Search Results (237)

CA, a major bioactive component of cinnamon essential oil, possesses broad-spectrum antibacterial, anti-inflammatory, and antioxidant activities and is widely recognized as a safe food additive. In the context of exploring natural plant extracts for the prevention and alleviation of bovine mastitis, it is noteworthy that research and clinical reports on the use of cinnamon essential oil for bovine mastitis are relatively limited. In this study, S. aureus isolates recovered from subclinical bovine mastitis were used to establish a mastitis infection model, and the antioxidant and anti-inflammatory effects of CA were preliminarily investigated in both mouse mastitis models and mammary epithelial cells (MAC-T). Studies have demonstrated that in a mouse model of mastitis, CA reduces inflammatory cell infiltration, decreases myeloperoxidase (MPO) activity, and inhibits S. aureus colonization in mammary tissue. CA alleviates oxidative stress by lowering malondialdehyde (MDA) levels, enhancing superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities, and activating the Nrf2/HO-1/xCT antioxidant pathway. In addition, CA suppresses the activation of the TLR4/NF-κB signalling pathway and modulates the expression of Bax and Bcl-2, accompanied by reduced levels of pro-inflammatory cytokines, including tumour necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6), ultimately ameliorating mastitis in mice. In the MAC-T cell infection model, CA significantly inhibited S. aureus adhesion and invasion and decreased the production of pro-inflammatory cytokines and mediators. CA also activated the Nrf2/HO-1/xCT antioxidant pathway, which led to decreased intracellular reactive oxygen species (ROS), concurrently inhibited the TLR4/NF-κB pathway, and downregulated apoptotic protein expression, ultimately attenuating inflammation in MAC-T cells. Collectively, these findings demonstrate that CA exerts protective effects against inflammation and oxidative damage in both mammary tissues and epithelial cells infected with S. aureus. Thus, this study provides a theoretical and practical basis for further exploring CA as a candidate therapy for bovine mastitis. Full article

Graves’ disease is an autoimmune hyperthyroid disorder in which thyroid peroxidase (TPO) plays a central role in excessive thyroid hormone production. However, current TPO-targeting drugs, such as propylthiouracil, have limited selectivity and can cause serious adverse effects. In this study, we established an integrated discovery pipeline to identify and optimize novel small-molecule TPO inhibitors. The pipeline began with high-throughput screening of approximately 7000 compounds from the KRICT chemical library for peroxidase inhibition, followed by cytotoxicity filtering and iterative medicinal chemistry to generate NE-2 derivatives. Lead compounds (NE-2-6, NE-2-7, and NE-2-8) were evaluated using enzymatic assays, selectivity profiling against myeloperoxidase (MPO) and lactoperoxidase (LPO), molecular docking, plasma pharmacokinetic profiling, and in vivo antithyroid pharmacodynamic testing in an Ad-TSHR289-induced thyroid hyperfunction model. NE-2-6 exhibited potent TPO inhibitory activity and relative selectivity within the tested peroxidase panel. UV–visible spectral scanning and H₂O₂-dependent inhibition assays suggested that NE-2-6 perturbs the heme-associated catalytic environment of TPO; however, these data do not establish a definitive binding mode or kinetic inhibition mechanism. In the Ad-TSHR289-induced thyroid hyperfunction model, NE-2-6 reduced serum T4 levels, supporting antithyroid pharmacodynamic activity. Because autoimmune endpoints and dose-matched PK–PD relationships were not fully assessed, the findings should be interpreted as preliminary preclinical evidence supporting further evaluation of NE-2-6 as a TPO-targeting antithyroid candidate. Full article

Background: Danshen–Gegen is a classic herb pair in traditional Chinese medicine, which has been used to treat cardiovascular and cerebrovascular diseases. Ischemic stroke (IS) is a prevalent cerebrovascular condition; ferroptosis is one of the contributing factors driving the progression of IS. This study aims to determine the underlying mechanism and examine if Danshen–Gegen (DG) extract may prevent cerebral ischemia–reperfusion injury by preventing ferroptosis. Methods: The comprehensive compositional characterization of DG was analyzed by ultra-high-performance liquid chromatography coupled with hybrid quadrupole-orbitrap high-resolution mass spectrometry (UPLC-Q-orbitrap MS). The experiments were conducted in middle cerebral artery occlusion/reperfusion (MCAO/R) rats and oxygen-glucose deprivation/re-oxygenation (OGD/R) cells. The neuroprotective effects of DG on IS were assessed by examining rat survival rates, infarct volume, behavioral scores, and cerebral water content. Then, we tested the accumulation of Fe²⁺ and lipid peroxidation products such as reactive oxygen species (ROS), glutathione (GSH), malondialdehyde (MDA), myeloperoxidase (MPO), and 4-hydroxynonenal (4-HNE) in rats and cells. The expression of nuclear factor erythroid-derived 2-like 2 (Nrf2), Solute Carrier Family 7 Member 11 (-xCT), Glutathione peroxidase 4 (GPX4), Cyclooxygenase-2 (COX-2), Transferrin Receptor 1 (TFR1), and Long-chain-fatty-acid–CoA ligase 4 (ACSL4) was also assessed in vivo and in vitro. Results: UPLC-Q-orbitrap MS analysis was performed to characterize the chemical profile of DG, and a total of 33 chemical constituents were successfully identified. DG significantly alleviated the ischemic damage to brain tissue, reduced infarct volume, and improved neurological dysfunction. The content of Fe²⁺ and lipid peroxidation products was markedly decreased. Furthermore, DG could restore the expression of Nrf2, -xCT, and GPX4 with the inhibition of COX-2, TFR1, and ACSL4, thus achieving a suppressive effect on ferroptosis. Conclusions: The regulatory influence of DG via the Nrf2/System xc-/GPX4 axis may play a crucial role in alleviating ferroptosis and enhancing recovery from cerebral ischemia injury. Full article

Experimental acute lung injury (ALI) models are widely used to investigate pulmonary inflammation and evaluate therapeutic strategies for acute respiratory distress syndrome (ARDS). Kaempferia marginata is a traditional medicinal plant used to treat fever and has been reported to possess anti-inflammatory properties in lipopolysaccharide (LPS)-activated macrophages. 6β-Acetoxysandaracopimaradien-1α,9α-diol (ASPD), a major isopimarane-type diterpenoid isolated from this plant, has not previously been investigated for its effects on ALI. This study employed an integrated network pharmacology, molecular docking, and experimental validation strategy to investigate the protective effects and potential mechanisms of ASPD against LPS-induced ALI. Network pharmacology analysis identified several inflammation-related hub targets associated with Src, MAPK, and PI3K/Akt signalling. In LPS-stimulated MLE-12 cells, ASPD reduced inflammatory cytokine production and inhibited the phosphorylation of JNK1/2, ERK1/2, p38 MAPK, Akt, and GSK-3β. In mice with LPS-induced ALI, ASPD alleviated histopathological lung injury, pulmonary oedema, and inflammatory cell infiltration while reducing IL-6, TNF-α, and myeloperoxidase activity without apparent toxicity. Immunohistochemical analysis demonstrated reduced Src and ERK1/2 expression in lung tissue. Molecular docking analysis predicted favourable binding affinities between ASPD and selected Src- and MAPK-related signalling proteins. These findings suggest that ASPD attenuates LPS-induced ALI and is associated with alterations in Src-, MAPK-, and Akt/GSK-3β-related signalling. Full article

Dental calculus is a highly prevalent oral condition in dogs and is widely recognized as an important risk factor for gingival inflammation and periodontal disease. Effective strategies for its prevention and treatment remain limited, highlighting the significance of exploring novel mechanisms underlying its formation. Neutrophil extracellular traps (NETs), a key component of innate immunity, have been found in various diseases. To investigate the relationship between NETs and canine dental calculus formation, NET-associated markers were assessed in the oral cavities of dogs with dental calculus and healthy controls. Based on previously published full-length 16S rRNA amplicon sequencing data of canine dental calculus, Porphyromonas gulae was selected as a candidate NET-inducing bacterium for subsequent validation experiments. Subsequent neutrophil stimulation experiments were conducted to explore the effects of NETs and related factors on dental calculus formation. Collectively, our findings demonstrate the presence of NETs within canine dental calculus and reveal that P. gulae present in canine dental calculus is capable of inducing NET formation. The level of myeloperoxidase–DNA complex in gingival crevicular fluid was significantly elevated in dogs with dental calculus. NETs promoted aggregation and microcrystal formation from calcium and phosphate ions under both physiological and supersaturated concentrations. By adhering to the surface of dental calculus, NETs facilitated calculus accumulation. This effect showed positive correlation with neutrophil counts and administration frequency, but was independent of the concentration of administered calcium and phosphate solutions. IL-1β promoted the formation of aggregated NETs but did not enhance calculus accumulation. DNase I inhibited this process by degrading NET-DNA. In conclusion, dental calculus and the calculus-inhabiting P. gulae could stimulate oral neutrophils to release NETs, which participate in and facilitate the initial formation, aggregation, and subsequent accumulation of canine dental calculus. Full article

The link between neutrophil extracellular traps (NETs) and hepatocyte ferroptosis in liver ischemia–reperfusion injury (LIRI) is unclear. Adipose-derived mesenchymal stem cell exosomes (ADSCs-Exo) hold therapeutic potential for LIRI. This study employed miniature pigs to investigate the NETs’ role and ADSCs-Exo’s protection in LIRI. In vitro, established hepatocyte oxygen-glucose deprivation/reoxygenation (OGD/R) model and Transwell co-culture system with polymorphonuclear neutrophils (PMNs). In vivo, a laparoscopic minimally invasive LIRI model was constructed in miniature pigs, followed by ADSCs-Exo intervention. Results demonstrated that NETs exacerbate OGD/R-induced hepatocyte ferroptosis via myeloperoxidase. ADSCs-Exo inhibited NET formation via the NADPH/MAPK pathway, thereby mitigating ferroptosis, and ultimately improved liver histopathology and function. This study is the first to demonstrate in a large animal model that ADSCs-Exo alleviate LIRI by inhibiting NET formation via the NADPH/MAPK pathway, consequently attenuating hepatocyte ferroptosis. These findings provide novel insights into LIRI pathogenesis, support the translational potential of ADSCs-Exo as a cell-free therapeutic strategy, and highlight the value of the miniature pig model in liver research. Full article

Background/Objectives: Ulcerative colitis is a chronic inflammatory disorder of the colon characterized by epithelial injury and excessive inflammatory responses. J2H-1802 is a newly synthesized hybrid molecule designed to combine the pharmacological properties of mycophenolate mofetil and 5-aminosalicylic acid. This study evaluated the protective and anti-inflammatory effects of J2H-1802 in a dextran sulfate sodium (DSS)-induced colitis mouse model and investigated its underlying mechanisms. Methods: Experimental colitis was induced in mice by administration of 2.5% (w/v) DSS for 7 days, followed by oral treatment with J2H-1802. Body weight, stool consistency, fecal bleeding, and disease activity index were assessed. Colon length and spleen weight were measured to evaluate macroscopic damage. Levels of tumor necrosis factor-α, interleukin-1β, interleukin-6, and myeloperoxidase in colon tissues were quantified, and the expression of phosphorylated nuclear factor-κB and cyclooxygenase-2 was analyzed by Western blotting. Results: J2H-1802 alleviated DSS-induced body weight loss, diarrhea, and fecal bleeding, resulting in reduced disease activity index scores. It also prevented colon shortening and attenuated splenomegaly. In addition, J2H-1802 significantly suppressed the elevated levels of tumor necrosis factor-α, interleukin-1β, interleukin-6, and myeloperoxidase in colon tissues. Western blot analysis further showed that J2H-1802 inhibited the DSS-induced upregulation of phosphorylated nuclear factor-κB and cyclooxygenase-2. Conclusions: J2H-1802 protected against DSS-induced colitis by reducing inflammatory responses and inhibiting the nuclear factor-κB/cyclooxygenase-2 signaling pathway. These findings suggest that J2H-1802 functions as a hybrid anti-inflammatory scaffold with in vivo pharmacological activity and may warrant further optimization and investigation in IBD models. Full article

Alveolar echinococcosis is a life-threatening zoonotic parasitic disease caused by infection of Echinococcus multilocularis larvae. To survive within the host’s immune milieu, E. multilocularis has evolved sophisticated immune evasion strategies, including the expression of immunomodulatory proteins that regulate the host immune response. Our previous studies have demonstrated that E. multilocularis calreticulin (EmCRT) possessed strong binding ability to human complement component C1q to inhibit C1q-initiated complement activation and biological functions. To further elucidate the mechanism by which EmCRT mediates C1q inactivation and immune evasion, the precise C1q-binding site on EmCRT was identified and analyzed in this study through expression of overlapping fragments and synthesis of overlapping peptides covering the identified functional fragment. The fragment expression and functional assay narrowed down the C1q-binding site to the EmCRT-S1 fragment located between amino acids 140 and 204 of EmCRT. The precise binding site was further pinpointed to the P5 peptide (EmCRT160–174 aa) by testing the synthetic peptides covering this region. The binding of peptide P5 to C1q markedly suppressed the activation of the C1q-mediated classical complement pathway and C1q-induced neutrophil chemotaxis, production of reactive oxygen species, cathepsin G, and myeloperoxidase. These findings suggest that the C1q-binding P5 peptide of EmCRT may serve as a potential target for the development of vaccines against echinococcosis or therapeutic drugs for complement-associated inflammatory or autoimmune diseases. Full article

Jacareubin (2), nujiangexanthone A, and α-mangostin display the highest anti-allergic effects among the active xantones through still not well-known mechanisms. This study investigates the SAR of jacareubin, its precursor xanthone V (1) and their peracetylated (1a and 2a), permethylated (1b and 2b) derivatives and their anti-allergic and anti-inflammatory effects. To characterize the inhibitory effect of jacareubin, 2a and 2b on the anaphylactic reaction, we first utilized in vitro models of bone marrow derived mast cells (BMMCs), determining their capacity of inhibiting the IgE/Antigen-induced degranulation, myeloperoxidase (MPO), and xanthine oxidase (XO) activation. Also, we utilized in vivo models of passive cutaneous anaphylaxis (PCA) and TPA-induced ear edema. In vitro tests showed that the compound 2b was more effective than jacareubin in the inhibition of BMMCs degranulation. Besides, in vivo models of PCA revealed that the fourth cyclized ring of jacareubin is the critical structural element for anti-allergic efficacy, as compound 1 was less effective. Additionally, hydroxyl groups were found to be essential for inhibiting MPO. Jacareubin was the only tested xanthone that directly inhibited XO, a result supported by molecular docking. Overall, jacareubin represents a promising multi-target scaffold that could be used for developing new treatments for inflammatory and allergic diseases. Full article

Circulating immune complexes (CICs) are frequently detected in the sera of patients with myeloperoxidase (MPO)–antineutrophil cytoplasmic antibody (ANCA)-associated glomerulonephritis (MPO-AAGN) and their role in activating the classical complement pathway has been suggested. However, the precise composition and functional characteristics of CICs in AAGN remain poorly understood. We analyzed serum samples from four patients with MPO-AAGN and confirmed CICs. An immunoadsorbent column was prepared using a monoclonal rheumatoid factor antibody that binds to CICs, enabling extraction via immunoprecipitation. CICs were analyzed by Western blotting to detect MPO and IgG. Their capacity to activate complement was assessed in vitro by measuring complement activation products using the enzyme-linked immunosorbent assay following incubation with normal human serum. Western blotting revealed distinct bands for both MPO and IgG in all samples. Incubation of extracted CICs with normal human serum resulted in elevated levels of complement activation products, including C5a and C5b-9. This increase was completely inhibited by the addition of EDTA or EGTA. In conclusion, CICs in the serum of patients with MPO-AAGN contain MPO and IgG (presumably MPO-ANCA). These CICs can activate the complement system, likely through the classical pathway. Our findings support the hypothesis that CICs composed of MPO and MPO-ANCA contribute to complement activation via the classical pathway and play a significant role in AAGN pathogenesis. Full article

Background/Objectives: The healing rate of wounds resulting from postoperative abdominal surgery interventions increases as the inflammation and infections in the wound are reduced. However, antibiotic resistance among microorganisms increases the incidence of surgical site infections (SSIs). Therefore, the need for new products that exhibit antimicrobial and anti-inflammatory activities, in addition to antibiotics, is increasing. Methods: Silver nanoparticles (CO-AgNPs) were obtained using the green synthesis technique with Cydonia oblonga L. leaves, which constitute a significant amount of waste, and the effects of the obtained nanoparticles on in vitro wound healing were determined. Results: It was observed that CO-AgNPs inhibited myeloperoxidase and collagenase, enzymes that negatively affect wound healing. Furthermore, they exhibited good antimicrobial activity against Pseudomonas aeruginosa, Escherichia coli, Bacillus subtilis, and Staphylococcus aureus, which are common hospital pathogens. The CO-AgNPs could exhibit enhanced wound-healing properties by inhibiting microorganisms. Conclusions: It was clear that the development of new, environmentally friendly, and biocompatible products containing CO-AgNPs could be feasible, particularly for wound healing following infected abdominal surgery. Full article

Escherichia coli O157:H7 is a pathogenic bacterium that causes severe intestinal infections characterized by inflammation and disruption of the intestinal barrier. Probiotic lactic acid bacteria (LAB) from milk can support intestinal health and combat enteric pathogens; however, the potential of feline milk-derived LAB against E. coli O157:H7 infection remains unclear. In this study, Pediococcus acidilactici (P. acidilactici) M22, isolated from feline milk, was evaluated for probiotic activity in vitro and in vivo in a C57BL/6 mouse model of Escherichia coli O157:H7 infection. In vitro assays demonstrated that M22 significantly inhibited the adhesion of Escherichia coli O157:H7 to intestinal epithelial cells. For in vivo assessment, C57BL/6 mice were orally administered M22 prior to infection with E. coli O157:H7. Protective effects were evaluated by monitoring body weight loss, colon length, disease activity index (DAI), myeloperoxidase (MPO) activity, cytokine levels, tight junction protein expression, oxidative stress markers, and gut microbiota composition. M22-treated mice exhibited significantly less body weight loss and lower DAI scores than infected controls. M22 also prevented colon shortening, indicating reduced colonic damage. Probiotic treatment attenuated neutrophil infiltration and mucosal inflammation, as evidenced by decreased colonic MPO activity, reduced levels of pro-inflammatory cytokines, and elevated anti-inflammatory IL-10. Additionally, M22 preserved intestinal barrier function by upregulating tight junction proteins and mitigating infection-induced histopathological changes. M22 supplementation enhanced antioxidant defenses in colonic tissue (lower malondialdehyde, higher superoxide dismutase and glutathione), indicating reduced oxidative stress. Furthermore, gut microbiota analysis (16S rRNA sequencing) revealed that M22 counteracted infection-induced dysbiosis, restoring microbial diversity and a healthy composition (enrichment of beneficial commensals and suppression of harmful bacteria). By safeguarding intestinal integrity and homeostasis, M22 emerges as a promising next-generation probiotic for improving intestinal health in companion animals. Full article

Background/Objectives: This study aims to investigate the effects of betanin, which possesses antioxidant and anti-inflammatory properties, against colitis, a disease in which inflammation and oxidative stress play a role in its pathophysiology, during the acute and subacute period. Methods: Thirty-two rats were included in the study and divided into four groups: control, colitis, 3-day betanin supplementation + colitis (bet3+colitis), and 15-day betanin supplementation + colitis (bet15+colitis). Experimental colitis was induced with trinitrobenzene sulfonic acid. Results: In the colitis group, malondialdehyde, myeloperoxidase, superoxide dismutase (SOD) inhibition rate, tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), interleukin-6, mucosal damage, and cell infiltration scores were higher than in the control group, while catalase and glutathione peroxidase (GPx) levels were lower. In the betanin supplementation groups, malondialdehyde, myeloperoxidase, TNF-α, IL-1β, mucosal damage, and cell infiltration scores were lower than in the colitis group, while GPx levels were higher. In addition, the SOD inhibition rate and interleukin-6 levels were lower in the bet15+colitis group than in the colitis group. TNF-α, IL-1β, interleukin-6, and GPx levels in the betanin groups were similar to the control group. Conclusions: Betanin supplementation demonstrated a significant anti-inflammatory effect by reducing inflammatory parameters and histopathological scores in both periods. Additionally, it exhibited a glutathione-related antioxidant effect by increasing GPx levels in both periods. However, although SOD inhibition rates decreased in the subacute period, no significant change in catalase levels was observed in either period. This indicates that it did not provide complete protection in terms of antioxidant effects in either period. Full article

Turmeric (Curcuma longa L., Zingiberaceae) is a widely consumed spice and functional food valued for its bioactive constituents. Using an activity-guided strategy, this study identified the dichloromethane fraction as the most potent anti-inflammatory fraction, exhibiting markedly stronger inhibition of reactive oxygen species (ROS) production than ibuprofen (IC₅₀ $\le$ 0.4 vs. 11.2 μg/mL). Bioassay-guided purification yielded bisacurone (1), didemethoxycurcumin (2), and β-turmerone (3), with compounds 1 and 2 reported here for the first time in this fraction. Among them, β-turmerone displayed the strongest anti-inflammatory activity (IC₅₀ = 4.7 μg/mL), consistent with in silico docking and molecular dynamics analyses, revealing greater binding affinity and complex stability with myeloperoxidase (ΔG_bind = −20.90 vs. −18.89 kcal/mol for ibuprofen). Gas chromatography–mass spectrometry (GC-MS) profiling revealed a phytochemical profile dominated by turmerones and curlone, correlating with the observed bioactivity. None of the fractions exhibited acute toxicity in brine shrimp lethality assays, indicating a favorable preliminary safety profile. Our findings demonstrate the value of activity-guided isolation combined with computational validation for identifying turmeric-derived bioactives with promising nutraceutical potential, warranting further in vivo evaluation. Full article

Cardiovascular diseases (CVDs) are increasingly being defined not only in terms of metabolic or purely vascular disorders, but also as complex immunometabolic disorders. One of the most groundbreaking discoveries in recent years is the role of neutrophil extracellular networks (NETs/NENs) as a key link between chronic vascular wall inflammation and thrombotic processes. In this article, we present a synthetic overview of the latest data on the biology of NETs/NENs and their impact on the development of atherosclerosis, endothelial dysfunction, and the mechanisms of immunothrombosis. We highlight how these structures contribute to the weakening of atherosclerotic plaque stability, impaired endothelial barrier integrity, platelet activation, and the initiation of the coagulation cascade. We also discuss the modulating role of classic risk factors such as hypertension, dyslipidemia, and exposure to tobacco smoke, which may increase the formation or hinder the elimination of NETs/NENs. We also focus on the practical application of this knowledge: we present biomarkers associated with the presence of NETs/NENs (cfDNA, MPO–DNA complexes, CitH3, NE), which may be useful in diagnostics and risk stratification, and we discuss innovative therapeutic strategies. In addition to classic methods for indirectly inhibiting NET/NEN formation (antiplatelet, anti-inflammatory, and immunometabolic agents), we present experimental approaches aimed at their neutralization and removal (e.g., DNase I, elastase, and myeloperoxidase inhibitors). We pay particular attention to the context of cardiac and cardiac surgical procedures (Percutaneous Coronary Intervention-PCI, coronary artery bypass grafting-CABG), where rapid NET/NEN bursts can increase the risk of acute thrombotic complications. The overall evidence indicates that NETs/NENs represent an innovative and promising research and therapeutic target, allowing us to view cardiovascular diseases in a new light—as a dynamic interaction of inflammatory, atherosclerotic, and thrombotic processes. This opens up new possibilities in diagnostics, combination treatment and personalisation of therapy, although further research and standardization of detection methods remain necessary. Full article