Search Results (1,348)

Fermentation represents a sustainable biotechnological approach for enhancing bioactive properties of plant-based foods, yet its anticancer effects remain underexplored. We evaluated the antiproliferative activity of fermented (with commercial probiotic lactic acid bacteria consortium) and unfermented plant-based beverages derived from tiger nut, carob, and rice using an in vitro model. Following INFOGEST 2.0 gastrointestinal digestion, bioaccessible fractions were applied to Caco-2 colorectal adenocarcinoma cells at 1:15 v/v dilution for 24 h. Analyses included cell viability, apoptosis detection, cell cycle distribution, reactive oxygen species production, glutathione content, mitochondrial membrane potential, and intracellular calcium levels. Fermented tiger nut achieved superior (p < 0.05) cytotoxicity compared to unfermented counterpart (39.6% vs. 77.4% cell viability) through dual mechanisms: depleting cellular antioxidant defenses (glutathione reduced to 55.9%) while inducing oxidative stress (180.3% ROS overproduction). This evoked irreversible apoptosis (76.9% early apoptosis) and extensive DNA fragmentation (84.8% SubG₁ population) via calcium-independent pathways. Fermented carob operated through cytostatic mechanisms, inducing G₀/G₁ cell cycle arrest (74.7% vs. 44.2% in blank digestion cells) without oxidative stress. Fermentation reduced (p < 0.05) rice beverage antiproliferative activity (90.2% vs. 71.9% unfermented beverage cell viability). These findings establish lactic acid fermentation as effective for developing plant-based beverages with anticancer mechanisms, offering dietary strategies for colorectal cancer prevention. Full article

Harmful cyanobacterial blooms (CyanoHABs) threaten freshwater ecosystems and human health. Inhibiting cyanobacteria through plant allelopathy is an effective and environmentally friendly approach for CyanoHAB control. In this study, we evaluated the inhibitory activities of several organic solvent extracts from Artemisia argyi against the common bloom-forming cyanobacterium Microcystis aeruginosa, explored the anti-algal mechanism of the active fraction, analyzed its secondary metabolites using liquid chromatography–high-resolution mass spectrometry (LC-HRMS), and screened the potential allelochemicals. The results showed that the crude extract of A. argyi leaves (CE) exhibited significant inhibitory effects on M. aeruginosa. Among several solvent fractions of CE, the dichloromethane extract (DE) demonstrated the strongest inhibitory effect, with a 7-day IC₅₀ of 70.43 mg/L. After treatment with DE, the contents of chlorophyll a (Chl a), carotenoids, and phycobiliproteins (PBPs) in M. aeruginosa were significantly reduced. Meanwhile, an excessive accumulation of reactive oxygen species (ROS), reduction of catalase (CAT) activity, increase in malondialdehyde (MDA) content, and shrinkage of the membrane were found in M. aeruginosa cells under DE treatments. There were 81 secondary metabolites annotated in DE by LC-HRMS. Among them, hispidulin, jaceosidin, 5,7,3′-trihydroxy-6,4′,5′-trimethoxyflavone, and eupatilin possessed strong inhibitory activities, with 7-day IC₅₀ values of 26.23, 27.62, 32.02, and 34.98 mg/L, respectively. These results indicated that the A. argyi extracts possess significant allelopathic activities on M. aeruginosa, and DE was identified as the primary active fraction. It inhibits algae growth by suppressing photosynthesis and inducing peroxidation, ultimately leading to cell death. Flavonoids in DE were the main allelochemicals responsible for the inhibition on algae of A. argyi extracts. Full article

Lipids, together with water and proteins, constitute the essential structure of cell membranes, and in the CNS, critically contribute to the production, function, and maintenance of the myelin sheath. Myelin produced by oligodendrocytes (OLs) acts as an electric insulator and assures proper conduction of information. Three major fractions of myelin lipids are cholesterol, phospholipids, and glycolipids. These lipids not only sculpt the myelin landscape as a structural support for proteins, but they also play a crucial role in molecular interactions underlying processes of protein trafficking and signal transductions. The high lipid content of myelin makes it susceptible to lipid metabolism disorders. Disorders in systemic and local lipid metabolism may lead to loss of myelin integrity and stability, and potentially to CNS demyelination seen in neurodegenerative diseases, notably progressive multiple sclerosis, for which there are few effective therapies. Precise interactions among disorders in lipid metabolism, function of oligodendrocytes, and demyelination/remyelination events, including de novo myelin formation and myelin remodeling processes, may lay the foundation for novel therapeutics for progressive MS and other demyelinating CNS conditions. Full article

Despite increasing evidence that cigarette smoke is a significant source of indoor fine particulate matter (PM_2.5), quantitative emission factors (EFs) for PM_2.5 and its toxic chemical composition in mainstream (MS) and sidestream (SS) smoke are still not well defined. In this study, we employed a custom-designed chamber to separately collect MS (intermittent puff) and SS (continuous sampling) smoke from eleven cigarette models, representing six brands and two product types, under controlled conditions. PM_2.5 was collected on quartz-fiber filters and analyzed for carbon fractions (using the thermal–optical IMPROVE-A protocol), nine water-soluble inorganic ions (by ion chromatography), and twelve trace elements (via ICP-MS). SS smoke exhibited significantly higher mass fractions of total analyzed species (84.7% vs. 65.9%), carbon components (50.6% vs. 44.2%), water-soluble ions (17.1% vs. 13.7%), and elements (17.0% vs. 7.0%) compared to MS smoke. MS smoke is characterized by a high proportion of pyrolytic organic carbon fractions (OC₁–OC₃) and specific elements such as vanadium (V) and arsenic (As), while SS smoke shows elevated levels of elemental carbon (EC1), water-soluble ions (NH₄⁺, NO₃⁻), and certain elements like zinc (Zn) and cadmium (Cd). The toxicity-weighted distribution indicates that MS smoke primarily induces membrane disruption and pulmonary inflammation through semi-volatile organics and elements, whereas SS smoke enhances oxidative stress and cardiopulmonary impairment via EC-mediated reactions and secondary aerosol formation. The mean OC/EC ratio of 132.4 in SS smoke is an order of magnitude higher than values reported for biomass or fossil-fuel combustion, indicative of extensive incomplete combustion unique to cigarettes and suggesting a high potential for oxidative stress generation. Emission factors (µg/g cigarette) revealed marked differences: MS delivered higher absolute EFs for PM_2.5 (422.1), OC (8.8), EC (5.0), Na⁺ (32.6), and V (29.2), while SS emitted greater proportions of NH₄⁺, NO₃⁻, Cl⁻, and carcinogenic metals (As, Cd, Zn). These findings provide quantitative source profiles suitable for receptor-oriented indoor source-apportionment models and offer toxicological evidence to support the prioritization of comprehensive smoke-free regulations. Full article

Background/Objectives: Aristolochia ringens, a medicinal plant widely used in traditional medicine, has shown potential therapeutic applications. This study aimed to investigate the anticancer mechanism of action of its crude extract against human colorectal adenocarcinoma cells (Caco-2 and HT-29). Methods: Cell viability was assessed using the MTT assay to determine IC₅₀ values. Immunofluorescence microscopy was used to examine nuclear morphology and microtubule integrity. Flow cytometry with PI staining was used for cell cycle analysis and Annexin V-FITC/PI staining for apoptosis detection. Mitochondrial membrane potential was evaluated using JC-1 dye. Bioactivity-guided fractionation was performed via HPLC, and GC–MS was used to profile active constituents. Results: The extract exhibited dose-dependent cytotoxicity with IC₅₀ values below 30 µg/mL in colon adenocarcinoma cell lines. Treated Caco-2 cells showed nuclear shrinkage and disrupted microtubules. PI-based flow cytometry revealed G₁ phase arrest, and Annexin V-FITC/PI staining indicated enhanced late apoptosis. JC-1 staining demonstrated mitochondrial depolarization. HPLC fractionation identified fractions 2 and 3 as active, and preliminary GC–MS analysis tentatively annotated the presence of alkaloids, sesquiterpenes/diterpenes, and steroidal compounds. Conclusions: A. ringens exerts anticancer effects through a mitochondria-mediated apoptotic pathway, involving G₁ checkpoint arrest and cytoskeletal disruption. These findings provide the first integrated cellular and mechanistic evidence of its anticancer potential in colorectal cancer, supporting its promise as a source of novel therapeutic lead compounds. Full article

Alcohol Use Disorder (AUD) poses global health challenges, and causes hematological alterations such as macrocytosis and oxidative stress. Disruption of protein structures by alcohol and/or its metabolites may exacerbate AUDs; proteomics can elucidate the underlying biological mechanisms. This study examined the proteins differentially expressed in the cytosol and membrane fractions of erythrocytes obtained from 30 male patients with AUD, comparing them to samples from 15 age- and BMI-matched social drinkers (SDs) and 15 non-drinkers (control). The analysis aimed to identify the molecular differences related to alcohol consumption. The AUD patient subgrouping was based on mean corpuscular volume (MCV), with 16 individuals classified as having a normal MCV and 14 having a high MCV. Proteins were separated via two-dimensional(2D)-gel electrophoresis, digested with trypsin, and identified via Matrix-Assisted Laser Desorption/Ionization Time-of-Flight (TOF) mass spectrometry (MALDI-TOF/TOF). Additionally, levels of malondialdehyde and 4-hydroxyalkenals (MDA + HAE), reduced glutathione (GSH), oxidized glutathione (GSSG), serum carbohydrate-deficient transferrin (%CDT), disialotransferrin (%DST), and sialic acid (SA) were analyzed. The results showed increased MDA + HAE and decreased total thiols in AUD patients, with GSSG elevated and the GSH/GSSG ratio reduced in the AUD MCV-high subgroup. Serum %CDT, %DST, and SA were significantly higher in AUD. Compared to the control profiles, the AUD group exhibited differential protein expression. Few proteins, such as bisphosphoglycerate mutase, were downregulated in AUD versus control and SD, as well as in the MCV-high AUD subgroup. Conversely, endoplasmin and gelsolin were upregulated in AUD relative to control. Cytoskeletal proteins, including spectrin-alpha chain, actin cytoplasmic 2, were overexpressed in the AUD group and MCV-high AUD subgroup. Several proteins, such as 14-3-3 isoforms, alpha-synuclein, translation initiation factors, heat shock proteins, and others, were upregulated in the MCV-high AUD subgroup. Under-expressed proteins in this subgroup include band 3 anion transport protein, bisphosphoglycerate mutase, tropomyosin alpha-3 chain, uroporphyrinogen decarboxylase, and WD repeat-containing protein 1. Our findings highlight the specific changes in protein expression associated with oxidative stress, cytoskeletal alterations, and metabolic dysregulation, specifically in AUD patients with an elevated MCV. Understanding these mechanisms is crucial for developing targeted interventions and identifying biomarkers of alcohol-induced cellular damage. The complex interplay between oxidative stress, membrane composition, and cellular function illustrates how chronic alcohol exposure affects cellular physiology. Full article

Microbial reduction in hexavalent chromium (Cr(VI)) is a well characterized bioremediation strategy, yet the mechanistic diversity among bacterial taxa necessitates detailed investigations into strain-specific pathways. Here, we report the isolation and characterization of Bacillus safensis BSF-4, a halophilic bacterium derived from saline-alkali soil, which demonstrates efficient Cr(VI) reduction capacity. Physiological assays showed that BSF-4 achieved 89.15% reduction of 20 mg/L Cr(VI) within 72 h, with Cr(III) identified as the primary extracellular end product. Resting cell assays and subcellular fractionation analyses confirmed that Cr(VI) reduction predominantly occurs in the extracellular milieu. X-ray photoelectron spectroscopy (XPS) further revealed soluble Cr(III) complexed with extracellular polymeric substances (EPS). Transcriptomic profiling indicated upregulation of membrane-associated transport systems (facilitating Cr(VI) exclusion) and quorum sensing (QS) pathways (mediating adaptive stress responses). These findings highlight a dual mechanism: (1) extracellular enzymatic reduction mediated by EPS-bound redox proteins, and (2) intracellular detoxification via QS-regulated defense pathways. Collectively, Bacillus safensis BSF-4 exhibits robust Cr(VI) reduction capacity under saline conditions, positioning it as a promising candidate for bioremediation of Cr(VI)-contaminated saline soils and aquatic ecosystems. Full article

Cu-BTC (HKUST-1) metal–organic framework (MOF) is widely recognized for its carbon capture capability due to its unsaturated copper sites, high surface area, and well-defined porous structure. This study developed mixed matrix membranes (MMMs) using cellulose triacetate (CTA), incorporating bimetallic Ni-Cu-BTC MOFs for CO₂/CH₄ separation, and benchmarked them against membranes fabricated with monometallic Cu-BTC. CTA was selected for its biodegradability, membrane-forming properties, and cost-effectiveness. The optimized membrane with 10 wt.% Ni-Cu-BTC achieved a CO₂ permeability of 22.9 Barrer at 25 °C and 5 bar—more than twice that of pristine CTA—with a CO₂/CH₄ selectivity of 33.8. This improvement stems from a 51.66% increase in fractional free volume, a 49.30% rise in the solubility coefficient, and a 51.94% boost in the diffusivity coefficient. Dual-sorption model analysis further confirmed enhanced solubility and adsorption mechanisms. These findings establish CTA/Ni-Cu-BTC membranes as promising candidates for high-performance CO₂ separation in natural gas purification and related industrial processes. Full article

Cotton Verticillium wilt is a disease that significantly impacts the cotton industry, severely affecting cotton quality and the economic well-being of farmers. Bacillus atrophaeus YL84 is a biocontrol bacterium with broad-spectrum antagonistic and growth-promoting characteristics, previously isolated by our laboratory. This study aimed to elucidate the antagonistic effects of sterilized fermentation filtrate from Bacillus atrophaeus YL84 on cotton Verticillium wilt pathogen Verticillium dahliae and its growth-promoting effects on cotton. The experiments were conducted in vitro and in vivo to assess these effects comprehensively. Using the dual culture method, it was found that Bacillus atrophaeus YL84 exhibited a high inhibition rate on mycelial growth of V. dahliae, with an inhibition rate of 84.11%. The undiluted YL84 sterilized fermentation filtrate and its 10% volume fraction dilution (fermentation filtrate diluted to 10%) exhibited inhibition rates of 80.25% and 72.16% for conidial germination and mycelial growth of V. dahliae, respectively. Scanning electron microscopy showed increased branching, swelling, and shortened internodes in the antagonized mycelia. Conductivity measurements revealed a significant enhancement caused by the YL84 filtrate, with conductivity increasing by 8.94 times compared to the control at a 250 μg/mL concentration. Similarly, protein leakage peaked at 9.47 times the control level at 250 μg/mL, demonstrating the filtrate’s potent impact on mycelial cell membrane permeability. The enzymatic activities of polygalacturonase (PG), cellulase (CL), and β-glucosidase (β-GC) were significantly reduced following treatment with YL84 sterilized fermentation filtrate, with reductions from control levels of 15.78, 10.11, and 5.01 U/mL to treatment levels of 11.81, 6.96, and 1.44 U/mL, respectively. Indoor pot experiments demonstrated that different concentrations of YL84 sterilized fermentation filtrate significantly suppressed the occurrence of cotton Verticillium wilt while promoting plant growth. Compared to the control group, application of 250 μg/mL YL84 sterilized fermentation filtrate resulted in a control efficacy of 66.69% for cotton Verticillium wilt, with increases in plant height, root length, fresh weight, and dry weight of 9.36–33.85%, 17.33–29.49%, 16.79–28.24%, and 25–58.33%, respectively. These findings underscore the potential of the YL84 filtrate as both a biocontrol agent and a promoter of cotton plant growth in agricultural settings. These results indicate that Bacillus atrophaeus YL84 sterilized fermentation filtrate possesses both disease-suppressing and growth-promoting activities, making it a promising candidate for development and use as a biocontrol agent and plant growth promoter. Full article

Mitochondrial fission and fusion appear to be relatively infrequent in cardiac cells compared to other cell types; however, the proteins involved in these events are highly expressed in adult cardiomyocytes (ACM). Therefore, these proteins likely have additional non-canonical roles. We have previously shown that DRP1 not only participates in mitochondrial fission processes but also regulates mitochondrial bioenergetics in cardiac tissue. However, it is still unknown where the DRP1 that does not participate in mitochondrial fission is located and what its role is at those non-fission spots. Therefore, this manuscript will clarify whether oligomeric DRP1 is located at the SR–mitochondria interface, a specific region that harbors the Ca²⁺ microdomains created by Ca²⁺ release from the SR through the RyR2. The high Ca²⁺ microdomains and the subsequent Ca²⁺ uptake by mitochondria through the mitochondrial Ca²⁺ uniporter complex (MCUC) are essential to regulate mitochondrial bioenergetics during excitation–contraction (EC) coupling. Herein, we aimed to test the hypothesis that mitochondria-bound DRP1 preferentially accumulates at the mitochondria–SR contacts to deploy its function on regulating mitochondrial bioenergetics and that this strategic position is modulated by calcium in a beat-to-beat manner. In addition, the mechanism responsible for such a biased distribution and its functional implications was investigated. High-resolution imaging approaches, cell fractionation, Western blot, 2D blue native gel electrophoresis, and immunoprecipitations were applied to both electrically paced ACM and Langendorff-perfused beating hearts to elucidate the mechanisms of the strategic DRP1 localization. Our data show that in ACM, mitochondria-bound DRP1 clusters in high molecular weight protein complexes at mitochondria-associated membrane (MAM). This clustering requires DRP1 interaction with β-ACTIN and is fortified by EC coupling-mediated Ca²⁺ transients. In ACM, DRP1 is anchored at the mitochondria–SR contacts through interactions with β-ACTIN and Ca²⁺ transients, playing a fundamental role in regulating mitochondrial physiology. Full article

Cytochrome P450 (CYP450) enzymes play an essential role in the metabolism of drugs, particularly in phase I metabolic reactions. In this article, we present a comprehensive review of fifteen selected enzymes belonging to the CYP450 family. The enzymes included in this analysis are CYP7A1, CYP3A4, CYP3A5, CYP2D6, CYP2E1, CYP2C8, CYP2C18, CYP2C9, CYP2C19, CYP2B6, CYP2A6, CYP2A13, CYP1B1, CYP1A1, and CYP1A2. We examined the influence of natural, polymorphic variations within their primary amino acid sequences on their enzymatic function and mechanisms of action. To begin, we compiled a dataset of naturally occurring polymorphic variants for these enzymes. This was achieved through a detailed analysis of entries in the UniProt database, as well as an extensive review of the current scientific literature. For each variant, we included commentary regarding its potential impact on enzyme activity or drug response, based on evidence observed in in vitro experiments, in vivo studies, or clinical trials. Particular emphasis was placed on how such polymorphisms might alter the metabolism of xenobiotics, thereby potentially affecting pharmacological outcomes. In this respect, the work represents the first comprehensive source in the scientific literature that systematically gathers and organizes data on CYP450 polymorphisms, including an assessment of their potential significance in processes mediated by these enzymes. A more detailed comparison of the polymorphism-related in vitro studies is devoted to CYP3A4, an enzyme that displays the largest fraction of clinically significant polymorphs. Secondly, we aimed to establish possible molecular explanations for why specific polymorphisms exhibit clinically or experimentally observable effects. To explore this, we performed a qualitative structural analysis of the enzymes, focusing on shared structural characteristics among the examined members of the CYP450 family. The results of this analysis demonstrate that there is no single universal mechanism by which polymorphisms influence the function of CYP450 enzymes. Instead, the mechanisms vary and may include alterations in the orientation of the enzyme within the lipid membrane, changes affecting the association or dissociation of substrates and products at the active site, structural stabilization or destabilization of the enzyme’s reactive centers, modifications in the way the enzyme interacts with its ligand, or alterations in the character of the interface involved in contact with its redox partner (electron transfer protein). Furthermore, among the polymorphisms that significantly impact enzyme function, mutations involving the substitution of arginine residues for other amino acids appear to be overrepresented. Full article

Mechanical and contractile forces in the vascular wall regulate smooth muscle cell migration. We previously demonstrated the presence of C3 complement products in atherosclerotic lesions of human aortas and showed that that C3-derived fragments promote key cellular processes, such as actin cytoskeleton organization and cell migration, in lipid-loaded human vascular smooth muscle cells (hVSMCs). In the present study, we aimed to investigate gene expression profiles related to cytoskeletal remodeling and cell adhesion in migrating hVSMCs with a particular focus on modulatory effect of the C3 complement pathway on these processes. We analyzed gene expression in migrating and non-migrating hVSMCs using real-time PCR and in silico network analysis. Additionally, we investigated cytoskeletal remodeling through Western blotting and confocal microscopy. PCR profiling revealed 30 genes with significantly altered expression in migrating hVSMCs compared to non-migrating control cells. In silico analysis identified six of these genes—PXN, AKT1, RHOA, VCL, CTNNB1, and FN1—as being associated with cytoskeletal remodeling and focal adhesion, with PXN occupying a central position in the interaction network. PXN expression was reduced at both the transcript and protein levels and showed altered subcellular localization in migrating lipid-loaded hVSMCs. Protein–protein interaction analysis using STRING predicted an association between PXN and the integrin complex αMβ2 (comprising ITGAM (CD11b) and ITGB2 (CD18)), which functions as receptors for the iC3b complement fragment. Confocal imaging of cell adhesion structures revealed that lipid-loaded hVSMCs stimulated with iC3b displayed a more diffuse PXN distribution and significantly increased PXN–F-actin colocalization in active cytoplasmic regions compared to lipid-loaded control cells. PXN–F-actin colocalization increased from 1.26% to 19.68%. Subcellular fractionation further confirmed enhanced PXN enrichment in the membrane fraction, with no significant changes observed in the cytosolic or cytoskeletal compartments. In conclusion, iC3b-mediated molecular signaling in lipid-loaded hVSMCs alters PXN distribution and enhances cytoskeletal remodeling, revealing novel molecular interactions in vascular remodeling and the progression of atherosclerotic lesions. Full article

Olive oil, a cornerstone of the Mediterranean diet, contains a saponifiable lipid fraction rich in oleic acid, and a non-saponifiable fraction composed of minor bioactive constituents such as squalene, vitamin E, oleuropein aglycone, hydroxytyrosol, oleocanthal, and oleacein, among other phenolic and triterpenic compounds. These components are well-documented for their cardiovascular, anti-inflammatory, antioxidant, and neuroprotective activities. This review explores the physiological relevance of olive oil lipids and their derivatives on cellular membranes and ion transport systems, by combining biochemical and electrophysiological insights. We discuss how oleic acid and its metabolites influence membrane lipid composition, modulate fluidity, and reorganize lipid rafts—key elements for the proper localization and function of ion channels. Additionally, we examine evidence showing that several olive oil components regulate ion channels such as TRP, potassium, calcium, and chloride channels, as well as other transporters, thereby influencing ionic homeostasis, oxidative balance, and signal transduction in excitable and non-excitable cells. By combining these findings, we propose a conceptual framework in which olive oil lipids and their derivatives act as multimodal regulators of bioelectrical signaling. By modulating cell membrane dynamics, these functional molecules help maintain cellular communication and homeostasis. This integrative view not only strengthens our understanding of olive oil’s health-promoting effects but also opens new avenues for targeting ion-regulatory mechanisms in metabolic, cardiovascular, and neurological diseases. Full article

Protein purification is often performed for various applications. However, enzyme purification processes typically involve multiple steps that reduce yield and increase production costs. To overcome these challenges, we developed a novel three-step process to purify a lipase from whole sardine viscera (WSV), leveraging protein properties and the structural affinity of lipases for dye ligands. A crude extract of the viscera (CEV) was obtained by grinding the whole viscera in 50 mM phosphate buffer (pH 7.0, Solution B) followed by centrifugation (6000× g; 30 min, 0 °C). Lipolytic activity (3.3 U/mg) was recorded only in the supernatant. The purification process began with ammonium sulfate fractionation (30–50% saturation), increasing lipolytic activity in the precipitate (PF30-50) to 32.9 U/mg. PF30-50 was then ultrafiltered using a 30 KDa MWCO membrane, where 5% of semi-purified lipases (SPLSV) was retained with an activity of 156.5 U/mg (UF30). Finally, the SPLSV was injected into a column packed with dye ligand affinity adsorbent, pre-equilibrated with 1.0 M ammonium sulfate in buffer A. The WSV lipase was eluted using a step gradient to progressively reduce salt concentration. SDS-PAGE analysis revealed a single band of purified lipase from sardine viscera (PLSV) corresponding to a molecular weight of 123.4 kDa, with a specific activity of 266.4 U/mg. The combination of ammonium sulfate precipitation, ultrafiltration, and dye-ligand affinity chromatography provides a scalable and reproducible approach with potential industrial relevance, particularly in biocatalysis and waste valorization contexts. Full article

The mechanisms underlying the effects of dapagliflozin in heart failure with reduced ejection fraction (HFrEF) are not yet fully understood. This study aims to evaluate the effect of the drug on cardiorespiratory function by assessing alveolar–capillary membrane characteristics, sleep apnea, pulmonary and cardiac performance in stable HFrEF patients. Seventy-three patients with stable HFrEF were enrolled, with 66 completing the six-month follow-up. Analyses included assessment of the alveolar–capillary membrane by diffusion capacity, including its membrane diffusion and capillary volume components and measurements of proSP-B in the blood, an emerging biomarker of alveolar–capillary membrane function. Pulmonary function tests, overnight respiratory monitoring, and echocardiographic parameter collection were also conducted. After 6 months, a reduction in circulating proSP-B levels was observed (32.65 ± 13.36 at baseline vs. 30.86 ± 12.45 AU at 6 months, p for trend 0.0092), accompanied by improvements in echocardiographic parameters (left ventricle ejection fraction and pulmonary pressures). Pulmonary function tests and overnight respiratory monitoring showed no significant changes in lung diffusion, spirometry, or obstructive sleep apnea (apnea hypopnea index from 5.0 [1.1–16.6] at baseline to 6.2 [0.7–13.8]/h; p = n.s.). A significant reduction in central sleep apnea (CSA) was noted in the 13 patients with at least one CSA at baseline (15 [3–48] vs. 0 [0–18.5]/h, p = 0.017). Dapagliflozin demonstrates both hemodynamic and non-hemodynamic effects, particularly improving alveolar–capillary membrane function. This study highlights the multifactorial benefits of dapagliflozin in patients with stable HFrEF and the potential of proSP-B as a sensitive marker for evaluating therapeutic response. Full article