Applied Biological Chemistry最新文献

Background

Nasopharyngeal carcinoma (NPC) is characterized by its aggressive behavior and a pronounced tendency for metastasis. Artesunate (ART), an antimalarial drug, has demonstrated anticancer properties; yet, the underlying mechanisms of its action in NPC are poorly defined.

Methods

The anti-tumor effects of ART on NPC cells (proliferation, apoptosis, migration, invasion, and angiogenesis) were assessed in vitro. An integrated approach combining network pharmacology (using NPC-related DEGs from dataset GSE118719 and predicted ART targets) and molecular docking identified potential targets. Key findings were validated through qRT-PCR, WB, gelatin zymography, loss/gain-of-function assays, and an in vivo xenograft model.

Results

ART significantly inhibited NPC cell proliferation, migration, invasion, and angiogenesis, while inducing apoptosis. Network pharmacology identified MMP2 as a core target, and molecular docking confirmed high-affinity binding between ART and MMP2. MMP2 was upregulated in NPC tissues and cells. Knockdown of MMP2 suppressed malignant phenotypes, while its overexpression reversed ART-induced inhibition. In vivo, ART treatment reduced tumor growth and downregulated the expression of MMP2 and Ki67.

Conclusion

ART exerts potent anti-NPC effects by directly targeting and inhibiting MMP2, highlighting its potential as a therapeutic agent for NPC.

A study was carried out to explore and address the challenges posed by rice blast disease (Magnaporthe oryzae B. C. Couch.). Keeping in view the unique qualities and cultural importance of this variety, the research introduces an innovative approach by integrating plant extracts into disease management practices through in-depth analyses and field experiments. Three concentrations of ethanol-based extracts each of the twenty-one were evaluated in vitro against the control. The percentage of mycelial inhibition was recorded and based on their effectiveness; six efficient botanicals were further evaluated in the field. The field experiments were carried out on the susceptible Mushk Budji cultivar of rice for two successive years. The results demonstrated that certain botanicals, especially extracts of Syzygium aromaticum and Podophyllum hexandrum, significantly reduced disease incidence and severity, and improved grain and straw yields, performing on par with the chemical control, Tricyclazole. Additionally, molecular docking was conducted to identify active compounds responsible for antifungal effects. The ligands for docking studiesquercetin, podophyllotoxin, and eugenolwere chosen based on literature mining for their reported antifungal activity. These phytochemicals displayed strong binding affinities ( -5.5 to − -3.8 kcal/mol) to the ToxB-like effector protein of M. oryzae, suggesting interference with fungal pathogenicity mechanisms. Among these, quercetin and podophyllotoxin were the most promising ligands. The integration of wet-lab assays and in silico analysis strengthens the case for deploying bioactive botanicals in integrated disease management. Our findings advocate the use of eco-friendly, plant-based fungicides as a viable strategy for blast disease control, promoting organic farming and preservation of indigenous rice varieties. The findings highlight the importance of developing eco-friendly, plant-based alternatives to chemical fungicides, aligning with the goals of sustainable agriculture and organic farming. Future research should focus on developing stable formulations of these botanicals, evaluating their synergistic potential with biocontrol agents, and validating their effectiveness through multi-location field trials. Additionally, further investigation into the molecular mechanisms of action and the long-term ecological and economic impacts of botanical fungicides is warranted.

Background

Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder. Granulosa cell dysfunction is an important factor causing PCOS.PLAU has been shown to be upregulated in granulosa cells of PCOS patients, but its potential mechanism in regulating the progression of PCOS has not been clarified. The present study focused on the mechanisms by which PLAU regulates granulosa cell injury and apoptosis in PCOS.

Methods

Establishment of a testosterone-induced granulosa cell model. Flow cytometry was performed to test granulosa cell viability and apoptosis. RT-qPCR was applied to measure the mRNA levels of hormone synthesis related genes. The expressions of PLAU, STAT3, and NLRP3 inflammasome as well as apoptosis-related proteins were assessed by western blot. Binding between STAT3 and NLRP3 was detected using ChIP-PCR assay. Inhibitors and agonists of STAT3 were utilized to validate the role of PLAU and STAT3 signaling in testosterone-treated granulosa cells.

Results

Upregulation of PLAU, STAT3, and NLRP3 signaling was observed in testosterone-treated granulosa cells. Both PLAU interference and STAT3 inhibitors inhibited testosterone-induced granulosa cell viability, impaired function injured, inflammation, and apoptosis. Additionally, downregulation of PLAU decreased STAT3 and NLRP3 levels. Further results showed that STAT3 bound to the NLRP3 promoter. STAT3 agonists partially counteracted the ameliorative effect of PLAU knockdown on testosterone-induced granulosa cell dysfunction.

Conclusion

These results suggest that downregulated PLAU may improve testosterone-induced granulosa cell dysfunction by inhibiting STAT3/NLRP3 signaling, providing a new molecular mechanism for PCOS progression.

Age–related cognitive decline is closely associated with impairments in synaptic structure and function, particularly in the hippocampus. Ecklonia cava, a brown seaweed rich in phlorotannins, has been suggested to possess neuroprotective properties; however, its effects on synaptic regulation and cognitive function in aging models remain unclear. In this study, we investigated the effects of phlorotannin supplement derived from E. cava (PS) on cognitive function and hippocampal synaptic integrity in senescence–accelerated mouse prone 8 (SAMP8) mice. In in vitro experiments, PS exerted neuroprotective effects in HT22 cells exposed to amyloid-β and L-glutamate. Moreover, PS inhibited the enzymatic activities of β-secretase and acetylcholinesterase, and exhibited notable ABTS-radical scavenging activity. Mice were orally administered PS at doses of 250 or 500 mg/kg/day for 17 weeks, and cognitive function was assessed using the Y-maze, passive avoidance, and Morris water maze tests. Hippocampal tissues were analyzed for dendritic spine density, synaptic protein expression, and activation of signaling pathways associated with synaptic plasticity and neurotrophic support. PS treatment significantly improved short–term, long–term, and spatial memory performance in SAMP8 mice. It also restored hippocampal spine density and upregulated synaptic proteins such as synapsin 1 and PSD95. Moreover, PS activated the NMDAR2B–CaMKII–CREB and BDNF–ERK pathway, accompanied by elevated BDNF immunoreactivity in the dentate gyrus. Collectively, these findings suggest that PS improves cognitive function during aging by restoring hippocampal synaptic density and reactivating key signaling pathways involved in synaptic plasticity and neurotrophic support. PS may serve as a promising dietary intervention for preventing or delaying age–related cognitive decline.

Three-prime repair exonuclease 1 (Trex1) prevents cytosolic DNA accumulation and immune activation, yet the physiological consequences of increased Trex1 expression in vivo remain unclear. In this study, we used Drosophila melanogaster, a model well suited for quantitative analyses of aging, sleep, and circadian rhythms, to generate flies that ubiquitously overexpress murine Trex1 under the Act5C-GAL4 driver, given that a clear Trex1 ortholog has not been identified in flies. Trex1 overexpression significantly extended the lifespan of male flies (40.55 ± 1.10 days in controls; n = 60 vs. 44.98 ± 1.59 days in Trex1; n = 57, p < 0.05, Bonferroni corrected), whereas female flies showed a modest but statistically non-significant increase in lifespan. Interestingly, Trex1 overexpressing flies exhibited more fragmented sleep and reduced circadian rhythm robustness compared with controls. These findings suggest that the mechanisms by which Trex1 overexpression influences lifespan and those regulating sleep and circadian stability may be functionally separable.

Programmed cell death protein-1 (PD-1) is a key immune checkpoint receptor and an important target for therapeutic antibody development. A critical factor in the development of therapeutic monoclonal antibodies is the selection and preparation of the target antigen. Here, we established a plant-based platform for human PD-1 production using Nicotiana benthamiana and evaluated its biochemical properties and immunogenic potential in comparison with bacterial (E.coli) and mammalian (HEK293T cells) expression systems. Codon optimization and promoter selection significantly enhanced transient PD-1 expression, with the pFM′M vector producing the highest yield. Although plant-derived PD-1 predominantly accumulated in the insoluble fraction and required denaturing purification, sufficient quantities were obtained for immunological evaluation. Mice immunized with plant-produced PD-1 exhibited strong splenic and lymph-node responses. ELISA and flow cytometry analyses demonstrated that antibodies generated from plant-derived PD-1 recognized native, cell-surface PD-1, although with lower potency than antibodies induced by mammalian cell-derived antigen. While further optimization is required to improve solubility and preserve native protein conformation, this study highlights the potential for generating antigens for therapeutic antibody discovery. Thus, N. benthamiana is a viable platform for producing immunogenic human PD-1 capable of eliciting functional antibodies.

Climate change imposes complex challenges on crop production, altering nutrient availability, soil dynamics, and plant nutrient uptake efficiency. Fertilization strategies calibrated under historical climate norms are insufficient to support nutrient uptake efficiency under stressors such as drought-induced evapotranspiration or flooding-induced denitrification. This study proposes a comprehensive framework to redefine fertilization practices based on interactions between climate factors, soil properties, and crop physiological responses. Through a review of recent research, we identify the critical effects of water-filled pore space on fertilizer use efficiency (FUE) and emphasize the need for moisture-sensitive fertilization management. Comparative strategies tailored to specific climate stresses are summarized, highlighting optimal timing, form, and fertilizer types for each scenario. We also present a research roadmap integrating experimental data collection, modeling, and adaptive fertilization technology development. This approach aims to establish climate-responsive fertilization systems capable of maintaining nutrient uptake and minimizing losses under variable environmental conditions. Our findings provide a scientific basis for precision fertilization strategies that align with future climate scenarios, offering practical insights for enhancing food security and agricultural resilience. Ultimately, the adoption of climate-adaptive fertilization techniques will be essential to sustain productivity and reduce environmental impacts in the face of ongoing climate variability.

Graphical Abstract

Bacillus subtilis is known for its probiotic and antioxidant potential. However, its functional properties at the molecular level remain underexplored. In this study, Bacillus subtilis isolated from Cheonggukjang, a Korean traditional fermented soybean paste (strain E5), was compared with its sporulation-deficient mutant (strain E5-M) obtained by disrupting the sigF gene and six consecutive downstream genes involved in sporulation (spoVAA, spoVAB, spoVAC, spoVAD, spoVAE, and spoVAF). Using RNA sequencing, we analyzed transcriptomic responses in human umbilical vein endothelial cell and hepatoblastoma cells pretreated with E5 or E5-M, followed by chemical induction of inflammation. Functional enrichment analysis revealed that E5 primarily regulated inflammatory pathways and intercellular signaling, while E5-M more prominently induced genes involved in tissue recovery, mitochondrial activity, and redox homeostasis. Real-time quantitative PCR validation confirmed a reduction in key inflammatory markers (tumor necrosis factor alpha, interleukin-1 beta, transcription factor p53, transcription factor p65 and BCL2-associated X protein), particularly in HepG2 cells. Whole genome sequencing showed no increase in allergenic potential due to the genetic modification. These findings suggest that suppression of sporulation in B. subtilis enhances its anti-inflammatory and oxidative stress-modulating activities, indicating its potential for safer and more functional applications in food and health industries.

The performance of wound dressings is closely linked to their ability to provide appropriate mechanical support, moisture retention, and antibacterial protection. Here, we report a covalently crosslinked alginate hydrogel dressing designed to overcome ion leaching associated with conventional Ca²⁺-crosslinked alginate and to enhance functional molecule efficacy through alginate oligosaccharide (AOS) incorporation. Covalent hydrogels were prepared using EDC/NHS chemistry with amino acid–based crosslinkers, and AOS was conjugated at varying concentrations. Compared with ionically crosslinked gels, covalent hydrogels exhibited reduced swelling and a compressive modulus of 2–5 kPa with ~ 40% hysteresis, approaching the lower range of skin elasticity. Although increasing AOS content further decreased re-swelling after drying, water-retention capacity was largely preserved. An antimicrobial peptide was covalently immobilized onto the hydrogel surface, imparting dose-dependent antibacterial activity. Notably, AOS incorporation significantly enhanced antibacterial efficacy, nearly doubling growth inhibition at lower peptide concentrations. Microstructural analysis revealed dense pore networks in ionically crosslinked gels, whereas AOS conjugation did not induce substantial structural changes in covalent hydrogels. This study demonstrates that AOS-conjugated, covalently crosslinked alginate hydrogels provide tunable mechanical properties and amplified antibacterial performance, offering a versatile platform for next-generation functional wound dressings.

Graphical abstract

Numerous risk factors were identified for the incidence of renal injury and fibrosis among which is environmental pollution whereas air pollution by diesel particulate matter (PM) represents one of the most serious factors especially in developing countries. The present study aimed to explore the potential renoprotective effects of fisetin; a natural flavonoid against diesel PM-induced renal injury and fibrosis and clarify the underlying molecular mechanisms. For this purpose, adult Wistar albino rats received diesel PM (0.064 mg/kg/via oral gavage, three times/week) and were co-treated with fisetin (2.5 mg/Kg/via oral gavage/ day) for six weeks and compared with age-matched normal control group (NC). Kidney functions were assessed in serum samples while kidneys were harvested for biochemical and histological examination. Compared to NC group, diesel PM-administrated rats displayed elevated level of lipocalin-2 (Lcn-2) and kidney dysfunction along with renal histological changes that were alleviated markedly by fisetin. Concurrent treatment with fisetin significantly lowered the renal levels of NF-κB, IL-23 in addition to other inflammatory mediators including IL-17, TNF-α and IL-6. Also, diesel PM-administrated rats co-treated with fisetin showed downregulation of mTOR/S6K-b1 signaling besides a significant decrease in the renal expression of caveolin-1. This was associated with marked attenuation of interstitial fibrosis in the renal tissues of diesel PM-administrated rats. In conclusion, these findings proved for the first time that fisetin could exert renoprotective effects against renal injury and fibrosis induced by diesel PM probably via downregulation of IL-23/mTOR/S6K-b1 signaling along with decreasing renal caveolin-1 expression.