Metabolism-related Enzymes Research Articles

The Metabolic Characteristics and Bioavailability of Resveratrol Based on Metabolic Enzymes.

The natural polyphenol resveratrol (RV) has garnered fame for its extensive pharmacological properties. Although clinical studies have shown some positive results, many contradictory outcomes remain. An important obstacle to the development of therapeutic applications for RV is its low bioavailability in vivo. This may be partially attributed to biotransformation mediated by phase I and II enzymes, such as cytochrome P450s, UDP-glucuronosyltransferases, and sulfotransferases. To date, more than 20 different types of metabolites have been detected after catalysis by these enzymes. Notably, RV and some of its metabolites serve as substrates for these enzymes. Conversely, RV can directly regulate the expression or activity of these enzymes. Given the increasing number of studies investigating the bioactivity of RV, this review summarizes its physicochemical and pharmacokinetic characteristics and describes the metabolism of RV and the bioactivities of its metabolites, with emphasis on the interaction between RV and its related metabolic enzymes. In addition to hepatic metabolism, the crucial roles of RV metabolism in multiple other tissues and organs cannot be overlooked, and they reveal the relationship between RV metabolism and its biological potential.

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.

Development and evaluation of a precision heat treatment device for delaying postharvest softening of muskmelon

To extend the limited commercial storage of muskmelons due to insufficient shelf life, a heat treatment device with precise temperature control was designed and applied to three muskmelon varieties: "Xizhou Mi 25" (XZM), "Jinmi 3" (JM), and "Jiashi" (JS). The effectiveness of this device and the effect of heat treatment on the softening of muskmelons during storage were evaluated. The results indicated no significant difference between the temperatures set by the control system and the actual measured temperatures (p > 0.05), demonstrating the high accuracy of the temperature control system. After optimization of the heat treatment device, no thermal damage was observed in the heat-treated group of muskmelons. The energy consumption of the heat treatment device was only half that of the insulated tank. By the 18th day of storage, the protopectin (PP) content in the heat-treated group was significantly lower than in the control group. Correlation analysis showed that hardness was positively correlated with PP content and negatively correlated with water-soluble pectin (WSP). Additionally, cell wall metabolism-related enzyme activities were negatively correlated with PP content. These results indicated that the heat treatment effectively delayed muskmelon softening during postharvest storage. This study provides technical support for the industrial application of heat treatment devices in muskmelon preservation.

Investigation of the preharvest arginine spraying mechanism on antioxidant system in postharvest broccoli via transcriptomics and metabolomics

According to our previous study, preharvest spraying of arginine for 1 day enhanced the antioxidant capacity of postharvest broccoli, but the underlying molecular mechanisms remain unclear. We here performed a comprehensive transcriptome and metabolome analysis to investigate the effects of arginine sprayed 1 day before harvest on the antioxidant capacity and yellowing of broccoli as well as the related action mechanisms. In total, 67 differentially expressed genes and 12 key differentially accumulated metabolites were identified in broccoli at 4 and 8 days during storage. Herein, the gene expression of superoxide dismutase, catalase, and the corresponding peroxisome protein Mpv17 (MPV17) was upregulated, consequently enhancing the scavenging of reactive oxygen species. Genes encoding glucosinolate synthesis- and metabolism-related enzymes were upregulated, and among them, the gene ST5b_c was upregulated by 8.23-fold, which promoted the accumulation of its metabolites. Expression of lignin- and flavonoid biosynthesis-related genes (CAD, POD, CHI, and CHS) was elevated. Flavonoid and lignin accumulation was promoted, with a greater antioxidant capacity at 4 and 8 days, respectively, in the arginine-sprayed broccoli. Additionally, the ascorbate–glutathione cycle was promoted in broccoli at 4 days, GLCAK and APX genes were upregulated by more than 2-fold, and broccoli exhibited a higher tolerance level because of the notably stimulated ascorbic acid and glutathione accumulation. This study offers new insights into the mechanisms through which preharvest spraying of arginine regulates the antioxidant capacity of postharvest broccoli.

Comparison of behavioral responses, respiratory metabolism-related enzyme activities, and metabolomics of the juvenile Chinese mitten crab Eriocheir sinensis with different tolerance to air exposure

Air exposure is a common stressor for Chinese mitten crab (Eriocheir sinensis) during rearing and transport, and air exposure tolerance can serve as a crucial indicator for assessing the quality of juvenile E. sinensis. In this study, juvenile E. sinensis were divided into two groups based on their behavioral responses: Group S, which exhibited strong tolerance to air exposure, and Group W, which exhibited weak tolerance. Immersed crabs, not exposed to air, served as a control group (Group C). Whole body morphological characteristics and enzyme activities related to respiratory metabolism in the hemolymph and anterior gills were compared among the three groups. Non-targeted LC-MS metabolomic analysis was conducted on anterior gills. The results showed that, independent of developmental stage, crabs that were larger and had higher condition factor were more tolerant to air exposure. Additionally, compared to Group W, air exposure had a relatively small effect on glycolysis and anaerobic respiratory metabolic processes in the hemolymph and anterior gills of Group S. In response to air exposure, E. sinensis experienced increased energy demand, and switched from aerobic to anaerobic respiration to increase energy supply. Simultaneously, air exposure induced oxidative stress in the hemolymph and anterior gills. This study enhances our understanding of the response mechanism of E. sinensis to air exposure and provides a theoretical reference for the identification of high-quality juvenile E. sinensis.

Exogenous glycine betaine maintains postharvest blueberry quality by modulating antioxidant capacity and energy metabolism

Blueberries are susceptible to decay and spoilage due to increased respiration rates and metabolism after harvest. The aim of this study was to examine the impact of postharvest application of exogenous glycine betaine (GB) on blueberry storage quality, antioxidant system, and energy metabolism in order to enhance understanding of the regulatory mechanisms of GB on blueberry quality. Fresh blueberries were soaked for 5 min in different concentrations of GB. Preliminary pre-tests revealed that 10 mmol L−1 GB treatment significantly retarded the quality deterioration, decay and dent rate, while increasing the firmness and soluble solid content. Concurrently, exogenous GB treatment resulted in an increase in the content of anthocyanosides, total flavonoids, total phenols, and reduced glutathione, also exhibited higher antioxidant enzyme activities, including superoxide dismutase, catalase, ascorbate peroxidase, polyphenol oxidase, phenylalanine ammonia-lyase, glutathione reductase, dehydroascorbate reductase, and monodehydroascorbate reductase. In GB-treated blueberries, endogenous GB content was significantly higher. The hydrogen peroxide value, superoxide anion, and malondialdehyde content, and higher levels of free radical scavenging rate (DPPH), which significantly improved antioxidant capacity. Meanwhile, succinate dehydrogenase, cytochrome C oxidase, H+-ATPase, and Ca2+-ATPase, decreased and slowed the senescence of blueberries, the ATP content and energy charge were observed to be elevated. In summary, exogenous GB treatment maintained the quality of post-harvest blueberries by increasing their antioxidant capacity and inhibiting the decrease in the activity of energy metabolism-related enzymes. The results indicate that 10 mmol L−1 GB treatment may be an efficacious approach for maintaining the quality and prolonging the shelf life of blueberries, offering novel insights into the preservation of this fruit.

Herb-drug interaction study of Yiqi Fumai lyophilized injection (YQFM) on pharmacokinetics of aspirin, nifedipine, and clopidogrel in rats.

Yiqi Fumai lyophilized injection (YQFM), a compound traditional Chinese medicine prescription derived from "Sheng Mai Powder," is approved for the treatment of cardiovascular diseases. YQFM is usually prescribed in combination with some Western medicines to treat patients, such as aspirin, nifedipine, and clopidogrel. However, the herb-drug interactions (HDIs) of YQFM are still unclear. We determined the effect of YQFM on drug metabolism-related CYP450 enzymes by in vitro assays. And the effects of YQFM on the pharmacokinetics of aspirin, nifedipine, or clopidogrel were analyzed in rats, as well as the effect of YQFM on the prothrombin time of aspirin or clopidogrel, to evaluate the safety and efficacy of co-administration. Our study indicated that the clinical dose of YQFM did not significantly influence the relevant CYP450 isoenzymes. Besides, YQFM had no effect on the pharmacokinetics of aspirin, nifedipine, or clopidogrel single and multiple administrations in rats. In pharmacodynamics study, YQFM also had no impact on prothrombin time of aspirin or clopidogrel. Based on the results of pharmacogenomics, pharmacokinetics, and pharmacodynamics, the HDIs of YQFM have a good safety profile, and the combination with the above three drugs might have synergistic effects due to the different efficacy of YQFM-quality markers.

Growth and physiological metabolic regulation mechanisms of the dominant plant Leymus secalinus in alpine meadow under nitrogen deposition

Nitrogen (N) deposition is an important pathway that affects the growth and development of alpine grassland plants. Under N deposition, Leymus secalinus has become the most dominant species in the alpine meadow of the Qinghai-Tibetan Plateau. However, its adaptive mechanisms to N deposition are still unknown. Therefore, we analyzed the physiological indices of Leymus secalinus under different N deposition levels (CK, 0 kg N ha−1 yr−1; N1, 8 kg N ha−1 yr−1; N3, 40 kg N ha−1 yr−1; N5, 72 kg N ha−1 yr−1) and focused on its growth and metabolism. The results indicated that the leaf carbon (C), N, amino acid (AA), and photosynthetic pigment contents in Leymus secalinus were significantly increased under N deposition, its endogenous hormone levels were regulated and the activities of N metabolism-related enzymes were enhanced. Metabolomics analysis further showed that the metabolites changed significantly and were mostly enriched in the amino acid metabolic pathway. Among them, glutamine and aspartic acid played key roles in N deposition for dominant growth of Leymus secalinus by regulating N and amino acid metabolism. These analyses unveiled the physiological and biochemical changes of dominant species in response to N deposition, identifying critical metabolites involved in this process. Furthermore, these findings provide substantial evidence explaining the ecological phenomenon of Leymus secalinus emerging as a dominant species under N deposition, serving as a data reference for understanding the physiological response and adaptation to N deposition in alpine grassland plants.

Expression patterns of folate metabolism-related enzymes in the bovine oviduct: estrous cycle-dependent modulation and responsiveness to folic acid

Folate metabolism is required for important biochemical processes that regulate cell functioning, but its role in female reproductive physiology in cattle during peri- and post-conceptional periods has not been thoroughly explored. Previous studies have shown the presence of folate in bovine oviductal fluid, as well as finely regulated gene expression of folate receptors and transporters in bovine oviduct epithelial cells (BOECs). Additionally, extracellular folic acid (FA) affects the transcriptional levels of genes important for the functioning of BOECs. However, it remains unknown whether the anatomical and cyclic features inherent to the oviduct affect regulation of folate metabolism. The present study aimed to characterize the gene expression pattern of folate cycle enzymes in BOECs from different anatomical regions during the estrous cycle and to determine the transcriptional response of these genes to increasing concentrations of exogenous FA. A first PCR screening showed the presence of transcripts encoding dihydrofolate reductase (DHFR), methylenetetrahydrofolate reductase (MTHFR), and methionine synthase (MTR) in bovine reproductive tissues (ovary, oviduct and uterus), with expression levels in oviductal tissues comparable to, or even higher than, those detected in ovarian and uterine tissues. Moreover, expression analysis through RT-qPCR in BOECs from the ampulla and isthmus during different stages of the estrous cycle demonstrated that folate metabolism-related enzymes exhibited cycle-dependent variations. In both anatomical regions, DHFR was upregulated during the preovulatory stage, while MTHFR and MTR exhibited increased expression levels during the postovulatory stage. Under in vitro culture conditions, ampullary and isthmic cells were cultured in the presence of 10, 50, and 100 μM FA for 24 h. Under these conditions, isthmus epithelial cells exhibited a unique transcriptional response to exogenous FA, showing a pronounced increase in MTR expression levels. Our results suggest that the expression of folate metabolism-related genes in BOECs is differentially regulated during the estrous cycle and may respond to exogenous levels of folate. This offers a new perspective on the transcriptional regulation of genes associated with the folate cycle in oviductal cells and provides groundwork for future studies on their functional and epigenetic implications within the oviductal microenvironment.

Study on the Mechanism of FOXA2 Activation on Glutathione Metabolic Reprogramming Mediated by ETV4 Transcription to Facilitate Colorectal Cancer Malignant Progression.

The metabolic imbalance of glutathione (GSH) has been widely recognized in most cancers, but the specific molecular mechanism of GSH metabolic regulation in the malignant progression of colorectal cancer (CRC) is unexplored. The objective of our project is to elucidate whether ETV4 affects the malignant progression of CRC through GSH metabolic reprogramming. Bioinformatics and molecular experiments measured the expression of ETV4 in CRC, and in vitro experiments explored the impact of ETV4 on CRC malignant progression. The Kyoto Encyclopedia of Genes and Genomes (KEGG) identified the pathway of ETV4 enrichment. The bioinformatics approach identified FOXA2 as an upstream regulatory factor of ETV4. The dual-luciferase assay, chromatin immunoprecipitation (ChIP) and co-immunoprecipitation (Co-IP) experiment verified the binding relationship between ETV4 and FOXA2. Cell viability, migration, and invasion abilities were determined by conducting CCK-8, wound healing, and Transwell assays, respectively. The expression levels of N-cadherin, E-cadherin, and vimentin were determined by utilizing immunofluorescence (IF). Metabolism-related enzymes GCLM, GCLC, and GSTP1 levels were detected to evaluate the GSH metabolism level by analyzing the GSH/GSSG ratio. In vivo experiments were performed to explore the effect of FOXA2/ETV4 on CRC progression, and the expression of related proteins was detected by western blot. ETV4 was highly expressed in CRC. Knocking down ETV4 suppressed CRC cell viability, migration, invasion, and epithelial-mesenchymal transition (EMT) progression in vitro. ETV4 was abundant in the GSH metabolic pathway, and overexpression of ETV4 facilitated CRC malignant progression through activation of the GSH metabolism. In addition, in vitro cellular experiments and in vivo experiments in nude mice confirmed that FOXA2 transcriptionally activated ETV4. Knocking down FOXA2 repressed the malignant phenotype of CRC cells by suppressing GSH metabolism. These effects were reversed by overexpressing ETV4. Our results indicated that FOXA2 transcriptionally activates ETV4 to facilitate CRC malignant progression by modulating the GSH metabolic pathway. Targeting the FOXA2/ETV4 axis or GSH metabolism may be an effective approach for CRC treatment.

Comparative Metabolomics Reveals Changes in the Metabolic Pathways of Ampicillin- and Gentamicin-Resistant Staphylococcus aureus.

Antibiotic resistance is a major global challenge requiring new treatments and a better understanding of the bacterial resistance mechanisms. In this study, we compared ampicillin-resistant (R-AMP) and gentamicin-resistant (R-GEN) Staphylococcus aureus strains with a sensitive strain (ATCC6538) using metabolomics. We identified 109 metabolites; 28 or 31 metabolites in R-AMP or R-GEN differed from those in ATCC6538. Moreover, R-AMP and R-GEN were enriched in five and four pathways, respectively. R-AMP showed significantly up-regulated amino acid metabolism and down-regulated energy metabolism, whereas R-GEN exhibited an overall decrease in metabolism, including carbohydrate, energy, and amino acid metabolism. Furthermore, the activities of the metabolism-related enzymes pyruvate dehydrogenase and TCA cycle dehydrogenases were inhibited in antibiotic-resistant bacteria. Significant decreases in NADH and ATP levels were also observed. In addition, the arginine biosynthesis pathway, which is related to nitric oxide (NO) production, was enriched in both antibiotic-resistant strains. Enhanced NO synthase activity in S. aureus promoted NO production, which further reduced reactive oxygen species, mediating the development of bacterial resistance to ampicillin and gentamicin. This study reveals that bacterial resistance affects metabolic profile, and changes in energy metabolism and arginine biosynthesis are important factors leading to drug resistance in S. aureus.

The Combined Application of Urea and Fulvic Acid Regulates Apple Tree Carbon and Nitrogen Metabolism and Improves Anthocyanin Biosynthesis

Improving apple peel color has been an important objective in apple production. To better understand the effect and mechanism of the combined application of urea and FA (fulvic acid) regulation of anthocyanin biosynthesis, a field experiment was performed in 2022 and 2023, respectively, under five treatments of urea + FA (CK, urea only; FA50, urea + 50 kg ha−1 FA; FA100, urea + 100 kg ha−1 FA; FA150, urea + 150 kg ha−1 FA; FA200, urea + 200 kg ha−1 FA), using isotope (13C and 15N) marking to analyze the changes in carbon (C) and nitrogen (N) nutrient distribution as well as anthocyanin biosynthesis in fruits. We observed that, under FA application conditions, anthocyanin content in the peel was elevated in both years, with increases of 15.98~52.88% in 2022 and 15.93~52.94% in 2023. The best promotion effects were observed under FA150 treatment. Apart from the expression levels of anthocyanin biosynthesis-related genes and transcription factors in the apple peel, this positive effect on anthocyanin content induced by FA addition was also found to be associated with the optimization of C and N distribution in leaves and fruits. On the one hand, the application of FA not only enhanced leaf photosynthetic-related indexes (such as Pn, Gs, and Rubisco activity) and influenced (increased) S6PDH, SPS, and SS activities in leaves, but also elevated fruit sugar metabolism-related enzyme (SDH, SS-c, AI, and NI) activity and upregulated fruit stalk sugar transporter (MdSOT1, MdSOT3, MdSUT1 and MdSUT4) gene expression, which ultimately promoted the synthesis and the leaf to fruit transport of photosynthates, thus promoting 13C-photosynthate accumulation in fruits. On the other hand, FA application elevated leaves’ N metabolism-related enzyme (GS and GOGAT) activity and optimized 15N distribution in leaves and fruits. Moreover, we also observed that FA application altered the fate of N fertilizer in apple orchards, showed an elevation in apple tree 15NUE and soil 15N residuals and showed a decrease in soil 15N loss. In summary, the appropriate application of FA150 (urea + 150 kg ha−1) synergistically optimized C and N nutrient distribution, and promoted anthocyanin biosynthesis in apple trees.

Repeated marine heatwaves aggravate the adverse effects of nano-TiO2 on physiological metabolism of the thick-shelled mussel Mytilus coruscus

Global climate change is a major trigger of unexpected temperature fluctuations. The impacts of marine heatwaves (MHWs) and nano-titanium dioxide (nano-TiO2) on marine organisms have been extensively investigated. However, the potential mechanisms underlying their interactive effects on physiological processes and metabolism remain poorly understood, especially regarding periodic MHWs in real-world conditions. In this study, the effects of nano-TiO2 (at concentrations of 0, 25, and 250 μg/L) and periodic MHWs on the condition index (CI) and underlying metabolic mechanisms were investigated in mussels (Mytilus coruscus). The results showed that mussels try to upregulate their respiration rate (RR) to enhance aerobic metabolism (indicated by elevated succinate dehydrogenase) under short-term nano-TiO2 exposure. However, even at ambient concentration (25 μg/L), prolonged nano-TiO2 exposure inhibited ingestion ability (decreased clearance rate) and glycolysis (inhibited pyruvate kinase, hexokinase, and phosphofructokinase activities), which led to an insufficient energy supply (decreased triglyceride, albumin, and ATP contents). Repeated thermal scenarios caused more severe physiological damage, demonstrating that mussels are fragile to periodic MHWs. MHWs decreased the zeta potential of the nano-TiO2 particles but increased the hydrodynamic diameter. Additionally, exposure to nano-TiO2 and periodic MHWs further affected aerobic respiration (inhibited lactate dehydrogenase and succinate dehydrogenase activities), metabolism (decreased RR, activities of respiratory metabolism-related enzymes, and expressions of PEPCK, PPARγ, and ACO), and overall health condition (decreased ATP and CI). These findings indicate that the combined stress of these two stressors exerts more detrimental impact on the physiological performance and energy metabolism of mussels, and periodic MHWs exacerbate the toxicological effects of ambient concentration nano-TiO2. Given the potential worsening of nanoparticle pollution and the increase in extreme heat events in the future, the well-being of mussels in the marine environment may face further threats.

Involvement of BcNAC083 in 1-MCP-induced delayed yellowing of pak choi (Brassica rapa subsp. Chinensis) through the regulation of reactive oxygen species metabolism and inhibition of major chlorophyll catabolic genes

Leaf yellowing is a typical senescence feature in postharvest pak choi. Plant-specific NAC transcription factors (TFs) are crucial regulators of plant senescence. In this study, a senescence-related NAC TF, BcNAC083, was investigated to identify its transcriptional regulation effects on the yellowing of pak choi by treatment with 1-methylcyclopropene (1-MCP). Applying 5.0 µL L–1 1-MCP alleviated yellowing and maintained a low respiration rate, high chlorophyll content, and relatively complete chloroplast structure in pak choi during storage. However, 10 μL L–1 ethylene accelerated yellowing, as demonstrated by the higher respiration rate, lower chloroplast structural integrity and chlorophyll content. In contrast to ethylene treatment, 1-MCP suppressed the activity of the reactive oxygen species (ROS) metabolism-related enzymes superoxide dismutase (SOD), catalase (CAT), ascorbic peroxidase (APX), and lipoxygenase (LOX), and correspondingly reduced superoxide anion (O2·–), hydrogen peroxide (H2O2), and malondialdehyde (MDA) accumulation. 1-MCP treatment also suppressed the transcript levels of ROS metabolic genes, including BcRBOHC, BcRBOHD, BcSOD, BcCAT, BcAPX, and BcLOX, and inhibited the activity of the chlorophyll catabolic enzymes chlorophyllase (CLH), Mg-de-chelatase, and pheophytin pheophorbide hydrolase (PPH) and transcript levels of associated genes, including BcCLH1/2, BcPPH1/2, BcPAO, BcNYC1, BcSGR1/2, BcHCAR, BcRCCR, and BcCAO. The expression of BcNAC083 was probably induced by ROS and was downregulated by 1-MCP and upregulated by ethylene. Molecular biology experiments determined that BcNAC083 bound directly to and activated the promoters of the chlorophyll metabolism-related genes BcCLH1, BcPPH1, BcNYC1, and BcHCAR. Importantly, transient overexpression of BcNAC083 stimulated the endogenous genes (NbCLH1, NbPPH1, NbNYC1, and NbHCAR) expression in tobacco leaves, resulting in rapid chlorophyll breakdown and leaf degreening. Treatment with 1-MCP may alleviated the yellowing of pak choi by inhibiting ROS accumulation and the transcriptional regulation of BcNAC083, which had positive regulatory effects on major chlorophyll metabolic genes.

Acute waterborne cadmium exposure induces liver ferroptosis in Channa argus

The impact of cadmium (Cd) toxicity on fish liver injury has received much attention in recent years. Currently, autophagy, apoptosis and endoplasmic reticulum stress were reported in Cd exposed fish liver, and if there are other mechanisms (such as ferroptosis) and relevant signaling pathways involved in fish remains unknown. An experiment was conducted to investigate Cd toxicity in Channa argus (Cantor, 1842) exposed to 0, 1.0, and 2.0 mg Cd/L of water for 96 h. Cd disrupted the structure of mitochondria in the liver. Besides, Cd induced ferroptosis by significantly increasing the level of Fe2+, ROS, MDA and significantly decreasing the level of Ferritin, GSH, GSH-Px, GPX4, GST and SOD (p < 0.05 in all cases). In addition, the mRNA expression of ferroptosis related genes, gpx4 and slc7a11, were significantly downregulated by Cd. Moreover, Cd exposure significantly inhibited the Nrf2/Keap1 signaling pathway, one of the pathways involved in ferroptosis, by upregulating the mRNA levels of keap1a and keap1b, and downregulating the mRNA levels of nrf2 and its target genes (ho-1, nqo1 and cat). Cd exposure also caused extensive accumulation of vacuoles and lipid droplets in liver, as well as an increase in triglyceride content. Cd significantly affected lipid metabolism related enzyme activity and gene expression, which were also regulated by Nrf2/Keap1 signaling pathway. In summary, these results indicate that ferroptosis is a mechanism in waterborne Cd exposed fish liver injury via the Nrf2/Keap1 signaling pathway and the Cd induced hepatic steatosis is also modulated by Nrf2/Keap1 pathway at the whole-body level in fish. These findings provide new insights into the fish liver injury and molecular basis of Cd toxicity.

Optimization of cultivation system for Bacillus Siamese strain and its biocontrol effect on postharvest Colletotrichum gloeosporioides

In this study, we established an effective cultivation system for Bacillus Siamese (N-1), identified key antibacterial substances, and explored the underlying mechanisms of optimized N-1 fermentation for controlling pathogens. Our results demonstrate that the optimized system significantly enhanced the inhibitory effect of N-1 on five types of pathogens affecting tropical fruits. Additionally, non-targeted metabolomics analysis and ultra-high performance liquid chromatography triple quadrupole mass spectrometer system (UPLC-MS/MS) revealed the presence of lipopeptides, including Surfactin and FengycinA, in the optimized fermentation. Furthermore, in vitro experiments indicated that lipopeptide treatment (LT) at a concentration of 3.125 mg mL−1 significantly distorted the hyphae, decreased the activity of antioxidant enzymes, increased the levels of reactive oxygen species (ROS), malondialdehyde (MDA) and hydrogen peroxide (H2O2) contents, and enhanced protein exosmosis rate of C. gloeosporioides. Moreover, LT markedly reduced the activities of energy metabolism-related enzymes, including succinate dehydrogenase (SDH), Na+K+-ATPase, and Ca2+Mg2+-ATPase, as well as ATP content, leading to apoptosis of C. gloeosporioides. In vivo experiments revealed that LT significantly inhibited the expansion of disease spots caused by C. gloeosporioides in mango fruit. These findings suggest that the established culture system could serve as a potential strategy to enhance biocontrol efficacy and prevent fruit decay.

Mitochondrial VDAC1 Silencing in Urethane-Induced Lung Cancer Inhibits Tumor Growth and Alters Cancer Oncogenic Properties.

Alterations in cellular metabolism are vital for cancer cell growth and motility. Here, we focused on metabolic reprogramming and changes in tumor hallmarks in lung cancer by silencing the expression of the mitochondrial gatekeeper VDAC1. To better mimic the clinical situation of lung cancer, we induced lung cancer in A/J mice using the carcinogen urethane and examined the effectiveness of si-m/hVDAC1-B encapsulated in PLGA-PEI nanoparticles. si-m/hVDAC1-B, given intravenously, induced metabolism reprogramming and inhibited tumor growth as monitored using MRI. Mice treated with non-targeted (NT) PLGA-PEI-si-NT showed many large size tumors in the lungs, while in PLGA-PEI-si-m/hVDAC-B-treated mice, lung tumor number and area were markedly decreased. Immunofluorescence staining showed decreased expression of VDAC1 and metabolism-related proteins and altered expression of cancer stem cell markers. Morphological analysis showed two types of tumors differing in their morphology; cell size and organization within the tumor. Based on specific markers, the two tumor types were identified as small cell (SCLC) and non-small cell (NSCLC) lung cancer. These two types of tumors were found only in control tumors, suggesting that PLGA-PEI-si-m/hVDAC1-B also targeted SCLC. Indeed, using a xenograft mouse model of human-derived SCLC H69 cells, si-m/hVDAC1-B inhibited tumor growth and reduced the expression of VDAC1 and energy- and metabolism-related enzymes, and of cancer stem cells in the established xenograft. Additionally, intravenous treatment of urethane-induced lung cancer mice with the VDAC1-based peptide, Retro-Tf-D-LP4, showed inhibition of tumor growth, and decreased expression levels of metabolism- and cancer stem cells-related proteins. Thus, silencing VDAC1 targeting both NSCLC and SCLC points to si-VDAC1 as a possible therapeutic tool to treat these lung cancer types. This is important as target NSCLC tumors undergo transformation to SCLC.

Exogenous leptin alleviates glutamate-excitotoxic injury caused by cerebral ischemia–reperfusion in mice by affecting the expression of glutamate transporters

Ischemic stroke is characterized by high morbidity and mortality and a lack of effective therapeutic interventions. Leptin plays an important role in regulating oxidative stress, angiogenesis, hematopoiesis, etc. Although recent studies have found a neuroprotective effect of leptin, little is known about its role in cerebral ischemia. This study explores the possible roles and potential preventative mechanisms of leptin in cerebral ischemia–reperfusion injury (CIRI). An in vivo middle cerebral artery occlusion/reperfusion (MCAO/R) mouse model was used to replicate the CIRI model, low (0.5 mg/kg), medium (1 mg/kg) and high (2 mg/kg) concentrations of leptin were injected intraperitoneally immediately after inserting the embolic line. After 1.5 h of ischemia and 24 h of reperfusion, we examined the neural function of the mice, collected brain tissue for histological examination, and screened for the optimal concentrations of leptin intervention. On this basis, we observed the changes of cortical apoptosis injury, intracellular calcium fluorescence intensity and astrocyte glial fibrillary acidic protein (GFAP) expression and morphological changes. In addition, we also tested the expression of transporters and metabolism-related enzymes (VGLUT-1, VGLUT-2, GLAST, GLT-1, GS, ATP1α1), the expression of inflammatory factors and the content of glutamate (Glu). Compared with the I/R group, we found that leptin improved neurological deficits, reduced the area of infarcts, maintained the normal morphology of astrocytes (AST), downregulated the expression of VGLUT-1, and upregulates the expression of GLT-1 and GLAST, thereby reducing the content of Glu in the synaptic cleft. Our studies suggest that leptin may have a neuroprotective effect by decreasing the excitotoxicity of glutamate.

Effects of fishing stress on fatty acid and amino acid composition and glycolipid metabolism in triploid rainbow trout

Triploid Oncorhynchus mykiss is an important economic fish worldwide. Fishing stress can affect its growth and meat quality. This study first explored the effects of fishing stress on fatty acid and amino acid in triploid O. mykiss. Results showed fishing stress significantly reduced the content of docosadienoic acid, Gly, Arg, and DAA (P < 0.05). Targeted lipidomics analysis furthered suggested that some lipid molecules belonging to TG, DG, PC, Cer, ChE, and So were significantly up-regulated; while some lipid molecules belonging to Cer, LPE, LPC, PS, PC, and SM were significantly down-regulated, suggesting an accelerated glycolipid metabolism. Eventually, the glycolipid metabolism-related enzyme activity and gene expressions were examined, and the results indicated that O. mykiss was anti-oxidative stress by affecting relevant glycolipid metabolism signaling pathways and participating in cellular redox homeostasis. Findings of this study provide a theoretical foundation for further investigation into the mechanisms through which fishing stress affects O. mykiss.

Multi-omics reveals the ecological and biological functions of Enterococcus mundtii in the intestine of lepidopteran insects

Insect guts offer unique habitats for microbial colonization, with gut bacteria potentially offering numerous benefits to their hosts. Although Enterococcus has emerged as one of the predominant gut commensal bacteria in insects, its establishment in various niches within the gut has not been characterized well. In this study, Enterococcus mundtii was inoculated into the silkworm (Bombyx mori L.) to investigate its biological functions. Genome-based analysis revealed that its successful colonization is related to adherence genes (ebpA, ebpC, efaA, srtC, and scm). This bacterium did not alter the activities of related metabolic enzymes or the intestinal barrier function. However, significant changes in the gene expressions levels of Att2, CecA, and Lys suggest potential adaptive mechanisms of host immunity to symbiotic E. mundtii. Moreover, 16S metagenomics analysis revealed a significant increase in the relative abundance of E. mundtii in the intestines of silkworms following inoculation. The intestinal microbiome displayed marked heterogeneity, an elevated gut microbiome health index, a reduced microbial dysbiosis index, and low potential pathogenicity in the treatment group. Additionally, E. mundtii enhanced the breakdown of carbohydrates in host intestines. Overall, E. mundtii serves as a beneficial microbe for insects, promoting intestinal homeostasis by providing competitive advantage. This characteristic helps E. mundtii dominate complex microbial environments and remain prevalent across Lepidoptera, likely fostering long-term symbiosis between the both parties. The present study contributes to clarifying the niche of E. mundtii in the intestine of lepidopteran insects and further reveals its potential roles in their insect hosts.