Hyaluronic acid engineered melanin-MOF nanoreactor synergistically remodeling redox and immune homeostasis for targeted acute lung injury therapy.

Nitric oxide as mediator, marker and modulator of microvascular damage in ARDS.

Baicalein ameliorates high-altitude hypoxic lung injury via macrophage polarization remodeling by downregulating ALOX15 pathway in ferroptosis.

Two of the usual suspects, platelet-activating factor and its receptor, implicated in acute lung injury.

To determine the role of matrix metalloproteinase-8 (MMP-8) in acute lung injury (ALI), we delivered LPS or bleomycin by the intratracheal route to MMP-8(-/-) mice versus wild-type (WT) mice or subjected the mice to hyperoxia (95% O(2)) and measured lung inflammation and injury at intervals. MMP-8(-/-) mice with ALI had greater increases in lung polymorphonuclear neutrophils (PMNs) and macrophage counts, measures of alveolar capillary barrier injury, lung elastance, and mortality than WT mice with ALI. Bronchoalveolar lavage fluid (BALF) from LPS-treated MMP-8(-/-) mice had more MIP-1alpha than BALF from LPS-treated WT mice, but similar levels of other pro- and anti-inflammatory mediators. MIP-1alpha(-/-) mice with ALI had less acute lung inflammation and injury than WT mice with ALI, confirming that MIP-1alpha promotes acute lung inflammation and injury in mice. Genetically deleting MIP-1alpha in MMP-8(-/-) mice reduced the increased lung inflammation and injury and mortality in MMP-8(-/-) mice with ALI. Soluble MMP-8 cleaved and inactivated MIP-1alpha in vitro, but membrane-bound MMP-8 on activated PMNs had greater MIP-1alpha-degrading activity than soluble MMP-8. High levels of membrane-bound MMP-8 were detected on lung PMNs from LPS-treated WT mice, but soluble, active MMP-8 was not detected in BALF samples. Thus, MMP-8 has novel roles in restraining lung inflammation and in limiting alveolar capillary barrier injury during ALI in mice by inactivating MIP-1alpha. In addition, membrane-bound MMP-8 on activated lung PMNs is likely to be the key bioactive form of the enzyme that limits lung inflammation and alveolar capillary barrier injury during ALI.

Acute lung injury (ALI) is a serious inflammatory lung disease. Imbalances in the polarization of classically activated (M1) and alternatively activated (M2) macrophages are closely related to ALI. Anisodamine has a promising therapeutic effect for septic shock. Nevertheless, the role of anisodamine in progression of ALI remains to be investigated. Our results showed that anisodamine significantly reduced lung damage, myeloperoxidase (MPO) activity, lung wet/dry ratio, total cell number, and protein concentrations in bronchoalveolar lavage fluid and decreased interleukin (IL)-6 level and the levels of M1 phenotypic markers, whereas it increased IL-10 level and the levels of M2 phenotypic markers in mice with a nasal instillation of lipopolysaccharide (LPS). Bone marrow-derived macrophages (BMDMs) were stimulated or transfected with LPS plus anisodamine or LPS plus G9a short hairpin RNA. Anisodamine and downregulation of G9a both promoted BMDM M2 polarization caused by IL-4 treatment and inhibited M1 polarization resulting from LPS treatment. Chromatin immunoprecipitation assay revealed that anisodamine inhibited G9a-mediated methylation and expression suppression on interferon regulatory factory 4 (IRF4). Overexpression of G9a or silence of IRF4 reversed the improvement effect of anisodamine on lung tissue injury, evidenced by an increase of MPO activity and the restoration of LPS-induced alterations of M1 and M2 polarization. In conclusion, anisodamine protected against LPS-induced ALI, during which anisodamine suppressed the LPS-stimulated alterations of macrophage M1 and M2 polarization through inhibiting G9a-mediated methylation of IRF4, suggesting that anisodamine was a potential therapeutic drug to alleviate ALI. SIGNIFICANCE STATEMENT: Anisodamine treatment was able to attenuate lung injury and pulmonary edema caused by lipopolysaccharide (LPS) stimulation, and the specific mechanism was that anisodamine reversed the LPS-induced alterations of M1 and M2 polarization by inhibiting G9a-mediated methylation and expression suppression of interferon regulatory factor 4, which suggests that anisodamine has the potential to alleviate acute lung injury.

Activation of NFkB and coagulation in lung injury by hyperoxia and excessive mechanical ventilation: one more reason “low and slow” is the way to go?

BackgroundHypervirulent Klebsiella pneumoniae (hvKp) infection-induced sepsis-associated acute lung injury (ALI) has emerged as a significant clinical challenge. Increasing evidence suggests that activated inflammatory macrophages contribute to tissue damage in sepsis. However, the underlying causes of widespread macrophage activation remain unclear.MethodsBALB/c mice were intravenously injected with inactivated hvKp (iHvKp) to observe lung tissue damage, inflammation, and M1 macrophage polarization. In vitro, activated RAW264.7 macrophage-derived exosomes (iHvKp-exo) were isolated and their role in ALI formation was investigated. RT-PCR was conducted to identify changes in exosomal miRNA. Bioinformatics analysis and dual-luciferase reporter assays were performed to validate MSK1 as a direct target of miR-155-5p. Further in vivo and in vitro experiments were conducted to explore the specific mechanisms involved.ResultsiHvKp successfully induced ALI in vivo and upregulated the expression of miR-155-5p. In vivo, injection of iHvKp-exo induced inflammatory tissue damage and macrophage M1 polarization. In vitro, iHvKp-exo was found to promote macrophage inflammatory response and M1 polarization through the activation of the p38-MAPK pathway. RT-PCR revealed exposure time-dependent increased levels of miR-155-5p in iHvKp-exo. Dual-luciferase reporter assays confirmed the functional role of miR-155-5p in mediating iHvKp-exo effects by targeting MSK1. Additionally, inhibition of miR-155-5p reduced M1 polarization of lung macrophages in vivo, resulting in decreased lung injury and inflammation induced by iHvKp-exo or iHvKp.ConclusionsThe aforementioned results indicate that exosomal miR-155-5p drives widespread macrophage inflammation and M1 polarization in hvKp-induced ALI through the MSK1/p38-MAPK Axis.Graphical

It is believed that reactive oxygen species (ROS) play a very important role in the pathogenesis of acute respiratory distress syndrome (ARDS), but the mechanism has not been so clear, owing to the absence of direct measurable (experimental) data. In majority of the medical studies on free radicals, the analysis of ROS has generally been done by the way of measuring their secondary and breakdown products. In our study, we used electron spin resonance (ESR), a sensitive and accurate technique to detect ROS directly and also used some other sensitive techniques including ultra-weak luminescence and chemical luminescence to identify the species and relative amount of ROS. Furthermore, superoxide dismutase (SOD) was pre-administrated in ARDS rats to verify the results from the above studies and explore the possibility of the clinical application of SOD in ARDS. The spectra of ESR showed that the concentration of ROS increased at 10 min and reached a summit at 30 min after injection of oleic acid (OA), then dropped gradually. The scavenging effects of different scavenging agents on ROS by the analysis of ultra-weak luminescence proved that superoxide anion was the main species of ROS in the development of OA-induced ARDS. Moreover, the results of quantified measure of superoxide anion by chemical luminescence also showed the accordant tendency exhibited in ESR measurement. The pre-treatment of SOD might distinctly inhibit the production of superoxide anion, obviously improve the blood gas status, lung wet/dry ratio and lung/body ratio in ARDS rats. It is suggested that ROS may play a key role in the initiation phase of ARDS, while superoxide anion may be a leading actor in this process and SOD could effectively protect rats from ARDS. These results may provide helpful information for the treatment and prevention of ARDS.

M1/M2 macrophage polarization plays a pivotal role in the development of acute lung injury (ALI). The hypoxia-inducible factor-1α/pyruvate kinase M2 (HIF-1α/PKM2) axis, which functions upstream of macrophage polarization, has been implicated in this process. The function of HIF-1α is known to be tightly regulated by SUMOylation. Upregulation of SUMO-specific peptidase 3 (SENP3), a deSUMOylation enzyme, is induced by reactive oxygen species (ROS), which are abundantly produced during ALI. To explore the links between SENP3, macrophage polarization, and lung injury, we used mice with Senp3 conditional knockout in myeloid cells. In the lipopolysaccharide (LPS)-induced ALI model, we found that in vitro and in vivo SENP3 deficiency markedly inhibited M1 polarization and production of pro-inflammatory cytokines and alleviated lung injury. Further, we demonstrated that SENP3 deficiency suppressed the LPS-induced inflammatory response through PKM2 in a HIF-1α-dependent manner. Moreover, mice injected with LPS after PKM2 inhibitor (shikonin) treatment displayed inhibition of M1 macrophage polarization and reduced lung injury. In summary, this work revealed that SENP3 promotes M1 macrophage polarization and production of proinflammatory cytokines via the HIF-1α/PKM2 axis, contributing to lung injury; thus, SENP3 may represent a potential therapeutic target for ALI treatment.

ARDS: progress unlikely with non-biological definition

The term acute respiratory distress syndrome (ARDS) was first coined in 1967 to define a clinical syndrome categorized by progressive hypoxemia, dyspnea, and increased work of breathing that is unresponsive to standard respiratory therapy.1 Acute respiratory distress syndrome presents as acute respiratory failure with noncardiogenic pulmonary edema and severe hypoxemia.2 Historically, this syndrome of potentially fatal pulmonary complications described in critically ill patients was further divided into acute lung injury (ALI) and ARDS, with ALI representing a more mild presentation of ARDS.2 More recently, the concept of ALI has been replaced with mild, moderate, and severe classifications for ARDS.3Despite the substantial progress made in understanding the mechanism of this disease process, ARDS continues to be a major clinical concern. The estimated prevalence in the intensive care unit (ICU) ranges from 4% to 9%, with up to 50% of patients with mild ARDS progressing to moderate or severe forms of the disease. Acute respiratory distress syndrome is associated with substantial mortality rates as high as 46%.2,4 In addition, ARDS is a major contributor to health care costs, with approximately $25 000 to $75 000 spent treating each episode.2 Prompt recognition of the clinical syndrome and initiation of management strategies are required to minimize mortality and improve outcomes in affected individuals.5 This article provides an overview of ARDS with a specific focus on current and evolving pharmacological treatment strategies for this potentially devastating disease state.The most recent representation of the ARDS diagnostic criteria is reflected in the Berlin definition, which was updated in 2011 from the original definition developed in 1994 by the American European Consensus Conference on ARDS.3 According to the Berlin definition, for a diagnosis of ARDS to be confirmed, the disease state must be characterized by acute onset of bilateral infiltrates evident on chest radiograph and a pulmonary artery wedge pressure of 18 mm Hg or less with no clinical evidence of left atrial hypertension. The degree of severity is categorized into 3 stages on the basis of hypoxemia as evidenced by the ratio of partial pressure of oxygen (PaO2) to fraction of inspired oxygen (FiO2): mild (200 mm Hg ≤ PaO2/FiO2 ≤ 300 mm Hg), moderate (100 mm Hg ≤ PaO2/FiO2 ≤ 200 mm Hg), and severe (PaO2/FiO2 ≤ 100 mm Hg). The Berlin definition addresses limitations provided in the American European Consensus Conference, resulting in a better predictor of validity of mortality.3The potential cause of ARDS can be attributed to direct or indirect causes. Direct risk factors for lung injury include pneumonia, aspiration of gastric contents, pulmonary contusion, fat or air emboli, inhalation injury, and reperfusion injury. Indirect lung injury occurs secondary to sepsis, severe trauma, cardiopulmonary bypass, toxic ingestions, acute pancreatitis, or transfusion of blood products.6 Pneumonia and sepsis are the most common causes of direct and indirect lung injury, respectively.2Acute respiratory distress syndrome can be characterized by 3 progressive clinical stages, the exudative, proliferative, and fibrotic phases (Table 1). The lungs move through this series of phases, regardless of the initial cause of the lung injury.7 The specific time frames associated with each phase vary considerably in the existing literature. In the initial exudative phase, also called the acute phase, widespread damage to the alveolar epithelium is present. The compromised epithelium increases the permeability of the alveolar-capillary barrier allowing fluid, rich in protein, to accumulate in the alveoli.8 Continued damage to the epithelium occurs secondary to release of proinflammatory mediators. Cytokines recruit neutrophils to the lungs, causing production of reactive oxygen species and proteases. Alveolar damage and impairment of surfactant production ultimately causes alveolar collapse and impaired gas exchange.2During the proliferative stage, also known as the subacute phase, the pulmonary edema seen in the exudative phase begins to resolve.4 During this time, the membranes within the alveoli have become overwhelmed by plasma proteins, fibrin, and cellular debris. Multiple detrimental pathways are activated causing impaired surfactant production, enhanced neutrophil activation, and stimulation of the complement and coagulation cascades.2,9 Type II alveolar cells proliferate during this phase, which is the body’s initial attempt to repair the lung injury.8 Unfortunately, the inflammatory mediators produced within the lungs commonly migrate into the bloodstream causing cellular apoptosis in other organs. Mortality is often not associated with hypoxemia or hypercapnia but rather from multisystem organ dysfunction.9The fibrotic, or chronic, phase is the final stage of ARDS. Not all patients progress to the later stages of the disease process, as some experience an uncomplicated course and recover quickly. Collagen deposition leads to pulmonary fibrosis, which is the hallmark feature of late-stage ARDS.2 Fibrosis can compromise the pulmonary vascular area by destroying alveoli and commonly leads to chronic inflammation.9 Recovery from ARDS begins to occur as a result of a rise in alveolar type II cells, which promote fluid removal. Type II cells begin to differentiate into type I cells returning the epithelial layer to normal and ultimately providing disease resolution.2Treatment of ARDS occurs through a combination of strategies that attempt to reduce pulmonary edema while reversing oxygen-exchange issues.10 The only treatment option that has shown to reduce mortality rates is lung-protective mechanical ventilation.7 A multitude of different pharmacological agents have been investigated for the treatment of ARDS with most failing to show consistent clinical benefit. Deciding on a management strategy for these patients can be challenging, because of the complexity of the disease and conflicting existing evidence.11 Although the most important aspect in the treatment of ARDS is lung-protective mechanical ventilation, this column focuses on the pharmacological options that can assist in the supportive management of this disease process (Table 2).Corticosteroids have been studied for years in the prevention and treatment of ARDS because of the anti-inflammatory and immunomodulatory characteristics they exhibit. Beneficial effects are due to inhibition of the inflammatory cascade at different stages of immune-mediated injury to the lung. Glucocorticoids inhibit the NF-κB signaling pathway, which in turn inhibits interleukin 1, interleukin 6, and tumor necrosis factor α, which are all inflammatory mediators in the process of ARDS.12 In addition, steroids inhibit interleukin 3, interleukin 5, and interleukin 8 while amplifying the antifibrotic properties of cortisol, promoting T-cell stimulation, impairing fibroblast proliferation, and minimizing collagen deposition. Potential benefit must be weighed against possible risks, including hyperglycemia, impaired wound healing, enhanced susceptibility to future infections, and extended muscle weakness.2Corticosteroids have been studied in the different phases of ARDS and are most efficacious in the early and late phases. Many randomized clinical trials have shown a decrease in ICU mortality rate, ventilation days, and inflammatory markers.13–16 In the latest study by Meduri et al,17 low-dose methylprednisolone administered in the first 72 hours after ARDS diagnosis resulted in reduced days on mechanical ventilation and in the ICU. In an earlier study, Steinberg et al16 used a moderate dose of methylprednisolone started 7 to 28 days after the diagnosis of ARDS. This study showed that although methylprednisolone increased the number of ventilator-free days, the mortality rates were comparable between groups. As clinical trials evolve, the trend continues to show beneficial results in ARDS with treatment of corticosteroids. In 2008, the Society of Critical Care Medicine published the latest recommendations for the use of corticosteroids in the treatment of ARDS.12 Moderate-dose glucocorticoid, defined as methylprednisolone 1 mg/kg as a load over 30 minutes followed by 1 mg/kg over 24 hours per day via continuous infusion for 14 days with slow-dose tapering, is recommended.12Neuromuscular blocking agents are commonly used in patients with severe gas-exchange deficiencies to facilitate mechanical ventilation when sedation is insufficient.18 Skeletal muscle inhibition produced through the use of neuromuscular blocking agents eliminates patient effort in the breathing process.19 Facilitation of paralysis helps reduce the risk of ventilator-induced injury by improving chest wall compliance, preventing patient-ventilator dyssynchrony, minimizing inflammatory mediator release, reducing overinflation of the lungs, and decreasing oxygen demand.18 Cisatracurium has the most extensive evidence supporting its short-term use with small-scale, randomized, controlled trials demonstrating improved 90-day survival, reduced mechanical ventilation time, and decreased days with documented organ failure.18 However, older studies have reported conflicting evidence describing that the use of neuromuscular blocking agents may potentially be associated with prolonged neuromuscular weakness that may make weaning ventilation more challenging.4Inhaled vasodilators, including nitric oxide and prostacyclins, selectively lead to vasodilation within the pulmonary vasculature that assists in improving oxygenation status without substantial adverse effects on systemic hemodynamics. Nitric oxide dilates the vasculature by increasing conversion to cyclic guanosine monophosphate resulting in smooth muscle relaxation.10 Prostacyclins, such as epoprostenol and alprostadil, act on the prostaglandin receptors to increase levels of cyclic adenosine monophosphate to cause relaxation of the vasculature.10 The vasodilation produced by these agents also may lead to other beneficial pulmonary and cardiovascular effects, including reduced pulmonary vascular resistance, minimized right ventricular afterload, and increased right ventricular stroke volume.10 In addition, nitric oxide may impede neutrophil and platelet activation seen in acute inflammation.7 Prostacyclins are increasingly becoming the vasodilator of choice due to lower cost compared with nitric oxide.4 Existing evidence demonstrates that inhaled vasodilators have not been associated with reduced ventilator days or mortality rates. However, this inhaled therapy may be beneficial in improving oxygenation, so it should be considered in patients with refractory hypoxemia.10,20The body’s natural surfactant supply is composed of 90% lipids and 10% protein and is responsible for preventing alveolar collapse by reducing surface tension within the lung. In addition, surfactant possesses anti-inflammatory properties, enhances phagocytic cell activity, and scavenges free oxygen radical species. Because of the disruption in the endogenous surfactant system in ARDS, the administration of exogenous surfactant appears to be a reasonable treatment strategy. Surfactant was originally administered as an aerosolized product, but tracheal instillation is now preferred because of concerns over low levels of alveolar deposition with the aerosolized formulation.9 Use of surfactant has not been shown to have beneficial effects on mortality rates or ventilator-free days, but oxygenation status was improved.20 Efficacy of exogenous surfactant may be questionable because synthetic products may be easily inactivated in the lungs and large volumes are needed to cover the lung surface area.11 Because of the widespread benefit seen in pediatric lung injury trials, further investigation of the use of exogenous surfactant in adult ARDS is warranted.20β2-Adrenergic agonists can enhance fluid clearance from the alveolar space.20 These agents increase sodium and chloride transport across the epithelial membrane via the Na+/K+ ATPase pump, which subsequently causes water to move in the direction of the electrolyte shift. Two potential mechanisms have been proposed to explain β2-agonists effect on fluid clearance, including augmentation of intracellular cyclic adenosine monophosphate upregulating the Na+/K+ ATPase pump and minimizing permeability between the alveolar-capillary barriers.9 Beneficial effects may also be seen due to cytoprotection and reduced endothelial permeability. β2-Agonists have been evaluated through both the intravenous and inhaled routes in clinical trials with inhaled formulations demonstrating a superior adverse effect profile. The place in therapy of these agents may be limited because of the risk of detrimental cardiac side effects, including tachyarrhythmias and cardiac ischemia.19In addition to antifungal properties, ketoconazole also may play a role in the treatment of ARDS because of its anti-inflammatory properties. Ketoconazole inhibits formation of thromboxane A2, which is responsible for strong vasoconstriction. In addition, ketoconazole impairs production of leukotrienes, which, when they accumulate, attracts other proinflammatory mediators to the site of injury.20 Release of cytokines from macrophages located within the alveoli also is hindered with the use of ketoconazole. Studies have shown that the use of ketoconazole may help prevent the development of ARDS, but it has no impact on the mortality rate in patients with established disease. Significant barriers impede routine use of ketoconazole in critically ill patients, including a multitude of drug interactions and the need for an acidic environment for absorption through the enteral route.9Antioxidant therapies have been proposed as a potential treatment strategy because reactive oxygen species, produced by neutrophils and macrophages, contribute to tissue damage in ARDS. N-acetylcysteine, which is traditionally used for acetaminophen overdoses, serves as a glutathione precursor. Glutathione is a natural antioxidant found in healthy lung tissue, but concentrations are depleted in patients with ARDS.9 The use of enteral nutrition high in omega-3 fatty acids may provide an additional source of antioxidant supplementation. Vitamins E and C also can be provided to minimize oxidative stress caused by ARDS.9 Overall, evidence suggests that antioxidant therapy likely offers no mortality benefit but may reduce the extent of lung injury.11HMG (3-hydroxy-3-methyl-glutaryl) CoA-reductase inhibitors, which are more commonly known as statins, possess multiple physiological benefits in addition to their ability to lower cholesterol level. Statins are thought to attenuate many underlying mechanisms that cause ARDS. In animal and human models, statin use has reduced local and systemic inflammatory processes as well as histological evidence of lung injury.21 Recent large-scale, randomized clinical trials have not found any clinical benefit to the use of statins in ARDS. Lack of improvement in outcomes combined with a questionable safety profile has led to statins no longer being recommended for the treatment of ARDS.21,22The emerging concept of regenerative medicine, including stem cell therapy and growth factors, can assist in the healing of damaged lung tissue. Contact between stem cells and the alveoli facilitates advantageous effects including anti-inflammatory and immunomodulatory properties and may improve integrity of the endothelial barriers.19,23 Mesenchymal stem cells are considered low in their immunogenicity, which opens up the option for use in different disease states. Both animal and human models have shown potential benefit of mesenchymal stem cell therapy for ARDS.23 The optimal route of delivery is still under investigation, with stem cells being administered directly into the lung in recent human models.19Keratinocyte growth factor (KGF) plays a pivotal role in repair of lung injury. Endogenous KGF assists in the proliferation of type II alveolar cells in ARDS, which are responsible for improving tissue repair. In addition, KGF may play a helpful role during the injury process of the disease by enhancing fluid clearance from the alveoli and minimizing endothelial permeability and edema.12 Palifermin, an intravenous formulation of KGF, is being investigated in a phase II clinical trial.24 Conversely, levels of vascular endothelial growth factor have been found to be higher than normal in patients with ARDS. This growth factor is associated with control of vascular permeability. Vascular endothelial growth factor inhibitors could be a potential future therapeutic option.19Since the initial recognition of ARDS in 1967, substantial advancement has occurred in understanding the pathophysiology and treatment strategies for this disease process. Declining mortality rates in the past decades can be attributed to lung-protective ventilation combined with enhanced supportive measures, particularly corticosteroids and neuromuscular blocking agents. Because of the persistence of relatively high mortality rates and serious complications associated with ARDS, there continues to be significant effort in discovering pharmacological agents that improve clinical outcomes. In the coming years, we can expect that new treatment strategies will be investigated and available for use in the management of ARDS.

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are devastating acute pulmonary conditions with high mortality rates and limited effective treatment options. This study aimed to investigate the therapeutic potential of XBJ on ALI and its potential mechanism. We developed an in vitro model of lipopolysaccharide (LPS)-induced ALI and evaluated the effects of XBJ pre-treatment on oxidative stress, inflammatory responses, and the polarization state of alveolar macrophages. LPS exposure significantly elevated the levels of reactive oxygen species (ROS) and oxidants 8-hydroxy-2'-deoxyguanosine (8-OHDG) and malondialdehyde (MDA) in alveolar macrophages. It also elevated the concentrations of pro-inflammatory cytokines, including tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and IL-23. XBJ and quercetin significantly mitigated the increase in these indicators. Moreover, XBJ and quercetin both downregulated the expression of key proteins in the NLRP3 inflammasome pathway in the ALI model. Similar to the ROS inhibitor N-acetylcysteine (NAC), XBJ and quercetin significantly decreased M1 polarization markers like CD86 and inducible nitric oxide synthase (iNOS), while increasing M2 polarization markers such as CD206 and arginase-1 (Arg-1). Notably, the overexpression of NLRP3 was able to reverse the inhibitory effect of XBJ on macrophage M1 polarization. XBJ inhibits the M1 polarization of alveolar macrophages by targeting ROS-mediated NLRP3 inflammasome signaling, thereby reducing the inflammatory response. These results indicate that XBJ may offer a novel therapeutic strategy for ALI/ARDS by modulating macrophage polarization and inflammation.

Acute lung injury (ALI) is marked by dysregulated inflammatory responses, disruption of the alveolar-capillary barrier, and defective tissue repair, in which macrophages function both as drivers of tissue damage and facilitators of resolution. Here, we explored the transcriptional regulation of macrophage polarization in ALI using integrative single-cell and bulk RNA sequencing approaches. Meis1 was identified as a critical transcriptional factor that promotes macrophage skewing toward the M2 phenotype, associated with anti-inflammatory and reparative functions. Mechanistic dissection revealed that Meis1 negatively regulates the MIF/CD74 signaling pathway, thereby limiting pro-inflammatory macrophage activity. Experimental validation in vitro confirmed that Meis1 enhances expression of M2-associated markers while suppressing inflammatory cytokine release. In vivo studies using a murine ALI model demonstrated that Meis1 overexpression alleviates lung inflammation, reduces tissue damage, and accelerates structural repair. These findings establish Meis1 as a key modulator of immune cell fate in injured lungs. Through inhibition of MIF/CD74 signaling, Meis1 constrains inflammatory toxicity and supports tissue repair, revealing a novel regulatory pathway that may be exploited for treating ALI and related inflammatory lung conditions.

Acute lung injury (ALI), a severe pulmonary disorder that poses a significant threat to life, is closely associated with macrophage ferroptosis and polarization. Lipocalin 2 (LCN2) has been previously reported to be implicated in the pathogenesis of ALI. However, the specific role of LCN2 in macrophage ferroptosis and polarization remains undetermined. Lipopolysaccharide (LPS) was used to establish a mouse model of ALI and also to stimulate mouse RAW264.7 cells. H&E staining was used for histopathologic evaluation, and immunohistochemistry analysis was used to determine the 4-HNE-positive cells. The secretion levels of TNF-α, IL-6, and IL-1β were assessed by ELISA. Gene and protein expression assays were performed using quantitative PCR and immunoblotting. The levels of MDA, GSH, ROS, and lipid ROS were detected to evaluate the alteration in ferroptosis. CD86+ and CD206+ cells were quantified by flow cytometry. The relationship between LCN2 and interferon regulatory factor 7 (IRF7) was confirmed by chromatin immunoprecipitation (ChIP) and luciferase reporter assays. LCN2 expression was upregulated in the lungs of LPS-induced ALI mice and LPS-stimulated RAW264.7 cells. In LPS-induced ALI mice, the depletion of LCN2 alleviated lung injury and ferroptosis, and also inhibited inflammation and macrophage M1 polarization. In LPS-stimulated RAW264.7 cells, the depletion of LCN2 suppressed ferroptosis, inflammation, and M1 polarization. Mechanistically, IRF7 enhanced LCN2 transcription in RAW264.7 cells by binding to its promoter region. More importantly, the silencing of IRF7 inhibited ferroptosis and M1 polarization in LPS-stimulated RAW264.7 cells by downregulating LCN2. Taken together, the IRF7/LCN2 cascade enhances the ferroptosis and M1 polarization of LPS-stimulated macrophages, thereby exacerbating ALI. Anti-IRF7 and anti-LCN2 therapies might potentially be exploited for the prevention and treatment in ALI.

Skin, our first interface to the external environment, is subjected to oxidative stress caused by a variety of factors such as solar ultraviolet, infrared and visible light, environmental pollution, including ozone and particulate matters, and psychological stress. Excessive reactive species, including reactive oxygen species and reactive nitrogen species, exacerbate skin pigmentation and aging, which further lead to skin tone unevenness, pigmentary disorder, skin roughness and wrinkles. Besides these, skin microbiota are also a very important factor ensuring the proper functions of skin. While environmental factors such as UV and pollutants impact skin microbiota compositions, skin dysbiosis results in various skin conditions. In this review, we summarize the generation of oxidative stress from exogenous and endogenous sources. We further introduce current knowledge on the possible roles of oxidative stress in skin pigmentation and aging, specifically with emphasis on oxidative stress and skin pigmentation. Meanwhile, we summarize the science and rationale of using three well-known antioxidants, namely vitamin C, resveratrol and ferulic acid, in the treatment of hyperpigmentation. Finally, we discuss the strategy for preventing oxidative stress-induced skin pigmentation and aging.