Differentially Expressed Metabolites Research Articles

LC-MS/MS-based metabolic profiling: unraveling the impact of varying degrees of curing on metabolite transformations in tobacco

The curing process regulates metabolite transformations of leaves and significantly influences the formation of tobacco quality. This study investigated the major physicochemical compositions and metabolic profiles under normal curing (NC), excessive curing (EC), and insufficient curing (IC) treatments. The results indicated that the contents of nicotine, nitrogen, potassium, and chlorine remained stable among treatments, while the sugar content in EC was significantly lower than in IC. LC-MS/MS identified 845 metabolites, with flavonoids as the most abundant class. Comparative analyses identified a series of differentially expressed metabolites (DEMs) among fresh leaf, NC, EC, and IC leaves at the end of 42°C, 54°C, and 68°C, respectively. At the end of 68°C, 256 up-regulated and 241 down-regulated common DEMs across treatments were isolated in comparison to fresh leaf, underscoring the consistency of metabolic changes during curing. Notably, nonivamide varied markedly across treatments, suggesting its potential as a key curing indicator. NC_68°C displayed 11 up-regulated and 17 down-regulated unique DEMs, differing from EC_68°C and IC_68°C, suggesting their potential availability in evaluating tobacco leaf quality. KEGG pathway analysis revealed temporal shifts in metabolic pathways, particularly those involved in secondary metabolite biosynthesis (such as flavonoids, flavones, flavonols) and amino acid metabolism, during the transition from yellowing to color-fixing. Correlation analysis isolated the top 25 DEMs correlated with curing degree and stage, which might play pivotal roles in the curing process and could serve as potential biomarkers for assessing curing degree and stage. Specifically, D-(+)-cellobiose displayed the strongest negative correlation with curing degree, while 5,7-dihydroxychromone exhibited the highest positive correlation coefficient. Furthermore, curcurbitacin IIa showed the highest positive correlation with curing stage, followed by hesperetin and 8-shogaol. Additionally, random forest analysis emphasized morellic acid as a core molecular metabolite across curing degrees, suggesting its potential as a biomarker. Debiased sparse partial correlation (DSPC) network analysis further pinpointed hispidulin as a key metabolite, underscoring its significance in elucidating flavonoid metabolism during the curing process. Collectively, this study enhances the understanding of metabolite transformations underlying tobacco curing processes and provides a valuable reference for optimizing curing strategies to achieve desired outcomes.

Integrative multi-omics analysis of chilling stress in pumpkin (Cucurbita moschata).

Pumpkin (Cucurbita moschata) is an important vegetable crop that often suffers from low-temperature stress during growth. However, the molecular mechanism involved in its response to chilling stress remains unknown. In this study, we comprehensively investigated the effect of chilling stress in pumpkin seedlings by conducting physiological, transcriptomic, and metabolomic analyses. Under chilling stress, there was an overall increase in relative electrical conductivity, along with malondialdehyde, soluble sugar, and soluble protein contents, but decreased superoxide dismutase and peroxidase activities and chlorophyll contents in seedling leaves compared with controls. Overall, 5,780 differentially expressed genes (DEGs) and 178 differentially expressed metabolites (DEMs) were identified under chilling stress. Most DEGs were involved in plant hormone signal transduction and the phenylpropanoid biosynthesis pathway, and ERF, bHLH, WRKY, MYB, and HSF transcription factors were induced. Metabolomic analysis revealed that the contents of salicylic acid (SA), phenylalanine, and tyrosine increased in response to chilling stress. The findings indicated that the SA signaling and phenylpropanoid biosynthesis pathways are key to regulating the responses to chilling stress in pumpkins. Overall, our study provides valuable insights into the comprehensive response of C. moschata to chilling stress, enriching the theoretical basis of this mechanism and facilitating the development of molecular breeding strategies for pumpkin tolerance to chilling stress.

Combined transcriptome and metabolome analysis reveals the mechanism of high nitrite tolerance in freshwater mussel Anodonta woodiana

Nitrite contamination and stress on aquatic organisms are increasingly emphasized in freshwater ecosystems. Freshwater bivalves exhibit high tolerance to nitrite; however, the underlying mechanism is unknown. Accordingly, this study investigated the tolerance mechanism of the globally occurring freshwater bivalve Anodonta woodiana. A. woodiana were exposed to nominal concentrations of 0, 250, 500, 1000, 2000, and 4000 mg/L nitrite for 96 h to calculate the 96-h median lethal concentration (96-h LC50). A combined transcriptome and metabolome analysis of the hemolymph (the most vital component of the bivalve immune system) was performed after exposing A. woodiana to 300 mg/L nitrite (approximately half the 96-h LC50) for 96 h. The 96-h LC50 of nitrite in A. woodiana was 618.7 mg/L. Transcriptome analysis identified 5600 differentially expressed genes (DEGs) primarily related to ribosomes, lysosomes, DNA replication, and nucleotide excision repair. Metabolome analysis identified 216 differentially expressed metabolites (DEMs) primarily involved in biosynthesis of amino acids, 2-oxocarboxylic acid metabolism, protein digestion and absorption, aminoacyl-tRNA biosynthesis, nucleotide metabolism, ABC transporters, and valine, leucine and isoleucine degradation. Combined transcriptome and metabolome analysis revealed that DEGs and DEMs were primarily associated with nucleotide (purine and pyrimidine) and amino acid metabolism (including aminoacyl-tRNA biosynthesis, cysteine and methionine metabolism, arginine and proline metabolism, and valine, leucine and isoleucine degradation) as well as the immune system (necroptosis and glutathione metabolism). This study is the first to describe the high tolerance of A. woodiana to nitrite and elucidate the molecular mechanisms underlying high nitrite tolerance in mussels.

Multi-omics analysis reveals the protective effects of Chinese yam polysaccharide against cisplatin-induced renal interstitial fibrosis

BackgroundChinese yam polysaccharide (SYDT) has been reported to protect renal function and mitigate renal fibrosis in mice with diabetic nephropathy. Based on a multi-omics analysis, the objectives of this study were to determine the effect of SYDT on cisplatin (CDDP)-induced chronic renal interstitial fibrosis (RIF) and the underlying molecular mechanisms using an in vivo model. MethodsRats were intraperitoneally injected with a single dose of CDDP and then treated with SYDT or amifostine (AMF). The levels of urinary N-acetyl-β-D-glucosaminidase (NAG), blood urea nitrogen (BUN) and serum creatinine (Scr) were detected to assess renal function. Renal tissue damage and fibrosis were evaluated using hematoxylin and eosin (H&E) and Masson's trichrome staining, respectively. In addition, this study applied transcriptomics and metabolomics to predict the possible mechanism of SYDT action, which was verified by several relevant examinations. ResultsSYDT significantly protected the renal function, alleviated renal tissue damage and fibrosis, as well as decreased the protein levels of vimentin, α-SMA and CTGF, whereas SYDT significantly increased MMP-1 protein level in renal tissues from rats treated with CDDP. There were 1130 differently expressed genes (DEGs) between the CDDP model and SYDT-M groups proved by transcriptome analysis, indicating that metabolic pathways were likely the primary targets of relevance. Consistent with the transcriptome analysis, metabolome analysis identified 276 differentially expressed metabolites (DEMs) between the SYDT-M and CDDP model groups, with predominant clustering within glycerophospholipid metabolism. Integrative analysis of the transcriptome and metabolome indicated that SYDT inhibited the glycerophospholipid metabolism pathway by regulating the target genes Gpd2, Gpam, Agpat3, Lcat, and Pla2g4b. Notably, integrative analysis showed that the Phospholipase D (PLD) signaling pathway may be the most relevant target. Moreover, related signaling pathway analysis confirmed that SYDT inhibited CDDP-induced RIF in rats by down-regulating the PLD pathway. ConclusionOur study showed that the alleviation of CDDP-induced RIF in vivo can be achieved through the inhibition of glycerophospholipid metabolism and PLD signaling pathways by SYDT.

Impact of Aerobic Exercise on Brain Metabolism: Insights from Spatial Metabolomic Analysis.

Exercise is acknowledged for its beneficial effects on brain health; however, the intricate underlying molecular mechanisms remain poorly understood. This study aimed to explore aerobic exercise-induced metabolic alterations in the brain. We conducted an eight-week treadmill running exercise program in two-month-old male C57/BL6J mice. Body weight, serum lipid, glucose levels, and spatial cognition were measured. Spatial metabolomic analysis was performed to compare the metabolomic profiles across different brain regions. Immunohistochemical methods were used to compare the expression of carnitine palmitoyltransferase 1c (CPT1c). Exercise induced significant changes in the analysed metabolomic profiles. There were 904 differentially expressed metabolites (DEMs) detected in the whole brain section. Notable alterations in lipid profiles were observed, and among the 292 lipids detected, there were 74 (25.34%), 85 (29.11%), and 78 (26.71%) lipids differentially expressed in the hippocampus, thalamus, and hypothalamus of the Exe group, respectively. Lipid metabolism related pathways and enzymes were also altered, with L-carnitine and CPT1c upregulated in the three regions (p<0.05), and epinephrine levels decreased in the hippocampus (p<0.05). Furthermore, the vitamin B6 metabolism pathway was altered in the hypothalamus. This study highlighted the significant changes in lipid metabolism induced by involuntary exercise in the brains of young male mice. Exercise also altered epinephrine levels and the vitamin B12 metabolic pathway in specific brain regions, which indicated the multifaceted effects of exercise on the brain.

Metabolite analysis of peach (Prunus persica L. Batsch) branches in response to freezing stress.

Cold resistance in fruit trees has a direct impact on food production and scientific studies. 'Donghe No.1' is an excellent cold-tolerant peach variety. Metabolomic changes under freezing stress were examined to understand the mechanisms of cold adaptation. The UPLC-MS/MS system was used to identify differentially expressed metabolites (DEMs) in branches of 'Donghe No.1' under freezing stress for 12 h at -5°C, -20°C, -25°C, or -30°C. In total, 1096 metabolites and 196 DEMs were obtained at -5°C vs -20°C, -25°C, and - 30°C, while 179 DEMs and eight shared DEMs obtained at -5°C vs -20°C, -20°C vs -25°C, and -25°C vs -30°C. KEGG enrichment identified 196 DEMs associated with amino acid metabolism, linoleic acid metabolism, alpha-linolenic acid metabolism, phenylpropanoid biosynthesis, and flavonoid biosynthesis under freezing stress. A metabolic network in 1-year-old peach branches under freezing stress is proposed. Moreover, these results enhance understanding of metabolite responses and mechanisms to freezing stress in peach and will help in future breeding of freezing-tolerant varieties and investigating tolerance mechanisms.

Immunocastration caused dysfunction of Père David’s deer (Elaphurus davidianus)

Abstract The population density of Père David’s deer (Elaphurus davidianus) in Jiangsu Dafeng Milu Nature Reserve was excessively high due to an overemphasis on population recovery, while ignoring the crucial relationship between population size and habitats. This study used immunocastration to capture four Père David’s deer and observed their behavior. Their blood was collected for sex hormone testing, transcriptomics, and metabolomics analysis. In addition, six uncastrated Père David’s deer were included as control group. The results showed that reproductive behavior was absent in castrated Père David’s deer, while castrated individuals exhibited a significant decrease in blood hormone testosterone concentration compared to uncastrated Père David’s deer. Transcriptome and metabolome analyses revealed 450 differentially expressed genes (DEGs) and five differentially expressed metabolites (DEMs), Combined analysis of transcriptome and metabolic data revealed a significant correlation between DEGs and DEMs. It shows that using immune castration to control the population of Père David’s deer may disrupt their cell apoptosis and disturb the cell cycle, leading to abnormal pathways and promoting the onset of many diseases. Therefore, a cautious approach should be adopted toward immune castration as a means of population control. This study can provide valuable references for the standardized management of Père David’s deer populations.

LC-MS metabolomics uncovers potential biomarkers of semen cryo-injury in goats.

Semen cryopreservation acts a crucial role in enhancing breed improvement and conserving genetic resources. However, it often leads to decreased sperm activity and reduced pregnancy rates. Despite significant advancements in semen freezing techniques for goats, the precise factors and mechanisms causing cryo-injury remain unclear. In this study, we examined the motility characteristics of fresh semen versus frozen-thawed semen and investigated changes in the metabolite profiles of seminal plasma using liquid chromatograph-mass spectrometry (LC-MS). A total of 364 differentially expressed metabolites (DEMs) were identified between fresh and frozen-thawed semen samples. Among these, 185 metabolites were significantly up-regulated, while 179 were down-regulated (p<0.05). The majority of these DEMs belonged to lipids and lipid-like molecules, as well as organic acids and derivatives. The Kyoto Encyclopedia of Genes and Genomes (KEGG) indicated that these DEMs were primarily involved in pathways related to amino acid synthesis and metabolism. Additionally, metabolite set enrichment analysis (MSEA) underscored the critical role of amino acid synthesis and metabolic pathways in semen cryopreservation. Specific metabolites such as alanine, proline, phenylalanine, tryptophan, tyrosine, adenosine, citric acid, flavin adenine dinucleotide (FAD), and choline emerged as potential biomarkers for sperm cryo-injury in goats. These findings provide valuable insights into enhancing the quality of semen cryopreservation in goats, contributing to improved breeding and genetic resource conservation efforts.

Recombinant leptins affect lipid metabolism in hepatocytes of Cynoglossus semilaevis

Leptin is a peptide hormone primarily produced by adipose tissue, which plays a crucial role in regulating energy balance by controlling appetite and energy expenditure. To better understand the intricate physiological functions of leptin in hepatocytes in tongue sole (Cynoglossus semilaevis), this study presents an integrated transcriptomic and metabolomic analysis of tongue sole hepatocytes following stimulation with lepA and lepB. Comparative analysis identified 2971 and 2900 upregulated differentially expressed genes (DEGs) and 1895 and 1821 downregulated DEGs in response to lepA and lepB stimulation, respectively. Notably, 5706 genes were commonly regulated by both stimulations, suggesting overlapping functional roles of these two leptin proteins. GO and KEGG enrichment analysis indicated significant enrichment in immune response (JAK-STAT signaling pathway, NF-κB signaling pathway, etc.) and lipid metabolism pathways, such as fatty acid synthesis, with similar findings for lepB. Metabolomic analysis demonstrated clear separation between treatment and control groups, with 34 medium- to long-chain fatty acids and numerous differentially expressed metabolites (DEMs) identified. KEGG analysis of the metabolome paralleled the transcriptomic findings, with shared pathways in lipid metabolism and immune function. Finally, joint analysis highlighted co-enrichment in linoleic acid metabolism and leishmaniasis pathways. Detailed mechanistic insights revealed the activation of JAK-STAT and NF-κB signaling pathways, modulation of arachidonic acid metabolism, and the influence on the MAPK signaling pathway. Additionally, lepA uniquely co-enriched in the fatty acid synthesis pathway, with specific gene regulation affecting the synthesis of various fatty acids. The study concluded that lepA and lepB exerted significant regulatory effects on lipid metabolism and immune responses in tongue sole hepatocytes through complex molecular networks. To our current knowledge, this represents the first direct evidence of leptin's role in immune function within teleost fish and offers valuable insights into the molecular mechanisms of lipid metabolism underlying the actions of lepA and lepB.

Identification of metabolic biomarkers in idiopathic pulmonary arterial hypertension using targeted metabolomics and bioinformatics analysis

Pulmonary arterial hypertension (PAH) is a life-threatening disease with a poor prognosis, and metabolic abnormalities play a critical role in its development. This study used metabolomics, machine learning algorithms and bioinformatics to screen for potential metabolic biomarkers associated with the diagnosis of PAH. In this study, plasma samples were collected from 17 patients diagnosed with idiopathic pulmonary arterial hypertension (IPAH) and 20 healthy controls. Plasma metabolomic profiling was performed by high-performance liquid chromatography-mass spectrometry. Gene profiles of PAH patients were obtained from the GEO database. Key differentially expressed metabolites (DEMs) and metabolism-related genes were subsequently identified using machine learning algorithms. Twenty differential plasma metabolites associated with IPAH were identified (VIP score > 1 and p < 0 0.05), and enrichment analysis revealed the arginine biosynthesis pathway as the most altered pathway. Using machine learning models, including least absolute shrinkage and selection operator (LASSO), random forest (RF) and support vector machine (SVM), we extracted key metabolites that correlated with clinical phenotypes. Our results suggested that five metabolites, kynurenine, homoserine, tryptophan, AMP, and spermine, are potential biomarkers for IPAH. Bioinformatics analysis also identified 3 metabolism-related genes, MAPK6, SLC7A11 and CDC42BPA, that are strongly correlated with pulmonary hypertension, demonstrating strong predictive power and clinical relevance. Our findings revealed some key genes associated with metabolism in PH, and provided crucial information about complex metabolic reprogramming signals and may lead to the identification of useful metabolic biomarkers for the diagnosis of PAH.

Unveiling the Molecular Mechanisms Regulating Muscle Elasticity in the Large Yellow Croaker: Insights from Transcriptomics and Metabolomics.

The large yellow croaker (Larimichthys crocea) is an important economic fish in China. However, intensive farming practices, such as high stocking densities, suboptimal water quality, and imbalanced nutrition, have led to a decline in muscle quality. Muscle elasticity is a key texture property influencing muscle quality. Herein, transcriptomic and metabolomic analyses were performed on four groups: male high muscle elasticity (MEHM), female high muscle elasticity (MEHF), male low muscle elasticity (MELM), and female low muscle elasticity (MELF), to explore the molecular regulation underlying muscle elasticity in the large yellow croaker. Transcriptomics identified 2594 differentially expressed genes (DEGs) across the four groups, while metabolomics revealed 969 differentially expressed metabolites (DEMs). Association analysis indicated that the valine, leucine, and isoleucine biosynthesis pathways were significantly enriched between the MELF and MEHF groups; 2-Oxoisovalerate and L-Valine were DEMs; and the gene encoding L-threonine ammonia-lyase was a DEG. In the MELM and MEHM groups, pathways such as arginine biosynthesis; arginine and proline metabolism; and valine, leucine, and isoleucine degradation were significantly enriched. 4-guanidinobutanoate, L-aspartate, N-acetylornithine, and L-leucine were among the DEMs, while the DEGs included glul, gls, srm, hmgcs, and aacs. These findings provide insights into the molecular mechanisms controlling muscle elasticity, representing a theoretical foundation to breed high-quality large yellow croakers.

Integrative multiomics analysis identifies key genes regulating intramuscular fat deposition during development

Intramuscular fat (IMF) content is an important indicator of livestock and poultry meat quality. Enhancing IMF deposition can significantly improve meat quality. Focusing on the core process of IMF deposition, this study used the Jingxing Yellow (JXY) chickens as a model organism and employed multi-omics approaches, including RNA-sequencing (RNA-seq), Whole-genome bisulfite sequencing (WGBS), and metabolomics, to identify the key genes influencing IMF deposition in chickens during development. The results indicated that the contents of triglycerides (TG) and phospholipids (PLIP) exhibited an upward trend. The TG content did not differ significantly between day 1 (D1) and day 7 (D7), but increased significantly after 35 days (D35) of age. The WGBS results revealed that CpG methylation was the predominant methylation type in the breast muscle tissue of JXY chickens. Integrative analysis of RNA-seq and WGBS identified 50 genes, including PLA2G4F, PALMD, PLSCR5, ARHGEF26, LUM, DCN, TNRC6B, CACNA1C, ROBO1, and MBTPS2, whose methylation levels were significantly negatively correlated with their expression levels. In addition, the combined Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of differentially-expressed metabolites (DEM) and differentially-expressed genes (DEG) converged on the glycerophospholipid metabolism pathway, which was significantly enriched in DEGs such as PLA2G4F, PLA2G15, LPIN1, MBOAT2, DGKH, AGPAT2, and CHKA, as well as DEM like glycerophosphocholine and phosphocholine. Notably, PLA2G4F was identified as a DEG by DNA methylation, suggesting that PLA2G4F could be a key candidate gene influencing IMF deposition during chicken development. These findings are expected to provide a solid theoretical foundation for improving meat quality through targeted genetic and epigenetic interventions.

Widely Targeted Metabolomics Analysis of the Roots, Stems, Leaves, Flowers, and Fruits of Camellia luteoflora, a Species with an Extremely Small Population

Camellia luteoflora is a rare and endangered plant endemic to China. It has high ornamental and potential economic and medicinal value, and is an important germplasm resource of Camellia. To understand the distributions and differences in metabolites from different parts of C. luteoflora, in this study, we used liquid chromatography–tandem mass spectrometry (LC–MS/MS) to examine the types and contents of chemical constituents in five organs of C. luteoflora: roots, stems, leaves, flowers, and fruits. The results showed that a total of 815 metabolites were identified in the five organs and were classified into 18 main categories, including terpenoids (17.1%), amino acids (10.4%), flavonoids (10.3%), sugars and alcohols (9.8%), organic acids (9.0%), lipids (7.1%), polyphenols (4.8%), alkaloids (4.8%), etc. A total of 684 differentially expressed metabolites (DEMs) in five organs were obtained and annotated into 217 KEGG metabolic pathways, among which metabolic pathways, ABC transporters, the biosynthesis of cofactors, and the biosynthesis of amino acids were significantly enriched. In DEMs, flowers are rich in flavonoids, polyphenols, organic acids, and steroids; fruits are rich in amino acids, alkaloids, vitamins, and xanthones; stems are rich in lignans; and leaves have the highest relative content of phenylpropanoids, ketoaldehydic acids, quinones, sugars and alcohols, terpenoids, coumarins, lipids, and others; meanwhile, the metabolite content is lower in roots. Among the dominant DEMs, 58 were in roots, including arachidonic acid, lucidone, isoliquiritigenin, etc.; 75 were in flowers, including mannose, shikimic acid, d-gluconic acid, kaempferol, etc.; 45 were in the fruit, including pterostilbene, l-ascorbic acid, riboflavin, etc.; 27 were in the stems, including salicylic acid, d-(-)-quinic acid, mannitol, (-)-catechin gallate, etc.; there was a maximum number of 119 dominant metabolites in the leaves, including oleanolic acid, l-glucose, d-arabitol, eugenol, etc. In sum, the rich chemical composition of C. luteoflora and the significant differences in the relative contents of metabolites in different organs will provide theoretical references for the study of tea, flower tea, edible oil, nutraceuticals, and the medicinal components of C. luteoflora.

An integrated metabolomics and proteomics analysis reveals copper‑zinc alloy surface contact-mediated metabolic disruption, lipid peroxidation, and osmotic imbalance of Cryptocaryon irritans tomonts

Cryptocaryon irritans cause massive losses in the mariculture industry and are among the most threatening pathogens affecting teleost species. Copper‑zinc alloy (CZA) surface contact effectively kills C. irritans within minutes. Thus, this study employed integrated metabolomics and proteomics analysis to investigate the killing mechanism of CZA against C. irritans. Moreover, malondialdehyde contents, CuZn-SOD, and ATPase activities were evaluated. Proteomics analysis identified 142 differentially expressed proteins (DEPs), including 91 (59 upregulated, 32 downregulated), 92 (42 upregulated, 50 downregulated), and 43 (17 upregulated, 26 downregulated) DEPs at 0.5 h, 1 h, and 0.5 h vs 1 h of contact treatment with the CZA surface, respectively. Similarly, the metabolomic analysis identified 377 differentially expressed metabolites (DEMs). There were 237 (94 upregulated, 143 downregulated), 264 (112 upregulated, 152 downregulated), and 193 (101 upregulated, 92 downregulated) DEMs at 0.5 h, 1 h, and 0.5 h vs 1 h of contact treatment with the CZA surface, respectively. KEGG-based integrative analyses revealed that CZA exposure significantly enriched proteins and metabolites involved in oxidative phosphorylation, purine, pyrimidine, cysteine, and methionine metabolism. Notably, CZA exposure inhibited the TCA cycle, disrupting energy metabolism, as evidenced by downregulation of key TCA proteins, including citrate synthase and malate dehydrogenase, and metabolites like succinic and malic acids at both 0.5 h and 1 h of CZA treatment. Prolonged CZA exposure also affected protein metabolism, with significant downregulation of ribosomal protein S3a (RPS3a) and proteasome activity. Additionally, CZA surface contact downregulated key proteins involved in pyrimidine (DHFR-TS), purine (RRM1), and cysteine and methionine metabolism (AHCY), leading to disturbances in the methionine cycle, suppression of glutathione metabolism, and alterations in cellular redox status. Quantitative polymerase chain reaction (qPCR) analysis confirmed the upregulation of citrate synthase, proteasome subunit alpha type, and adenosylhomocysteinase with prolonged CZA exposure. Additionally, extended CZA exposure significantly decreased the MDA content due to the increased CuZn-SOD activity. Moreover, CZA exposure reduced Ca2+/Mg2+-ATPase and Na+/K+-ATPase activities. Altogether, CZA surface contact caused metabolic disruption, lipid peroxidation, and osmotic imbalance of C. irritans tomonts, highlighting the broad molecular impact of CZA on cellular processes.

LC-HRMS-based global metabolomics profiling unravels the distinct metabolic signature of lapatinib-resistant and trastuzumab-resistant HER2+ breast cancer cells

The effectiveness of lapatinib (LAP) and trastuzumab (TRZ), the first-line therapies for HER2+ breast cancer, has been limited owing to the development of acquired resistance in patients with HER2+. This study aimed to investigate the alterations in metabolic signatures in LAP-resistant HCC1954 and TRZ-resistant HCC1954 and pathways in human HER2+ breast cancer cells using liquid chromatography–high-resolution mass spectrometry (LC-HRMS) and enrichment analysis. The HCC1954 parental cells were sequentially treated 13 rounds with LAP or TRZ to develop resistant cells and then tested for their cytotoxicity using the MTT assay. Metabolites were prepared from HCC1954 parental (MBXWT), HCC1954-LAP (MBXLAP), and HCC1954-TRZ (MBXTRZ) cells prior to LC-HRMS, chemometric, enrichment, and joint pathway analyses. LAP- and TRZ-resistant cells were successfully developed from HCC1954, and 29 and 17 differentially expressed metabolites (DEMs) were identified between MBXWT-MBXLAP and MBXWT-MBXTRZ, respectively. The analysis of DEMs between MBXWT and MBXLAP revealed significant enrichment in D-amino acid metabolism, while MBXWT and MBXTRZ identified valine, leucine, isoleucine biosynthesis, ascorbate, and aldarate metabolism. Joint pathway enrichment analysis of LAP-resistant DEMs and differentially expressed genes (DEGs) showed enrichment in glutathione metabolism, while that of TRZ-resistance and DEGs showed enrichment in carbohydrate metabolism, namely pentose and glucuronate interconversions, starch and sucrose metabolism, and galactose metabolism. The findings from this study indicate considerable metabolic changes in LAP- and TRZ-resistant HCC1954 cells, which are crucial for understanding the resistance mechanisms and developing strategies to overcome these problems.

Tire rubber-derived contaminant 6PPD had the potential to induce metabolism disorder in early developmental stage of zebrafish

The increasing release of tire-derived particles, particularly those containing N-(1,3-dimethylbutyl)-N′-phenyl-p-phenylenediamine (6PPD), into the environment has raised concerns regarding their ecological impact. This study aims to elucidate the toxicological effects of 6PPD on the metabolism in early developmental stage of zebrafish. Larval zebrafish were exposed to 10 and 100 μg/L 6PPD, and some endpoints in biochemical parameters, gene expression, and metabolism were analyzed. The results showed that 6PPD exposure disrupted glucolipid metabolism in zebrafish larvae, evidenced by increased triglyceride (TG) levels and decreased glucose content. Nile red staining indicated significant lipid accumulation in the liver and intestines. Additionally, RT-qPCR analysis revealed the upregulation of genes involved in lipid synthesis and metabolism, such as ppar-γ and fas, and downregulation of glycolysis-related genes like pk and gk. Furthermore, the untargeted metabolomics technique was used to identify a total of 220 differentially expressed metabolites (DEMs) with changes in amino acid metabolism, lipid metabolism, and the TCA cycle. KEGG pathway enrichment analysis highlighted disruptions mainly in Taurine and hypotaurine metabolism, Arginine and proline metabolism, and Histidine metabolism, which played very important roles on energy metabolism in zebrafish. The results provided some critical insights into the ecological risks associated with 6PPD.

Mitigating the impact of Erysiphe Lonicerae on honeysuckle through preventive use of potassium dihydrogen phosphate and a comparative analysis of floral composition across three developmental stages

Honeysuckle, a key traditional Chinese medicinal herb, holds significant economic and therapeutic value. However, its three flowering stages lack clarity in component differentiation, leading to resource waste. Furthermore, the plant is consistently challenged by powdery mildew caused by Erysiphe lonicerae during cultivation. This study demonstrates that E. lonicerae reduces the flowering period from 12.4 to 8.6 days and alters the expression of two key flowering genes. However, foliar application of 0.1 % KH₂PO₄ three times weekly effectively mitigates the reduction in flowering duration. Initiating treatment at least one month prior to E. lonicerae inoculation significantly reduces the disease's impact, with earlier treatments proving more effective. Further, A comprehensive analysis of the three floral stages revealed significant metabolite and enzyme activity differences. We identified 175 differentially expressed metabolites (DEMs) between gold flowers and buds, 60 between gold and silver flowers, and 162 between buds and silver flowers, particularly in lipids, phenylpropanoids, and polyketides. Enzyme activities of PPD, SOD, PPO, and PAL, along with flavonoid content and antioxidant capacity, varied significantly across these stages. Notably, a total of 13 medicinal compounds were uniquely distributed across the floral stages, indicating that these compounds are not concentrated solely in the buds.In summary, The prophylactic use of KH₂PO₄ can effectively reduce the damage caused by E. lonicerae.The distinct metabolic profiles of the buds, silver flowers, and gold flowers highlight the need for further research into their optimal medicinal applications.

Integrating lipidomics and metabolomics to reveal biomarkers of fat deposition in chicken meat

Local chicken breeds in China are highly regarded for their superior meat flavor. This study utilized lipidomics and non-targeted metabolomics to identify biomarkers influencing intramuscular fat (IMF) deposition in the breast muscle of 42- and 180-day-old Jingyuan chickens. Results revealed that IMF content was higher in the breast muscle of 180-day-old Jingyuan chickens compared to 42-day-old chickens (P < 0.01). We identified 248 differentially expressed lipids (DELs) and 1042 differentially expressed metabolites (DEMs). The breast muscle of 180-day-old chickens contained higher levels of TG, fatty acid (FA) and cholesteryl ester (CE), with C16:1 and C18:1 being particularly abundant. Integration of non-targeted metabolomic analyses emphasized glycerolipid metabolism and vitamin digestion and absorption as the main pathways distinguishing between 42- and 180-day-old chickens. Additionally, the differential metabolites LysoPS 18:1, LysoPC 20:3, LysoPC 18:2, LysoPI 20:3, and Pantothenic acid contributed to enhanced meat flavor in Jingyuan chickens.

Molecular and metabolic insights into the mechanism of exogenous methyl jasmonate in enhancing the postharvest resistance of kiwifruit to Botrytis cinerea

Gray mold, caused by Botrytis cinerea infection, significantly impacts postharvest kiwifruit, leading to spoilage and food safety issues. Meanwhile, methyl jasmonate (MeJA) induces defense responses in plants. This study aimed to identify the optimal MeJA concentration to induce disease resistance in kiwifruits. Notably, various plant defense enzymes were detected in MeJA-treated kiwifruit. Moreover, 0.01 mol/L MeJA was found to enhance B. cinerea resistance in kiwifruit by increasing antioxidant enzyme activity. Widely targeted metabolomics analysis revealed 62–64 differentially expressed metabolites (DEMs) between the different treatment groups. Additionally, RNA-seq analysis revealed 930–2900 differentially expressed genes (DEGs), between these groups. Subsequently, combined DEG and DEM analysis highlighted phenylpropanoid biosynthesis and plant hormone signaling as key transduction pathway associated with MeJA-induced resistance. Overall, these findings identified the mechanism of MeJA-induced resistance in kiwifruit, providing a theoretical basis for the safe and effective control of postharvest diseases in kiwifruit.

Metabolic changes induced by heavy metal copper exposure in human ovarian granulosa cells

Copper (Cu) is a common heavy metal and a hazardous environmental pollutant. Emerging epidemiological evidence suggests that Cu exposure is associated with female infertility, especially ovarian dysfunction. However, the mechanisms underlying ovarian toxicity remain poorly understood. Granulosa cells play crucial roles in follicle development and are the main target cells of environmental pollutants for ovarian toxicity. In this study, we investigated the effects of Cu exposure on human granulosa (KGN) cells by using cell biology and metabolomics methods, and explored the molecular mechanisms of Cu-induced cytotoxicity. We found that Cu reduced cell viability in a dose- and time-dependent manner. Then, metabolomic analyses led to the identification of 279, 368 and 466 differentially expressed metabolites (DEMs) in KGN cells exposed to 10, 60 and 240 μM Cu, respectively. Pathway enrichment analysis revealed that high Cu led to disturbances of glutathione metabolism, nucleotide metabolism, glycerophospholipid and ether lipid metabolism. Using cell biological assays, we found that exposure to high Cu significantly decreased the GSH/GSSG ratio and altered the activities of the antioxidant enzymes SOD and CAT. Exposure to high Cu significantly increased the level of mitochondrial ROS. These findings further supported the results revealed by metabolomic analysis and provided clues for elucidating the mechanism by which Cu interferes with the development of ovarian follicles.