Mechanistic Insights Research Articles

Modular Synthesis of Polycarbonyl Compounds via Regioselective Hydration of Oxo-Alkynes.

A novel protocol has been developed for the modular synthesis of polycarbonyl compounds by catalytic hydration of 1,3-diketone-tethered alkynes. The hydration process exhibits good regioselectivity and high yields at room temperature, avoiding the use of strong acids and noble metals and the requirement for elevated temperatures. Mechanistic insights suggest that the hydration proceeds through a concerted process of alkyne protonation and remote carbonyl participation. This approach provides direct access to tandem heterocycle dyads and triads.

Electrochemical oxidation-driven formation of nickel/nickel-based compounds on hollow carbon shells: Mechanistic insights and energy storage applications

Hydrangea-like nickel/nickel-based compounds decorated hollow carbon shells were synthesized through low-temperature calcination and a facile electrochemical oxidation process. This three-dimensional hollow hierarchical structure ensures intimate contact between the electrically conductive nickel (Ni) substrate and uniformly distributed electrochemically active nickel-based compounds. This hierarchical structure offers abundant active sites and accessible pathways, maximizing energy storage, particularly during rapid charge–discharge cycles. With 30 min of electrochemical oxidation, the optimized Ni-compound-based electrode exhibits a specific capacity of 643 C g−1 at 1 A/g. When assembled into a nickel-zinc battery cell with a zinc foil anode, the cell demonstrates swift current responses, with full capacity recovery even after a twentyfold increase in current density, followed by a return to 1 A/g. Density functional theory computations reveal that the electrochemical oxidation, conducted for an optimized duration, results in partial oxidation of Ni(OH)2, reducing the surface adsorption energy of OH− from the electrolyte and improving charge storage capacity.

Protective effect of 6'-Sialyllactose on LPS-induced macrophage inflammation via regulating Nrf2-mediated oxidative stress and inflammatory signaling pathways.

Macrophages play a central role in cardiovascular diseases, like atherosclerosis, by accumulating in vessel walls and inducing sustained local inflammation marked by the release of chemokines, cytokines, and matrix-degrading enzymes. Recent studies indicate that 6'-sialyllactose (6'-SL) may mitigate inflammation by modulating the immune system. Here, we examined the impact of 6'-SL on lipopolysaccharide (LPS)-induced acute inflammation using RAW 264.7 cells and a mouse model. In vivo, ICR mice received pretreatment with 100 mg/kg 6'-SL for 2 h, followed by intraperitoneal LPS injection (10 mg/kg) for 6 h. In vitro, RAW 264.7 cells were preincubated with 6'-SL before LPS stimulation. Mechanistic insights were gained though Western blotting, qRT-PCR, and immunofluorescence analysis, while reactive oxygen species (ROS) production was assessed via DHE assay. 6'-SL effectively attenuated LPS-induced p38 MAPK and Akt phosphorylation, as well as p65 nuclear translocation. Additionally, 6'-SL inhibited LPS-induced expression of tissue damage marker MMP9, IL-1β, and MCP-1 by modulating NF-κB activation. It also reduced ROS levels, mediated by p38 MAPK and Akt pathways. Moreover, 6'-SL restored LPS-suppressed Nrf2 and HO-1 akin to specific inhibitors SB203580 and LY294002. Consistent with in vitro results, 6'-SL decreased oxidative stress, MMP9, and MCP-1 expression in mouse endothelium following LPS-induced macrophage activation. In summary, our findings suggest that 6'-SL holds promise in mitigating atherosclerosis by dampening LPS-induced acute macrophage inflammation.

Stepwise increasing TMC concentration in NF membranes fabrication for controlled PA layer thickness, compactness, and charge distribution: Maintaining high permeance, enhancing desalination, and mechanistic insights

Piperazine (PIP)-based polyamide (PA) nanofiltration (NF) membranes exhibit limited rejection of divalent cations due to their negatively charged surfaces and insufficient compactness, restricting their broader application in various water treatment processes. To overcome this challenge, a novel strategy—stepwise increasing TMC concentration (SITC) during the interfacial polymerization (IP)—was proposed to optimize the PA layer structure. Conventional PA layer demonstrates a vertical diffusion structure: a compact barrier region underneath a thick and loose region. The regulatory goal of SITC is to enhance compactness while minimizing the increase in hydraulic resistance. The regulatory mechanism suggested SITC could regulate the IP reaction in both over time (throughout the IP process) and across the reaction region. During the initial IP stage, SITC reduced the reaction rate and increased PIP/TMC ratio in the bottom reaction zone, thereby improving the compactness and potential of final bottom barrier region. Thus, SITC can effectively restrict the permeation of cations, significantly increasing MgCl2 rejection from 33.70 % to 92.12 %. In the subsequent diffusion-limited stage, SITC enhanced the restriction on PIP diffusion, decreasing the thickness of the final upper loose region and surface potential. Those advantageous characteristics allowed the membrane to maintain a high rejection for Na2SO4 (>98.44 %). Concurrently, the membrane maintained a high water permeance (13.00 L·m−2·h−1·bar−1), attributed to the thin (16.69 nm) and optimized compactness distribution of its PA layer. This study provided a novel mechanism insight into the functional customization of NF membranes by decoupling the synthesis-property-performance relationship.

Decoupling of sulfation reactions to enhance zinc extraction from electric arc furnace dust: Unveiling the significance of O2 activation and mechanistic insights

Electric arc furnace dust (EAFD) is a hazardous solid waste generated during steelmaking, boasting significant concentrations of zinc and iron. Currently, a large amount of EAFD is disposed of in landfills, causing severe environmental pollution and resource wastage. To address this issue, the sulfation roasting followed by water leaching process was proposed, offering efficient zinc extraction, iron separation, low cost, and environmental friendliness advantages. This study presented a novel experimental design that decoupled the sulfation process for EAFD into gas–solid and solid–solid processes to uncover the sulfation mechanisms. During these processes, Zn-bearing phases were successfully converted into zinc sulfate (ZnSO4), while Fe-bearing phases transformed into iron oxide (Fe2O3). The resulting compounds were then separated using a water leaching process, achieving a high zinc extraction rate of 97.21% and a low iron residue of 0.61%, respectively, for the gas–solid sulfation process, and 98.76% and 0.81% for the solid–solid sulfation process. Comprehensive characterizations, in situ diffusion Fourier transform infrared spectroscopy (DRIFTS) experiments, and density functional theory (DFT) calculations provide a deeper understanding of the sulfation restructuring mechanisms in gas–solid and solid–solid roasting processes, highlighting the significant role of O2 activation. In the solid–solid sulfation process, Fe2(SO4)3, derived from the oxidation of FeSO4 by O2, was identified as the predominant compound responsible for sulfation. In the gas–solid sulfation process, sulfite species (SO32-) played a critical role as intermediates, eventually transforming into sulfate species (SO42-) with the assistance of O2. Moreover, adsorbed SO2 underwent oxidation and activation by adsorbed O2, forming the sulfate functional group. Migration of this functional group to the adjacent Zn atom through a transition state, established a Zn-SO4 bond, disrupting the Zn-O bond on ZnFe2O4 and completing the sulfation of ZnFe2O4. This study provides a fresh perspective on the application of sulfation roasting for the extraction of valuable metals, contributing to resource sustainability and effective management of zinc-bearing resources.

Control of Staphylococcus aureus infection by biosurfactant derived from Bacillus rugosus HH2: Strain isolation, structural characterization, and mechanistic insights

Novel antimicrobials are urgently needed to combat methicillin-resistant Staphylococcus aureus (MRSA) infections. This study explores the potential of biosurfactants derived from Bacillus rugosus HH2 as a novel antibacterial agent against MRSA. The biosurfactant, identified as surfactin, demonstrated surface-active properties, reducing surface tension to 37.63 mN/m and lowering contact angles in a concentration-dependent manner. It remained stable across a wide range of pH (4−10), temperatures (30–80 °C), and salinity levels (3–18 %). The biosurfactant inhibited the growth of both methicillin-sensitive S. aureus and MRSA, with minimum inhibitory concentrations ranging from 128 to 256 μg/mL. Additionally, it showed anti-biofilm activity, preventing biofilm formation and dispersing established biofilms. Field-emission scanning electron microscopy revealed that the biosurfactant disrupted bacterial cell membranes, leading to leakage. Furthermore, it reduced the production of virulence factors in S. aureus, including hemolysin and lipase. Transcriptomic analysis indicated downregulation of genes associated with quorum sensing and cell adhesion in MRSA. Molecular docking studies showed strong interactions between surfactin and key MRSA proteins, underscoring its potential to overcome antibiotic resistance. Biocompatibility was confirmed through in vitro cytotoxicity and in vivo phytotoxicity tests. In summary, this study presents surfactin as a promising novel antibacterial agent against MRSA, providing insights into its mechanisms of action.

Deficiency of PvDRAM2 increased the nitrite sensitivity of Pacific white shrimp (Penaeus vannamei) by inhibiting autophagy

Autophagy is an essential response mechanism to environmental stress during the evolution of organisms. DRAM2 (Damage-regulated autophagy regulator 2) is recognized as necessary for the process of p53-mediated cell apoptosis. Although the role of DRAM2 in apoptosis has been confirmed, the mechanism of its relationship with autophagy is still unclear. Here we describe PvDRAM2 features and functions. We found that nitrite stress induced autophagy accumulation and ROS production. A novel DRAM-homologous protein, DRAM2, was cloned, and its expression is significantly up-regulated under nitrite stress conditions. PvDRAM2 primarily localizes within the cytoplasmic lysosome.Loss of PvDRAM2 increased sensitivity response to nitrite stress of Pacific white shrimp. And silenced of PvDRAM2 promoted ROS production and inhibited autophagy accumulation. In addition, silenced of PvDRAM2 decreased the autophagy-related protein of p62, Beclin 1, and LC3 expression under nitrite stress of Pacific white shrimp. Collectively, these studies uncover a novel critical role for PvDRAM2 in regulating autophagy under nitrite stress. Specifically, PvDRAM2 is essential for the induction of autophagy, enabling Pacific white shrimp to adapt to environmental stress. This provides mechanistic insight into how autophagy functions as a way for Pacific white shrimp to cope with environmental challenges.

Enhancing electrocatalytic oxidation via spatial confinement in Ti4O7 Nanopore Membranes: Synthesis, Characterization, and Mechanistic insights

Enhancing electrocatalytic oxidation via spatial confinement in Ti4O7 Nanopore Membranes: Synthesis, Characterization, and Mechanistic insights

MXene/TiO2 NAs on CC flexible electrode for electrocatalytic tigecycline degradation: Performance, Mechanistic insights, and toxicological assessment

In this study, a composite electrode (MXene/TiO2 NAs@CC) based on TiO2 nanotube arrays (NAs) and two-dimensional MXene-coated carbon cloth (CC) was prepared. The electrode greatly improved the catalytic activity by capturing protons through the abundant active sites of TiO2 nanotube arrays and promoting proton reduction with the high conductivity of MXene. Moreover, the composite electrode possessed the largest specific surface area (9.1660 m2·g−1) according to the results of the BET test. Under optimal conditions, the composite electrode degraded tigecycline (TGC) up to 86.66 % in 20 min. Furthermore, the electrocatalytic performance was not affected after 10 cycling experiments and 60 days of exposure to air, indicating its excellent stability. In addition, the cathodic co-catalytic reaction mechanism involving anode ·OH, HClO, and cathode H* was proposed. The degradation pathways of TGC and the toxicity of possible intermediates were also analyzed. Finally, a two-stage continuous wastewater reactor with high efficiency and low consumption (90.46 % removal rate, 0.061 kWh/m3) was designed. This study provides a new idea for designing an electrocatalytic reactor with high efficiency, stability, and low consumption, enriching the field of electrocatalytic treatment strategies for antibiotic wastewater. It is of great significance for the protection of the water environment and human health.

Synergistic Action of Rutin-Coated Zinc Oxide Nanoparticles: Targeting Biofilm Formation Receptors of Dental Pathogens and Modulating Apoptosis Genes for Enhanced Oral Anticancer Activity.

Oral diseases are often associated with bacterial and fungal pathogens such as Staphylococcus aureus, Streptococcus mutans, Enterococcus faecalis, and Candida albicans. This research explored a novel approach to addressing these pathogens by synthesizing zinc oxide nanoparticles (ZnO NPs) coated with rutin (RT), a plant-derived compound. The synthesized ZnO-RT NPs were comprehensively characterized using UV-Vis spectrophotometer, SEM, and EDAX techniques to confirm their structural composition. The antioxidant potential was assessed through free radical scavenging assays. Additionally, the antimicrobial activity of ZnO-RT NPs was evaluated using a zone of inhibition assay against oral pathogens. Molecular docking studies with the Autodock tool were performed to elucidate the interactions between RT and the receptors of oral pathogens. The findings demonstrated that ZnO-RT NPs exhibited robust free radical scavenging activity. Furthermore, they showed significant antimicrobial activity with a minimal inhibitory concentration of 40 μg/mL against oral pathogens. ZnO-RT NPs also displayed dose-dependent anticancer effects on human oral cancer cells at concentrations of 10, 20, 40, and 80 μg/mL. Mechanistic insights into the anticancer activity on KB cells revealed the upregulation of apoptotic genes. This study underscores the promising potential of ZnO-RT NPs for dental applications due to their strong antioxidant, anticancer, and antimicrobial properties. These nanoparticles offer a hopeful prospect for addressing oral pathogen challenges and enhancing overall oral health.

Mechanistic insights into melanoidins-induced hydrophilicity of thermal hydrolyzed sludge and its impact on dewaterability

Although thermal hydrolysis pretreatment enhances disposal efficiency of sludge, it inevitably leads to melanoidins formation, which will negatively impact the subsequent wastewater treatment processes. However, their effect on the dewaterability of thermal hydrolyzed sludge (THS) remains poorly understood. This study aimed to uncover the underlying mechanisms of how melanoidins affecting dewaterability of THS. Using resin-adsorption method to reduce melanoidins content by 50% led to 21% and 6% decreases in capillary suction time and specific resistance to filtration, respectively, and a 14% increase in sludge cake solid content. Conversely, accumulating melanoidins to 200% worsened THS dewaterability, altering surface morphology and reducing floc stability, which increased the content of bound water by 9%. Additionally, a higher melanoidins level increased the hydrophilic components in extracellular polymeric substances while reducing hydrophobic sites and structures. These findings indicate that melanoidins impair THS dewaterability by altering flocs spatial properties and increasing hydrophilic structures and components.

Advancements in Contemporary Pharmacological Innovation: Mechanistic Insights and Emerging Trends in Drug Discovery and Development

Developing a new drug and bringing it to the market is a complex and time-consuming process that involves multiple phases of drug discovery and development. However, recent advancements in various technologies, such as multi-omics, genome editing, Artificial Intelligence (AI), and Machine Learning (ML), have significantly improved this process. These technologies have made the process more accurate, less time-consuming, and cost-effective compared to the conventional methods of drug discovery and development. In the current age, discovering and developing drugs is a collaborative effort that involves scientific breakthroughs, technological advancements, and regulatory oversight. The pharmaceutical industry is constantly innovating new techniques, fostering interdisciplinary collaboration, and prioritizing patient-centered approaches. In this review, we explore the latest and most updated information about using advanced technologies in drug discovery. The review begins by briefly explaining the conventional drug discovery and development process, and then delves into the applications of multi-omics, genome editing technology, systems biology, artificial intelligence, and machine learning.

Type III interferons induce pyroptosis in gut epithelial cells and impair mucosal repair.

Tissue damage and repair are hallmarks of inflammation. Despite a wealth of information on the mechanisms that govern tissue damage, mechanistic insight into how inflammation affects repair is lacking. Here, we investigated how interferons influence tissue repair after damage to the intestinal mucosa. We found that type III, not type I or type II, interferons delay epithelial cell regeneration by inducing the upregulation of Z-DNA-binding protein 1 (ZBP1). Z-nucleic acids formed following intestinal damage are sensed by ZBP1, leading to caspase-8 activation and the cleavage of gasdermin C (GSDMC). Cleaved GSDMC drives epithelial cell death by pyroptosis and delays repair of the large or small intestine after colitis or irradiation, respectively. The type III interferon/ZBP1/caspase-8/GSDMC axis is also active in patients with inflammatory bowel disease (IBD). Our findings highlight the capacity of type III interferons to delay gut repair, which has implications for IBD patients or individuals exposed to radiation therapies.

Assessment of Cerium Adsorption Potential of Phosphoric acid Activated Biochar in Aqueous System: Modelling and Mechanistic Insights

Cerium pollution in waterbodies by improper industrial waste disposal is a major concern due to its detrimental impacts on the environment. Therefore, treatment of cerium-contaminated water is inevitable. Hence, this study is focused on the remediation of cerium pollution using phosphoric acid-activated biochar (PPMB) as an adsorbent, synthesized upon pyrolytic activation of palmyra palm male flower-based pristine biochar (PMFB) with H3PO4 at 500°C. The physico-chemical surface properties of PMFB and PPMB were evaluated through various microscopic and spectroscopic analyses. The key parameters such as biochar dosage, pH, temperature, contact time and initial cerium concentration were optimized as 0.5 g/L, 5.0, 303 K, 180 min and 50 mg/L respectively via batch adsorption. Pseudo-second order kinetic and Toth isotherm are the best-fitted models. The thermodynamic parameters including ΔG◦ (-30.4707±0.7618 kJ/mol at 303 K), ΔH◦ (16.1499±0.78 kJ/mol), and ΔS◦ (153.617±3.8404 J/mol/K) conveying that cerium adsorption onto PPMB was spontaneous, endothermic, and highly disordered at PPMB-bulk adsorption medium interface. Precipitation, electrostatic attraction, and surface complexation are predicted to be the predominant mechanisms for the chosen PPMB-cerium adsorption system. Moreover, cerium phytotoxicity on Vigna radiata explains the real-time applicability and feasibility of cerium adsorption using PPMB. Thus, the key findings of this study specified that the higher adsorption capacity of PPMB (141.3484±6.9856 mg/g) contributed by the incorporated phosphate groups, predominant mesoporosity, SSABET of 230.559 m2/g and anionic surface at a wider pH range (pH>3.08) make PPMB as efficient, economically feasible and environmentally friendly adsorbent for cerium adsorption in aqueous system.

Organometallic Ru(III) Catalysts for α-Alkylation of Carbonyl Compounds using Alcohols: Mechanistic Insights via Detection of Key Intermediates.

Three novel cyclometalated ruthenium complexes ([Ru.L(9)] [Ru.L(10)] and [Ru.L(11)]) featuring azo functionalities were synthesized and characterized using a variety of spectroscopic techniques, namely FT-IR, electronic absorption spectroscopy, and ESI-MS. Representative solid-state structures of the acquired complexes were determined through X-ray crystallography. These complexes were evidenced to be efficient catalysts for the synthesis of various α-alkylated compounds utilizing simple acetophenone derivatives with easily affordable and economically viable alcohols, which were isolated and characterized via 1H and 13C NMR spectroscopy. The optimum reaction conditions were found by employing toluene as solvent, potassium tert-butoxide as base at 115 oC temperature utilizing 0.8 mol% of catalyst [Ru.L(10)]. The yield of the desired compounds found to be in the range of 83 - 97%. Additionally, mass spectrometry provided insights into the in-situ generated ruthenium hydride and ruthenium alkoxy intermediates, shedding light on the catalytic mechanism.

Sodium butyrate activates the extrinsic and intrinsic apoptotic processes in murine cementoblasts

Background/purpose: The metabolic by-product butyric acid of Gram-negative anaerobic bacteria can invoke pathological effects on periodontal cells resulting in inflammation and further destruction of periodontium. However, limited researches on the effects of butyric acid on cementoblasts were reported. Therefore, this study aimed to investigate the type of cell death in murine cementoblast (OCCM.30) caused by adding the different concentrations of sodium butyrate to the cell culture. Materials and methodsOCCM.30 cells were exposed to sodium butyrate (0, 2, 4, 8, 16 mM) for 48 h. Cell viability was determined by microculture tetrazolium assay. Cell cycle distribution and cell death were analyzed by flow cytometry. Caspase-mediated apoptotic cascade was evaluated by Western blot. ResultsThe concentrations of sodium butyrate≧4 mM were found to inhibit cell viability of OCCM.30 cells in a dose-dependent manner (P < 0.05). Sodium butyrate elevated sub-G1 cell population which exhibited cell apoptosis in OCCM.30 cells (P < 0.05). In addition, early and later apoptotic cells were found in sodium butyrate-induced cell death. Sodium butyrate significantly stimulated the degradation of procaspases-3, -8, and -9 levels, respectively (P < 0.05). Simultaneously, sodium butyrate corresponded to augment the levels of cleaved forms of caspases-3, -8, and -9, respectively (P < 0.05). ConclusionTaken together, sodium butyrate is a cytotoxic agent and can induce apoptosis on cementoblasts. The pathway involved in apoptosis is activated by caspase family signaling pathways. These evidences may provide a new mechanistic insight into the mechanism of damage of cementoblasts during the development and progression of periodontitis.

New mechanistic insights into soil ecological risk assessment of arsenite (III) and arsenate (V)：Cellular and molecular toxicity responses in Eisenia fetida

Inorganic arsenic (iAs) is a persistent bioaccumulation carcinogen that is most abundant in soils in the form of arsenite-As (III) and arsenate-As (V). However, there is currently very little explicit evidence about cytotoxicity of As on soil organisms. Moreover, toxicological data for iAs and proteotoxicity is shortage. The purpose of the present work is to elucidate the cytotoxicity mechanism of As (III) and As (V) to earthworms, a soil ecological sentinel species, and the molecular mechanisms by which As (III)/As (V) directly bind to antioxidative enzyme Cu/Zn-superoxide dismutase (Cu/Zn-SOD). Results indicate that iAs triggered cell membrane injury and genotoxicity. As (V) (56.15%) induced lower cell viability than As (III) (61.88%). Higher ROS and lipid peroxidation level in As (V) support greater cytotoxicity. Differences in cellular uptake due to valence induced diverse levels of oxidative stress and cytotoxicity. At the molecular level, As (III) (129.33%) induced higher Cu/Zn-SOD activity than As (V) (110.75%). Changes in backbone, secondary structure, amino acid microenvironment and particle size of Cu/Zn-SOD further revealed the mechanisms of differential molecular toxicity of As (III) and As (V). Binding reactions with Cu/Zn-SOD explain differences in molecular toxicity. Collective research showed that iAs-induced oxidative stress and binding reactions determine the difference of SOD activity between As (III) and As (V) at the cellular level. This work offers new insights into the health risk assessment of As.

Multifunctional carbonyl-rich compounds for constructing “corrosion-resistant electrodes and electrolyte interface” in high-voltage sodium-metal batteries

In recent years, there has been notable advancement in the development of high-performance materials for sodium-ion batteries, particularly in layered oxide cathodes. However, research on enhancing the interface between electrolytes and cathodes remains nascent. One promising approach involves the addition of electrolyte additives, recognized for their cost-effectiveness and efficiency in bolstering the electrode/electrolyte interface to enhance material stability during high-voltage cycling. In this work, we investigate the use of glutaric anhydride (GA) as an electrolyte additive to improve the cycling stability of NaNi1/3F1/3M1/3O2/Na (NFM/Na) high-voltage sodium-metal batteries. Batteries using GA-containing electrolytes demonstrate impressive capacity retention: 80.51 % after 200 cycles at 4.0 V and 76.01 % after 150 cycles at 4.1 V. The research delves into the mechanistic insights behind GA’s effectiveness in enhancing cyclic stability. Theoretical calculations reveal that GA plays a crucial role in modifying the structure of the first solvation shells, promoting the formation of an elastic cathode-electrolyte interface (CEI) film enriched with OCO groups. This uniform and stable CEI film effectively preserves the material’s crystal structure and mitigates the dissolution of transition metals, thereby prolonging battery lifespan under demanding extremely cycling conditions.

Trifolin Attenuates Hypertension-Mediated Cardiac Injury by Inhibiting Cardiomyocyte Apoptosis: Mechanistic Insights and Therapeutic Potential

BackgroundHypertension-induced cardiac disease is a common complication and a significant contributor to mortality in hypertensive patients, largely due to cardiomyocyte apoptosis. Although Trifolin has been identified as a potential antihypertensive compound, its therapeutic role in hypertension-induced cardiac injury remains uncertain. PurposeThis study aims to evaluate the protective effects of Trifolin and explore the underlying mechanisms of its action against hypertension-induced cardiac injury. MethodsIn vivo, mice were infused with Angiotensin II (AngII, 500 ng/kg/min) or saline via osmotic pumps and treated with Trifolin (0.1, 1.0, or 10.0 mg/kg/day) or Valsartan (10 mg/kg/day) for four weeks. In vitro, H9C2 cells were stimulated with AngII (1 μM) and treated with Trifolin (25, 50, or 100 μM). Various assays, including echocardiography, hematoxylin and eosin staining, TUNEL assay, Annexin-V/propidium iodide staining, and JC-1 staining, were used to assess Trifolin's therapeutic effects on hypertension-related cardiac injury and cardiomyocyte apoptosis. Potential pharmacological mechanisms were analyzed through network pharmacology and confirmed via Western blotting. ResultsTrifolin treatment improved cardiac function by increasing left ventricular ejection fraction and fractional shortening while reducing tissue disorganization in AngII-treated mice. It also reduced cardiomyocyte apoptosis, reversing the upregulation of Bax and cleaved caspase-3 and the downregulation of Bcl-2. Network pharmacology identified 314 common targets of Trifolin linked to hypertensive heart disease, with involvement in apoptosis, MAPK, PI3K/AKT, and HIF-1 signaling pathways. Trifolin treatment increased p-PI3K/PI3K and p-AKT/AKT ratios while decreasing p-ERK/ERK, p-p38 MAPK/p38 MAPK, and p-JNK/JNK ratios in both mouse and cell models. ConclusionTrifolin alleviates AngII-induced cardiac injury and cardiomyocyte apoptosis, potentially through the regulation of MAPK, PI3K/AKT, and HIF-1 signaling pathways.

Mechanistic insights into antihypertensive activity of mushroom-derived protein-peptides via metabolomic and proteomic approaches

Stropharia rugosoannulata mushroom peptides have well-defined sequences, specific structures, and excellent antihypertensive activity. In this study, metabolomic and proteomic analyses revealed the antihypertensive mechanism of mushroom-derived protein-peptides, GQEDYDRLRPL (GL-11P) and KSWDDFFTR (KR-9P), to demonstrate their potential antihypertensive buck-regulation systems and pathways. Protein-peptides were predominantly down-regulated expression of endogenous metabolites in antihypertensive regulation, and bile acid analogs were key endogenous differentially expressed metabolic markers. Antihypertensive treatment with protein-peptides activated the immune and signal transduction systems, affected extracellular regions, metal ion binding, and receptor binding ability. The ECM-receptor interaction pathway, with differentially expressed proteins in antihypertensive treatment, distinguished the protein-peptides from blank control and was also a critical pathway that distinguished the two protein-peptides. Up-regulatory pathways of protein expression dominated by GL-11P's antihypertensive regulation, while down-regulatory pathways of protein expression dominated by KR-9P's antihypertensive regulation. Protein-peptides exerted antihypertensive regulatory effects by enhancing immune responses, up-regulating protein and enzyme binding capacity, and down-regulating inflammatory mediator release. Differentially expressed protein interactions produced in protein-peptides antihypertensive therapy mostly up-regulated the Reactome pathway and were dominated by immune system regulation. This study provided data support for utilizing mushroom-derived protein-peptides in antihypertensive therapy, and enriched the theoretical basis of protein-peptides' antihypertensive regulation mechanisms.