New thiazine derivatives with dual potential for lipoxygenase inhibition and hemolytic activity: Synthesis, characterization and molecular docking studies.

Background: Inflammation is a complex biological response of vascular tissues to harmful stimuli such as pathogens, damaged cells or irritants and plays a crucial role in the pathogenesis of various chronic diseases including arthritis, cardiovascular disorders and cancer. Despite the availability of synthetic anti-inflammatory drugs, their long-term use is often associated with adverse effects, necessitating the search for safer and more effective alternatives. Anti-inflammation refers to mechanisms that suppress or regulate inflammation to protect tissues from damage. It includes natural body controls, anti-inflammatory cytokines, Lipoxygenase and therapeutic agents like NSAIDs, corticosteroids, and antioxidants. Effective anti-inflammatory action restores balance, promotes healing, and prevents progression of chronic illnesses. This thesis investigates the anti-inflammatory potential of selected natural and synthetic compounds through In-vitro model. The present study aims to investigate the anti-inflammatory activity of ethanolic extract of leaves of Putranjiva roxburghii wall. Methodology: The anti-inflammatory activity of the test samples was evaluated using egg albumin denaturation inhibition assay and lipoxygenase (LOX) inhibition assay. In the egg albumin assay, varying concentrations (50 μg/ml,100 μg/ml,200 μg/ml,400 μg/ml,800 μg/ml) of the test sample were incubated with egg albumin solution at pH 7.4 and 37 °C for 30 minutes. Protein denaturation was induced by heating the mixture at 70 °C for 15 minutes. After cooling, absorbance was measured at 280 nm, and the percentage inhibition of protein denaturation was calculated using diclofenac sodium as the reference. For the lipoxygenase inhibition assay, lipoxygenase was used to catalyze the oxidation of linoleic acid in the presence of the test compounds. The reaction mixture was incubated at 25°C for 5 minutes, and the formation of conjugated dienes was monitored by measuring absorbance at 234 nm. The percentage inhibition of LOX activity was calculated and compared with a standard anti-inflammatory agent. Result:It is anticipated that the ethanolic leaf extract of Putranjiva roxburghii wall. will exhibit significant in-vitro anti-inflammatory activity, with considerable inhibition of albumin denaturation and lipoxygenase enzyme activity, demonstrating its potential as a natural anti- inflammatory agent. Conclusion: Putranjiva roxburghii Wall. leaf extract showed positive anti-inflammatory activity by inhibiting egg albumin denaturation and lipoxygenase, confirming its potential as a natural anti-inflammatory agent.

CarvenS™ is a herbal syrup formulated with standardized extracts of Thyme (Thymus vulgaris L., Lamiaceae, herba) and Licorice (Glycyrrhiza glabra L. root, Fabaceae) in a Stevia sp.-base, commonly used in Türkiye for the relief of cough, cold, and flu symptoms. This study assessed its in vitro inhibitory potential against angiotensin-converting enzyme 2 (ACE2), transmembrane protease serine 2 (TMPRSS2), neuraminidase (NA), and selected pro-inflammatory mediators including tumor necrosis factor-alpha (TNF-α), cyclooxygenase (COX), and lipoxygenase (LOX), respectively. In addition, the phytochemical profile was determined using high-performance liquid chromatography (HPLC), which revealed the presence of 1.07%(w/v) rosmarinic acid and 0.4%(w/v) glycyrrhizic acid. Enzyme inhibition assays were performed on a microplate system using commercial kits at a final concentration of 20µg/mL. The syrup showed strong inhibitory effects on ACE2 (81%), TMPRSS2 (89%), and NA (85%). Anti-inflammatory activity was also evident, with inhibition of TNF-α (74.2%), COX-1 (76.3%), COX-2 (78.9%), and 5-LOX (83.1%), respectively. As an overall conclusion, the experimental findings indicate that CarvenS™ demonstrates substantial enzyme inhibitory activity relevant to both antiviral and anti-inflammatory pathways. Further in vivo studies are needed to validate these results and clarify the underlying mechanisms.

Lipoxygenases are enzymes found in plants, mammals, and other organisms that catalyse the hydroperoxidation of polyunsaturated fatty acids, such as arachidonic, linoleic, and linolenic acids. They have attracted a lot of attention as molecular targets for industrial and biomedical applications, due to their implication in key biological processes, such as plant development and defence, cell growth, as well as immune response and inflammation. Soybean (Glycine max) lipoxygenase (LOX) is a versatile biocatalyst used in biotechnology, pharmaceutical, and food industries. sLOX1, a soybean LOX isoform, is central in various industrial applications; thus, it is of particular interest to develop an efficient sLOX1 isolation process, control its activity, and leverage its potential as an effective industrial biocatalyst, tailoring it to a specific desired outcome. In this study, sLOX1 was extracted and purified from soybean seeds using an optimized protocol that yielded an enzyme preparation with higher activity compared to the commercially available lipoxygenase. Comprehensive biophysical characterization employing dynamic and electrophoretic light scattering, fluorescence, and Fourier-transform infrared spectroscopies revealed that sLOX1 exhibits remarkable structural and functional stability, particularly in sodium borate buffer (pH 9), where it retains activity and integrity up to at least 55 °C and displays minimal aggregation under thermal, ionic, and temporal stress. In contrast, sLOX1 in sodium phosphate buffer (pH 6.8) remained relatively stable against ionic strength and time but showed thermally induced aggregation above 55 °C, while in sodium acetate buffer (pH 4.6), the enzyme exhibited a pronounced aggregation tendency under all tested conditions. Overall, this study provides physicochemical and stability assessments of sLOX1. The combination of enhanced catalytic activity, high purity, and well-defined stability profile across diverse buffer systems highlights sLOX1 as a promising and adaptable biocatalyst for industrial applications, offering valuable insights into optimizing lipoxygenase-based bioprocesses.

The arachidonic acid pathway plays a pivotal role in the biosynthesis of important inflammatory and signal transducing agents such as prostaglandins, leukotrienes and thromboxanes. When this pathway is deregulated, it leads to pathological conditions such as cardiovascular diseases, metabolic diseases, and cancer. Two key enzymes of the pathway are cyclooxygenases (COXs) and lipoxygenases (LOXs), which are responsible for the production of prostaglandins and leukotrienes, respectively. Consequently, these enzymes have long been recognized as key therapeutic targets for the treatment and management of inflammatory disorders and other pathological conditions associated with inflammation. In this review, we describe the new evidence over the last 4 years regarding the arachidonic acid pathway. Moreover, we will pay attention to the structure and function of the COX-2 and 5-LOX enzymes and their role in inflammation, as well as define their active sites. Later, we will discuss the most potent, dual inhibitors of COX-2 and 5-LOX enzymes, based on in vitro and in vivo experiments, from 2020-2024. Structure-activity relationship (SAR) analysis of these compounds revealed four key structural features required for potent dual inhibition of cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX). We refer to these criteria as "The Rule of Four for Inflammation".

Nociceptive pain is a cardinal feature of traumatic and inflammatory bone diseases. However, whether and how nociceptors actively regulate the immune response during bone regeneration remains unclear. Here, we found that neutrophil-triggered nociceptive ingrowth functioned as negative feedback regulation to inflammation during bone healing. A unique Il4ra+Ccl2high neutrophil subset drove intense postinjury TRPV1+ nociceptive ingrowth, which in return dissipated inflammation by activating the production of pro-resolving mediator lipoxin A4 (LXA4) in osteoblasts. Mechanistically, osteoblastic autophagy activated by nociceptor-derived calcitonin gene-related peptide (CGRP) suppressed the nuclear translocation of arachidonate 5-lipoxygenase (5-LOX) to favor the LXA4 biosynthesis. Moreover, in alveolar bone from patients with Type II diabetes, we found diminished nociceptive innervation correlated with reduced autophagy, increased inflammation, and impaired bone formation. Activating nociceptive nerves by spicy diet or topical administration of a clinical-approved TRPV1 agonist showed therapeutic benefits on alveolar bone healing in diabetic mice. These results reveal a critical neuroimmune interaction underlying the inflammation-regeneration balance during bone repairing and may lead to novel therapeutic strategies for inflammatory bone diseases.

Pre harvest spraying of benzothiadiazole (BTH) induces grape disease resistance and regulates the quality of wine grapes. This study investigated the mechanism of spraying 50 mg/L BTH during the pea-size stage to regulate the metabolic pathway of grape lipoxygenase (LOX). The results indicated that BTH up-regulated the expression of VvFAD21, facilitating the conversion of saturated fatty acids into unsaturated fatty acids. Additionally, BTH treatment enhanced the expression levels of VvLOXA, VvLOXB, VvLOXC, VvHPL1, VvHPL2, VvADH1, and VvADH2, resulting in increased production of C6/C9 aldehydes and alcohols. Antibacterial assays of LOX metabolites demonstrated that 40-160 μg/mL of hexanal and E-2-hexenal significantly inhibited Botrytis cinerea, the causal agent of grape gray mold. This study clarifies the mechanisms by which BTH regulates LOX metabolism to enhance grape health and berry quality, providing a theoretical basis for its application in improving grape disease resistance and controlling berry quality.

In the present study, novel Schiff base ligands were synthesized via the condensation of salicylaldehyde derivatives with o-aminophenol, and their metal complexes with Co(II), Ni(II), Cu(II), Zn(II), Cd(II), and UO₂(VI) metal actetates were successfully prepared. The compounds were comprehensively characterized using FT-IR, 1H-NMR, 13C-NMR, UV-Vis spectroscopy, X-ray diffraction (XRD), scanning electron microscopy (SEM), and magnetic susceptibility measurements. FT-IR spectra confirmed azomethine (C = N) coordination through characteristic bands at 1612-1630cm-1, while 1H-NMR spectra exhibited diagnostic azomethine proton signals at δ 8.84-8.62ppm. XRD analysis revealed a semi-crystalline nature with prominent reflections at 2θ = 18.02° and 20.87°, and SEM images demonstrated nanorod-like morphologies. Magnetic moment values supported the proposed coordination geometries of the paramagnetic complexes. The biological activity of the synthesized compounds was investigated through in vitro inhibition studies against lipoxygenase (LOX), a ~ 97kDa enzyme purified from Chenopodium quinoa Willd. with 77.89% purity and 7.24% yield. All compounds exhibited competitive inhibition toward LOX, with inhibition constants (Ki) in the range of 0.014 ± 0.002 to 0.858 ± 0.194μM, indicating remarkable inhibitory potency. Molecular docking studies further substantiated the experimental results by revealing key hydrogen-bonding and π-cation interactions between the most active compounds and essential amino acid residues within the LOX active site. Overall, the results demonstrate that these Schiff base metal complexes represent promising candidates for the development of potent LOX inhibitors.

We have recently demonstrated that 7-O-methylpunctatin (MP), a novel homoisoflavonoid, suppresses inflammation-induced arterial pathogenesis. However, the precise biochemical mechanisms underlying its atheroprotective effects remain elusive. In this study, we employed various in silico studies to elucidate MP's plausible potential and the specific molecular pathways through which it exerts its influence on atherosclerosis. Our analysis of MP's pharmacokinetic, physicochemical, and toxicological properties revealed a profile characterized by favorable absorption, efficient metabolism and excretion, and minimal toxicity. Through target identification and protein-protein interaction analyses, we identified ALOX5 as a pivotal hub gene-an enzyme critically involved in the pathogenesis of atherosclerosis. Furthermore, we identified ten transcription factors and four kinases as potential targets. Molecular mechanics/generalized-born surface area calculations, complemented by time-scale molecular dynamics simulations, revealed that MP binds to ALOX5 with high affinity, modulating its structural stability, rigidity, compactness, overall folding pattern, and residual correlations and motions. These findings corroborate previous in vitro and in vivo investigations that underscore the anti-atherosclerotic effects of ALOX5 inhibition, thereby positioning MP as a promising therapeutic candidate for combating atherosclerosis.

Lipoxygenase expression and oxylipin analysis in human THP-1 monocyte-derived macrophages.

ALOX15: a key regulator in the pathogenesis and therapy of chronic rhinosinusitis with nasal polyps.

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

Copper(II) nitrate reacts with tris(m-tolyl)phosphine (m-MePh)3P in 1 : 2 and 1 : 3 molar ratios to form [Cu((m-MePh)3P)2(NO3)] (1) which contains two isomers (1A and 1B) in the crystal lattice and [Cu((m-MePh)3P)3(NO3)] (2). The reaction of copper(II) nitrate with tris(p-tolyl)phosphine (p-MePh)3P in 1 : 3 molar ratio results in the {[Cu((p-MePh)3P)3(NO3)]·(DMF)·(H2O)} (3) complex. The complexes were characterized in the solid state using melting point (m.p), X-ray fluorescence spectroscopy (XRF), and attenuated total reflectance-Fourier transform infra-red (ATR-FT-IR) spectroscopy, and in solution using cryoscopy, ultraviolet-visible (UV-vis) and nuclear magnetic resonance (1H-NMR) spectroscopy. Their crystal structures were determined using single crystal X-ray crystallography in the solid state while the molecular weight was calculated using cryoscopy. A CCDC search shows that 1A and 1B are the first known chiral Cu(I) complexes of the CuP2O2 core. Binding affinity of complexes 1-3 toward calf thymus DNA (CT-DNA) was investigated ex vivo using UV-vis and fluorescence spectroscopy, viscosity measurements, and DNA denaturation assays. Their lipoxygenase (LOX) inhibitory activity was also studied. In silico computations further rationalized the DNA and LOX interactions with 1-3. In vitro assays were conducted to evaluate the activity of 1-3 against human breast adenocarcinoma (MCF-7) cells, and the results are presented herein.

NnLOX1-mediated aroma formation in lotus (Nelumbo nucifera) through multi-omics integration and machine learning.

Nano-selenium alleviates cadmium-induced ferroptosis in chicken liver by modulating the TRPM2 channel.

Lipidomic analysis of phospholipids and transcript expression of lipid metabolism genes in the liver and muscle of Atlantic salmon fed microbial oil and canola oil.

Enzymes, as biological catalysts, are widely used for their excellent selectivity and sensitivity, playing a crucial role in biosensors, offering significant potential for monitoring food products and health conditions. However, enzyme activity can be compromised by environmental factors like high humidity, pH, and temperature variations. This study explores the feasibility of improving the stability of the unstable enzyme lipoxygenase (LOX) through immobilization using silk, a naturally occurring fibrous protein. A linoleic acid biosensor is designed by combining silk‐entrapped LOX with poly(3,4‐ethyloxy thiophene):poly(styrene sulfonate) (PEDOT:PSS) and horseradish peroxidase (HRP). The findings demonstrate that immobilizing LOX with different molecular weight silks extends the shelf‐life of the freeze‐dried PEDOT:PSS/HRP/LOX more than tenfold, from less than 1 day to 10 days. The results of the characterization tests (e.g., Fourier transform infrared spectroscopy, rheology) highlighted the interdependency between silk properties, such as its molecular weight, the silk conformation, and the pore shape and size, and enzyme stability. Low molecular weight silk resulted in the best performance, enabling storage at room temperature. It can therefore be concluded that immobilization with silk is an efficient method to enhance the shelf‐life of enzyme‐based biosensors, for diagnostic and other applications.

The growing demand for medicinal plants in herbal medicine highlights their therapeutic value, yet heavy metal contamination, such as cobalt (Co), poses potential health risks. Cobalt’s dual role as an essential micronutrient and a toxic pollutant necessitates a more profound understanding of its impact on medicinal plants like Adhatoda vasica. We investigated how varying Co concentrations affect A. vasica shoot growth, leaf anatomy, antioxidant enzyme activity, and secondary metabolite profiles. Additionally, molecular docking was performed to assess the interaction of the metabolites with the skin cancer-related protein anti-ssDNA antigen-binding fragment (PDB code: 1P7K). Low Co concentrations (50 µM) enhanced shoot dry weight by 41.45%, while higher levels (100–1000 µM) reduced it by up to 66.86%. Cobalt exposure increased hydrogen peroxide (H2O2) and lipoxygenase (LOX) activity, indicating reactive oxygen species (ROS)-induced oxidative stress. Higher Co levels increased superoxide dismutase (SOD), catalase (CAT), phenylalanine ammonia-lyase (PAL), and polyphenol oxidase (PPO) but decreased peroxidase (POD) and ascorbate peroxidase (APX) activity. HPLC-UV and GC-MS data showed that Co altered the secondary metabolites of A. vasica, including phenolics, flavonoids, alkaloids, and terpenoids, in both qualitative and quantitative ways. Molecular docking shows that naringin has a higher binding affinity (-9.2 kcal/mol) to PDB: 1P7K than phenolics (-4.8 to -6.4 kcal/mol). Cobalt stress significantly impacts A. vasica, altering its leaf structure, growth, and antioxidant defenses. These effects extend to secondary metabolites in a dose-dependent manner. These findings highlight the plant’s potential for Co tolerance and its metabolites’ therapeutic promise, particularly naringin, for skin cancer applications.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12896-025-01088-9.

To observe the effect of "Shugan Tiaoshen" (regulating mental activities by relieving depressed liver) acupuncture on hippocampal iron metabolism, antioxidant system and lipid peroxidation related factors in chronic inflammatory pain and depression comorbidity (CIPDC) rats and its protective effect on hippocampal neurons, so as to explore its mechanisms underlying improvement of CIPDC. A total of 48 male SD rats were randomly divided into control, model, electroacupuncture (EA) and medication (paroxetine) groups, with 12 rats in each group. The CIPDC model was established by injection of complete Freund's adjuvant into the toe surface of the left hind paw. From 15 to 28 days after modeling, EA was applied to "Yintang" (GV24+), "Baihui" (GV20), "Hegu" (LI4) and "Taichong" (LR3) for 30 min, once a day for 14 days. Rats in the medication group were treated with intragastric administration of paroxetine (10 mg/kg), once a day for 14 days. Paw withdrawal mechanical threshold (PWMT) and paw thermal withdrawal latency (PTWL) were detected for evaluating the rats' pain response behavior in each group at 0, 7, 14, 21 and 28 days after modeling. The sucrose preference test, open field test (OFT) and forced swimming test (FST) were conducted to calculate the sucrose preference value, and to record the total distance traveled in 30 min and the time spent in the central zone of the open field during the first 5 min, and the immobility (floating) time during the last 4 min for evaluating the rats' depression-like behavior at 0, 14 and 28 days after modeling. H.E. staining was used to observe the histopathological changes of hippocampal neurons. Prussian blue staining was used to observe the iron deposition in the hippocampus tissue. A transmission electron microscopy was used to observe the ultrastructure of mitochondria of hippocampal neurons. The content of Fe2+ in the hippocampus tissue was detected by colorimetry method. ELISA method was used to measure the contents of glutathione (GSH) and malondialdehyde (MDA) in the hippocampus tissue. The expression levels of ferritin heavy chain 1 (FTH1), ferritin light chain (FTL), solute carrier family 7 member 11 (SLC7A11), glutathione peroxidase 4 (GPX4), acyl-coa synthetase long chain family member 4 (ACSL4), and lipoxygenase (LOX) proteins and mRNAs of hippocampus were detected by Western blot and real-time fluorescence quantitative PCR, separately. Compared with the control group, the PWMT and PTWL at 7, 14, 21 and 28 days after modeling, sucrose preference value (SPV), total motion distance of OFT, time spent in the central zone, GSH content, and expression levels of FTH1, FTL, SLC7A11 and GPX4 proteins and mRNAs were significantly decreased (P<0.01), while immobility time of FST, the contents of Fe2+ and MDA, and the expression levels of ACSL4 and LOX protein and mRNAs were considerably increased (P<0.01) in the model group. In contrast to the model group, both EA and medication group had an obvious increase in the PWMT and PTWL at 21 and 28 days after modeling, SPV, total motion distance, time spent in the central zone, GSH content, and the expression levels of FTH1, FTL, SLC7A11 and GPX4 proteins and mRNAs (P<0.01, P<0.05), and a striking decrease in the immobility time, contents of Fe2+ and MDA, and the expression levels of ACSL4 and LOX protein and mRNAs (P<0.01, P<0.05). No significant differences were found between the EA and medication groups in all the indexes mentioned above. Morphological observation showed a large number of yellow-brown iron deposition in the hippocampus, neuronal degeneration, disordered arrangement and necrosis of neurons, shrinkage and deformation of mitochondria, increased membrane density, blurred boundary, and decrease and disappearance of mitochondrial cribriae in the model group, which was obviously milder in the injury state in both EA and medication groups, including reduction of yellow-brown iron deposition, regular arrangement of neurons, obvious nucleolus and complete structure of mitochondria, etc. "Shugan Tiaoshen" acupuncture can improve the inflammatory pain and depression in rats with CIPDC , which may be related to its functions in activating antioxidant activity and promoting iron metabolism balance, reducing lipid peroxidation, and inhibiting ferroptosis, thereby promoting the repair and remodeling of hippocampal neurons and ameliorating the disease process of CIPDC.

As an important branch of the lipoxygenase (LOX) metabolism pathway, hydroperoxide lyase (HPL) is involved in regulating plant development and defense responses. However, the upstream regulatory mechanism of HPL remains unclear in soybean. In the present study, by analyzing the upstream promoter region of the GmHPL gene, cis-elements such as MYB motifs, G-box motifs, ERE motifs and W-box motifs were predicted, which were related to the stress response. Yeast one-hybrid was employed and two transcription factors were identified, GmERF36 and GmILR3. The orthologs of ERF36 and ILR3 in Arabidopsis were involved in pathogen stress. A dual-luciferase reporter assay verified the yeast one-hybrid results and indicated that GmERF36 and GmILR3 suppressed the expression of the GmHPL protein. The qRT-PCR results indicated that GmHPL and GmERF36 initially displayed inverse expression patterns within 24 h after Colletotrichum truncatum treatment (GmERF36 was upregulated while GmHPL was downregulated); then, both of them were upregulated before decreasing. The results indicated that the response of GmHPL to pathogen stress partially depended on GmERF36. Our study gives rise to new insights into the upstream regulatory network of the GmHPL gene.