Viticis Fructus (VF) is an herbal medicine widely applied in both raw and processed forms. To elucidate the pharmacokinetic differences among raw VF, stir-fried Viticis Fructus (F-VF) and wine-processed Viticis Fructus (W-VF), an ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was established to simultaneously determine the concentrations of 16 compounds in rat plasma. Method validation showed that this method exhibited good linearity, precision, accuracy, stability, extraction recovery, and matrix effect. Results showed that compared with the VF, F-VF significantly increased the Cmax and AUC of chlorogenic acid, agnuside, isoorientin and luteolin (p < 0.01) and prolonged the T1/2 of agnuside (p < 0.01). W-VF shortened the Tmax of vanillic acid and protocatechuic acid, but significantly reduced the Cmax and AUC of p-hydroxybenzoic acid, casticin and agnuside (p < 0.01). This study will provide guidance for optimising the selection of VF and its processed products, so as to achieve personalised therapeutic effects.

ETHNOPHARMACOLOGICAL RELEVANCE:Suxiao Jiuxin Pills (SJP), as a classic Chinese patent medicine formula, is clinically used to treat angina pectoris and myocardial ischemia (MI). However, its pharmacodynamic basis and potential molecular mechanisms remain incompletely elucidated. Bioinformatics analysis identified S100A9 as a key MI target, and ligustilide (Lig) (primary active component of SJP) as its specific ligand. AIM OF THE STUDY:This study aims to elucidate the molecular mechanism by which Lig targets S100A9 to inhibit neutrophil activation and alleviate MI. MATERIALS AND METHODS:Bioinformatics analysis of human MI datasets and murine MI model validation identified S100A9 as SJP's potential target for alleviating early post-MI inflammatory injury. Molecular docking, SPR, MD simulations, and CETSA were used toscreened SJP's S100A9-targeting components. In MI models, S100A9 expression (infarcted myocardium/peripheral blood) was quantified by qRT-PCR, Western blot, and ELISA. Cardiac function was evaluated by echocardiography, TTC staining, enzymatic and histopathological assays. Flow cytometry assessed post-MI myocardial neutrophil/monocyte infiltration and neutrophil S100A9 expression. NET formation and NLRP3 inflammasome-associated molecules expression were assessed by Western blot and qRT-PCR, respectively. The mRNA expression of inflammatory cytokines was analyzed by qRT-PCR. Serum IL-1β levels were quantified by ELISA. Flow cytometry analyzed bone marrow (BM)/spleen neutrophil/monocyte infiltration and BM cell subset proportions. qRT-PCR determined mRNA levels of BM proliferation- and differentiation-related factors. An ischemia-reperfusion (I/R) model further confirmed the cardioprotective effect of Lig targeting S100A9 to inhibit neutrophil activation. RESULTS:S100A9 was the key target mediating SJP's cardioprotection, with its major bioactive component Lig as a specific S100A9 ligand. Post-MI, Lig downregulated S100A9, attenuated inflammation and apoptosis, reduced infarct size, and improved cardiac function. It suppressed myocardial neutrophil/monocyte infiltration, NET formation, and NLRP3 inflammasome activation, and decreased cardiac TNF-α/IL-1β and serum IL-1β levels. Furthermore, it reduced inflammatory cell proportions in BM and spleen, increased BM Lin- and LSK cells, decreased megakaryocyte-erythroid progenitor ratios, and downregulated granulocyte proliferation/differentiation-related mRNA. In the I/R model, Lig exerted consistent cardioprotection via targeting S100A9. CONCLUSION:Lig, the principal bioactive component of SJP, confers cardioprotection by targeting S100A9 to inhibit neutrophil activation and attenuate acute myocardial inflammatory injury post-MI.

Mitochondria are metabolic hubs that house their own genomes (mitochondrial DNA [mtDNA]), which encode components of the oxidative phosphorylation (OXPHOS) machinery. The mitochondrial central dogma not only governs compartmentalised metabolism but also intensively intertwines with multiple biological processes, and its dysregulation is a hallmark of cancer and metabolic diseases. In this review, we highlight recent advances in mitochondrial biogenesis from a metabolic perspective, with a particular emphasis on cancer. Metabolites act as donors for diverse chemical modifications, which have been systematically identified on mtDNA, rRNA, and tRNA. Besides, post-translational modifications of proteins involved in mtDNA replication, transcription, and translation has been revealed to connect metabolic signals with mitochondrial biogenesis. A comprehensive landscape of the mitochondrial central dogma has deepened our understanding of how mitochondria coordinate OXPHOS with other organelle-specific processes to obtain a flexible metabolic network, which potentiates tumor growth. Notably, non-canonical products and biological functions of the mitochondrial central dogma further reshape our concepts of cancer initiation and progression. Given that dysregulated mitochondrial biogenesis is found in multiple human disorders, including cancer, targeting this pathway offers new therapeutic opportunities. Genome-wide studies and drug screens have identified metabolic nodes and small molecules with potential to correct mitochondrial dysfunction in cancer, while emerging tools such as mtDNA editing enable precise intervention. Despite a maturing picture of mitochondrial biogenesis, many hidden players and functions remain to be uncovered to fully decipher mitochondrial biology in cancer.

Erzhi Pills (EZP), a traditional Chinese herbal formula, has demonstrated potential aging-modulating properties, while its mechanisms in modulating immunosenescence remain incompletely understood. Two complementary aging murine models were employed to investigate the anti-immunosenescence efficacy of EZP, providing experimental validation for its translational application in delaying age-related immune decline. Morphological and physiological parameters were monitored and thymic/splenic organ coefficients were calculated. Histopathological evaluation of thymic involution was performed via hematoxylin–eosin (H E) staining. Flow cytometry quantified splenic T cell subsets (naïve/memory CD4+ and CD8+ T cells). Reverse transcription quantitative PCR (RT-qPCR) analyzed mRNA expression of key immunosenescence markers (Lin28a, GDF-11, Sirt1, IL-2, IL-17), while enzyme-linked immunosorbent assay (ELISA) measured serum levels of pro-inflammatory cytokines (TNF-α, IFN-γ). Metabolomic profiling further elucidated EZP's bioactive pathways. EZP administration significantly attenuated age-related degeneration in both murine models by restoring thymic and splenic architecture, as evidenced by increased organ coefficients and reduced histopathological damage. EZP rebalanced T cell homeostasis through selective expansion of naïve T cells and contraction of memory T cell subsets, with a pronounced increase in CD8+ T cell populations. At the molecular level, EZP upregulated Lin28a, Sirt1, and IL-2 expression while modulating systemic cytokine profiles-reducing TNF-α and augmenting IFN-γ in the natural aging cohort. These findings suggest EZP mitigates chronic inflammatory aging and enhances immune responsiveness of effector T cells. EZP's anti-aging mechanism was mediated by fatty acid metabolism modulation. This study provides evidence supporting EZP's potential as a novel therapeutic strategy for immunosenescence and warrants further investigation into its clinical translation for geriatric populations.

The NOD-like receptor family pyrin domain-containing protein 3 (NLRP3) inflammasome is an intracellular protein complex containing a nucleotide-binding oligomerization domain, leucine-rich repeats, and a pyrin domain. It is a key regulator of inflammation in viral pneumonia (VP). Small-molecule inhibitors targeting various NLRP3 binding sites are advancing into early clinical trials, but their therapeutic utility is incompletely established. Xuanfei Baidu Formula (XF), clinically used for VP treatment, attenuates NLRP3 activation by hampering caspase-11 to impede polarization of pro-inflammatory macrophages in a model of lipopolysaccharide (LPS)-induced lung injury inmice. Herein, we demonstrate that XF attenuated influenza A virus (IAV)-induced lung inflammation as well as lung injury in immunocompetent (but not in macrophage-depleted) mice. RNA sequencing of sorted lung macrophages from IAV-infected mice revealed that XF inhibited activation of the NLRP3 inflammation and interleukin (IL)-1β production. Quantitative nuclear magnetic resonance of XF enabled us to develop XF-Comb1, a fixed-ratio combination of five bioactive compounds that recapitulated the bioactivity of XF in suppressing NLRP3 activation in macrophages in vitro and in vivo. Interestingly, XF-Comb1 inhibited assembly of the NLRP3 inflammasome through multi-site interactions with functional residues of NLRP3, apoptosis-associated speck-like protein containing caspase recruitment domain (ASC), and caspase-1. Taken together, this work advances the development of NLRP3 inhibitors by translating a complex herbal formula into defined bioactive compounds.

Cardiovascular diseases (CVDs), characterized by a high incidence rate and high mortality, have become the leading cause of death globally. CVDs include coronary heart disease, stroke, hypertension, and peripheral vascular diseases. In China, the death rate of CVDs ranks the first in all major diseases. At present, the main methods to treat ischemic heart disease are drug therapy, intervention and operation. These methods only alleviate symptoms of heart failure and myocardial ischemia and improve patients' quality of life by partially restoring myocardial reperfusion. Due to the extensive irreversible necrosis of myocardial cells caused by ischemia and hypoxia, these methods cannot reverse the damage, resulting in suboptimal long-term outcomes. Although mature cardiomyocytes have been proved not to be terminally differentiated cells, they have very limited ability of regeneration and proliferation, so they can not completely replace the damaged myocardium and restore the contractile function. Although heart transplantation can replace the damaged heart, its clinical application and promotion are limited by the source of donor, expensive cost, immune rejection, and ethical problems. It has become an urgent task for clinical medicine to seek new and better treatment. The main content of this paper is to explore the application of stem cells and gene technology in the treatment of myocardial infarction (MI).

Background: Peripheral artery disease (PAD) is a high-risk vascular condition, and vascular remodeling has become a promising therapeutic approach. Paeoniflorin (PF) is the main bioactive compound in the roots of Paeonia lactiflora Pall, which is commonly used to treat a range of cardiovascular disorders. However, the mechanisms underlying the ameliorating effects of PF on PAD remain unclear. Therefore, the purpose of this study was to explore the therapeutic efficiency of PF on PAD and determine its mechanisms. Methods: The blood flow of mice was detected with a laser Doppler dot scanning imaging system. HE staining was used to observe the morphological changes of ischemic muscle. The changes in the serologic indexes were detected with an automatic biochemical assay, and the capillary density of ischemic gastrocnemius was detected with a Lectin immunofluorescence assay. The expression of angiogenesis-related proteins in ischemic gastrocnemius was detected with Western blotting, and the proportion of macrophages and neutrophils in total cells was detected with flow cytometry. Results: PF significantly increased blood flow, capillary density and protein expressions of vascular endothelial growth factor A (VEGFA), matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 2 (MMP9), and estrogen receptor α (ERα) in mouse ischemic tissue in a PAD model. PF enhances the migration of endothelial cells and promotes the formation of tubular structures, involving the ERα/ROCK2 signaling pathway. Furthermore, PF was found to promote the phenotypic transformation of macrophages and alleviated grave inflammatory responses during vascular remodeling. Conclusions: We determined that PF as a potent compound in promoting angiogenesis and mitigating inflammatory responses during revascularization.

BACKGROUND:Viral pneumonia is a common clinical infectious disease in which host immunity plays a pivotal role in its onset and progression. A compromised adaptive immune response, particularly involving CD8+ T cells, is a fundamental pathophysiological feature of viral pneumonia. The Xuanfei Baidu Formula (XFBD), recognized as one of the "three medicines and three prescriptions" for the effective clinical management of Coronavirus Disease 2019 (COVID-19) in China, has been shown to enhance lymphocyte counts in patients with pneumonia. However, the pharmacological mechanisms and active compounds of XFBD in modulating T-cell therapy for viral pneumonia remain unclear. PURPOSE:This study aims to elucidate the pharmacological mechanisms and therapeutic components of XFBD in modulating adaptive immune CD8+ T cells, thereby providing an experimental basis for its clinical application. METHODS:A murine model of pneumonia was established through intranasal administration of Influenza A Virus (IAV) and the Delta variant of SARS-CoV-2. Treatment with XFBD or oseltamivir was conducted over a period of 5 or 7 days. Lung protection was evaluated by measuring pulmonary viral titers, inflammatory factors, and conducting histopathological examinations. Clinical data analysis involved assessing alterations in the quantity and functionality of T cells in patients infected with influenza and COVID-19. The therapeutic potential of XFBD in modulating T cell activity was ascertained in TCRbeta-delta- mice. Flow cytometry was employed to analyze changes in T cells and functional factors of CD8+ T cells in both lung tissue and blood samples. Transcriptomic analysis and western blot were utilized to identify XFBD targets involved in the regulation of CD8+ T cells. High-performance liquid chromatography (HPLC) was used to characterize XFBD and its principal components, while molecular docking studies elucidated the molecular mechanisms of key compounds. Primary CD8+ T cells were stimulated with CD3/CD28 to develop an in vitro activation model. The active compounds in XFBD that influence CD8+ T cell proliferation via the MAPK pathway were evaluated using Carboxyfluorescein Succinimidyl Ester (CFSE) assay. RESULTS:Our research findings indicate that XFBD can alleviate lung inflammation and ameliorate damage caused by immune imbalances induced by viral infections. XFBD significantly improves the condition of viral pneumonia by enhancing the infiltration and functionality of T cells, particularly CD8+ T cells, within the lungs. The therapeutic efficacy of XFBD is substantially reduced in T cell-deficient mice. XFBD primarily influences CD8+ T cell proliferation, which is associated with the mitogen-activated protein kinase (MAPK) signaling pathway. Tissue distribution and molecular binding experiments identified polydatin and naringenin as the key compounds that facilitate CD8+ T cell proliferation through the MAPK signaling pathway. CONCLUSION AND IMPLICATIONS:In conclusion, we found that XFBD regulates CD8+ T cell proliferation through the MAPK signaling pathway, thereby combating viral infection, which helps explain the theory and scientific principle of drug action and provides a scientific basis for the clinical application of XFBD.

For this investigation, a UHPLC-QqQ-MS/MS method was developed for the simultaneous determination of 15 components (chrysin, chrysin-7-O-β-D-glucuronide, hispidulin, wogonoside, quercetin, quercetin-7-O-β-D-glucoside, baicalin, baicalein, acacetin, oroxin A, oroxin B, oroxylin A, oroxylin A-7-O-β-D-glucuronide, apigenin, and scutellarein) in rat plasma. An ACQUITY UPLC BEH C18 column was utilized for separation, and simultaneous detection of the analytes was achieved through the multiple reaction monitoring mode. Method validation results were all within acceptable ranges for biological sample determination. Subsequently, the method was applied to pharmacokinetic studies in rats following oral administration of Oroxyli seed (OS) extract. The results indicated that baicalin, oroxylin A, and oroxylin A-7-O-β-D-glucuronide exhibited significant bimodal phenomena. The maximum concentration of baicalin was 13376.96 ± 2232.32 ng/mL, indicating a relatively high blood concentration. This may be related to the high content of oroxin A and oroxin B in OS extract being metabolized to baicalin. The present study may provide guidance for the further application of OS.

BACKGROUND:Spinal cord injury (SCI) is a central nervous system (CNS) disorder, and it often results in severe neuronal damage. However, there are still no effective treatments for SCI, so it is important to explore and identify effective therapeutic strategies. As a natural compound extracted from walnuts, juglone has been previously reported to exhibit various biological activities, including anti-tumor and anti-inflammatory effects, but the effects on central nerve system are still blank. PURPOSE:This study aims to evaluate the therapeutic effects of juglone in spinal cord injury and elucidate its molecular mechanisms in neuroprotection. STUDY DESIGN:In vitro neuronal OGD/R experiments and in vivo spinal cord injury model experiments were employed to investigate the neuroprotective effects of juglone on neurons. METHODS:Neuronal OGD/R models and spinal cord injury models were used to detect the neuroprotective effect of juglone. Immunofluorescence, Western Blot and ELISA were employed to investigate the effects of juglone on necroptosis and pyroptosis of neurons. RNA-Seq analysis, immunoprecipitation, Western Blot, qRT-PCR, immunofluorescence and ChIP-qPCR were utilized to elucidate the molecular mechanism of its neuroprotective effect. RESULTS:Juglone can inhibit necroptosis and pyroptosis both in vivo and in vitro, thereby exerting neuroprotective effects. Mechanistically, we identify a novel FOS/USP53 signaling axis, in which juglone suppresses the expression of FOS that directly regulates the deubiquitinating enzyme USP53. Reduced FOS expression leads to the downregulation of USP53, thereby promoting the ubiquitination and degradation of MLKL and GSDMD. This cascade ultimately alleviates necroptosis and pyroptosis in injured neurons. CONCLUSION:Juglone is a potential neuroprotective drug that exerts its effect by inhibiting necroptosis and pyroptosis through the FOS/USP53/ubiquitination signaling axis. These findings provide a novel potential drug target for the treatment of spinal cord injury.

α-Cyperone (C15H22O), a critical bioactive sesquiterpene, serves as a representative chemical compound of Cyperi Rhizoma—a classical functional food. To investigate the pharmacokinetic characteristics of α-cyperone, a quantified method was developed in plasma, bile, urine, and feces by ultra-high-performance liquid chromatography tandem triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS). After being validated, the developed method was applied in a plasma pharmacokinetic study as well as biliary, urinary, and fecal excretion kinetics studies. It revealed poor absolute bioavailability (F = 1.36%) and rare excretion (total cumulative excretion = 0.022%) of α-cyperone, which suggested extensive first-pass metabolism. This study provided crucial insight into explaining the in vivo process and promoting the further development of α-cyperone.

Cancer remains a leading cause of mortality worldwide, necessitating the continuous exploration of novel therapeutic strategies. Natural polysaccharides derived from medicinal plants have emerged as a promising source of anti-cancer agents due to their diverse biological activities and generally favorable toxicity profiles. In this study, aiming to identify novel anti-tumor polysaccharides, a homogeneous arabinan polysaccharide, designated HJBP85-1, was isolated and purified from Rhodiola rosea L. rhizomes using alkaline extraction and sequential chromatographic purification techniques. Comprehensive chemical characterization, utilizing techniques such as high-performance gel permeation chromatography (HPGPC), multi-angle laser light scattering (MALLS), fourier transform infrared spectroscopy (FT-IR), methylation analysis, and nuclear magnetic resonance (NMR) spectroscopy, revealed that HJBP85-1 is a branched arabinan, primarily composed of α-linked arabinose residues, with an average molecular weight of approximately 13 kDa. In vivo anti-tumor experiments conducted using a zebrafish xenograft model demonstrated that HJBP85-1 significantly inhibited tumor progression. Specifically, treatment with HJBP85-1 at a concentration of 400 μg/mL achieved remarkable inhibition rates of up to 93 % for tumor proliferation and 85 % for metastasis. Mechanistically, HJBP85-1 exhibited its anti-tumor effects by inhibiting tumor cell migration and angiogenesis. Furthermore, it modulated the immune response by promoting macrophage M1 polarization and inducing dendritic cell maturation and activation, as evidenced by increased nitric oxide (NO) production and the upregulation of specific surface markers. These findings highlight the promising anti-tumor properties of HJBP85-1 and strongly suggest its potential as a novel therapeutic agent for cancer treatment, thus warranting further investigation.

Spinal cord injury is characterized by high incidence and high disability, and the specific targets and drugs have not yet been explored. Lipid droplet is a type of organelles that regulates lipid metabolism and oxidative stress. And the regulatory mechanisms of lipid droplets on spinal cord injury remain unclear. Herein, it is found that GTPase activation of Annexin A7 (ANXA7) promotes the up-regulation of genes related to lipid droplet formation. ANXA7 can interact with peroxisome proliferator-activated receptor gamma (PPARγ) to enhance the stability of PPARγ, and promote lipid droplet formation and interaction with mitochondria through promoting Perilipin 5 expression. Then, oxidative stress and lipid peroxidation are inhibited due to the promotion of nuclear factor erythroid 2-related factor 2 (NRF2) nuclear translocation and expression of glutathione peroxidase 4 (GPX4). ANXA7 activation promotes lipid droplet formation and mitochondria-lipid droplet interaction by enhancing nuclear translocation of PPARγ, which contributes to inhibiting lipid peroxidation and neuron damage. Furthermore, activation of PPARγ can promote neural function recovery and spinal cord repair in mice. The focus of this study is to investigate the effects of lipid droplets regulated by ANXA7/PPARγ, providing new targets and strategies for spinal cord injury.

This study first compared the protective effects of different parts of Perilla frutescens (L.) Britton leaves (PFo), stems (PCa), and seeds (PFr) on LPS/ D-GalN-induced acute liver injury (ALI) in rats, to screen the most effective parts. The results indicated that PFo can effectively improve the histopathological changes of the liver, significantly reduce ALT, AST, LDH, TBil, TNF-α, and IL-1β levels, and increase GSH levels in ALI rats. Moreover, PFo inhibited apoptosis and ROS production in D-GalN-stimulated HepG 2 cells, and suppressed TNF-α and IL-6 production in LPS-elicited RAW 264.7 cells. Subsequently, based on the optimal results of PFo's anti-ALI effects, comprehensive analyses of liver metabonomics and lipidomics, and network pharmacology indicated that TNF, AKT1, ALB, STAT3, ESR1, EGFR, and PTGS2 might be the major targets for the anti-ALI of PFo, demonstrating that PFo exerts therapeutic properties against ALI might be related to suppressing oxidative stress, inflammation, and apoptosis.

BackgroundCervical cancer (CC) remains a major malignancy threatening women’s health, with high-grade squamous intraepithelial lesions playing a critical role in the progression toward CC. Exploring the molecular characteristics of epithelial cells (EPCs) as high-stage intraepithelial neoplasia evolves into CC is essential for the development of effective targeted drugs for cervical cancer. Single-cell RNA sequencing technology can fully understand the immune response at each molecular level, providing new ideas and directions for the precise treatment of CC.MethodsSingle-cell RNA sequencing was employed to comprehensively map EPCs characteristics. The differentiation trajectory of EPCs was inferred using Slingshot, while enrichment analysis highlighted the biological functions of EPCs. Cellchat visualized cell-cell interactions, and SCENIC was used to infer transcription factor regulatory networks in EPCs. CCK-8, colony formation, and EDU experiments were used to verify cell proliferation changes. Scratch assays and transwell assays were used to verify cell migration and invasion.ResultsA distinct EPCs subpopulation with high TOP2A expression was identified, predominantly originating from tumor tissues. This subpopulation exhibited disrupted mitosis and cell cycle regulation, along with features of high proliferation, high energy metabolism, and matrix plasticity. It played a key role in shaping the tumor microenvironment via the LAMC1-(ITGA3-ITGB1) signaling pathway. FOXM1, a key transcription factor in this cell subpopulation, significantly inhibited the proliferation and invasion of cervical cancer cells.ConclusionThrough in-depth analysis of EPCs, this study provides promising insights and potential therapeutic targets for precision targeted treatment strategies for CC.

Bacillus subtilis is the model Gram-positive and industrial chassis bacterium; it has blossomed as a robust and promising host for enzyme, biochemical, or bioflocculant production. However, synthetic biology and metabolic engineering technologies of B. subtilis have lagged behind the most widely used industrial chassis Saccharomyces cerevisiae and Escherichia coli. CRISPR (an acronym for clustered regularly interspaced short palindromic repeats) enables efficient, site-specific, and programmable DNA cleavage, which has revolutionized the manner of genome editing. In 2016, CRISPR technology was first introduced into B. subtilis and has been intensely upgraded since then. In this Review, we discuss recently developed key additions to CRISPR toolkit design in B. subtilis with gene editing, transcriptional regulation, and enzyme modulation. Second, advances in the B. subtilis chassis of efficient biochemicals and proteins with CRISPR engineering are discussed. Finally, we conclude with perspectives on the challenges and opportunities of CRISPR-based biotechnology in B. subtilis, wishing that B. subtilis can be comparable to traditional industrial microorganisms such as E. coli and S. cerevisiae someday soon.

BACKGROUND:Adverse ventricular remodeling following myocardial infarction (MI) is a critical factor in the progression of heart failure (HF). However, clinical interventions are limited by the lack of reliable biomarkers and targeted therapies. Soluble suppression of tumorigenicity 2 (sST2) is recognized as a prognostic biomarker in HF, with elevated levels associated with poorer outcomes, while its reduction is indicative of successful therapeutic intervention in mitigating remodeling. Tetramethylpyrazine (TMPZ), a bioactive alkaloid derived from Ligusticum chuanxiong Hort., has demonstrated potential in the treatment of MI, with our previous research linking it to the suppression of sST2. Nevertheless, the mechanisms by which TMPZ regulates sST2-mediated remodeling remain inadequately understood. PURPOSE:This study aims to elucidate the molecular mechanisms through which TMPZ modulates sST2 to alleviate ventricular remodeling post-MI. METHODS:MI was induced in mice, followed by TMPZ treatment for 1, 3, 7, and 14 days​​. Cardiac function was monitored via echocardiography pre-surgery and on days 3, 7, and 14 post-MI. Serum myocardial enzymes were quantified automatically. Cardiac tissue fibrosis and histopathology were evaluated using Masson's trichrome and HE staining, while immunohistochemistry detected ventricular remodeling-associated proteins. sST2 secretion was measured by ELISA. Bioinformatic analyses (KEGG and GO) were performed on the overlapping targets between the cardiac transcriptomic dataset at 3 days post-MI (GSE217268) and Ligusticum chuanxiong Hort.. Primary cardiomyocytes were subjected to oxygen-glucose deprivation (OGD) and treated with TMPZ. Measurement of cell viability using the CCK-8 assay. qRT-PCR and Western blotting evaluated mRNA and protein expression of Sirt1, p300, Yy1, and sST2. Molecular docking and SPR analysis characterized TMPZ-Sirt1 binding. Sirt1 activity in cardiac tissue was measured colorimetrically. Co-immunoprecipitation (Co-IP) assessed p300 acetylation and Yy1 interaction. The Sirt1 inhibitor EX527 confirmed TMPZ-mediated sST2 regulation via Sirt1 in the MI model. RESULTS:TMPZ administration markedly improved cardiac function, reduced circulating biomarkers of myocardial injury, attenuated histopathological damage, and inhibited ventricular remodeling in a murine model of MI. Treatment with TMPZ also significantly lowered serum levels of sST2. Integrative bioinformatic analysis of GEO datasets and network pharmacology suggested that Ligusticum chuanxiong Hort. influences biological processes related to protein deacetylation, specifically via the differentially expressed gene Sirt1. Subsequent in vivo and in vitro experiments established that TMPZ mediates the downregulation of sST2 through the Sirt1/p300/Yy1/sST2 signaling cascade. Molecular docking and SPR analysis suggested a potential interaction between TMPZ and Sirt1. Based on this interaction, we further verified that TMPZ enhances Sirt1's deacetylase activity. Co-IP experiments showed that TMPZ treatment reduces acetylation of the transcriptional coactivator p300, which is a downstream effector of Sirt1, resulting in inhibition of Yy1 activation and consequent suppression of sST2 synthesis and secretion. Finally, in vivo inhibition of Sirt1 with EX527 confirmed that the cardioprotective effects of TMPZ against post-MI ventricular remodeling are dependent on Sirt1-mediated reduction of sST2 secretion. CONCLUSIONS:We found that increased serum sST2 levels were significantly correlated with the progression of adverse ventricular remodeling following MI. Furthermore, our study provides the first preclinical evidence demonstrating that TMPZ attenuates this remodeling process by targeting the Sirt1/p300/Yy1/sST2 signaling axis. Collectively, these findings not only highlight sST2 as a promising therapeutic target but also establish a mechanistic rationale for developing novel interventions through pharmacological modulation of the Sirt1/sST2 pathway.

Natural medicines play an indispensable role in treating thrombotic-related diseases and a thorough investigation of their material basis is crucial for medicine development. The rapid advancement in medicine-active component screening technologies has paved new avenues for studying natural medicines, holding significant theoretical and practical value. This review focuses on the application progress of multimodal screening technologies, including high-throughput screening, chip technology, molecular biology methods, fluorescence sensors, and computational biology, in the screening of anticoagulant medicines. The aim is to provide a reference framework for screening and validating active components in natural medicines. The early application of these technologies can swiftly assess the safety and efficacy of medicines, accelerating the medicine development process and reducing the failure rate in clinical trials. Nonetheless, the overall mechanisms of action of natural medicines and the correlation between their chemical components and thrombotic diseases remain challenging areas that require further in-depth exploration and technological innovation.

The microglia, displaying diverse phenotypes, play a significant regulatory role in the development, progression, and prognosis of Parkinson’s disease. Research has established that glycolytic reprogramming serves as a critical regulator of inflammation initiation in pro-inflammatory macrophages. Furthermore, the modulation of glycolytic reprogramming has the potential to reverse the polarized state of these macrophages. Previous studies have shown that Levistilide A (LA), a phthalide component derived from Angelica sinensis, possesses a range of pharmacological effects, including anti-inflammatory, antioxidant, and neuroprotective properties. In our study, we have examined the impact of LA on inflammatory cytokines and glucose metabolism in microglia induced by lipopolysaccharide (LPS). Furthermore, we explored the effects of LA on the AMPK/mTOR pathway and assessed its neuroprotective potential both in vitro and in vivo. The findings revealed that LA notably diminished the expression of M1 pro-inflammatory factors induced by LPS in microglia, while leaving M2 anti-inflammatory factor expression unaltered. Additionally, it reduced ROS production and suppressed IκB-α phosphorylation levels as well as NF-κB p65 nuclear translocation. Notably, LA exhibited the ability to reverse microglial glucose metabolism reprogramming and modulate the phosphorylation levels of AMPK/mTOR. In vivo experiments further corroborated these findings, demonstrating that LA mitigated the death of TH-positive dopaminergic neurons and reduced microglia activation in the ventral SNpc brain region of the midbrain and the striatum. In summary, LA exhibited neuroprotective benefits by modulating the polarization state of microglia and altering glucose metabolism, highlighting its therapeutic potential.

The aim of this study was to investigate the potential mechanism by which cryptotanshinone(CTS) may exert its anti-myo-cardial ischemic effect through the regulation of macrophage polarization via the dendritic cell-associated C-type lectin 1(Dectin-1) signaling pathway. Male C57BL/6 mice, aged six weeks, were utilized to establish myocardial ischemia models and were subsequently divided into five groups: sham, model, CTS low-dose(21 mg·kg~(-1)·d~(-1)), CTS high-dose(84 mg·kg~(-1)·d~(-1)), and dapagliflozin(0.14 mg·kg~(-1)·d~(-1)). The cardiac function, serum enzyme levels, Dectin-1 expression, macrophage polarization, and neutrophil infiltration in the myocardial infarction area were assessed in each group. An in vitro model of M1-type macrophages was constructed using lipopolysaccharide/interfe-ron-γ(LPS/IFN-γ) stimulated RAW264.7 cells to investigate the impact of CTS on macrophage polarization and to examine alterations in key proteins within the Dectin-1 signaling pathway. In the CTS group, compared to the model group mice, there was a significant improvement in the cardiac function and myocardial injury, along with a notable increase in the ratio of M2/M1-type macrophages in the myocardial infarcted area and a decrease in neutrophil infiltration. Additionally, Dectin-1 exhibited low expression. The results of in vitro experiments demonstrated that CTS can decrease the expression of M1-type marker genes and increase the expression of M2-type marker genes. Besides, it can decrease the levels of Dectin-1 and the phosphorylation of its associated proteins, including spleen tyrosine kinase(Syk), protein kinase B(Akt), nuclear factor-kappaB p65(NF-κB p65), and extracellular signal-regulated protein kinases(ERK1/2). Additionally, CTS was found to enhance the phosphorylation of signal transducer and activator of transcription-6(STAT6). The above results suggest that CTS exerts its anti-myocardial ischemic injury effect by regulating macrophage polarization through the Dectin-1 signaling pathway.