Quercetin Antagonizes Doxorubicin-Induced Cardiotoxicity via HO-1/PGC-1α-ALOX5 Axis: Arachidonic Acid Metabolism-Ferroptosis Crosstalk as a Therapeutic Target.

Gastrodia elata Blume (GEB) is widely used to treat cardio-cerebrovascular disease in China and in traditional Chinese medicine it is considered to be a dispelling wind and dredging collateral. However, the mechanism and active components of the plant in treating ischemic stroke (IS) remain unclear. The present study aimed to identify the active components and mechanism of GEB in treating IS using network pharmacology and molecular docking technology. Network analysis predicted 752 potential targets from 14 compounds in GEB, sharing 32 key targets with IS-associated targets. Gene Ontology analysis of key targets showed that 'oxidative stress', 'immune response' and 'regulation of blood circulation' were significantly enriched. Kyoto Encyclopedia of Genes and Genomes pathway analysis indicated that the key targets regulated 11 representative pathways including 'arachidonic acid metabolism', 'lipid and galactose metabolism'. In the protein-protein interaction network, five core targets, including toll-like receptor agonist, STAT3, myeloperoxidase (MPO), prostaglandin-endoperoxide synthase and matrix metalloproteinase (MMP)9, were identified and successfully docked with four active components: Palmitic acid, alexandrin, para-hydroxybenzaldehyde and gastrodin. Alexandrin, para-hydroxybenzaldehyde, and gastrodin are closely related to brain ischemia/reperfusion damage and repair. Therefore, to further verify the mechanism of action of three active components in the second part, we established the HT22 oxygen-glucose deprivation-reperfusion (OGD/R) model. Cell Counting Kit-8 assay and western blot analysis demonstrated that these three active components of GEB regulated core targets of molecular docking, such as STAT3, MPO and MMP9. In vitro experiments showed that OGD/R decreased cell survival, while this effect was reversed by the three active components of GEB. In addition, western blot analysis indicated that alexandrin upregulated expression of phosphorylated-STAT3, para-hydroxybenzaldehyde downregulated MPO and gastrodin downregulated MMP9. Therefore, the present study showed that GEB may prevent and treat IS via interaction between the active components and the main targets, which is key for investigating the efficacy of traditional Chinese medicine.

Although Siraitia grosvenorii (abbreviated as S.g.) is frequently used to prevent and cure diabetes problems, the precise mechanism underlying its ability to do so remains unknown. Through network pharmacology and molecular docking techniques, we studied the early molecular mechanisms of S.g in the treating of proliferative diabetic retinopathy (PDR) in this study. The Traditional Chinese Medicine Systems Pharmacology (TCMSP) database was used to screen the active compounds and related targets of S.g. Oral bioavailability (OB) 30% and drug likeness (DL) 0.18 were used as screening criteria. The active compounds without knowledge of a probable target were excluded. The Uniprot database included converted symbols for the associated targets. GEO2R was used to explore several genes related to PDR. Using jvenn web service to intersect targets of S.g and PDR. The Xiantao Academic Online website was used to examine the expression patterns of intersect targets in PDR samples. The STRING database was used to create a protein-protein interaction (PPI) network of intersecting targets. Cytoscape software was used to show the PPI network, MCODE software was used to evaluate the network’s core proteins, and CytoHubba software was used to extract the important networks of the top three targets. Omicshare platform carried a functional analysis using the Gene Ontology (GO) and pathway enrichment analysis using the Kyoto Encyclopedia of Genes and Genomes (KEGG). Pymol, AutoDock Vina software, Schrödinger Software were used to conduct molecular docking experiments or pockets search on the top three targets. The results showed that 85 targets were matched to six active compounds of S.g. 18 intersect targets were found. Seven DEGs were up-regulated and eleven genes were down-regulated when these targets were divided into two groups. TNF, PTGS2, and CASP3 were the main targets, according to the PPI network. The intersect targets were mostly related to angiogenesis, cell proliferation, oxidative stress, inflammatory response, and metabolism. It was discovered that the core targets TNF, PTGS2, and CASP3 had various levels of affinity for their respective compounds. Interestingly, multiple good drug-forming pockets for CASP3 and PTGS2 targets were identified through Schrödinger software. In particular, six compounds bind to the top three core targets to inhibit IL-17 signaling pathway, AGE-RAGE signaling pathway in diabetic complications, Pathways in cancer and 14 other signaling pathways to inhibit inflammation, apoptosis, oxidative stress, arachidonic acid metabolism, and angiogenesis to prevent and treat PDR. The study’s findings, which served as a guide for the widespread use of S.g in PDR clinical practise, included multi-substances and targets of S.g to prevent and cure PDR.

Myocardial infarction (MI) triggers oxidative stress, mitochondrial dysfunction, and cardiomyocyte apoptosis. At present, it remains urgently needed to develop novel therapies specifically suppressing cardiomyocyte apoptosis via improving mitochondrial dysfunction following MI. Jiedu Huoxue Decoction (JDHXD) may have the effect of ameliorating myocardial injury after MI. This study examined the protection exerted by JDHXD against myocardial injury post-MI and investigated the underlying mechanisms of action. UHPLC/Orbitrap-MS, network pharmacology and transcriptome analysis were used to study the effective components and potential targets of JDHXD for treating MI. In vivo: The MI mice received JDHXD (12.74/25.48g/kg/day) or captopril treatment for 28days. Later, cardiac function (tested by echocardiography and histopathology), apoptosis, oxidative stress, mitochondrial ultrastructure, mitochondrial fission/fusion and PTEN/AKT/GSK3β pathway protein levels were evaluated. In vitro: TBHP-induced cardiomyocytes (H9C2 cells and NCMs) were exposed to JDHXD treatment (50-200μg/mL), with or without PTEN inhibitor Bpv (2μM) or PTEN overexpression (through adenoviral transduction). Afterwards, cell apoptosis, oxidative stress, mitochondrial membrane potential, and relative proteins were assayed. Experiments such as molecular docking and surface plasmon resonance imaging (SPRi) were conducted to verify the effective components of JDHXD in preventing myocardial mitochondrial injury after MI. The results of network pharmacology and transcriptomics suggest that JDHXD may ameliorate myocardial injury after MI through modulating PTEN for activating the PI3K/AKT/GSK3β signaling pathway. In vivo: JDHXD dose-dependently improved left ventricular function, improved the oxidative stress-induced imbalance of mitochondrial fission/fusion, and inhibited cardiomyocyte apoptosis post-MI in association with suppressing the PTEN/AKT/GSK3β pathway. In vitro: JDHXD suppressed the TBHP-induced cardiomyocyte apoptosis, attenuated oxidative stress, preserved mitochondrial potential, and restored mitochondrial dynamics. PTEN inhibitor did not augment JDHXD's effects, whereas PTEN overexpression partially abolished JDHXD's protection against myocardial injury induced by oxidative stress. UHPLC/Orbitrap-MS, molecular docking, SPRi and experiments in vitro confirmed that puerarin is one of the main components of JDHXD in regulating the PTEN/AKT/GSK3β pathway to improve mitochondrial function after MI and inhibit cardiomyocyte apoptosis. JDHXD against oxidative stress-induced cardiomyocyte apoptosis post-MI through ameliorating mitochondrial dysfunction, which is partially mediated by suppressing the PTEN/AKT/GSK3β pathway to inhibit excessive mitochondrial fission and promote mitochondrial fusion.

Angiotensin-converting enzyme 1 (ACE1) is a peptide involved in fluid and blood pressure management. It regulates blood pressure by converting angiotensin I to angiotensin II, which has vasoconstrictive effects. Previous studies have shown that certain compounds of natural origin can inhibit the activity of angiotensin-converting enzymes and exert blood pressure-regulating effects. Surface Plasmon Resonance (SPR) biosensor technology is the industry standard method for observing biomolecule interactions. In our study, we used molecular simulation methods to investigate the docking energies of various herbal metabolites with ACE1 proteins, tested the real-time binding affinities between various herbal metabolites and sACE1 by SPR, and analyzed the relationship between real-time binding affinity and docking energy. In addition, to further explore the connection between inhibitor activity and real-time binding affinity, several herbal metabolites' in vitro inhibitory activities were tested using an ACE1 activity test kit. The molecular docking simulation technique's results and the real-time affinity tested by the SPR technique were found to be negatively correlated, and the virtual docking technique still has some drawbacks as a tool for forecasting proteins' affinities to the metabolites of Chinese herbal metabolites. There may be a positive correlation between the enzyme inhibitory activity and the real-time affinity detected by the SPR technique, and the results from the SPR technique may provide convincing evidence to prove the interaction between herbal metabolites and ACE1 target proteins.

Perillaldehyde ameliorates sepsis-associated acute kidney injury via inhibiting HSP90AA1-mediated ferroptosis and pyroptosis: Molecular structure and protein interaction of HSP90AA1.

Study on the mechanism of Trichosanthes kirilowii Maxim. against COPD based on serum chemical composition analysis, network pharmacology, and experimental study.

Machine learning prediction of myocardial ischaemia‒reperfusion injury: Clinical features and Qishen Yiqi dripping pills mechanism.

Integrating serum pharmacochemistry, network pharmacology, and metabolomics to explore the protective mechanism of Hua-Feng-Dan in ischemic stroke.

Background: Hydroxysafflor Yellow A (HSYA), the major bioactive component from Carthamus tinctorius L., exerts significant protective effects against myocardial ischemia-reperfusion injury (MIRI). Mitophagy is pivotal in the pathological process of MIRI, yet the specific molecular mechanism underlying HSYA-mediated mitophagy regulation remains unclear. Objective: This study aimed to investigate the association between HSYA treatment and mitochondrial autophagy in murine MIRI and to explore the potential mechanistic role of the SIRT1-FOXO3-BNIP3 signaling pathway using functional loss-of-function and rescue experiments. These findings may provide preliminary evidence supporting the clinical translational potential in MIRI therapy. Methods: Mouse myocardial ischemia-reperfusion injury (MIRI) model and oxygen-glucose deprivation/reoxygenation (OGD/R)-induced AC16 cardiomyocyte injury models were established. Metabolomics, molecular docking, and surface plasmon resonance (SPR) techniques were combined to screen the potential targets of HSYA. The SIRT1 inhibitor EX527 and SIRT1 siRNA were used to verify the underlying mechanism. Cardiac function, myocardial infarct size, mitochondrial function, the expression of autophagy-related proteins, and protein–protein interaction were detected and analyzed. Results: Compared with the MIRI group, HSYA significantly improved cardiac function in mice, as evidenced by increased left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) (p < 0.01), attenuated ST-segment elevation, and improved myocardial perfusion. HSYA also markedly reduced myocardial infarct size (p < 0.01) and serum levels of CK-MB, LDH, and cTnI (all p < 0.01) and ameliorated myocardial histopathological damage and mitochondrial ultrastructural integrity. Mechanistic studies revealed that HSYA significantly upregulated the expression of SIRT1, FOXO3, BNIP3, Beclin-1, and the LC3II/I ratio while downregulating p62 expression (p < 0.01), consistent with enhanced mitophagy-related activity. Furthermore, these protective effects were markedly attenuated upon SIRT1 inhibition or siRNA-mediated silencing, whereas HSYA intervention partially reversed these alterations. Additionally, co-immunoprecipitation (Co-IP) and pull-down assays demonstrated that HSYA promoted protein–protein interactions between SIRT1-FOXO3, FOXO3-BNIP3, and BNIP3-LC3B. Conclusions: These findings highlight that HSYA is associated with improved cardiac function, enhanced mitophagy-related activity, and upregulated SIRT1-FOXO3-BNIP3 signaling, providing robust experimental evidence for its clinical translational application in MIRI treatment.

Myocardial infarction (MI) is a severe cardiovascular disease with complex pathogenesis, in which inflammatory responses and oxidative stress play critical roles. Yixinyuan Gaofang (YXYGF) is a novel traditional Chinese medicine (TCM) formulation, and its effects and mechanisms in MI remain incompletely understood. MI was induced in rats via ligation of left anterior descending (LAD) coronary artery. Rats received intragastric administration of YXYGF at different dosage, and cardiac function was evaluated using echocardiography and hemodynamic measurements. Histological assessments included 2,3,5-triphenyltetrazolium chloride (TTC), hematoxylin-eosin (HE), and Masson staining to observe infarct size, tissue injury, and fibrosis. Serum biomarkers of myocardial injury were measured. Enzyme-linked immunosorbent assay (ELISA) and biochemical assays quantified myocardial inflammatory cytokines and oxidative stress markers. Ultra-high-performance liquid chromatography–mass spectrometry (UHPLC-MS) identified YXYGF chemical constituents, and network pharmacology analysis predicted potential signaling pathways. Tumor necrosis factor/nuclear factor-κB (TNF/NF-κB) pathway-related proteins were examined using Western blotting. YXYGF at different dosage all significantly improved left ventricular function in MI rats and markedly reduced infarct size, alleviated myocardial injury and fibrosis. In addition, YXYGF also inhibited inflammatory cytokine production and oxidative stress. UHPLC-MS identified 365 chemical components of YXYGF, of which 179 met the pharmacodynamic screening criteria. Mechanistic analysis suggested that the protective effects of YXYGF were closely associated with inflammatory response, hypoxia response, coagulation, and cardiovascular-related signaling pathways, with the TNF/NF-κB pathway identified as a key mechanism. Further mechanistic verification revealed that the NF-κB activator PMA could partially reverse the protective effects of YXYGF on cardiac function, myocardial function, histopathological damage, and oxidative inflammatory microenvironment in rats with myocardial infarction. YXYGF effectively improves cardiac dysfunction, reduces infarct size, fibrosis, and inflammatory and oxidative responses in MI rats, at least partially through inhibition of the TNF/NF-κB signaling pathway. These findings provide experimental evidence supporting the therapeutic application of YXYGF in cardiovascular disease prevention and treatment. However, these findings are based on an animal model, and further clinical studies are required to validate the therapeutic potential of YXYGF in humans.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-026-46845-8.

Hydroxysafflor yellow A mitigates lipopolysaccharide-induced acute lung injury by enhancing pulmonary microvascular endothelial barrier function via Calpain-1/HIF-1α inhibition.

Berberine (BBR) possesses varied pharmacological properties, including anti-apoptotic and potent neuroprotective effects, and can ameliorate cognitive impairments associated with diverse diseases. Despite the noted potential of BBR in mitigating cognitive deficits associated with chronic cerebral hypoperfusion (CCH), the precise mechanisms underlying its therapeutic effects remain inadequately defined. To explore these mechanisms, a CCH rat model was developed using a refined micro-spring method for bilateral common carotid artery stenosis (BCAS). For the experimental setup, rats were systematically divided into six groups: a Sham group (n = 15), a Sham + BBR group (n = 15), a BCAS group (n = 15), a BCAS + BBR group (n = 15), a BCAS + BBR + Colivelin group (with Colivelin serving as a STAT3 activator, n = 15), and a BCAS + AG490 group (AG490 acting as a JAK2 inhibitor, n = 15). Cognitive performance was evaluated through the Morris water maze and novel object recognition (NOR) tests. Additionally, neuronal integrity was assessed by Nissl and TUNEL staining within the hippocampal region. The study further examined the protein expressions of JAK2, STAT3, phosphorylated JAK2, phosphorylated STAT3, and cleaved caspase-3 using western blot analysis. Interaction targets of BBR were predicted through the STITCH database, and its binding affinity to STAT3 was confirmed using molecular docking and surface plasmon resonance (SPR) techniques. The findings indicated an increase in apoptosis and a decline in cognitive abilities among the hippocampal neurons of the BCAS model rats. These deleterious effects, however, were substantially alleviated following treatment with BBR. The study posits that BBR primarily exerts its neuroprotective effects through the inhibition of the JAK2/STAT3 pathway. Notably, while the activation of this pathway by Colivelin exacerbated neuronal damage and cognitive decline, its inhibition via AG490 markedly decreased apoptosis and improved cognitive outcomes. Therefore, this research suggests that BBR enhances cognitive functions in BCAS rats predominantly by reducing apoptosis in hippocampal neurons through the modulation of the JAK2/STAT3 pathway.

Phosphodiesterase 10A (PDE10A) has been confirmed to be an important target for the treatment of central nervous system (CNS) disorders. The purpose of the present study was to identify PDE10A inhibitors from herbs used in traditional Chinese medicine. Pharmacophore and molecular docking techniques were used to virtually screen the chemical molecule database of Sophoraflavescens, a well‑known Chinese herb that has been used for improving mental health and regulating the CNS. The pharmacophore model generated recognized the common functional groups of known PDE10A inhibitors. In addition, molecular docking was used to calculate the binding affinity of ligand‑PDE10A interactions and to investigate the possible binding pattern. Virtual screening based on the pharmacophore model and molecular docking was performed to identify potential PDE10A inhibitors from S.flavescens. The results demonstrated that nine hits from S.flavescens were potential PDE10A inhibitors, and their biological activity was further validated using literature mining. A total of two compounds were reported to inhibit cyclic adenosine monophosphate phosphodiesterase, and one protected against glutamate‑induced oxidative stress in the CNS. The remaining six compounds require further bioactivity validation. The results of the present study demonstrated that this method was a time‑ and cost‑saving strategy for the identification of bioactive compounds from traditional Chinese medicine.

Disruption of TAB1/p38α Interaction Using a Cell-permeable Peptide Limits Myocardial Ischemia/Reperfusion Injury

SuHeXiang Wan in the treatment of stroke: Prediction potentially active metabolites using a combination of in silico analysis and experimental viability assessment.