Abstract Establishing a unified physical and molecular coordinate system from fragmented multi-modal data is a longstanding challenge in biology. Here, we present MAPS, a modality-agnostic platform for spatial biology comprising (1) MAPS-alignment for ultrafast alignment of any modality, (2) MAPS-integration for both anchored and unanchored integration across orthogonal modalities for 3D multi-modal reconstruction, and (3) MAPS-Explorer for large-scale interactive 3D analysis. MAPS outperformed existing methods across extensive benchmarks on 34 datasets spanning 16 technology platforms and 6 modalities, while delineating fine-grained multi-modal tissue architectures across diverse biological systems in mouse and human. At cross-consortium scale, MAPS integrated 434 slices comprising 21 million cells from 18 atlases and 5 modalities to construct the most comprehensive 3D multi-modal mouse brain atlas. At individual laboratory scale, MAPS empowered routine 2D spatial assays to reconstruct continuous 3D multi-modal landscapes of human hepatocellular carcinoma, revealing the limitations of 2D spatial relationships and uncovering depth-dependent immune-state transitions.

Immune evasion limits the efficacy of cancer immunotherapy, yet the metabolic basis underlying differential immune sensitivity remains poorly explored. Here, we established an integrated platform that couples mass spectrometry imaging (MSI) with a patient-derived tumor organoid/immune cell coculture system to delineate metabolic adaptations driving tumor immune resistance. By optimizing a pretreatment workflow that preserves morphological integrity, we achieved in situ MSI mapping of phospholipids, fatty acids, taurine, glutathione, and other metabolites within tumor organoids. Colorectal cancer (CRC) organoids segregate into immune-sensitive and immune-resistant subtypes according to their responses to cocultured peripheral blood mononuclear cells (PBMCs), and the resistant fraction exhibits significantly elevated B7-H3 expression. MSI of these organoids further revealed that phospholipid metabolism is markedly reprogrammed in immune-resistant organoids. This metabolic signature was corroborated by MSI of human CRC tissues, and dynamic MSI of organoids exposed to PBMCs showed that immune-resistant variants preserved a significantly larger phospholipid pool than did their sensitive counterparts, indicating superior membrane-lipid conservation under immune pressure. This integrated MSI-organoid immunity platform offers a broadly applicable framework for dissecting immune-resistance mechanisms and highlights phospholipid metabolism as a potential targetable axis to restore antitumor immunity in cancer treatment.

Tumor-associated macrophages, pivotal regulators of antitumor immunity, exert dual functions through their tumoricidal M1 and tumor-promoting M2 phenotypes, which are closely linked to their metabolic states. While conventional mass spectrometry imaging (MSI) can characterize the metabolic features of macrophages, it fails to capture dynamic metabolic activity and real-time substrate utilization within individual cells. In this research, we present an integrated approach that couples cell-resolved matrix-assisted laser desorption/ionization (MALDI)-MSI with stable isotope tracing to visualize dynamic metabolic heterogeneity across individual macrophage phenotypes in situ. Using isotopically labeled fatty acids as metabolic tracers, we revealed that M1 macrophages exhibit significantly enhanced synthesis of phospholipids, including phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), and phosphatidic acid (PA), compared to M2 macrophages, highlighting a polarization-specific metabolic signature linked to their antitumor function. Moreover, we observed that coculture with tumor cells markedly downregulated the levels of newly labeled phospholipids in M1 macrophages. Critically, the pharmacological inhibition of cPLA2, a key enzyme in the phospholipid metabolic pathway, significantly impaired the antitumor efficacy of M1 macrophages. These findings collectively demonstrate the functional importance of phospholipid metabolism in sustaining macrophage-mediated antitumor immunity. We envision that this spatially resolved metabolic tracing strategy will open new avenues for investigating cell-resolved metabolic crosstalk in complex biological environments.

Lonicera japonica Flos (LJF) is widely used in pharmaceuticals and functional foods, with its bioactive constituents significantly influenced by processing methods. This study characterized the dynamic changes in chemical components in LJF under different maceration and decoction durations. Using UPLC-Q-TOF-MS and molecular networking, a total of 260 metabolites were unambiguously identified or tentatively characterized, including 66 iridoids, 42 flavonoids and 49 phenolic acids. Among these, 11 phenolic acids and 3 flavonoids were absent in the macerated samples. Twenty-two representative compounds were quantified using calibration curves. Most secondary metabolites, particularly phenolic acids, exhibited lower levels in the macerated samples than the decocted samples (e.g., 5-O-caffeoylquinic acid: 65.67–106.41 μg/g during maceration vs. 32,783.05–55,754.68 μg/g during decoction). The decoction process significantly enhances the extraction of active constituents. Notably, certain iridoids (e.g., 7-O-methyl morroniside: 92.91–354.59 μg/g during maceration vs. 50.43–171.40 μg/g during decoction) were better preserved under maceration, highlighting its advantage for retaining heat-sensitive bioactive components. During the decoction process, 5-hydroxycinnamoylquinic acids tended to transform into 3- and 4-hydroxycinnamoylquinic acid isomers. Most di-hydroxycinnamoylquinic acids and flavonoids significantly decreased after 30 min. Nitrogen-containing seco-iridoids declined rapidly after 15 min. To balance extraction efficiency with the preservation of heat-sensitive bioactive components, a decoction time of 15–30 min is recommended. The study systematically elucidates the dynamic changes in bioactive components under two preparation methods, offering critical insights and a scientific foundation for the precision utilization of LJF in pharmaceutical and functional food industries.

Reliable ion annotation and identification remain persistent challenges in mass-spectrometry-based untargeted metabolomics. Here, we elucidate the identities, sources, and formation mechanisms of numerous previously unexplained ions by showing that microdroplets formed during electrospray ionization can promote a wide array of chemical transformations. These include redox, addition, condensation, reductive amination, decarboxylative coupling, and radical reactions, many of which are facilitated at the gas-aqueous interface by reactive oxygen and nitrogen species. Activation of metabolite chemical bonds generates cations, anions, and radical intermediates through the loss of protons, electrons, or functional groups, ultimately leading to bond formation and the generation of artifactual ions or false positive ions. These artifacts are frequently misassigned as endogenous metabolites and account for hundreds of thousands of previously unidentified features. As an example, we show that in a recently published untargeted metabolomics analysis of 1969 ions, the annotation rate was substantially improved to over 50% from the previous value of 9%, showing the importance of this new form of identification. Finally, we describe practical strategies to minimize artifactual ion formation, thereby improving the reliability of metabolomic analyses.

Ferroptosis, a major mechanism of non-apoptotic programmed cell death, critically regulates the homeostasis and functionality of peripheral CD4+ and CD8+ T cells1-6. Here we demonstrate that in mouse, resistance of T cells to ferroptosis depends critically on the composition of standard rodent diets, and that dietary effects on ferroptosis (DEFs) have a crucial role in regulation of T cell homeostasis and immune responses. DEFs are microbiota-independent and are driven by variations in dietary polyunsaturated and monounsaturated fatty acids (PUFAs and MUFAs) that lead to variations in abundance of lipid species in lymphoid tissues and T cells. Consistently, ferroptosis resistance of human T cells also correlated with plasma lipid profiles across multiple healthy cohorts, exhibiting negative associations with PUFA/MUFA ratios in major lipid classes. DEFs dictate T cell resilience in the absence of the essential lipid peroxide scavenger GPX4 and broadly modulate T cell-dependent humoral immunity and T cell-mediated anti-tumour immunity, including in chimeric antigen receptor T cell therapy. Mechanistically, ACSL4, which preferentially biosynthezises PUFA-containing phospholipids7, is highly expressed in T cells and underpins DEF-mediated regulation of follicular helper T (TFH) cell generation and function. Our findings reveal the physiological significance of lipid metabolism in driving DEFs in immunity and suggest strategies targeting lipid metabolism to enhance vaccine efficacy and T cell-mediated immunotherapy.

This study focuses on the potential hazards of borneol (BO) to aquatic organisms and human health. BO has antibacterial, anti-inflammatory and antioxidant activities, and is widely used in medicine, cosmetics, and detergents. In this study, zebrafish was used as a model organism to systematically evaluate the effects of BO on the heart, liver, kidney, and nervous system. The effects of BO on metabolites of zebrafish were studied using MALDI-MSI. The results showed that a high concentration of BO (500 μM) could induce morphological abnormalities (swim-bladder loss, spinal curvature, body-length shortening), cardiotoxicity (decreased heart rate, increased SV-BA distance), hepatotoxicity (reduced liver area index), and neurotoxicity (impaired behavioral ability, and dopamine neuron development deficits), but there was no renal toxicity observed in zebrafish. Additionally, MALDI-MSI analysis showed that BO exposure significantly altered the levels of metabolites, including phospholipids, fatty acids, choline, and amino acids. The contents of PC-34:1, PC-34:2, PI-36:4, PE-36:1, LysoPE-22:5, LysoPC-18:1, FA-18:2, phenylalanine, lysine and glutathione were significantly increased, while the contents of PC-38:6 and PC-40:6 were significantly decreased. Notably, BO elicited a significant alteration in the mRNA expression levels of genes associated with phospholipid metabolism, fatty acid metabolism, choline metabolism, and amino acid metabolism (such as elovl5, chpt1, chka, setd7, hgd). This study revealed that BO exerted toxicity on multiple organs and demonstrated that BO causes metabolic dysregulation in zebrafish. These findings provide a novel insight into the toxicity of BO.

Phenylthiourea (PTU) is a well-known inhibitor of melanin synthesis that has been extensively utilized in ecotoxicological studies involving zebrafish. Although there are reports suggesting that PTU may influence the toxicity of various compounds, the underlying mechanisms of its action remain unclear. Bavachalcone (BavaC) has a wide range of applications in agriculture and medicine, and it can enter groundwater through a variety of pathways that may pose a risk to aquatic ecosystems. We found that PTU enhanced the hepatotoxicity of BavaC in zebrafish, but the mechanism was unclear. In this study, the interactive effects of 200 μM PTU and 2.5 μM BavaC on the toxicity of zebrafish larvae were evaluated after 72 h of exposure. PTU significantly increased BavaC-induced hepatotoxicity, which was characterized by liver hypoplasia, hepatocyte vacuolation, and lipid accumulation. Matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI) revealed that the contents of fatty acids, phosphatidylcholine and glutathione significantly increased. The results of RNA sequencing (RNA-seq) and Real-time PCR (RT-qPCR) analyses indicated that several metabolic pathways, including xenobiotic biodegradation and metabolism, amino acid metabolism, lipid metabolism and carbohydrate metabolism, were differentially regulated in the PTU+BavaC group compared to the BavaC group. Our findings indicate that PTU-induced metabolic disorders exacerbate BavaC hepatotoxicity and highlight the need to reconsider the use of PTU in zebrafish-based toxicity assessments of environmental pollutants.

Metabolic crosstalk among diverse cellular populations contributes to shaping a competitive and symbiotic tumor microenvironment (TME) to influence cancer progression and immune responses, highlighting vulnerabilities that can be exploited for cancer therapy. Using a spatial multiomics platform to study the cell-specific metabolic spectrum in hepatocellular carcinoma (HCC), we map the metabolic interactions between different cells in the HCC TME and identify a unique tumor-immune-cancer-associated fibroblast (CAF) "interface" zone, where cell-cell interactions are enhanced and accompanied by significant upregulation of lactic acid and long-chain polyunsaturated fatty acids. Further combining single-cell mass spectrometry imaging of patient-derived tumor organoids, cocultured CAFs, and macrophages, we demonstrate that CAFs increase glycolysis and secrete lactic acid to the surrounding microenvironment to drive immunosuppressive macrophage M2 polarization. These findings facilitate the understanding of cancer-associated metabolic interactions in complex TME and provide clues for targeted clinical therapies.

INTRODUCTION:Angelica sinensis is one of the most popular traditional Chinese medicines (TCM) and has been extensively used to treat various diseases. Hundreds of endogenous ingredients have been isolated and identified from this herb, but their spatial distribution within the plant root is largely unknown. OBJECTIVES:In this study, we tried to investigate and map within-tissue spatial distribution of metabolites in Angelica sinensis roots. MATERIAL AND METHODS:After optimization of experiment conditions, the 1,5-diaminonaphthalene (1,5-DAN) was chosen as the matrix and was sprayed on the surface of root sections. Then matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) was employed to perform in situ detection and obtain detail spatial distribution information of metabolites in Angelica sinensis roots. RESULTS:The spatial distributions of a wide range of metabolites including organic acids, amino acids, oligosaccharides, and phospholipids were characterized and visualized in Angelica sinensis roots. Majority of these metabolites were located in the phloem and xylem, while ferulic acid was mainly present in the cork layer. The results revealed a dramatic metabolic heterogeneity among different regions of the roots and distinct spatial distribution patterns of different metabolites. Additionally, the metabolic pathways involved in the biosynthesis of choline were also successfully localized and visualized. CONCLUSION:This study comprehensively characterized the spatial distribution of metabolites in Angelica sinensis roots, which would prompt the understanding of its chemical separation, biosynthesis, and pharmacological activities.

Organs collaborate to maintain metabolic homeostasis in mammals. Spatial metabolomics makes strides in profiling the metabolic landscape, yet can not directly inspect the metabolic crosstalk between tissues. Here, we introduce an approach to comprehensively trace the metabolic fate of 13C-nutrients within the body and present a robust computational tool, MSITracer, to deep-probe metabolic activity in a spatial manner. By discerning spatial distribution differences between isotopically labeled metabolites from ambient mass spectrometry imaging-based isotope tracing data, this approach empowers us to characterize fatty acid metabolic crosstalk between the liver and heart, as well as glutamine metabolic exchange across the kidney, liver, and brain. Moreover, we disclose that tumor burden significantly influences the host's hexosamine biosynthesis pathway, and that the glucose-derived glutamine released from the lung as a potential source for tumor glutamate synthesis. The developed approach facilitates the systematic characterization of metabolic activity in situ and the interpretation of tissue metabolic communications in living organisms.

Cancer cells are marked by metabolic reprogramming. Mapping metabolites and their related functional proteins contributes to a comprehensive understanding of metabolic regulation during cancer progression. Mass spectrometry imaging enables the visualization of metabolites in heterogeneous cancer tissues. However, direct mass spectrometry analysis of functional proteins is challenging due to their high molecular weight and low ionization efficiency. Herein, we developed three mass probes for matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) analysis of Ki67, glutaminase, and arginase on biological samples. Mass probes were created by assembling thiolated mass tags (MTs) and specific antibodies on gold nanoparticles (AuNPs). The probes were able to target Ki67, glutaminase, and arginase for MALDI-MSI analysis. Thiolated MTs were dissociated online during MALDI-MSI. This empowered the conversion of Ki67, glutaminase, and arginase signals to MT ions, realizing functional protein analysis by MALDI-MSI. Additionally, MALDI-MSI was able to detect related metabolites directly in adjacent tissue sections, providing complementary data on metabolites and their related functional proteins. This helped to deepen our understanding of cancer metabolism from multiple perspectives.

INTRODUCTION:Neobavaisoflavone (Neo), a major bioactive compound in Psoralea corylifolia L., exhibits potent antibacterial, anti-inflammatory, anticancer, and antioxidant activities; however, the clinical use of P. corylifolia is limited by its potential hepatotoxicity. OBJECTIVE:This study aims to systematically elucidate the hepatotoxic effects of Neo, unravel its underlying molecular mechanisms, and explore potential strategies for prevention and therapeutic intervention. METHODS:In this study, a multi-model experimental strategy-including zebrafish larvae, high-fat diet (HFD)-fed mice, primary mouse hepatocytes, and human hepatocyte cell lines-was employed to investigate Neo-induced hepatic steatosis. Subsequently, the hepatotoxicity effects and molecular mechanisms of Neo's hepatotoxicity were investigated by ex vivo and in vivo experiments such as Mass spectrometry imaging (MSI), untargeted lipidomic, Transcriptomic analysis, co-immunoprecipitation and crystallographic. RESULTS:MSI revealed conserved hepatic accumulation of Neo and its lipid metabolites in both zebrafish and mouse models, with marked deposition of long-chain fatty acids (LCFAs). These findings were corroborated by untargeted lipidomic. Transcriptomic analysis further implicated disruptions in PPARα-mediated fatty acid metabolism and oxidative stress pathways. Mechanistically, co-immunoprecipitation and crystallographic studies demonstrated that Neo binds to the ligand-binding domain of PPARα, specifically at residues Ala333, Tyr334, Met220, Asn219, and Glu286, thereby impairing its nuclear translocation. Importantly, PPARα overexpression or pharmacological activation with fenofibrate (Tricor) significantly attenuated Neo-induced hepatic steatosis and oxidative stress. CONCLUSIONS:Collectively, these findings uncover a novel mechanism whereby Neo disrupts lipid homeostasis and induces oxidative stress via PPARα inhibition, and highlight PPARα activation as a potential strategy to mitigate Neo-associated hepatotoxicity, offering valuable insights for its safer therapeutic application.

OBJECTIVE:To clarify the impact of X-ray irradiation combined with PD-1 immune checkpoint inhibitor treatment on lung tissue in a mouse model of radiation-induced lung injury (RILI) and to investigate its underlying mechanisms. METHODS:A mouse RILI model was established by a single 16 Gy dose of whole-thorax irradiation. Mice were then treated with either an PD-1 inhibitor or a control. Lung injury and fibrosis were assessed by histological staining. Inflammatory cytokine levels (TNF-α, IL-6) were measured by ELISA. Macrophage M1 polarization was analyzed by flow cytometry and immunohistochemistry. The activation of the NF-κB signaling pathway was evaluated by Western blot. RESULTS:The lung injury indices in the group treated with irradiation combined with PD-1 inhibitor were higher than those in the irradiation-only group. Mechanistic studies found that PD-1 inhibitors promoted the activation of the NF-κB signaling pathway and simultaneously regulated macrophage polarization in lung tissue, promoting the differentiation of M1-type pro-inflammatory cells. CONCLUSION:Whole-thorax X-ray irradiation combined with PD-1 inhibitors can exacerbate lung injury and pulmonary fibrosis in mice, and the mechanism of action may be through the regulation of NF-κB signaling pathway activation to promote macrophage polarization towards the M1 type.

Tumor microenvironment (TME) is characterized by complex cellular composition and high molecular heterogeneity. Characterizing the metabolic interactions between different cells in the TME is important for understanding the molecular signatures of tumors and identifying potential metabolic vulnerabilities for tumor treatment. In this research, we develop a single-cell spatial metabolomics method to profile cell-specific metabolic signatures and cell-cell metabolic interactions using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI). Different low-molecular-weight metabolites and lipids including glutamate, aspartate, glutamine, taurine, phenylalanine, glutathione, fatty acids, phospholipids, etc. were successfully detected and imaged after optimizing cell culture conductive slides, cell washing, and fixation procedures. Subsequently, we carried out single-cell spatial metabolomics on H460 large-cell lung cancer cells, HT-29 colorectal cancer cells, A549 lung cancer cells, HUH-7 liver cancer cells, and cancer-fibroblasts coculture system. We revealed that the metabolic profiles of both cancer cells and fibroblasts were altered after cell coculture. Glutamate and aspartate significantly increased in fibroblasts after coculture with cancer cells, corresponding to their indispensable roles in the creation of pro-cancer microenvironment. In addition, we discovered that the expressions of fatty acids and phospholipids in tumor cells and fibroblasts were also changed after cell coculture, which is closely related to the competition for energy and nutrient metabolites between different cells. We anticipate this single-cell analysis method to be broadly used in the investigations of diverse cellular models and cell-cell metabolic interactions.

Colorectal cancer is the second leading cause of cancer-related deaths worldwide, and its development typically involves complex metabolic reprogramming. By mapping the spatial distributions of metabolites and N-glycans in heterogeneous colorectal cancer tissues, we can elucidate cancer-associated metabolic and N-glycan changes. Herein, we combine mass spectrometry imaging-based metabolomics and N-glycomics to characterize the spatially resolved reprogramming of metabolites and N-glycans in colorectal cancer tissues. The metabolic characteristics of different regions of colorectal cancer were evaluated through the utilization of orthogonal partial least squares discriminant analysis. In combination with metabolic pathway enrichment analysis, significant alterations were identified in the fatty acid metabolism, arginine and proline metabolism of colorectal cancer. Cancer cell regions exhibited a marked upregulation of saturated fatty acids, monounsaturated fatty acids, polyamines, and histidine. Additionally, we discovered that the high-mannose N-glycans were predominantly distributed in tumor tissue regions, whereas complex N-glycans were more commonly found in the normal tissue regions adjacent to the tumor. Such findings provide new insights into the spatial signatures of metabolites and N-glycans in colorectal cancer, thereby offering a crucial basis for the diagnosis of colorectal cancer and potential vulnerabilities that might be targeted for cancer therapy.

Exploring the metabolic characteristics of different plant organs and tissues at a spatial level can help us to better understand the functional mechanisms of biological tissues and cells. Mass spectrometry imaging (MSI) provides a reliable tool for this purpose. However, its application for high-resolution metabolic mapping across various plant organs remains a significant challenge due to the intrinsic biological properties of plant samples and unfavorable analysis conditions. This study aimed to develop a novel MSI platform that can expand more diverse plant samples in spatial metabolomics research and enhance the detection efficiency of plant metabolites. The platform (AMG-LDI-MSI) based on an Au nanoparticles-loaded MoS2 and doped graphene oxide (Au@MoS2/GO) flexible film substrate combined with laser desorption/ionization (LDI)-MSI was established to enhance the detection and visualization of metabolites in various plant tissues. It has a non-sectioning, matrix-free, dual-ion mode imaging strategy, enabling high-throughput detection of metabolites and high-resolution molecular imaging within a micrometer scale. The Au@MoS2/GO as a new substrate can offer high sensitivity and molecular coverage for diverse plant metabolites (10 classes) under the positive and negative ion modes. Moreover, the AMG-LDI-MSI platform overcomes the limitations of plant tissues (e.g., fragile leaf, water-rich fruit, or lignified roots) for in situ imaging. We successfully applied the platform to map the metabolite spatial dynamics in different types of fresh tissues (rhizome, main root, branch root, fruit, leaf, and nodule) from medicinal plants, obtained the high-quality mass spectral imaging data, and demonstrated the universality and applicability of the platform to multiple plant tissues. These results demonstrate the significant advantages of enhancing the detection of multiple tissue metabolites in plants and their high-resolution imaging. It has overcome the limitations of previously reported MSI methods, suggesting that it could become a widely used tool for deciphering metabolic networks in plant biology.

Colorectal cancer (CRC) is a frequently lethal disease, with stage II/III CRC accounting for ≈70%. Metabolic reprogramming plays a pivotal role in deciphering cancer heterogeneity and progression. Here, 9 datasets and 83 machine learning algorithm combinations are leveraged‌ to develop the Machine Learning-based Metabolic gene Prognostic Signature (MALMPS) model. The MALMPS model outperformed traditional clinical traits and molecular features in predicting prognosis for stage II/III CRC patients across training and validation datasets. COX7B, a key gene in MALMPS, is shown to promote CRC malignancy through multi-omics analysis and in vitro assays. CRC patients are stratified into high- and low-risk groups based on the median cutoff of MALMPS. Notably, the high-risk subgroup exhibited poor prognosis, activated inflammation, and enriched carbohydrate, glycosaminoglycan, and lipid metabolism, with therapeutic potential for IGF-1R and Wnt/β-catenin inhibitor. In contrast, the low-risk group displayed a TGF-β pathway inactivating mutation and enriched in nucleotides, cofactors, and amino acids metabolism. Metabolite profiling in the in-house SDCRC dataset validated the distinct metabolic alterations between the two groups. These findings indicate that MALMPS is a valuable instrument for predicting the recurrence risk of stage II/III colorectal cancer, particularly for identifying individuals at high risk.

Curcumin is widely recognized for its diverse antitumor properties, ranging from breast cancer to many other types of cancers. However, its role in the tumor microenvironment remains to be elucidated. In this study, we established a 3D tumor spheroids model that can simulate the growth environment of tumor cells and visualized the antitumor metabolic alteration caused by curcumin using mass spectrometry imaging technology. Our results showed that curcumin not only exerts a profound impact on the growth and proliferation of breast cancer cells but in situ multivariate statistical analysis also reveals the significant effect on the overall metabolic profile of tumor spheroids. Meanwhile, our visualization map characterized curcumin metabolic processes of reduction and glucuronidation in tumor spheroids. More importantly, abnormal metabolic pathways related to lipid metabolism and polyamine metabolism were also remodeled at the metabolite and gene levels after curcumin intervention. These insights deepen our comprehension of the regulatory mechanism of curcumin on the tumor metabolic network, furnishing powerful references for antitumor treatment.

Eclipta prostrata L. has been used in traditional medicine and known for its liver-protective properties for centuries. Wedelolactone (WEL) and demethylwedelolactone (DWEL) are the major coumarins found in E. prostrata L. However, the comprehensive characterization of these two compounds on non-alcoholic fatty liver disease (NAFLD) still remains to be explored. Utilizing a well-established zebrafish model of thioacetamide (TAA)-induced liver injury, the present study sought to investigate the impacts and mechanisms of WEL and DWEL on NAFLD through integrative spatial metabolomics with liver-specific transcriptomics analysis. Our results showed that WEL and DWEL significantly improved liver function and reduced the accumulation of fat in the liver. The biodistributions and metabolism of these two compounds in whole-body zebrafish were successfully mapped, and the discriminatory endogenous metabolites reversely regulated by WEL and DWEL treatments were also characterized. Based on spatial metabolomics and transcriptomics, we identified that steroid biosynthesis and fatty acid metabolism are mainly involved in the hepatoprotective effects of WEL instead of DWEL. Our study unveils the distinct mechanism of WEL and DWEL in ameliorating NAFLD, and presents a “multi-omics” platform of spatial metabolomics and liver-specific transcriptomics to develop highly effective compounds for further improved therapy.