Combinatorial chemistry & high throughput screening最新文献

Introduction: Depression is a common comorbidity in epilepsy, significantly impacting patients' quality of life. The hippocampus, linked to depression and neurodegeneration, is vulnerable in epilepsy. Epileptogenesis involves inflammation, oxidative stress, and neuronal damage, with the PI3K/AKT pathway playing a key role. Apigenin (API), a flavonoid in fruits and vegetables, shows neuroprotective, anti-inflammatory, and anti-apoptotic effects. This study investigates API's mechanisms in a LiCl-pilocarpine epileptic rat model, focusing on hippocampal neurogenesis and PI3K/AKT signaling as potential therapeutic targets.

Methods: We studied the effects of API and valproate (VPA) on depressive behavior and astrocytes in Lithium chloride (LiCl)-pilocarpine-induced epileptic rats. Additionally, we predicted the potential molecular targets of API for treating epilepsy using network pharmacology. Finally, we conducted in vivo experiments to validate the predicted mechanism.

Results: In the API and VPA groups, there was a reduction in seizure frequency and seizure severity compared with the control group. The model group showed more depressive behavior than the control (CON) group, and these behaviors improved significantly after VPA and API treatment. HE staining showed that both API and VPA treatment improved LiCl-pilocarpine-induced nuclear contraction and cell swelling. Nissl staining demonstrated that Nissl vesicles in the CA3 region of the hippocampus were decreased in the model group, but the neurons were larger, more abundant, and more neatly arranged after API and VPA treatment. In the model group, the p-PI3K/PI3K and p-AKT/AKT protein ratios and PI3K, AKT mRNA expression were reduced, while brain-derived neurotrophic factor (BDNF) and glial fibrillary acidic protein (GFAP) were markedly increased. API and VPA treatment effectively reversed these changes.

Discussion: API reduces seizures and depressive behaviors in LiCl-pilocarpine-induced epileptic rats, comparable to VPA API mitigates hippocampal neuronal damage, preserves Nissl bodies, and suppresses astrocyte activation via the PI3K/AKT pathway, suggesting neuroprotective and anti-inflammatory effects. While API shows promise as an antiepileptic and antidepressant agent, further studies are needed to confirm its direct modulation of PI3K/AKT and efficacy in other epilepsy models.

Conclusion: Our study suggests that API improves depression in rats and has anti-epilepsy activity, which may be involved in activating the PI3K/AKT pathway to protect astrocytes.

Background: Some studies have shown a link between Alzheimer's disease (AD) and COVID-19. This includes a Mendelian randomization study, which suggests that Alzheimer's disease and COVID-19 may be causally linked in terms of pathogenic mechanisms. However, there are fewer studies related to the two in terms of common pathogenic genes and immune infiltration. We conducted this study to identify key genes in COVID-19 linked to Alzheimer's disease, assess their relevance to immune cell profiles, and explore potential novel biomarkers.

Methods: The RNA datasets GSE157103 and GSE125583 for COVID-19 and Alzheimer's disease, respectively, were acquired via the GEO database and subsequently processed. Through the utilization of differential expression analysis and Weighted Gene Co-expression Network Analysis (WGCNA), genes associated with Alzheimer's disease and COVID-19 were identified. The immune cell signatures were estimated using the xCell algorithm, and correlation analysis identified links between key genes and significantly different immune cell signatures. Finally, we conducted transcription factor (TF) analysis, mRNA analysis, and sensitivity drug analysis.

Results: Differential analysis identified 3560 (2099 up-regulated and 1461 down-regulated) and 1456 (640 up-regulated and 816 down-regulated) differential genes for COVID-19 and AD compared to normal controls, respectively. WGCNA analysis revealed 254 key module genes for COVID-19 and 791 for AD. We combined the differential genes and WGCNA key module genes for each disease to obtain two gene sets. The intersection of these two gene sets was examined to obtain intersecting genes. Subsequently, PPI network analysis was conducted, leading to the identification of 12 hub genes. Then, 12 immune-related hub genes were further identified. Immune infiltration patterns and the correlation between 12 hub genes and 64 immune cell types were analyzed. The analysis revealed a significant positive correlation between the two diseases under study. The relationship network between Transcription Factors and mRNA, as well as the predictions of drugs, further illustrate the strong association between the two diseases. This provides valuable information for further target exploration and drug screening.

Conclusion: Our study suggests potential shared genes, signalling pathways, and common drug candidates that may be associated with COVID-19 and AD. This may provide insights for future studies of AD patients infected with SARS-CoV-2 and help improve diagnostic and therapeutic approaches.

Objective: This study aimed to characterize the bioactive compounds in Guang hawthorn (Malus doumeri) leaf extracts and evaluate their potential anti-obesity effects.

Methods: 70% Ethanol and ethyl acetate extracts from M. doumeri leaves were administered to high-fat diet-induced obese rats. Serum levels of total cholesterol (TC), triglycerides (TG), highdensity lipoprotein (HDL-C), and low-density lipoprotein (LDL-C) were assessed. The chemical composition of the leaf extracts (HML) was analyzed using multi-chromatography, NMR, and mass spectrometry. Network pharmacology approaches were employed to explore the interactions between the identified compounds and obesity-related targets.

Results: Rats receiving a high dose of the leaf extracts exhibited significantly lower TC, TG, and LDL-C levels compared to controls (p < 0.05), with a concurrent increase in HDL-C (p < 0.05). Three bioactive compounds-phlorizin, sieboldin, and kumatakenin β-7-O-glucoside-were identified in HML. Through database screening, 373 unique targets for these compounds and 8484 obesity-related targets were identified. Using R and Cytoscape software, 272 overlapping targets and 32 core targets were determined. Functional enrichment analysis revealed that the anti- obesity effects of HML were related to phosphorylation, cell proliferation, apoptosis, and kinase activities. Notably, the 32 core targets were implicated in cancer-related proteoglycans, progesterone- mediated oocyte maturation via FoxO, and the PI3K-Akt signaling pathway.

Conclusion: This research demonstrates the anti-obesity potential of sieboldin and kumatakenin β-7-O-glucoside in HML. These compounds, along with phlorizin, modulated the PI3K-Akt and FoxO signaling pathways and proteoglycan synthesis, contributing to the reversal of obesityrelated effects. These findings suggest that M. doumeri leaf extracts may offer a promising natural therapeutic approach for obesity management.

Objective: The objective of this investigation was to examine the mechanism through which Radix isatidis operates, utilizing network pharmacology and molecular docking techniques.

Methods: Active components and associated targets of Radix isatidis were identified using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) alongside the GeneCards database. A protein-protein interaction (PPI) network connecting the targets of the active ingredients with those related to febrile diseases was constructed through STRING. The analysis of core nodes was carried out using Cytoscape software, followed by a further exploration of the PPI network using the DAVID database. Lastly, the underlying mechanism of the antipyretic action was also examined utilizing the DAVID database. Functional annotation through Gene Ontology (GO) and enrichment analysis of pathways from the Kyoto Encyclopedia of Genes and Genomes (KEGG) were conducted utilizing the DAVID database, with essential components chosen for molecular docking through Pymol software. Mice were injected intraperitoneally with lipopolysaccharide (LPS) and treated by continuous gavage with Radix isatidis. They were then evaluated using temperature monitoring, blood tests, organ index calculation, PI3K-AKT pathway protein assays, and inflammatory factor reverse transcription polymerase chain reaction (RT-PCR) assays.

Results: Twelve active components of Radix isatidis were screened, and 107 genes were identified at the intersection of Radix isatidis and fever. These genes were found to be involved in the PI3K-AKT signaling pathway, proteoglycans in cancer, and mechanisms related to blood lipids and atherosclerosis. The top nine targets screened by constructing a PPI network were IL6, AKT1, EGFR, STAT3, CASP3, ESR1, PTGS2, PPARG, and MAPK3, which indicated that Radix isatidis may play a protective role by affecting the PI3K/AKT-related signaling pathway. In in vivo experiments, Radix isatidis inhibited the activation of PI3K/AKT-related pathways, reduced inflammatory responses, and improved symptoms in mice treated with LPS. Conclusion The molecular mechanism of action of the antipyretic effect of Radix isatidis was predicted through network pharmacology, and it was also verified in vivo, as Radix isatidis reduced the inflammatory response, lowered the body temperature of mice, and protected LPStreated mice via effects on the PI3K-AKT-related signaling pathway.

Objective: To elucidate the therapeutic mechanisms of Shenling Baizhu Powder (SLBZ) in treating ulcerative colitis (UC) through an integrated approach combining network pharmacology and experimental validation, highlighting its potential as an alternative or complementary therapy.

Methods: The active constituents and corresponding targets of SLBZ were identified using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). UC-related targets were obtained from DrugBank and the Comparative Toxicogenomics Database (CTD). Overlapping targets were visualized using a Venn diagram, and protein- protein interactions were mapped using the STRING database. The "compound-componentdisease- target" network was constructed through GO function analysis and KEGG pathway enrichment analysis. Molecular docking was performed using Discovery Studio 4.5 software. A UC mouse model was induced with dextran sodium sulfate (DSS), and mice were divided into normal, UC, SLBZ (low, medium, high doses), and mesalazine groups, treated for 14 days. Histopathological changes in colon tissues were assessed, and serum levels of IL-4, TNF-α, and HIF-1α were measured by immunohistochemistry to verify SLBZ's targets in UC treatment.

Results: A total of 408 active ingredients and 2118 corresponding targets were identified in SLBZ, with 2052 targets associated with UC, resulting in 610 overlapping targets. Further refinement identified 54 active ingredients and 86 targets, with quercetin, betulin, catharanthamine, and glyasperin B as key components. Core targets included TP53, AKT1, JUN, and HSP90AA1. KEGG analysis suggested SLBZ's involvement in PI3K/Akt, JAK2/STAT3, and TNF signaling pathways. Molecular docking confirmed strong binding affinities between key targets and main components. Histological analysis showed that SLBZ alleviated DSS-induced colonic tissue injury, while immunohistochemistry revealed reduced expression of TNF-α and STAT3, and upregulated IL-4, indicating modulation of the JAK/STAT3 pathway.

Conclusion: SLBZ exerts therapeutic effects on UC by targeting key proteins (e.g., TP53, AKT1, JUN) and modulating critical pathways (e.g., JAK2/STAT3). This study provides a theoretical basis for the potential of SLBZ as a multi-targeted therapeutic agent for UC, which can be further explored through clinical trials and multi-omics analyses to validate its efficacy and safety.

Objective: The objective of this study is to develop and characterize a Crocus sativus (saffron)-based solid lipid nanoparticle (SLN) nasal spray for treating depression by enabling direct nose-to-brain delivery and evaluating its antidepressant potential in a Drosophila melanogaster model.

Materials and methods: Phytochemical screening, antioxidant assays, and HPLC quantification of picrocrocin were performed on Crocus sativus extract. The SLN-based nasal spray was formulated and characterized for particle size, zeta potential, polydispersity index (PDI), drug entrapment efficiency, in vitro drug release, and stability over 4 weeks. The antidepressant efficacy was assessed via a climbing assay in Drosophila melanogaster.

Results: Phytochemical analysis revealed phenolic content (11-36 μg GAE/mg), flavonoid content (43-56 μg QE/mg), and carotenoid content (1.9-30 μg βC/mg). HPLC analysis quantified picrocrocin at 6.3 mg/g, confirming its presence. The SLNs exhibited a particle size of 110-225 nm, a zeta potential of -1 to -0.8 mV, a PDI of 1, and a drug entrapment efficiency of 99.76%. Drug release reached 37% over 270 minutes, and the nasal spray maintained a pH of 5.8, a viscosity of 23.1 cP, and stability over 4 weeks. In vivo, the climbing assay demonstrated improved locomotor activity, indicating significant antidepressant potential.

Conclusion: The Crocus sativus SLN nasal spray exhibits high entrapment efficiency, stability, and promising in vivo antidepressant effects, supporting its potential as a novel therapy for treatment-resistant depression via direct nose-to-brain delivery.

Background: Soaps are vital for preserving our health and personal hygiene since they not only eliminate germs but also rid the body of pollutants.

Method: The current study aims to determine the physicochemical and antibacterial properties of Eucalyptus camaldulensis leaves using the agar disc diffusion technique and assess the effectiveness of different branded liquid soaps (25 mg/ml, 50 mg/ml, 75 mg/ml, and 100 mg/ml) with the Eucalyptus leaf extract against skin-infecting human pathogenic bacteria.

Results: The combined antimicrobial susceptibility of E. camaldulensis and five liquid soaps showed an inhibition zone of 17.67±0.58, 13.33±0.58, 12.67±0.58, and 15.67±0.58 against Staphylococcus aureus, Streptococcus pyogenes, Pseudomonas aeruginosa, and Escherichia coli. The antibacterial properties of Av soap by itself did not work against S. pyogenes. Nevertheless, the extract and DI together showed a detrimental effect against S. aureus and P. aeruginosa, with no halo forming.

Conclusion: Antimicrobial activity was observed to increase with higher concentrations of the soap-extract combinations. Although liquid soap (seve) was effective against bacterial isolates, a combination of eucalyptus and aqua vera was shown to be more effective.

Background: Obstructive sleep apnea syndrome [OSAS] is a common sleep breathing disorder accompanied by multiple organ intermittent hypoxemia. Our previous study has suggested that the expression of a lncRNA termed ENST00000592016 [lnc2016 for short] derived from plasma exosomes is remarkably elevated in OSA patients compared to the normal population, and lnc2016 can improve the diagnostic efficiency of OSA.

Objective: To unmask the role of the lnc2016 in vascular endothelial cells, targeted hypoxia is the goal of the current research.

Methods: Primary human ADSCs and HUVEC cells were cultured. CCK-8, cytometric assay, transwell, and tubular formation assay were used to determine cell viability, cell apoptosis, cell cycle, cell migration, as well as tubular formation ability.

Results: we found that adipose-derived stem cells [ADSCs]-derived exosomes contained robust lnc2016. After co-culture with human umbilical vein endothelial cells [HUVECs], exosomal lnc2016 could enhance cell proliferation, DNA synthesis, migration, and tubular formation, whereas suppress cell apoptosis of HUVECs against hypoxic conditions.

Conclusion: Our data have revealed that ln2016 can promote the cell growth, migration, DNA synthesis, and tubular formation as well as suppress the cell apoptosis of vascular endothelial cells against hypoxia in vitro.

Objectives: Population with Latent tuberculosis infection (LTBI) is the principal source of active tuberculosis (ATB) cases. The identification of reliable diagnostic biomarkers is critical for the prevention and control of the progression from LTBI to ATB. The aim of this study is to screen biomarkers that can distinguish LTBI from ATB patients by using a comprehensive bioinformatics analysis strategy.

Methods: The transcriptomic datasets were obtained from the GEO database. Hub genes and critical signal pathways for differentiating latent and active TB, were identified by a comprehensive bioinformatics analysis strategy comprising Weighted Gene Co-Expression Network Analysis (WGCNA), Differentially Expressed Gene (DEG), Protein-Protein Interaction (PPI), Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, and hub genes were verified by RT-qPCR in this study.

Results: The transcriptome profiles of GSE193777, GSE157657, GSE168519, GSE107991, and GSE107992 were extracted from the GEO database, in which a total of 18,397 protein-coding genes from 206 samples were included in the bioinformatics analysis. Combined with Weighted Gene Co-Expression Network, differentially expressed gene, functional enrichment, and proteinprotein interaction analyses, six hub genes were identified. The results of RT-qPCR confirmed that the expression levels of four hub genes (HLA-DOA, ECH1, PARN and TRAPPC4) were downregulated in the LTBI group compared with the ATB group.

Conclusion: Our findings may provide crucial clues to potential biomarkers that can distinguish patients with LTBI from those with ATB, aiding the understanding of the mechanism underlying the progression of LTBI to ATB.

Background: ARL6IP1 has been linked to cancer progression, but its precise role in BC, particularly in metabolism and its interaction with an OLFM4, remains unclear.

Aims: This study aimed to investigate the role of ADP-ribosylation factor-like 6 interacting protein 1 (ARL6IP1) in breast cancer (BC) cell behavior and metabolism and explore its interaction with an olfactomedin-4 (OLFM4) as a potential therapeutic target.

Objective: The objective of this study was to determine the effects of ARL6IP1 knockdown on BC cell proliferation, invasion, migration, apoptosis, oxidative stress, and glycolysis. Additionally, this study also explored the interaction between ARL6IP1 and OLFM4 and their combined role in BC progression and metabolism.

Methods: Key gene modules in the GSE73540 dataset were identified through weighted gene co-expression network analysis (WGCNA). Three BC-related datasets (GSE73540, GSE22820, and GSE36295) and The Cancer Genome Atlas (TCGA) were applied for additional examination of differentially expressed genes (DEGs). Intersection analysis selected ARL6IP1 as a hub gene for prognostic analysis. In vitro experiments investigated how ARL6IP1 knockdown influences BC cell proliferation, invasion, migration, apoptosis, epithelial-mesenchymal transition (EMT), oxidative stress, and glycolysis. The connection between ARL6IP1 and an OLFM4 was confirmed using Co-immunoprecipitation (Co-IP), and their roles in BC tumor progression and glycolysis were evaluated.

Results: ARL6IP1 was elevated in BC datasets and linked with poor BC prognosis. Experiments demonstrated that knockdown of ARL6IP1 significantly reduced BC cell growth while promoting apoptosis and oxidative stress. Besides, ARL6IP1 knockdown reduced glycolysis, as manifested by decreased extracellular acidification rate (ECAR), glucose consumption, adenosine triphosphate (ATP) levels, and lactate production while increasing mitochondrial respiration (OCR). Co-IP validated the connection between ARL6IP1 and OLFM4, and OLFM4 overexpression partially counteracted the suppression of glycolysis and cell behavior resulting from ARL6IP1 knockdown.

Conclusion: ARL6IP1 is a critical regulator of BC progression, influencing glycolysis, mitochondrial function, and key cellular behaviors. Targeting the ARL6IP1-OLFM4 axis offers a promising therapeutic strategy for managing BC.