Differences In Metabolic Pathways Research Articles

Integrated Transcriptomic and Metabolomic Analysis of Rat PASMCs Reveals the Underlying Mechanism for Pulmonary Arterial Hypertension.

Pulmonary arterial hypertension (PAH) is a kind of pulmonary vascular lesion characterized by vasoconstriction and reshaping of small pulmonary arteries, ultimately resulting in increased pulmonary artery pressure and pulmonary vascular resistance, and eventually leading to right ventricular failure and death. This study was aimed to construct a platelet-derived growth factor BB (PDGF-BB)-induced rat pulmonary artery smooth muscle cells (PASMCs) model and conduct a combined transcriptomic and metabolomic analysis to identify proliferation-related targets, thereby enhancing understanding of the pathogenesis underlying PAH. Rat PASMCs were isolated and cultured in the presence or absence of PDGF-BB for 24 hours. Cells were collected for transcriptomics and metabolomics investigations. A total of 1288 differentially expressed genes (572 up-regulated and 716 down-regulated) were identified in PDGF-BB-treated rat PASMCs compared to control cells. Subsequently, Gene ontology (GO) enrichment analysis revealed that 791 enriched GO terms were significantly enriched in PDGF-BB treated cells. Similarly, 294 differential metabolic pathways were enriched in PDGF-BB treated cells according to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, Gene Set Enrichment Analysis (GSEA) were performed on the differentially expressed genes (DEGs). It turned out that 7219 gene sets were more enriched in PDGF-BB treated cells. In addition, a total of 28 secondary differential metabolites were identified in PDGF-BB-treated rat PASMCs compared to control cells (p-value < 0.05 and VIP > 1). We speculate that Mylk, Pla2g4a, Gucy1b1, Adcy8, Adcy4, Gucy1a2, Col3a1, and Plcb4 are potential targets for the treatment of PAH.

Analysis of ischemic stroke biomarkers based on non-targeted metabolomics

Biomarkers for ischemic stroke (IS) are yet to fulfill clinical requirements. This study used non-targeted metabolomics to investigate differential metabolites and metabolic pathways in plasma and brain tissue following IS, with the aim of identifying new potential biomarkers and therapeutic targets. Twelve Tibetan miniature pigs were randomly assigned to a model- or sham-operation group. An electrocoagulation-based anterior temporal approach was employed to occlude the middle cerebral artery, thereby creating a model for IS. Plasma and brain tissue samples were collected 36 h post-surgery and analyzed using liquid chromatography-mass spectrometry. Principal component and partial least squares discriminant analyses were used to screen for differential metabolites and exclude exogenous metabolites at p<0.05. Compounds were classified according to the HMDB (Human Metabolome Database), and subjected to KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway and VIP (variable importance in projection) analyses. Plasma metabolomics revealed that 86 metabolites were upregulated while 149 were downregulated, with (Z)-3-oxo-2-(2-pentenyl)-1-cyclopentylacetic acid, trans-cinnamic acid and cinnamoylglycine determined to be significant metabolites. Fifty-eight differential metabolites were upregulated in brain tissue and 53 were downregulated, with 2,3-dihydroflavon-3-ol, guanidinoacetic acid (GAA), N-acetyl-D-tryptophan, oxidized glutathione, 2-hydroxyquinoline, and N-acetyl-L-aspartate (NAA) identified as significant metabolites. Organic acids and derivatives, lipids and lipid-like molecules, organoheterocyclic compounds, and organic oxygen compounds were found to be common compound categories among the top five types of compound in both plasma and brain tissue. Common metabolic pathways in plasma and brain tissue include amino acid metabolism, digestive system, cancer overview, and lipid metabolism pathways, with the (Z)-3-oxo-2-(2-pentenyl)-1-cyclopentylacetic acid, GAA, oxidized glutathione, and NAA metabolites serving as potential biomarkers. This study provides a theoretical foundation for the early screening and development of clinical treatment strategies for IS.

Comparative metabolomic analysis of Haematococcus pluvialis during hyperaccumulation of astaxanthin under the high salinity and nitrogen deficiency conditions.

Revealing the differences of metabolite profiles of H. pluvialis during hyperaccumulation of astaxanthin under the high salinity and nitrogen deficiency conditions was the key issues of the present study. To investigate the optimum NaCl and NaNO3 concentration and the corresponding metabolic characteristic related to the astaxanthin accumulation in H. pluvialis, a batch culture experiment was conducted. The results indicated that 7.5g·L- 1 and 0g·L- 1 (nitrogen deficiency) were the optimum NaCl and NaNO3 levels for the astaxanthin accumulation respectively, under which the highest astaxanthin contents reached up to 7.51mg·L- 1 and 5.60mg·L- 1. A total of 132 metabolites were identified using LC-MS/MS technique, among which 30 differential metabolites with statistical significance were highlighted. Subsequently, 18 and 10 differential metabolic pathways in the high salinity (HS) and nitrogen-deficient (ND) treatments were extracted and annotated respectively. The values of Fv/Fm, Yield and NPQ were all at the highest level in the ND group during the experiment. The levels of the metabolites in the ND group were almost lower than those both in the control (CK) and HS group, while which in the HS group were substantially at the higher or close levels compared to the CK group. Finally, 7 metabolic markers related to the astaxanthin accumulation were highlighted in the HS and ND group respectively. L-Proline, L-Aspartate, Uridine 5'-monophosphate (UMP), Succinate, L-2-Hydroxygluterate, L-Valine and Inosine 5'-monophosphate (IMP) were identified as the metabolic markers in the HS group, whose fold change were 0.85, 4.14, 0.31, 0.66, 3.10, 1.32 and 0.30. Otherwise, the metabolic markers were Glyceric acid, Thymine, sn-Glycerol 3-phosphate, Glycine, Allantoic acid, L-Valine and IMP in the ND group, with the fold change 0.23, 2.11, 0.38, 0.41, 0.50 and 2.96 respectively. The results provided the comparative metabolomic view of astaxanthin accumulation in H. pluvialis under the different cultivation conditions, moreover showed a novel insights into the astaxanthin production.

Integrating Metabolomics and Genomics to Uncover the Impact of Fermented Total Mixed Ration on Heifer Growth Performance Through Host-Dependent Metabolic Pathways.

With the increasing demand for enhancing livestock production performance and optimizing feed efficiency, this study aimed to investigate the effects of fermented total mixed ration (FTMR) containing different proportions of rice straw and sheath and leaves of Zizania latifolia on systemic nutrient metabolism and oxidative metabolism under host genetic regulation and on growth performance of heifers. A total of 157 heifers aged 7-8 months were selected, and their hair was collected for whole-genome sequencing. They were randomly assigned into four groups of 18 to 21 cattle each and fed FTMR containing varying levels of rice straw (21% in LSF, 28% in MSF, 35% in HSF) or 31% sheath and leaves of Zizania latifolia (ZF) for a two-month period. At the end of trial, blood and urine samples were collected to measure biochemical indexes and metabolomics. The results showed that high rice straw content and ZF diets could increase blood glucose and non-protein nitrogen in heifers, that is, blood glucose and urea nitrogen levels in HSF and ZF groups were higher than those in LSF and MSF groups (p < 0.05). Meanwhile, the two diets could improve the antioxidant level of heifers. Urine metabolomics analysis between the groups identified three differential metabolic pathways, including 11 metabolites. Among them, l-homoserine and o-acetylserine had significant SNPs associated with them, which promoted glutathione metabolism. Although there was no significant effect of diet on heifers' average daily gain (ADG) in body weight (p > 0.05), there was substantial inter-individual variation in metabolites among all animals, as further correlation analyses illustrated. Twenty-eight metabolites were significantly associated with ADG (R > 0.3, p < 0.05). Four of them were identified as biomarkers, primarily regulating energy metabolism and oxidative balance. In conclusion, feeding HSF and ZF FTMR enhances glutathione metabolism and antioxidant capacity in heifers, positioning key metabolites as candidates for ADG markers. This integrative omics approach underscores the potential for enhancing livestock productivity and promoting sustainable agricultural practices.

Exploring changes in metabolites and fecal microbiota of advanced gastric cancer based on plasma metabolomics and 16S rDNA sequencing.

Based on plasma metabolomics and 16S rDNA sequencing to evaluate the changes in metabolites and fecal microbiota of advanced gastric cancer. Firstly, plasma metabolomics was used to screen for differential metabolites and metabolic pathways in gastric cancer. Then, 16S rDNA sequencing was performed on fecal samples to study the differential intestinal microbiota in gastric cancer patients. Finally, conduct a correlation analysis between them. A total of 152 differential metabolites were identified, and we screened 10 of them. All metabolites were enriched into 42 differential metabolic pathways, of which 13 have P values less than 0.05. 16S rDNA sequencing showed significant differences in 4 microbial communities at the phylum level. There are significant differences in 23 communities at the genus level. We focus on Lactobacillales, Lactobacillus, Streptococcus, Veillonella, Bacilli and Megasphaera. Correlation analysis shows that the intestinal microbiota and plasma metabolites jointly affect the occurrence and development of gastric cancer. For the first time, we comprehensively used plasma metabolomics and 16S rDNA sequencing to reveal the changes and correlations between metabolites and intestinal microbiota in advanced gastric cancer. We have discovered new potential biomarkers for gastric cancer. This deepens our understanding of the physiological and pathological mechanisms of advanced gastric cancer and helps to improve the diagnosis and treatment of advanced gastric cancer.

Microbiota dynamics and metabolic mechanisms in fermented sausages inoculated with Lactiplantibacillus plantarum and Staphylococcus xylosus.

Lactiplantibacillus plantarum and Staphylococcus xylosus are common starters for fermented sausages. Several studies have demonstrated the impact of these two strains on the quality of fermented sausages. However, the mechanism underlying the effects of these two microorganisms on co-cultivation in sausages remains unclear. This study aimed to investigate the effects of inoculation with various combinations of starters on the microbial communities and metabolic profiles of fermented sausages. High-throughput sequencing revealed that, during sausage fermentation, Firmicutes was the dominant bacterial phylum, and the primary microorganisms were Lactococcus, Staphylococcus, Lactobacillus, and Pseudomonas. On the last day of fermentation, the highest abundance of Staphylococcus was observed in the co-inoculation group. Furthermore, inoculated fermentation effectively inhibited the growth of pathogenic and spoilage bacteria. Metabolomic analysis of the four groups of samples identified 208 metabolites in positive ion mode and 109 in negative ion mode. A total of 31 differential metabolites were identified (P<0.05, variable importance in the projection >1.5), primarily benzene and substituted derivatives, carboxylic acids and derivatives, and fatty acyls. Five crucial differential metabolites (subaphylline, naringenin, 1-hexadecanol, beta-alanyl-L-lysine, and 3'-AMP) were identified as potential biomarkers for fermented sausages. Key differential metabolite metabolic pathways indicated that L. plantarum YR07 dominated in metabolite regulation during sausage fermentation, and S. xylosus Y-18 downregulated the fatty acid degradation pathway, which also affected the metabolism of fermented sausages. Co-cultivation of the two bacteria exhibited a synergistic effect on the metabolism of the fermented sausages. This study offers further insights into improving the quality of fermented sausages, thereby establishing a theoretical foundation for the production of excellent fermenters.

Untargeted metabolite profiling reveals metabolic disorders in livers of rats with mepiquat exposure.

Mepiquat is a contaminant produced in thermal-processed food. It can induce spleen and liver injury. However, the mechanism that mepiquat induced hepatotoxicity remains unclear. In this study, a ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS)-based metabolomic analysis of livers in rats was performed to explore metabolic alterations. Our results demonstrated that there were 36 differential metabolites. Eleven major disordered metabolic pathways were found. Network analysis of differential metabolites and metabolic pathways indicated that glutamic acid was a possible key upregulated metabolite. It participated in nine metabolic networks. Glutathione and l-proline may be key downregulated metabolites. They were involved in eight and seven metabolic pathways, respectively. Compared with the control group, serum concentrations of alanine aminotransferase, aspartate aminotransferase, and total bile acid in high dosage group increased significantly. In addition, histopathological analysis showed liver injury in rats with mepiquat exposure. These data were consistent with results of metabolomics. Our results offered new insights for molecular mechanism of liver toxicity induced by mepiquat. PRACTICAL APPLICATION: Mepiquat is a contaminant formed in thermal-processed food. It can induce spleen and liver injury. Our results offered new insights for molecular mechanism of liver toxicity induced by mepiquat. It will provide important information for official limits of mepiquat in foods.

Buyang huanwu decoction inhibits the activation of the RhoA/Rock2 signaling pathway through the phenylalanine metabolism pathway, thereby reducing neuronal apoptosis following cerebral ischemia-reperfusion injury.

Buyang Huanwu Decoction (BYHWD) exerts its anti-cerebral ischemia effects through multiple pathways and targets, although its specific mechanisms remain unclear. Ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF-MS) metabolomics and other methods were employed to investigate the role of BYHWD in inhibiting neuronal apoptosis following cerebral ischemia-reperfusion by modulating the RhoA/Rock2 pathway. A rat model of exhaustion swimming combined with middle cerebral artery occlusion (ES+I/R) was established to evaluate the intervention effects of Buyang Huanwu Decoction on cerebral ischemia-reperfusion. This was assessed using neurological function scores, Qi deficiency and blood stasis syndrome scores, HE staining, Nissl staining and TT staining. Differential metabolites and metabolic pathways associated with cerebral ischemia-reperfusion were identified using UPLC-QTOF-MS metabolomics, with key differential metabolites validated through ELISA. Molecular docking techniques were employed to predict interactions between the key differential metabolite, hippuric acid, and its primary downstream pathways. Finally, the levels of neurocellular apoptosis, as well as key molecules in the RhoA/Rock2 signaling pathway and the mitochondrial apoptosis pathway, were measured. The results indicated that the primary differential metabolites associated with BYHWD's protective effects against ischemia-reperfusion injury were hippuric acid, lysophosphatidic acid, and lysophosphatidylethanolamine, with the main metabolic pathway being phenylalanine metabolism. Molecular docking studies demonstrated that malonic acid exhibited a strong affinity for proteins related to the RhoA/Rock2 signaling pathway and the mitochondrial apoptosis pathway.Furthermore, we found that BYHWD treatment significantly decreased the apoptosis rate of cells following cerebral ischemia-reperfusion and inhibited the expression of key molecules in both the RhoA/Rock2 signaling pathway and the mitochondrial apoptosis pathway in brain tissue. BYHWD ameliorated brain tissue injury after cerebral ischemia/reperfusion in rats with qi deficiency and blood stasis. The underlying mechanism may involve BYHWD's inhibition of the RhoA/Rock2 signaling pathway activation through modulation of the phenylalanine metabolism pathway, thereby reducing neuronal apoptosis mediated by the mitochondrial apoptosis pathway.

Regulation of ROS metabolism in macrophage via xanthine oxidase is associated with disease progression in pulmonary tuberculosis.

Pulmonary tuberculosis (PTB) exacerbation can lead to respiratory failure, multi-organ failure, and symptoms related to central nervous system diseases. The purpose of this study is to screen biomarkers and metabolic pathways that can predict the progression of PTB, and to verify the role of the metabolic enzyme xanthine oxidase (XO) in the progression of PTB. To explore the biomarkers and mechanisms underlying the progression of PTB, plasma metabolomics sequencing was conducted on patients with severe PTB, non-severe PTB, and healthy individuals. Screening differential metabolites and metabolic pathways that can predict the progression of PTB, and verifying the function and mechanism of action of XO through experiments. The purine metabolism, sphingolipid metabolism, and amino acid metabolism between the three groups differ. In patients with severe PTB, the levels of xanthosine and hypoxanthine are increased, while the levels of D-tryptophan, dihydroceramide and uric acid are decreased. Inhibition of XO activity has been observed to reduce the levels of tumor necrosis factor (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6), as well as to suppress the production of reactive oxygen species (ROS) and the activation of the NF-κB pathway, while also promoting the growth of MTB within cells. D-tryptophan, xanthosine, and dihydroceramide can be utilized as biomarkers for progression of PTB, assisting in the evaluation of disease progression, and XO stands out as a potential therapeutic target for impeding the progression of PTB.

Antioxidative and Cytoprotective Effects of Rosa Roxburghii and Metabolite Changes in Oxidative Stress-Induced HepG2 Cells Following Rosa Roxburghii Intervention.

Rosa Roxburghii (RR), a traditional Chinese medicinal fruit, is rich in bioactive substances that make it a potential natural antioxidant resource. This research aimed to study the antioxidant properties of RR by in vitro experiments and through intracellular assessment in H2O2-induced HepG2 cells. A non-targeted metabolic analysis was conducted to indicate changes in intracellular and extracellular metabolites. Differential metabolites and metabolic pathways were explored using PCA, PLS-DA, and KEGG pathway analysis. The results showed that RR rich in bioactive substances exhibited a significant antioxidative property in vitro and intracellularly. This property may be achieved by scavenging free radicals, increasing the activity of catalase (CAT), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and the levels of bicinchoninic acid (BCA) while reducing the reactive oxygen species (ROS) generation. This study identified 13 differential metabolites intracellularly and 7 extracellularly, among which the key differential metabolites included D-glucopyranose, D-mannose, fructose, citric acid, malic acid, cholesterol, and cholestenone. These key metabolites primarily regulated glucose-related metabolism, the citrate cycle, and the primary bile acid biosynthesis pathway in H2O2-induced HepG2 cells. These findings provide potential application evidence of RR in the development of natural resources for functional foods.

Differences in toxicity and serum metabolomics of ethyl acetate fraction of Euphorbiae Pekinensis Radix before and after vinegar processing

This study aimed to investigate the toxicity differences of the ethyl acetate fraction of Euphorbiae Pekinensis Radix before and after vinegar processing and explore the detoxification mechanism of vinegar processing using non-targeted metabolomics. The changes in terpenoid components in the ethyl acetate fraction before and after vinegar processing were analyzed using UFLC-Q-TOF-MS. Normal rats were orally administered the raw and vinegar-processed ethyl acetate fractions of Euphorbia Pekinensis Radix. The toxicity differences in ethyl acetate fractions of Euphorbia Pekinensis Radix before and after vinegar processing were evaluated by pathological morphology, serum liver and kidney function, oxidative damage, and inflammatory injury indicators, and apoptosis factors. Serum metabolomics technology was utilized to identify changes in endogenous metabolites. Principal component analysis(PCA) and partial least squares discriminant analysis(PLS-DA) were employed to identify differential metabolites and metabolic pathways related to the detoxification of vinegar-processed Euphorbia Pekinensis Radix. The content of terpenoid components in the ethyl acetate fraction of Euphorbia Pekinensis Radix significantly decreased after vinegar processing. Histopathological sections and various indicators revealed that both the raw and vinegar-processed ethyl acetate fractions of Euphorbia Pekinensis Radix could induce toxicity in the liver, stomach, and intestine, with a reduction after vinegar processing. The toxicity was associated with oxidative damage, inflammatory injury, and apoptosis. A total of 13 differential metabolites and 5 main metabolic pathways related to Euphorbia Pekinensis Radix toxicity were identified by serum metabolomics. PCA and PLS-DA score plots indicated that both the raw and vinegar-processed ethyl acetate fractions disrupted the endogenous metabolic profiles in rats, mainly concentrating on lipid metabolism, primary bile acid biosynthesis, and arachidonic acid metabolism, with vinegar processing alleviating these metabolic disruptions. Therefore, the ethyl acetate fraction of Euphorbia Pekinensis Radix possesses liver, stomach, and intestinal toxicity, and vinegar processing reduces its toxicity by decreasing the content of terpenoid components. The detoxification mechanism may be related to alleviating oxidative damage, inflammatory injury, apoptosis, and improving lipid metabolism.

Physiology and transcriptome profiling reveal the drought tolerance of five grape varieties under high temperatures

The evaluation of plant stress tolerance and the screening of key regulatory genes under the combined stresses of high temperature and drought are important for the study of plant stress tolerance mechanisms. In this study, the drought tolerance of five grape varieties was evaluated under high-temperature conditions to screen key genes for further exploration of resistance mechanisms. By comparing and analysing the morphological characteristics and physiological indicators associated with the response of grapevines to drought stress and integrating them with the membership function to assess the strength of their drought tolerance, the order of drought tolerance was found to be as follows: 420A>110R>CS>fercal>188-08. To further analyse the mechanism of differences in drought tolerance, transcriptomic sequencing was performed on the drought-tolerant cultivar 420A, the drought-sensitive cultivar 188-08 and the control cultivar CS. The functional analysis of differential metabolic pathways indicated that the differentially expressed genes were enriched mainly in biological that 420A had higher antioxidant activity. Moreover, the transcription factors which differentially expressed were also analysed in the five grape varieties, and several genes, such as VvAGL15, VvLBD41, and VvMYB86, appeared to be closely related to drought tolerance, suggesting their potential involvement in the regulation of grapevine drought tolerance and their value in drought tolerance research.

Study on serum metabolomics characteristics of obese patients with erectile dysfunction.

Erectile dysfunction (ED) is a common male sexual health problem that can be associated with obesity. This study aimed to identify serum metabolic differences and pathways related to ED in obese men using non-targeted metabolomics techniques. We included 54 obese male patients with (n = 27) and without (n = 27) ED. We collected 5 mL of fasting elbow vein blood and analyzed serum metabolites using ultra-high-performance liquid chromatography-mass spectrometry. Multivariate statistical methods (principal component analysis and orthogonal partial least squares discriminant analysis) were used to identify differential metabolites between the groups. Finally, pathway analysis using the Kyoto encyclopedia of genes and genomes database identified 4 differential metabolic pathways in obese men with ED compared to obese men without ED. A total of 77 differential metabolites were identified in obese men with ED compared to the control group (obese men without ED) using a threshold of variable importance in the projection > 1 and P < .05. Pathway analysis revealed 4 main differences: glycine, serine and threonine metabolism, glycerophospholipid metabolism, aminoacyl-tRNA biosynthesis, and D-glutamine and D-glutamate metabolism. Specific metabolites associated with these pathways included betaine aldehyde, choline, L-threonine, phosphatidylcholine, L-serine, and D-glutamine. Our findings suggest abnormalities in fatty acid metabolism, phospholipid metabolism, and amino acid metabolism between obese men with and without ED. Metabolites such as betaine aldehyde, choline, L-threonine, phosphatidylcholine, L-serine, and D-glutamine may be potential biomarkers for distinguishing obese men with ED.

Metabolomic analysis of the browning and browning inhibition pulp of Fuji apple at different ripening stages

Apples at different ripening stages display distinct processing properties and browning characteristics in products. This study aimed to identify differential metabolites and metabolic pathways of browning pulp and browning inhibition pulp of Fuji apple across four ripening stages (M1–M4), corresponding to days after full bloom (DAFB: 148, 155, 162, and 171). We investigated the physicochemical characteristics, browning-related enzyme activities, and metabolomic profiles of apples at the four stages, revealing defensive metabolic changes associated with browning throughout ripening. Specifically, the browning rate in apples increased with ripening, peaking before declining in the late stages, and exhibited positive correlations with activities of polyphenol oxidase (PPO), peroxidase (POD), and catalase (CAT), and negative correlations with superoxide dismutase (SOD), phenylalanine ammonia-lyase (PAL), and H2O2 levels. Metabolomic profiling identified 315, 393, and 243 distinct metabolites in fresh control (FC), browning pulp (BR), and browning-inhibited pulps (CM) of apple at the four ripening stages, respectively. KEGG enrichment analysis revealed that FC and BR of the four ripening apples were differentiated by phenylpropanoid biosynthesis, while CM was featured by flavonoid biosynthesis. The characterized differential metabolites in BR were changed to α-linolenic acid metabolism with ripening, while those in CM were from flavonoid and phenylpropanoid biosynthesis. Furthermore, the in-depth analysis revealed 40 significantly differential metabolites associated with amino acid, nucleotide, coenzyme, and vitamin metabolism, potentially modulating the stress-responsive cellular redox balance. This research provides critical insights for browning prevention in the processing of fruits at various ripening stages.

Integration of metabolomics and transcriptomics to reveal metabolic characteristics and key targets associated with lncRNA Vof-16 in H19-7 cells

Cognitive disorders represent one of the most common chronic complications of diabetes. Our previous study has demonstrated that long non-coding RNA (lncRNA) Vof-16 is upregulated in the hippocampal tissue of streptozotocin (STZ)-induced diabetic rats. Despite this finding, the specific roles and underlying mechanisms of lncRNA Vof-16 in diabetes-related cognitive dysfunction remain largely unexplored. To elucidate the mechanism involved, lncRNA Vof-16 was overexpressed in rat hippocampal cell line H19-7 through lentivirus transfection. We integrated metabolomics and transcriptomics approaches to identify potential targets and metabolic pathways influenced by lncRNA Vof-16. Key proteins and pathways were subsequently validated using western blotting and immunofluorescence staining. Transcriptomics indicated that lncRNA Vof-16 overexpression may modulate autophagic activity in H19-7 cells. Metabolomic profiling revealed that the primary differential metabolic pathways included trehalose degradation, tryptophan metabolism, vitamin B6 metabolism, glycolysis, pterine biosynthesis, and the pentose phosphate pathway. Ingenuity Pathway Analysis (IPA) of gene-metabolite networks demonstrated that the high lncRNA Vof-16 expression group exhibited a significantly higher association with autophagy compared to the low lncRNA Vof-16 expression group. Western blot results confirmed that lncRNA Vof-16 overexpression led to decreased protein expression levels of ATG3 and ATG12. Specifically, lncRNA Vof-16 reduces autophagy in hippocampal neurons by targeting the elevated levels of phospho-p70S6K, a downstream effector of mTORC1, potentially contributing to the pathogenesis of diabetic cognitive impairment. The construction of gene-metabolite networks may offer promising new strategies for addressing the growing issue of diabetic cognitive impairment.

Insights into flavor quality and metabolites profiles of fresh cheese with different probiotics by SPME-GC-MS and untargeted metabolomics

In this study, fresh cheeses produced with four novel probiotics (Lactobacillus casei PB-LC39, Lactobacillus rhamnosus PB-LR76, Lactobacillus helveticus HH-LH17, and Lactobacillus plantarum HH-LP56) were named as LC, LR, LH, and LP, respectively. SPME-GC-MS and untargeted metabolomics were used to compare and analyze the flavor quality, metabolites and metabolic pathways of LC, LR, LH and LP, and the potential function of differential metabolites was emphasized. The results demonstrated that the incorporation of probiotics resulted in a significant increase in the number of volatile flavor compounds and varying flavor profiles within the cheese. Especially LC, exhibited aromas reminiscent of wine, fruit and rose, and displayed the most favorable flavor qualities among all probiotic cheeses. The results of differential metabolite screening and metabolic pathway demonstrated that probiotic cheese could result in the production of 146–192 differential metabolites, mainly carbohydrates, proteins and acids. Arginine biosynthesis was a key differential metabolite pathway made by probiotics. The LC, LR, LH, and LP groups contained 22, 27, 23, and 24 functional metabolites, including L-carnitine, naringenin, and turanose et al., which might confer anti-inflammatory and improve lipid metabolism functions. These findings provide a theoretical basis for the further functional evaluation and development of probiotic cheese.

Influence of Varied Phosphorus Fertilizer Ratios on the Rhizosphere Soil Microbial Community in Idesia polycarpa Seedlings

Phosphorus (P) is crucial for tree growth and development, and it significantly influences the rhizosphere microbial community. However, the effects of phosphorus addition on the microbial communities in the rhizosphere soil of Idesia polycarpa remain understudied. In this study, two-year-old “Yuji” Idesia polycarpa seedlings were used to investigate the effects of phosphorus fertilization at four different levels of 0 g (control, CK), 0.92 g (low phosphorus, LP), 1.83 g (medium phosphorus, MP), and 2.75 g (high phosphorus, HP) per plant. The fertilizers were applied every 40 days over 120 days. MiSeq high-throughput sequencing of the 16S rRNA and ITS1 genes was employed to analyze the microbial community composition and diversity of bacteria and fungi in the rhizosphere soil under different phosphorus levels. The results showed that compared with CK treatment, the application of phosphorus fertilizer changed the physicochemical properties of the soil. The LP treatment significantly increased the soil pH, while the HP treatment group exhibited the highest soil-available phosphorus (AP) content. LP treatment significantly increased the number of microbial OTUs in the early and rapid growth stages and the richness and diversity of microbial communities. In addition, the bacterial community structure was significantly correlated with soil pH and AP, while the fungal community had no significant effect. The primary metabolic pathway function of bacteria in the rhizosphere soil of Idesia polycarpa seedlings is mainly metabolism, while fungi are mainly biosynthesis. Compared with CK treatment, 20 differential metabolic pathways were screened out in the bacterial community. Only two differential metabolic pathways were screened out in the fungal community by LP treatment at 120 days. In summary, applying low-level phosphate fertilizer is conducive to promoting the diversity of rhizosphere soil microorganisms. Therefore, potted planting of Idesia polycarpa seedlings is more suitable for applying low phosphorus levels.

Metabolic phenotypes and vitamin D response in the critically ill: A metabolomic cohort study

Background & AimsAlthough vitamin D deficiency is common in critically ill patients, randomized controlled trials fail to demonstrate benefits of supplementation. We aimed to identify distinct vitamin D3 responsive metabolic phenotypes prior to trial intervention of high-dose vitamin D3 by applying machine learning clustering method to metabolomics data from the Correction of Vitamin D Deficiency in Critically Ill Patients (VITdAL-ICU) trial. MethodsIn the randomized, placebo-controlled VITdAL-ICU trial, critically ill adults received placebo or high-dose vitamin D3. To distinguish vitamin D3 responsive metabolic phenotypes prior to intervention, we implemented consensus clustering with partitioning around medoids algorithm to the plasma metabolome data before randomization. Individual metabolite differences were determined utilizing linear mixed-effects regression models stratified for metabolomic phenotypes with false discovery rate adjustment. The association between vitamin D3 supplementation and 180-day mortality was evaluated in each metabolic phenotype, applying multivariable logistic regression analysis. ResultsIn 453 critically ill adults, the study identified 4 distinct metabolic phenotypes (clusters A. N=134; B. N=123; C. N=92; D. N=104). We found differential metabolic pathway patterns in the four clusters. Specifically, branched chain amino acid catabolic metabolites, long-chain acylcarnitines and diacylglycerol species are significantly increased in a specific metabolic phenotype (cluster D) following high-dose vitamin D3. Further, in cluster D high-dose vitamin D3 supplementation had a significantly lower adjusted odds of 180-day mortality after controlling age, sex, Simplified Acute Physiology Score II, admission diagnosis, and baseline 25-hydroxyvitamin D (OR 0.28 (95%CI, 0.09–0.89); P=0.03). In metabotype A, B, and C, high-dose vitamin D3 supplementation was not significantly associated with lower 180-day mortality following multivariable adjustment. ConclusionIn this post-hoc cohort study of the VITdAL-ICU trial, the clustering analysis of plasma metabolome data identified biologically distinct metabolic phenotypes. Among clusters, we found the different associations between high-dose vitamin D3 supplementation and specific metabolite pathways as well as 180-day mortality. Our findings facilitate further research to validate metabolic phenotype-targeted strategies for critical illness treatments.

Analysis of the characteristics of intestinal microbiota in patients with different severity of obstructive sleep apnea

Intestinal microbiota imbalance plays an important role in the progression of obstructive sleep apnea (OSA), and is considered to be the main mediator that triggers metabolic comorbidities. Here, we analyzed the changes in intestinal microbiota in patients with different severities of OSA based on apnea hypopnea index (AHI) classification, and explored the role of intestinal microbiota in the severity of OSA. This study included 19 healthy volunteers and 45 patients with OSA [5 ≤ AHI < 15 (n = 14), 15 ≤ AHI < 30 (n = 13), AHI ≥ 30 (n = 18)]. Relevant sleep monitoring data and medical history data were collected, and microbial composition was analyzed using 16S rRNA high-throughput sequencing technology. The diversity analysis of intestinal microbiota among different groups of people was conducted, including alpha diversity, beta diversity, species diversity, and marker species as well as differential functional metabolic pathway prediction analysis. With the increase of AHI classification, the alpha diversity in patients with OSA significantly decreased. The results revealed that the severity of OSA is associated with differences in the structure and composition of the intestinal microbiota. The abundance of bacteria producing short-chain fatty acids (such as Bacteroides, Ruminococcacea, and Faecalibacterium) in severe OSA is significantly reduced and a higher ratio of Firmicutes to Bacteroidetes. Random forest analysis showed that Parabacteroides was a biomarker genus with important discriminatory significance. The differential metabolic pathway prediction function shows that the main function of maintaining intestinal microbiota homeostasis is biosynthetic function. Our results show that the differences in the composition of intestinal microbiota in patients with different severities of OSA are mainly related to short-chain fatty acid-producing bacteria. These changes may play a pathological role in OSA combined with metabolic comorbidities.

Effects of exposure to 17α-methyltestosterone on hepatic lipid metabolism in Gobiocypris rarus

This study aimed to investigate the effects of 17α-Methyltestosterone (MT) on hepatic lipid metabolism in Gobiocypris rarus. G. rarus was exposed to varying concentrations of MT (0, 25, 50, and 100 ng/L) for durations of 7, 14, and 21 d. Biochemical and transcriptomic analyses were conducted using methods, such as ELISA, RT-qPCR, Western Blotting, and RNA-seq, to decipher the key signals and molecular mechanisms triggered by MT in vivo. The results revealed that MT induced hepatomegaly in G. rarus and markedly increased the hepatic steatosis index (HSI). After 14 d of exposure, significant increase in PPARγ mRNA expression was observed, whereas after 21 d, PPARα mRNA expression was significantly reduced. The expression pattern of SREBP1C mRNA initially decreased before increasing, mirroring the trend observed for SREBP1C protein expression. Furthermore, MT increased the levels of key lipid synthesis enzymes, including HSL, CPT1, GPAT, and FAS, thereby fostering lipid accumulation. RNA-seq analysis revealed that MT modulated hepatic bile acid metabolism via the PPAR pathway, consequently influencing cholesterol and lipid metabolism. Considering the differential metabolic pathways of MT across genders, it is postulated that MT may undergo aromatization to estrogen within G. rarus, thereby exerting estrogenic effects. These findings provide crucial experimental insights into the detrimental effects of MT in aquatic settings, underscoring its implications for safeguarding aquatic organisms and human health.