Applied Biochemistry and Biotechnology最新文献

Objective This study aimed to explore the possible mechanism of adiponectin on the insulin signaling in streptozotocin (STZ)-induced gestational diabetes mellitus (GDM) rats. Methods The GDM rats were induced by injection of 40 mg/kg STZ, and then orally treated with adiponectin (5, 10, 20 mg/kg) every day from gestation day (GD)7 to GD20. The body weight and fasting blood glucose (FBG) were observed every 3 days from GD9 to GD18. Meanwhile, the insulin tolerance test (ITT), homeostasis model assessment insulin resistance (HOMA-IR), and adiponectin expression were observed in rats. Moreover, the insulin signaling-related factors, sex hormone-binding globulin (SHBG), PPAR-α, phospho-AMPK alpha, glucose transporter-3 (GLUT3), and phospho-insulin receptor substrate-1 (p-IRS1) in placental tissues were measured in rats. In vitro, HTR-8-Svneo cells were induced by 30 mM glucose and then treated with adiponectin (50, 100, 200 ng/mL) to observe the changes in cell viability, SHBG expression, and the insulin signaling-related factors. Moreover, silencing or overexpression of SHBG was achieved via transfection with siRNA or pEX-4 SHBG, respectively. The effects of SHBG on the insulin signaling-related factors were observed in cells. Results Adiponectin treatment significantly improved the STZ-induced decrease in body weight, increase in the FBG, ITT, and HOMA-IR with increasing doses. Specifically, adiponectin treatment upregulated the SHBG expression and the insulin signaling-related factors (PPAR-α, p-AMPK, GLUT3, and p-IRS1) in the placental tissues of GDM rats with increasing doses. In line with the results of animal experiments, adiponectin treatment alleviated the high glucose-induced decrease in cell viability, SHBG expression, and the levels of GLUT3 and p-IRS1 in HTR-8-Svneo cells with increasing doses. Moreover, the levels of GLUT3 and p-IRS1 were regulated by the SHBG expression. SHBG silencing weakened the effect of adiponectin in improving the levels of GLUT3 and p-IRS in the high-glucose-induced cells. Conclusion The study showed that adiponectin upregulated the SHBG expression and improved the insulin signaling in STZ-induced GDM rats, as well as adiponectin upregulated the insulin signaling-related factors via SHBG in high-glucose-induced trophoblast cells. This study suggests that adiponectin may be a potential therapeutic target in GDM.

1,8-cineole, a bicyclic monoterpenoid compound with significant application value, is widely used in the fields of flavoring, pharmaceuticals, and biofuels. Although recent advances have enabled its biosynthesis in model microorganisms (Escherichia coli and Saccharomyces cerevisiae), the inherent cytotoxicity of terpenoids and the low catalytic efficiency of terpene synthases remain major bottlenecks limiting further improvement in biosynthetic efficiency. To address these limitations, we employed Serratia marcescens HBQA7-a non-model microorganism with high terpene tolerance, previously isolated in our lab-as a chassis for constructing an efficient 1,8-cineole biosynthetic system. Heterologous expression and functional screening of six 1,8-cineole synthase (CinS) genes from different sources were performed, among which SoCinS from Salvia officinalis exhibited the highest catalytic activity. Rational enzyme engineering through site-directed mutagenesis further improved SoCinS activity. In addition, fusion with high-expression tag proteins significantly improved enzyme expression levels, leading to increased 1,8-cineole titers. Under 72-hour shake flask fermentation conditions, the 1,8-cineole concentration reached 2.1 g/L. Finally, fed-batch fermentation in a 5-L bioreactor yielded a final 1,8-cineole titer of 10.2 g/L, demonstrating a higher productivity and greater industrial application potential than current model microbial systems. This work highlights the promising potential of non-model microorganisms in the efficient biosynthesis of terpenoid natural products.

Plant-mediated synthesis is a most recent approach toward green synthesis of copper-based nanoparticles. In this approach nanoparticles are capped with phytochemicals, suggesting that plants with unique biological properties might transfer these benefits to the nanoparticles. However, there is no experimental evidences to directly corelate bioactivities of nanoparticles to their capping phytochemicals. This study aims to assess the impacts of herbal antimicrobial compounds on the antimicrobial potency of resulted nanoparticles. Cu(OH)₂ nanoparticles (CuNPs) were synthesized by leaf extracts of eucalyptus and tobacco. Resulted particles were characterized to exhibit similar physicochemical properties which are determinative for antimicrobial activity. Antimicrobial effects of the leaf extracts and CuNPs were tested against Staphylococcus aureus and Escherichia coli. Eucalyptus leaf extract was found effective against S. aureus with growth inhibition zone of 13.1 ± 0.6 mm. CuNPs exhibited efficacy against both bacterial strains with inhibition zones exceeding 15 mm. Notably, no significant difference in antimicrobial effectiveness of CuNPs was observed, suggesting that the antimicrobial superiority of eucalyptus extract was not transferred to nanoparticles. These results could reshape our understanding about the impacts of herbal bioactive molecules on the bioactivity of plant-mediated synthesized nanoparticles.

Ferroptosis is a critical contributor to ischemia-reperfusion (I/R) injury and subsequent organ failure. While ENPP2 has been implicated in regulating ferroptosis in cardiomyocytes, its specific role in myocardial I/R injury remains unclear. This study aims to elucidate the function of ENPP2-mediated ferroptosis in myocardial ischemia-reperfusion injury (MI/RI), to provide novel insights into potential therapeutic strategies. A mouse model of MI/RI was established and subjected to interventions with ENPP2 overexpression and/or SIRT1 knockdown. In vitro, cardiomyocytes were treated with palmitate and subjected to hypoxia/reoxygenation (H/R) to simulate I/R injury. These cells received treatments with ENPP2 overexpression (oe-ENPP2), SIRT1 overexpression (oe-SIRT1), PGC-1α silencing (si-PGC-1α), and/or SIRT1 knockdown (sh-SIRT1). Additionally, Erastin-induced ferroptosis in cardiomyocytes was used to assess the protective effects of oe-ENPP2. Ferroptosis was assessed through the lipid peroxidation (MDA, 4-HNE), iron and Fe²⁺ assays, GPx4 and SLC7A11 expression, and transmission electron microscope. Overexpression of ENPP2 significantly alleviated myocardial infarction in MI/RI mice, as indicated by the upregulation of GPx4 and SLC7A11 protein levels. In cardiomyocytes subjected to hypoxia/reoxygenation (H/R) or erastin-induced ferroptosis, oe-ENPP2 reduced apoptosis rates, preserved Fe²⁺ content, and restored GPx4 and SLC7A11 expression. Silencing PGC-1α blocked the protective effect of oe-ENPP2 against H/R-induced ferroptosis in HL-1 cells. Additionally, SIRT1 overexpression inhibited PGC-1α acetylation, whereas SIRT1 knockdown similarly reversed the anti-ferroptotic effects of oe-ENPP2 in H/R-treated HL-1 cells. SIRT1 silencing blocked the protective effects of oe-ENPP2 against myocardial infarction and fibrosis in MI/RI mice via the PGC-1α/NRF1 pathway. ENPP2 overexpression protects the mouse myocardium from I/R-induced ferroptosis injury via the SIRT1/PGC-1α/NRF1 pathway. These findings suggest a novel gene therapy strategy for mitigating myocardial I/R injury.

Hypertrophic cardiomyopathy (HCM) is a complex genetic disorder characterized by left ventricular hypertrophy and impaired cardiac function. Although progress has been made in understanding its genetic basis, the discovery of novel biomarker candidates and therapeutic targets remains crucial for improving diagnosis and treatment. Recent studies have demonstrated that copper metabolism plays an important role in various diseases, suggesting that copper-related genes (CRGs) may be of significant importance in HCM. In this study, we analyzed bulk RNA-seq and single-cell RNA-seq (scRNA-seq) data from healthy controls (HC) and HCM patients. Differentially expressed genes (DEGs) were identified through differential expression analysis, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to identify associated pathways. Weighted Gene Co-expression Network Analysis (WGCNA) was used to identify gene modules related to the HCM phenotype, and a diagnostic model was constructed based on these modules. Additionally, cell type-specific expression patterns were explored through single-cell analysis, and Gene Set Enrichment Analysis (GSEA) and MR analyses were conducted to evaluate the causal relationships between genes and HCM risk. The study found that DEGs associated with HCM were significantly enriched in pathways related to immune responses. WGCNA identified gene modules highly correlated with HCM, among which the blue module exhibited the strongest correlation with HCM. The diagnostic model constructed based on DEGs and WGCNA module genes demonstrated good diagnostic performance, with FCN3, TIPARP, and PROM1 emerging as potential diagnostic biomarker candidates for HCM. Additionally, single-cell analysis revealed the expression characteristics of different cell types in HCM, and causal relationships between key genes and HCM risk were confirmed through GSEA and MR analyses. This study identified FCN3, TIPARP, and PROM1 as cuproptosis-associated biomarker candidates that showed reproducible expression patterns across cohorts and experimental validation. These findings are associative and hypothesis-generating; they suggest potential diagnostic utility that warrants prospective clinical evaluation and mechanistic studies (e.g., perturbation of copper homeostasis and gene manipulation) before any therapeutic inference can be made.

Diatoms are unicellular eukaryotic algae, renowned for their intricately patterned silica cell walls, which exhibit remarkable morphological precision and nanostructural complexity. They possess a unique and rich biochemical profile and are prolific producers of biologically active polysaccharides, broadly categorized as intracellular, extracellular (primarily sulfated), and cell wall-associated types. These polysaccharides play vital roles in biofilm formation, carbon cycling, nutrient storage, and ecosystem dynamics, while also holding substantial promises in commercial and biotechnological fields. This review provides an integrated overview of diatom polysaccharide chemotypes-storage β-glucans, cell-wall uronic- and sulfate-rich scaffolds, and extracellular exopolymers-and evaluates the conventional versus emerging extraction and purification techniques, discussing trade-offs in yield, selectivity, and polymer integrity. The diverse structural characterization methods for elucidating monosaccharide linkages and functional modifications have been reviewed. The genomic and metabolic insights into polysaccharide biosynthesis have been elaborated along with elucidation of the relationship between extracellular polymeric substances and bacterial community assembly. The multifaceted applications of diatom-derived polysaccharides in carbon sequestration, biomedicine (e.g., anticoagulant, antioxidant, antiviral, anticancer, immunomodulatory agents), materials science, and environmental remediation has been discussed along with the current challenges-species variability, efficient frustule disruption, and scalable processing. The-genomics-guided strain optimization and sustainable bioprocess design holds immense future potential for diatom derive polysaccharides.

The 5-methylcytosine (5mC) epigenetic modification is a prominent pattern of DNA methylation. Nonetheless, its function and relationship with the immune microenvironment (IME) in heart failure (HF) are unclarified. Firstly, seven microarrays and two high-throughput sequencing datasets were downloaded from GEO for this research. Secondly, random forests, LASSO logistic regression, and SVM-RFE were leveraged for screening hub genes. Quantitative PCR, Western blot, and immunofluorescence staining were conducted in a male rat model of HF after myocardial infarction for validation. Thirdly, 5mC-related HF samples were allocated into two distinct categories using consensus clustering algorithms. Finally, single-sample gene-set enrichment analysis and CIBERSORT deconvolution algorithm were utilized for further exploring the IME in HF, including infiltrating immune cell abundance score, human leukocyte antigen (HLA), and immune checkpoints (ICPs). Among these 5mC regulators, four hub genes were uncovered, and the diagnostic model exhibited an outstanding ability to distinguish HF and healthy samples (0.969, 95%CI, 0.953-0.985). DNMT3B was identified as greatly influencing cardiac function. DNMT3B and MBD2 were finally identified as hub genes in the HF model. Two 5mC subtypes manifested different modification patterns, infiltrating immunocytes, ICPs, and HLA gene expression. Furthermore, 305 DEGs were found in 5mC subtypes, and many functions were associated with important pathophysiological mechanisms of HF. The HF diagnostic model was established using 5mC robust core biomarkers. 5mC methylation regulators may offer novel perspectives for understanding the mechanisms, accurate diagnosis, and effective interventions for HF.

Breast cancer (BC) remains the most commonly diagnosed cancer among women worldwide and a leading cause of cancer-related mortality. Despite advances in early detection and treatment modalities, challenges such as recurrence, metastasis, and therapy resistance persist, necessitating the development of novel therapeutic strategies. Benzyl isothiocyanate (BITC), a naturally occurring isothiocyanate derived from cruciferous vegetables, has emerged as a promising anticancer agent due to its potent chemopreventive and therapeutic properties. This review comprehensively explores the investigational potential of BITC in the management of breast cancer, emphasizing both its molecular mechanisms and its targeted delivery through nanotechnological approaches. The chemistry and pharmacology of BITC are discussed, highlighting its ability to modulate key signaling pathways, induce apoptosis, inhibit metastasis, and arrest the cell cycle in breast cancer cells. Special attention is given to its pharmacokinetic profile, including bioavailability and metabolic stability, which are crucial for clinical translation. Furthermore, the review examines preclinical findings that demonstrate BITC's suppressive effects on tumor growth and its synergistic interactions with conventional therapies. Innovative strategies for BITC delivery, such as nanoparticle-based systems, are also evaluated for their potential to enhance therapeutic efficacy and reduce systemic toxicity. The safety profile and toxicity considerations of BITC are critically assessed based on current preclinical evidence. By synthesizing findings from a broad range of preclinical studies, this review underscores the multifaceted anticancer potential of BITC and its promise as a complementary or alternative approach in breast cancer treatment. Continued research into its mechanistic actions, delivery optimization, and translational applications is essential to harness BITC's full potential in improving patient outcomes.