Combinatorial chemistry & high throughput screening最新文献

Introduction: Metabolic Dysfunction-Associated Steatohepatitis (MASH) is a growing global health concern, with only one FDA-approved therapy currently available. Acetyl-CoA carboxylase (ACC) inhibition has emerged as a promising strategy, yet effective and clinically translatable inhibitors remain limited. This study aimed to identify potential ACC inhibitors for MASH via drug repurposing.

Methods: A small-molecule library was screened using structure-based virtual screening, and candidate compounds were validated in a free fatty acid-induced MASH cell model. Intracellular triglyceride (TG) and aspartate aminotransferase (AST) levels were measured, while quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to evaluate lipid metabolism- related gene expression. Molecular dynamics simulations were conducted to further evaluate binding stability.

Results: Lanatoside C was identified as the most potent candidate. In vitro studies revealed significant reductions in TG and AST levels, downregulation of lipogenesis-related genes (SREBP1, FASN, ACC), and upregulation of fatty acid oxidation genes (CPT1A, ACOX1, FABP1). Molecular dynamics simulations confirmed the stable binding of Lanatoside C to ACC.

Discussion: These findings indicate that Lanatoside C exerts dual regulatory effects on lipid metabolism by suppressing fatty acid synthesis and enhancing oxidation. As an FDA-approved cardiac glycoside, Lanatoside C's known pharmacological profile supports its potential repositioning for MASH, although further in vivo studies and mechanistic validation are warranted.

Conclusion: Lanatoside C demonstrates promise as a repurposed ACC inhibitor for MASH treatment, offering a cost-effective repurposing strategy to advance therapeutic options for MASH.

Introduction: The rising demand for sustainable, nutritionally rich food sources has sparked interest in non-traditional protein sources, including gastropods. In northeastern India, freshwater gastropod species are widely consumed, particularly by certain tribes and communities, both as a traditional delicacy and for their nutritional value. This study delves into the biochemical composition of three commonly consumed freshwater gastropod species in Assam, namely Pila globosa, Pila scutata, and Brotia costula, to evaluate their potential as alternative food sources, with a focus on nutritional value and palatability.

Methods: In the present study, proximate analysis was done by using standard methods given in the Association of Official Analytical Chemists (AOAC, 2015). Mineral analysis was carried out using the atomic absorption spectrophotometer (AAS). High Performance Liquid Chromatography (HPLC) (method QA.16.5.10/AOAC 19th edition) was employed for the determination of amino acid contents of the samples.

Results: Among the species analyzed, P. scutata and P. globosa displayed the highest protein content, whereas B. costula exhibited greater levels of ash and carbohydrates. Mineral analysis showed that P. globosa contained the highest concentrations of calcium (Ca), magnesium (Mg), iron (Fe), zinc (Zn), and copper (Cu). Amino acid profiling revealed lysine and serine as the key essential and non-essential amino acids, with P. globosa and P. scutata showing higher concentrations, respectively.

Discussion: This study highlights the nutritional richness of three freshwater gastropod species, with Pila globosa and Pila scutata showing high protein and mineral content, and Brotia costula contributing to energy and mineral intake. Their amino acid profiles further enhance their dietary value. Gastropods in local culture embody the use of snail meat as a part of traditional food source, highlighting different ethnomedicinal beliefs. Despite their benefits, these species remain underutilized, suggesting the need for broader awareness and integration into mainstream diets to support food security and nutrition in northeastern India.

Conclusion: Given the current scenario of food disparity, incorporating gastropods into human diets could provide a sustainable and reliable source of protein and micronutrients, particularly in areas where food security is a concern.

Introduction: Gynostemma pentaphyllum is commonly used in southern China to treat hyperlipidemia and tumors, primarily due to its active saponin compounds. This study aimed to analyze the inhibitory effect of gypenoside LI (Gyp LI) on clear cell renal cell carcinoma (ccRCC), which is mediated through DUSP1-related lipid metabolism.

Materials and methods: The concentration of Gyp LI required to inhibit the ccRCC cell lines 769-P and ACHN was determined using a CCK8 assay, and DUSP1 upregulation was confirmed by transcriptomic sequencing, RT-qPCR, and Western blot analysis. The roles of DUSP1 and Gyp LI in regulating cell proliferation were further validated using CCK-8, colony formation, and flow cytometry assays. . Additionally, we established a nude mouse xenograft tumor model using ACHN cells that overexpress DUSP1. We assessed the effects of Gyp LI and DUSP1 on tumor growth through histopathological examinations. We employed untargeted tissue metabolomics to identify metabolic pathways influenced by Gyp LI via upregulation of DUSP1.

Results: CCK-8 experiments showed that Gyp LI inhibited the proliferation of ccRCC cells. Additionally, transcriptome sequencing revealed that DUSP1 expression increased following Gyp LI intervention. We transfected cells with siDUSP1 and ovDUSP1 and assessed cell proliferation using CCK8, colony formation, flow cytometry, and WB analyses of apoptosis-related proteins, confirming that both ovDUSP1 and Gyp LI inhibited ccRCC cell proliferation. Animal studies revealed that both ovDUSP1 and Gyp LI inhibited tumor growth. Non-targeted metabolomics analysis revealed that Gyp LI affected seven metabolites associated with ccRCC treatment by upregulating DUSP1.

Conclusion: Gyp LI upregulates DUSP1 to suppress the metabolism of choline, linoleic acid, and alpha-linolenic acid, ultimately suppressing the incidence and progression of ccRCC.

Introduction: Polygonatum sibiricum (P. sibiricum) possesses antioxidant and antiinflammatory activities. We explored the multi-target mechanisms of P. sibiricum against hepatocellular carcinoma (HCC), aiming to improve its poor prognosis.

Materials and methods: Active compounds and disease targets of P. sibiricum were retrieved from the TCMSP and CTD databases. A PROTEIN-PROTEIN INTERACTION (PPI) network was constructed using the STRING database, and functional enrichment was performed with the clusterProfiler package. A compound-target-pathway network was developed in Cytoscape. Immune infiltration was assessed via CIBERSORT and ESTIMATE algorithms, while ligand-target binding was evaluated by molecular docking and 100-ns molecular dynamics (MD) simulations. In vitro experiments were performed to explore the expression and functions of the key genes.

Results: We screened 9 active components, 87 putative targets, and 240 HCC-related genes. Twenty overlapping targets were used to construct a PPI network. Network analysis identified baicalein and five core targets (FOS, MMP9, AKT1, TP53, and PTGS2). Molecular docking and 100-ns MD simulations confirmed stable ligand-protein binding. Immune profiling showed that higher expression of the core targets was related to higher StromalScore, ImmuneScore, and lower tumor purity. Enrichment analysis revealed that these genes were involved in critical pathways, including angiogenesis, EMT, and inflammation response. Functionally, MMP9 knockdown suppressed HCC cell proliferation, migration, and invasion.

Discussion: P. sibiricum, particularly through baicalein targeting FOS/MMP9/AKT1/ TP53/PTGS2, inhibited HCC development by modulating EMT/angiogenesis pathways and immune milieu. However, these findings required further verification.

Conclusion: Baicalein was identified as an active compound targeting five crucial genes to suppress HCC progression, uncovering a new anti-HCC mechanism of P. sibiricum.

Introduction: Longlutong Decoction (LLTD) is a Chinese traditional prescription used for coronary heart disease (CHD). The present study aimed to illuminate the mechanisms of LLTD treatment on CHD.

Methods: The therapeutic effect of LLTD on CHD was investigated using a CHD rat model. The chemical components of LLTD were identified, following which network pharmacology approaches were utilized to identify active components and disease-related targets. GO and KEGG analyses were conducted to explore potential molecular mechanisms. Finally, the molecular mechanism of LLTD treatment of CHD was verified.

Results: Histopathological assessment revealed markedly attenuated myocardial injury severity in the medicated groups when compared to the model group. Moreover, 81 potential active ingredients were identified in LLTD, with 645 overlapping targets between component targets and disease targets. Network analysis identified Pinocembrin, Magnoflorine, Jatrorrhizine as key active ingredients, and AKT1, TNF, IL-6, STAT3, and Bcl-2 as primary core targets. A total of 1792 biological processes were affected according to GO analysis, and 187 pathways were identified through KEGG analysis. Finally, molecular docking and experimental results validated that LLTD could alleviate cardiomyocyte injury in CHD by regulating the primary core targets.

Discussion: This study indicates that LLTD may achieve systematic modulating of the signaling network through a "network pharmacology" model, which provides valuable insights for the development of multi-target therapies targeting the complex pathological mechanism underlying CHD.

Conclusion: LLTD may exert cardioprotective effects by regulating inflammatory responses, apoptosis, and oxidative stress.

Introduction: Hyperglycemia-induced renal inflammation and the subsequent activation of inflammatory cytokines play critical roles in Diabetic Kidney Disease (DKD) progression. However, the underlying molecular mechanisms, particularly those involving microRNAs, remain poorly understood.

Methods: Serum samples from DKD patients (n = 10) and healthy controls (n = 10) were subjected to microRNA transcriptome profiling. An in vitro DKD model was established by treating Human Renal Tubular Epithelial Cells (RTECs and HK-2) with High Glucose (HG). The levels of pro-inflammatory cytokines, Interleukin (IL)-1β, IL-6, and Tumor Necrosis Factor (TNF)-α, were assessed using quantitative reverse transcription-PCR and enzyme-linked immunosorbent assay. Dual-luciferase reporter assays evaluated the relationship between miR-509-3p and Yesassociated protein 1 (YAP1). In situ hybridization and immunohistochemistry were performed to validate the expressions of miR-509-3p and YAP1 in kidney tissues from DKD patients (n = 38) and controls (n = 38).

Results: MiR-509-3p was identified as one of the most significantly dysregulated microRNAs in the serum of DKD patients. Its expression was significantly downregulated in a dose-dependent manner in HG-induced RTECs and HK-2 cells (p <0.01). Overexpression of miR-509-3p significantly suppressed HG-induced pro-inflammatory cytokine expression (p <0.01). Suppression of miR-509-3p further enhanced HG-induced IL-1β, TNF-α, and IL-6 expression, whereas this effect was reversed by silencing YAP1 (p <0.01). Mechanistically, miR-509-3p directly targeted the 3'-UTR of YAP1 mRNA. Consistently, DKD renal tissues exhibited reduced miR-509-3p and elevated YAP1 expression, with a significant negative correlation (p = 0.002). Furthermore, miR-509-3p and YAP1 were significantly correlated with estimated glomerular filtration rate and urine albumin-creatinine ratio (p <0.001).

Conclusion: The study preliminarily suggests that miR-509-3p/YAP1 axis may act as a crucial regulator of hyperglycemia-induced tubular inflammation, offering a potential therapeutic target for DKD.

Introduction: Cartilage defects (CDs) are orthopedic conditions with limited regenerative potential. This study aimed to identify endoplasmic reticulum (ER) stress-related biomarkers and construct a diagnostic model to enhance the early detection of CD.

Methods: This study analyzed the transcriptomic dataset GSE129147 to identify ER stressrelated differentially expressed genes (ERSRDEGs) between CD and control tissues using the limma package (version 3.58.1). Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analyses were employed for functional enrichment. Immune infiltration was assessed using cell-type identification, which involved estimating the relative subsets of RNA transcripts and single-sample gene set enrichment analysis. Diagnostic models were constructed using logistic regression, support vector machine, and least absolute shrinkage and selection operator regression.

Results: Twenty ERSRDEGs were identified, with CYBB, ATP6V1A, and TNFRSF12A significantly upregulated in CD samples. GO and KEGG analyses highlighted oxidative stress response and extracellular matrix remodeling as key mechanisms in CD pathogenesis. Immune analysis revealed an increase in regulatory T cells and a reduction in CD8⁺ T cells. TNFRSF12A showed strong immune associations and, together with TWIST1 and ATP6V1A, formed the final preliminary diagnostic model. The preliminary LASSO model achieved satisfactory predictive accuracy (AUC: 0.7-0.9).

Discussion: These findings suggest that ER stress and immune imbalance jointly contribute to cartilage degeneration. The identified genes, particularly TNFRSF12A, TWIST1, and ATP6V1A, not only serve as potential biomarkers but also provide preliminary evidence for new mechanistic insights into stress-immune crosstalk in CD.

Conclusion: This study reveals the key roles of ER stress and immune dysregulation in CDs. Moreover, the ERSRDEG-based diagnostic model provides preliminary bioinformatics evidence and potential molecular indicators for targeted diagnostics and therapies.

Introduction: All-trans retinoic acid (ATRA), a therapeutic mainstay for acute promyelocytic leukemia, is associated with off-target effects on skeletal development, including premature growth plate closure. However, the molecular mechanisms underlying ATRA-induced growth plate senescence remain poorly understood.

Methods: Using Sprague-Dawley rats, ADTC5 chondrocyte cell lines, and integrated multiomics approaches (transcriptome sequencing, weighted gene co-expression network analysis, molecular docking, and functional assays), we investigated how ATRA modulates growth plate development. Animal models were treated with graded ATRA doses, while in vitro studies included cell viability assays, RNA interference, and Western blot analysis to validate interactions in the signaling pathway.

Results: ATRA induced dose-dependent growth plate thinning (high-dose: 59.79 μm vs. control: 511.35 μm) and skeletal growth retardation in rats. Transcriptomic analysis identified ITGB2 as a pivotal gene, with molecular docking revealing a strong binding interaction (-240.25 kcal/mol) between ITGB2 and YAP mediated by hydrogen bonds/salt bridges. Functional experiments revealed that ATRA upregulated ITGB2, which activated YAP, a Hippo pathway effector, thereby suppressing Wnt/β-catenin signaling by inhibiting β-catenin. This led to downregulation of osteogenic markers (Runx2/SOX9) and enhanced growth plate closure. YAP knockdown reversed these effects, restoring β-catenin and downstream target gene expression (c-myc, cyclin D).

Conclusion: ATRA accelerates growth plate closure through the ITGB2-YAP axis, disrupting Wnt/β-catenin signaling. These findings establish a mechanistic framework for developing therapeutic strategies targeting ITGB2 or YAP to delay premature growth plate senescence in pediatric disorders.

Introduction: This study aimed to investigate the therapeutic mechanism of Duzhong Bushen Formula (DBF) in ameliorating lumbar Intervertebral Disc Degeneration (IDD) via regulation of the MAPK/AP-1 pathway.

Methods: Based on a circular puncture-induced rat model of IDD, network pharmacology and molecular docking analysis were employed to identify key therapeutic targets and pathways for DBF in treating IDD. In vivo validation of these targets was conducted via MRI, histopathology, ELISA, Western blot, and qPCR, with therapeutic efficacy further evaluated.

Results: DBF administration significantly reduced inflammatory markers (CRP, IL-6, TNF-α), increased serum levels of Albumin (ALB) and Estrogen (E2), and downregulated key components of the MAPK/AP-1 pathway (p-p38, c-FOS, c-JUN). High-dose DBF demonstrated the most pronounced therapeutic effects.

Discussion: The results suggest that DBF attenuates IDD by suppressing inflammatory responses and oxidative stress through modulation of the MAPK/AP-1 signaling axis.

Conclusion: DBF may be a promising multi-target therapeutic agent for the treatment of IDD, particularly in cases involving kidney deficiency and blood stasis.