressively increasing daily doses of a viral mimetic, polyinosinic:polycytidylic acid [poly(I:C)] into the airways of mice over the course of 1 week. Some mice also received increasing daily doses of ovalbumin to mimic virus-encoded protein accumulation. Animals receiving both poly(I:C) and ovalbumin displayed particularly high cytokine levels and neutrophilia, suggestive of both innate and antigen-specific, adaptive immune responses. The extent of these responses was diminished by genetic deletion (P2Y14R, P2X7R) or pharmacologic modulation (P2Y14R antagonists, A3AR agonists) of purinergic receptors. These results suggest that pharmacologic modulation of select purinergic receptors might be therapeutically useful in treating COVID-19 and other pulmonary infections.

Berberine ameliorates pulmonary inflammation in mice with influenza viral pneumonia by inhibiting NLRP3 inflammasome activation and gasdermin D-mediated pyroptosis.

Viral pneumonia is a common complication caused by Influenza A virus infection and is characterized by severe pulmonary inflammation. A previous study showed that berberine (BBR) significantly ameliorated the pulmonary inflammation in mice with influenza viral pneumonia but its underlying mechanism is not entirely understood. In this study, we reproduced the mouse model of influenza viral pneumonia through intranasal infection of A/Puerto Rico/8/34 (H1N1), to further investigate the anti-inflammatory mechanism of BBR based on nucleotide-binding oligomerization domain-like receptor protein (NLRP) 3 inflammasome activation and Gasdermin D (GSDMD)-mediated pyroptosis. Consistent with MCC950 (10 mg/kg, a specific NLRP3 inflammasome inhibitor), BBR (10 mg/kg) obviously improved the weight loss and survival rate of infected mice, alleviated their pulmonary inflammation, and suppressed the accumulation of tumor necrosis factor and interleukin (IL)-6 in lungs without obvious inhibition on viral multiplication (hemagglutinin titer and nucleoprotein messenger RNA). Moreover, BBR (10 mg/kg) reduced the expressions of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), and cysteinyl aspartate-specific proteinase (Caspase)1 (Caspase1 precursor [Pro-caspase1] + Caspase1p20 subunit) and the ratio of Caspase1p20 subunit to Caspase1, thus inhibiting the NLRP3 inflammasome activation and resulting in the decreased contents of mature IL-1β and IL-18 in lungs. The GSDMD expression (GSDMD precursor [Pro-GSDMD] + GSDMD-N terminal [NT]) and the ratio of GSDMD-NT to GSDMD were also declined by BBR (10 mg/kg). These evidence indicate that BBR may ameliorate pulmonary inflammation in mice with influenza viral pneumonia through inhibiting NLRP3 inflammasome activation, as well as depressing GSDMD-mediated pyroptosis via declining GSDMD expression and restraining NLRP3 inflammasome-mediated GSDMD activation.

Interleukin-37 ameliorates cigarette smoke-induced lung inflammation in mice

Cigarette smoking (CS) is the leading cause of chronic obstructive pulmonary disease, and its severity is closely related to lung inflammation. Interleukin (IL)-37 is a newly discovered member of the IL-1 family with anti-inflammatory activity. Our study aimed to elucidate the effect of IL-37 on CS-induced lung inflammation in mice. In this study, mice were exposed to six cigarettes for 1 h three times daily (4 h smoke-free intervals) for 10 consecutive days. Mice were treated intranasally with IL-37-expressing lentivirus and empty lentivirus particles 1 day before the first CS or sham exposure. Mice were sacrificed on day 11 to evaluate the effect of IL-37 on CS-induced pulmonary inflammation in mice. Administering IL-37-expressing lentivirus significantly reduced CS-induced weight loss in mice compared to empty lentivirus controls (P < 0.05). Histological analysis showed that IL-37 significantly alleviated inflammatory cell recruitment, alveolar septum enlargement, alveolar wall attenuation, mucus hypersecretion, and goblet cell metaplasia in mouse lungs (P < 0.001). IL-37 expression also significantly inhibited CS-induced increases in inflammatory cells (including lymphocytes, neutrophils, and macrophages) in mouse lungs (P < 0.05), as well as pro-inflammatory cytokines such as IL-1β, IL-6, IL-17, monocyte chemotactic protein-1 and tumor necrosis factor-α production (P < 0.05). IL-37 also significantly reduced myeloperoxidase activity in mouse serum (P < 0.01) and lung tissues (P < 0.001). Therefore, IL-37 can ameliorate CS-induced pulmonary inflammation in mice and IL-37 may be a potential therapeutic strategy for CS-induced lung inflammatory diseases.

Network pharmacology study to reveal active compounds of Qinggan Yin formula against pulmonary inflammation by inhibiting MAPK activation

Ethnopharmacological relevancePneumonia is common and frequently-occurred disease caused by pathogens which predisposes to lung parenchymal inflammation leading pulmonary dysfunction. To prevent and alleviate the symptoms of pneumonia, Qinggan Yin formula (QGY) was composed based on clinical experience and four classical traditional Chinese medicine prescriptions which frequently applied to treat infectious diseases. Aim of the studyTraditional Chinese medicine is a complex mixture and it is difficult to distinguish the effective component molecules. The aim of this study is to identify the compounds of QGY with anti-inflammatory effects and investigate the molecular mechanism. Materials and methodsThe high-resolution mass spectrometry and molecular networking were performed for comprehensive chemical profiling of QGY. Network pharmacology was used to generate “herbal-target-pathway” network for target predictions. The anti-inflammation effects of QGY were evaluated in mice model of lipopolysaccharide (LPS) induced acute inflammation. Tail transected zebrafish was also employed to validate macrophage migration reversed effect of QGY. Based on the molecular enrichment analysis, the active substances of QGY with anti-inflammatory effects were further identified in cellular model of macrophage activation. The mechanisms of active substances were investigated by testing their effects on the expression of correlated proteins by Western blot. ResultsIn total, 71 compounds are identified as major substances of QGY. According to the results of network pharmacology, QGY shows moderate anti-inflammatory effects and inhibit pulmonary injury. MAPK signaling pathway was predicted as the most related pathway regulated by QGY. Moreover, QGY significantly inhibit LPS-induced pulmonary inflammation in mice, and reversed macrophage migration toward the injury site in zebrafish. We also validate that some major compounds in QGY significantly attenuated the release of IL-1β, IL-6 and TNF-α in LPS-stimulated macrophage. Those active substances including acacetin and arctiin can inhibit the phosphorylation of ERK/JNK and down-regulated the protein expression of BCL-2. ConclusionCollectively, QGY possessed pronounced anti-inflammation effects. The integration of network pharmacology and experimental results indicated arctiin, iridin, acacetin, liquiritin, and arctigenin are major active substances of QGY with anti-inflammatory effects. The underlying mechanism of QGY involves MAPK signaling pathway and oxidative stress pathway.

A Combined Transcriptomic and Proteomic Approach to Reveal the Effect of Mogroside V on OVA-Induced Pulmonary Inflammation in Mice.

Mogroside V is a bioactive ingredient extracted from the natural food Siraitia grosvenorii which possesses functions that stimulate lung humidification and cough relief activities, but its underlying mechanisms were rarely studied. To estimate its potential protective effect on ovalbumin (OVA)-induced pulmonary inflammation and understand its system-wide mechanism, integrated omics was applied in this study. Mogroside V effectively reduced the levels of IgE, TNF-α, and IL-5 in OVA-induced mice. The results of RNA-seq and data-independent acquisition proteomics approach revealed that 944 genes and 341 proteins were differentially expressed in the normal control group (NC) and ovalbumin-induced control group (OC) and 449 genes and 259 proteins were differentially expressed between the OC and the group treated with 50 mg/kg mogroside V (MV). After a combined analysis of the transcriptome and the proteome, 93 major pathways were screened, and we discovered that mogroside V exerts an anti-inflammation effect in the lung via NF-κB and JAK-STAT, both of which are among the signaling pathways mentioned above. In addition, we found that the key regulatory molecules (Igha, Ighg1, NF-κB, Jak1, and Stat1) in the two pathways were activated in inflammation and inhibited by mogroside V. Thus, mogroside V may be the main bioactivity component in S. grosvenorii that exerts lung humidification and cough relief effects.

Ncf1 Governs Immune Niches in the Lung to Mediate Pulmonary Inflammation in Mice.

Neutrophil cytosolic factor 1 (Ncf1) is a major genetic factor associated with autoimmune diseases and has been identified as a key player in autoimmune mediated inflammation. We addressed the role of Ncf1 in an antigen-induced pulmonary inflammation model, and found that the Ncf1m1j mutation, causing a deficient reactive oxygen species response, alleviated disease. The Ncf1m1j mutation was associated with a reduced inflammatory cell infiltration in airways, but had limited effect on mucus secretion, antibody production and lung fibrosis. The disease remission in the Ncf1 mutated mice was reversed when functional Ncf1 was transgenically expressed in alveolar macrophages, suggesting that the cellular inflammation was depended on functional Ncf1 in alveolar macrophages. By determining cytokine and chemokine profiles in lung and serum, we found that Ncf1 deficiency allowed an increased expression of Th1 cytokines, including TNF-α, IFN-γ and IL-12. Since also epithelial cytokines were found to be regulated by Ncf1, we tested the effect of Ncf1 in IL-33 and IL-25 induced lung inflammation models. Mice with the Ncf1m1j mutation showed less sensitivity to IL-33, but not IL-25, induced lung inflammation, in a macrophage independent manner. The mice with deficient Ncf1 showed a reduced eosinophil infiltration and group 2 innate lymphoid cell (ILC2) activation. The production of IFN-γ in CD4+ T cells was increased, whereas IL-5 and IL-13 in ILC2 were decreased. Importantly, anti-IFN-γ antibody treatment of Ncf1 deficient mice increased eosinophil infiltration and rescued ILC2 activation in the lung. We conclude that Ncf1 deficiency enhances Th1 response, deactivates ILC2, and protects against pulmonitis.

Cardamonin attenuates phorbol 12-myristate 13-acetate-induced pulmonary inflammation in alveolar macrophages

Pulmonary inflammation involves complex immune responses in which alveolar macrophages release pro-inflammatory proteins and cytokines. Cardamonin is a spice component that exerts anti-inflammatory and anti-oxidative properties against pulmonary inflammation. Herein, the aim of this research is to investigate the effects of cardamonin on pulmonary inflammation and its mechanism. Pulmonary inflammation in mice was induced by intratracheal administration of PMA. PMA-stimulated acute fibrosis, pulmonary edema, and inflammatory responses were ameliorated by oral administration of cardamonin in vivo. In MH-S alveolar macrophages, PMA-induced pro-inflammatory responses, including iNOS, COX-2, MMP-9 and cytokines expressions were reduced by cardamonin. The anti-oxidative Nrf2/HO-1 axis was also provoked by cardamonin in MH-S alveolar macrophages. In addition, MMP-9 expression induced by PMA is also decreased by the down-stream metabolites of HO-1, indicating that HO-1 expression partially contributes to the anti-inflammatory effect exerted by cardamonin. In this study, cardamonin demonstrates anti-inflammatory and anti-oxidative effects on PMA-induced pulmonary inflammation and activating Nrf2/HO-1 axis in alveolar macrophages. Cardamonin also ameliorates pulmonary inflammation, rapid fibrosis in vivo, suggesting powerful health benefits.

Tomatidine Improves Pulmonary Inflammation in Mice with Acute Lung Injury.

Tomatidine, which is isolated from green tomato, can ameliorate inflammation and oxidative stress in cells and animal experiments and has been shown to improve airway inflammation in a murine model of asthma. Here, we investigated whether tomatidine can ameliorate acute lung injury in mice. Mice were given tomatidine by intraperitoneal injection for 7 consecutive days, and then, lung injury was induced via intratracheal instillation of lipopolysaccharide (LPS). Tomatidine reduced inflammatory cytokine expressions in bronchoalveolar lavage fluid (BALF), attenuated neutrophil infiltration in the BALF and lung tissue, increased superoxide dismutase activity and glutathione levels, and alleviated myeloperoxidase expression in the lung tissue of mice with lung injury. Tomatidine also decreased inflammatory cytokine and chemokine gene expression in inflammatory lungs and attenuated the phosphorylation of mitogen-activated protein kinase and nuclear factor kappa B. Furthermore, tomatidine enhanced the production of heme oxygenase-1, decreased the secretion of inflammatory cytokines and chemokines in LPS-stimulated lung epithelial cells, and attenuated THP-1 monocyte adhesion. Our findings suggest that tomatidine attenuates oxidative stress and inflammation, improving acute lung injury in mice.

Role of experimental periodontitis in inducing pulmonary inflammation in mice.

The aim of the study was to explore the potential role of experimental periodontitis in pulmonary inflammation in mice. Mice were divided into control, ligature-induced periodontitis (L) and ligature plus Porphyromonas gingivalis (P. gingivalis)-induced periodontitis (LPG) groups. Alveolar bone resorption, pulmonary function, lung tissue histology and cytokine expression were examined at 2, 4 and 8weeks. Then cytokines and neutrophils in the peripheral blood and lung tissue were further assessed at 8weeks to determine the role of cytokines induced by LPG periodontitis, and the effect of P. gingivalis was evaluated using P. gingivalis-IgG and P. gingivalis gingipain. Alveolar bone resorption was more severe in the L and LPG groups. However, pulmonary inflammation was observed only in the LPG group at 8weeks when cytokines and neutrophils in the peripheral blood and lung tissue were the most significant elevation, along with higher levels of P. gingivalis-IgG and P. gingivalis gingipain. Cytokine levels were also increased in the gingival tissue, peripheral blood and lung tissue in the L group, accompanied by elevated peripheral blood neutrophils, but not as significantly as that in the LPG group. LPG periodontitis can trigger pulmonary inflammation over the long term, in which cytokines and P. gingivalis play an important role.

Platelets orchestrate the resolution of pulmonary inflammation in mice by T reg cell repositioning and macrophage education.

Beyond hemostasis, platelets actively participate in immune cell recruitment and host defense, yet their potential in the resolution of inflammatory processes remains unknown. Here, we demonstrate that platelets are recruited into the lung together with neutrophils during the onset of inflammation and alongside regulatory T (T reg) cells during the resolution phase. This partnering dichotomy is regulated by differential adhesion molecule expression during resolution. Mechanistically, intravascular platelets form aggregates with T reg cells, a prerequisite for their recruitment into the lung. This interaction relies on platelet activation by sCD40L and platelet P-selectin binding to PSGL-1 on T reg cells. Physical platelet-T reg cell interactions are necessary to modulate the transcriptome and instruct T reg cells to release the anti-inflammatory mediators IL-10 and TGFβ. Notably, the presence of platelet-T reg cell aggregates in the lung was also required for macrophage transcriptional reprogramming, polarization toward an anti-inflammatory phenotype, and effective resolution of pulmonary inflammation. Thus, platelets partner with successive immune cell subsets to orchestrate both the initiation and resolution of inflammation.