Combinatorial Chemistry & High Throughput Screening - Online First

This study aimed to investigate the mechanism of Juanbi Lijieqing Decoction (JLD) in alleviating acute gouty arthritis (AGA) by modulating PPARγ expression to suppress the TLR4/NF-κB pathway.

A total of 84 male SD rats were divided into 7 groups of 12 rats. One group was randomly selected as the normal control group (Group A), while the remaining 72 rats were used to establish an acute gouty arthritis model through intraperitoneal injection of potassium oxonate combined with MSU ankle joint injection. These rats were randomly assigned to the model group (Group B), the high-dose Juanbi Lijieqing Decoction group (Group C), the medium-dose group (Group D), the low-dose group (Group E), the etoricoxib group (Group F), and the pioglitazone group (Group G), with 12 rats per group. The acute gouty arthritis model was established by intraperitoneal injection of potassium oxonate, followed by monosodium urate (MSU) injection into the ankle joint, and then by pharmacological intervention in each group. The ankle swelling index, pain threshold changes, and serum uric acid levels were observed in each group of rats. The pathological state of synovial tissue in each group was evaluated by hematoxylin-eosin (HE) staining. The levels of TNF-α, IL-6, and IL-1β were detected by enzyme-linked immunosorbent assay (ELISA). The protein expressions of TLR4, NF-κB, and PPARγ were detected in vivo and in vitro using Western blot.

JLD effectively reduced local swelling, relieved pain, and lowered serum uric acid levels in rats with AGA. Both in vivo and in vitro experiments demonstrated that the Chinese medicine groups showed a significant reduction in TNF-α, IL-1β, and IL-6 levels. Moreover, in in vivo experiments, the expression of PPARγ protein was significantly upregulated in the JLD and pioglitazone groups, whereas the expressions of TLR4 and NF-κB p65 proteins were significantly downregulated, a pattern not observed in the etoricoxib group. In vitro experiments demonstrated significant increases in PPARγ protein expression in the pioglitazone and medicated serum groups, accompanied by significant decreases in TLR4 protein expression. Meanwhile, the NF-κB inhibitor group only exhibited a downregulation of TLR4 protein expression.

Our findings demonstrated that JLD alleviated acute gouty arthritis by upregulating PPARγ expression, which subsequently inhibited the TLR4/NF-κB signaling pathway. This mechanism effectively reduced inflammatory cytokine production (TNF-α, IL-1β, and IL-6), explaining the observed anti-swelling and analgesic effects.

JLD mitigates AGA symptoms by promoting PPARγ, which in turn inhibits TLR4/NF-κB signaling, thereby reducing inflammation, uric acid, and joint swelling. This highlights the therapeutic potential of JLD for gout management, though long-term effects and molecular targets warrant further study.

Luohuazizhu granules (LHZZG) are made of Callicarpa nudiflora Hook. (CN), which is used to treat ulcerative colitis (UC). The anti-inflammatory effects of CN on UC have been previously reported. However, the biological effects of LHZZG on bile acids (BAs) in UC and the underlying mechanisms remain unexplored.

Integrated metabolomics were used to explore the regulatory mechanisms of LHZZG for BA metabolism in UC mice. Both 16S rDNA sequencing and flow cytometry analyses were combined to comprehensively assess gut microbiota (GM) and immune responses.

Twenty-five differential biomarkers were identified in the untargeted metabolomic analysis, most of which were correlated with BA metabolism. UC signs were significantly alleviated after LHZZG treatment. The targeted metabolomics analysis revealed BA metabolic disorders to be significantly improved following LHZZG treatment. Additionally, the imbalances in the GM and immune cells related to BA metabolism were restored.

This study not only confirmed significant dose-dependent protective effects of LHZZG in UC mice, but also performed the first investigation into the underlying mechanisms related to BA metabolism and immune function. Nevertheless, the limitations precluded a definitive mechanistic explanation for the observed changes. Consequently, in-depth mechanistic investigations will be prioritized in subsequent research to experimentally validate this hypothesis.

BAs could serve as biomarkers for evaluating the therapeutic effects of LHZZG on UC. This study has provided the first detailed explanation of the mechanism underlying the effects of LHZZG from a BA metabolic perspective, providing a foundation for their clinical application in UC.

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.

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.

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.

This study identified immune-related hub genes and demonstrated their association with natural killer T cell dysfunction in AD and COVID-19, suggesting the existence of common neuroinflammatory pathways. These findings provide molecular evidence for immunological crosstalk between the two diseases.

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.

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

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 the core nodes was conducted using the Cytoscape software, followed by further exploring the PPI network using the DAVID database. Lastly, the underlying mechanism of the antipyretic action was also examined utilizing the DAVID database. Mice were injected intraperitoneally with lipopolysaccharides (LPS) and treated by continuous gavage with Radix isatidis. The mice were then evaluated using temperature monitoring, blood tests, organ index calculations, PI3K-AKT pathway protein assays, and reverse transcription polymerase chain reaction (RT-PCR) assays for inflammatory factors.

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 identified by constructing a PPI network were IL6, AKT1, EGFR, STAT3, CASP3, ESR1, PTGS2, PPARG, and MAPK3, indicating that Radix isatidis may play a protective role by affecting the PI3K/AKT-related signaling pathway.

In the in vitro experimental validation, a fever model was established using LPS, while Radix isatidis was used for treatment, and the PI3K/AKT/NF-κB pathway was validated by temperature monitoring, observation of pathological tissue sections, western blotting, immunohistochemistry, RT-PCR, and other technical means. In vivo experiments were conducted to verify the method in multiple mediums, and both the genetic changes and related pathway proteins are consistent with the KEGG prediction.

The PI3K/AKT pathway was identified through PPI network analysis, key target identification, and KEGG pathway enrichment. Subsequent in vivo studies in mice confirmed that Radix isatidis could alleviate inflammation and body fever caused by LPS by affecting the PI3K/AKT pathway.

To explore the therapeutic mechanisms of Shenling Baizhu Powder (SLBZ) in ulcerative colitis (UC) using network pharmacology and experimental validation, assessing its potential as an alternative therapy.

Active constituents and targets of SLBZ were identified using TCMSP, DrugBank, and CTD. A UC mouse model was induced with DSS and treated with SLBZ for 14 days. Histopathological changes and serum levels of IL-4, TNF-α, and HIF-1α were measured.

SLBZ contained 408 active ingredients with 2118 targets, 610 of which were associated with UC. Key components included quercetin, betulin, catharanthine, and glyasperin B. Core targets were TP53, AKT1, JUN, and HSP90AA1. SLBZ modulated PI3K/Akt, JAK2/STAT3, and TNF pathways. Histological analysis showed SLBZ alleviated DSS-induced colonic tissue injury, reduced TNF-α and STAT3, and upregulated IL-4.

SLBZ targets key proteins and pathways in UC, suggesting its potential as a multi-targeted therapeutic agent. Further studies are needed to validate its efficacy and safety.

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.

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.

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.

The favorable physicochemical characteristics of the nasal spray, along with the observed behavioral improvements in the fly model, suggest that Crocus sativus SLNs effectively cross the nasal-brain barrier and exert antidepressant-like effects. These findings support its potential for non-invasive management of treatment-resistant depression.

Transcriptome-level insights into electroacupuncture (EA)’s mechanisms for alleviating intestinal mucosal barrier damage in diarrhea-predominant irritable bowel syndrome (IBS-D) are limited. This study aimed to construct ceRNA networks and elucidate EA's role in restoring barrier integrity via lncRNA-miRNA-mRNA regulation in IBS-D rats.

The IBS-D model was established by neonatal maternal separation (NMS), 4% acetic acid enema and restrain stress (RS). Rats were randomized into control, model, and EA groups. After 2-week EA treatment, colonic morphology was assessed by HE staining and TEM; intestinal barrier biomarkers were analyzed via ELISA and WB. RNA-seq identified differentially expressed RNAs (DE RNAs) to construct ceRNA networks. GO and KEGG analyzed EA-modulated DE mRNAs. RT-qPCR validated RNA-seq; WB and IF confirmed mast cell (MC) involvement in EA-regulated pathways.

RNA-seq identified 426 up-regulated and 429 down-regulated DE mRNAs, 342 up-regulated and 362 down-regulated DE lncRNAs, and 10 up-regulated and 48 down-regulated DE miRNAs following EA. Constructed ceRNA networks included 7 DE lncRNAs-miR-139-3p-Bid and -miR-378b-Slc4a5. GO analysis linked EA to defense response, hormone regulation, and cytokine function pathways. KEGG implicated antigen processing/presentation, neuroactive ligand-receptor interaction, PPAR signaling, and glutathione metabolism. RT-qPCR validated RNA-seq results.

This RNA-seq study reveals EA mitigates IBS-D intestinal mucosal barrier damage by regulating genes and ceRNA networks, providing novel transcriptomic insights into its therapeutic mechanisms.

The objective of this study is to evaluate the role of mycoplasma resistance genes in pediatric mycoplasma pneumonia and to identify the factors that necessitate antibiotic adjustments.

We retrospectively analyzed clinical data from children diagnosed with mycoplasma pneumonia at Chongqing Medical University Children's Hospital (January-October 2023). We categorized patients based on antibiotic treatment adjustments: the antibiotic adjustment group and the no adjustment group. We compared demographic characteristics, clinical outcomes, and the gene resistance rate that point mutations A2063G and A2064G in the 23S rRNA between groups. Logistic regression was employed to determine the factors prompting a switch from macrolides to alternative antibiotics.

The study included 551 cases, with 341 in the no adjustment group and 210 in the antibiotic adjustment group (54 switched to doxycycline, 156 to levofloxacin). There was no significant difference in the prevalence of resistance genes between the groups (71.8% vs. 71.4%; P=0.916). Significant differences were observed in hospital stay duration, C-reactive protein (CRP), D-dimer, fibrinogen, procalcitonin levels, and lung consolidation (P<0.05). Logistic regression identified elevated CRP and procalcitonin levels as independent predictors of the need for alternative antibiotics.

Resistance genes do not predict the need for second-line antibiotics in pediatric mycoplasma pneumonia; however, elevated CRP, and procalcitonin levels significantly correlate with this necessity.

Despite the global burden of chronic obstructive pulmonary disease (COPD), its pathogenesis remains elusive, and current therapies fail to halt disease progression. Calycosin, a bioactive isoflavone from Traditional Chinese Medicine (TCM), exhibits anti-inflammatory and antioxidant properties, yet its therapeutic potential in COPD remains unexplored.

We integrated transcriptomic data of human lung tissue from the Gene Expression Omnibus (GEO) database (GSE8581) and human serum metabolomic data from a published research paper to identify COPD-associated pathways. Network pharmacology, including target screening, analysis, and molecule docking, was employed to elucidate the mechanisms of calycosin for COPD therapy.

Multi-omics analysis revealed significant activation of the pyruvate metabolism pathway and glyoxylate/dicarboxylate metabolism pathway in COPD patients, with 590 differentially expressed genes (DEGs) and 116 differentially expressed metabolites (DEMs) identified. Calycosin targets six key regulators (NME1, ALDH2, PGAM1, LDHA, PCNA, RASD1) with binding affinities (−5.1 to −10.2 kcal/mol) validated via molecular docking, implicated in these pathways.

This study bridges TCM-derived natural products with modern omics-driven drug discovery, revealing calycosin as a promising COPD intervention by targeting metabolic hubs to mitigate inflammation and metabolic dysfunction, and pioneering an integrated multi-omics and network pharmacology framework for elucidating TCM mechanisms. However, the lack of direct experimental validation in COPD models and the use of data from different biological sources limit the extrapolation of the results. Further in vitro and in vivo experiments are needed.

This integrated analysis highlights distinct metabolic pathway perturbations, specifically in pyruvate and glyoxylate/dicarboxylate metabolism, as key components of COPD pathophysiology and proposes calycosin as a mechanistically grounded candidate for modulating these pathways. This work shifts focus towards metabolic dysregulation as a central therapeutic target, providing a foundation for developing novel strategies to manage COPD beyond symptom control.

Pulsatillae radix (PR), a medicinal root plant and a well-known Chinese herbal remedy, is primarily used for its heat-clearing, detoxifying, blood-cooling, and antiinflammatory properties.

The constituents of PR were systematically analyzed using UHPLC-Q-TOF-MS/MS. Potential targets of active components were identified via the SwissTargetPrediction and PharmMapper databases, while (ulcerative colitis) UC-related disease targets were retrieved from GeneCard and other relevant databases. Overlapping targets between PR and UC were determined using Venn analysis. The STRING database was employed to generate a PPI network for the intersecting targets, and core targets were identified using the CytoNCA plugin. GO and KEGG pathway enrichment analyses were conducted using the DAVID platform. Lastly, molecular docking of key components with target proteins was carried out using PyMOL.

A total of 27 active compounds, 237 drug targets, and 4622 disease targets were identified. Intersection analysis revealed 141 shared targets, while the PPI network identified 10 hub targets. GO and KEGG enrichment analyses indicated that the hub targets were primarily associated with phosphorylation, cytoplasmic functions, nuclear receptor activity, as well as pathways related to the AGE-RAGE products signaling, TCR signaling, lipid and cholesterol metabolism, and various cancer-related pathways. Molecular docking experiments demonstrated that (+)- pinoresinol, cichoric acid, β-ecdysone, pulsatilla saponin D, 23-HBA, and AB4 exhibited stable binding to PIK3R1, TLR4, and ESR1, with AB4 forming the most stable complex with ESR1.

PR exerts therapeutic effects on UC through the synergistic actions of multi-components (AB4, 23-HBA), multi-targets (ESR1, TLR4, PIK3R1), and multi-pathways (AGE-RAGE, TCR).

Observational studies have shown a link between constipation (CN) and psychiatric disorders, including Schizophrenia (SP), Bipolar disorder (BD), Schizoaffective disorder (SD), and Parkinson’s disease (PD). However, it is still unknown whether CN affects the occurrence and development of psychiatric disorders or whether psychiatric disorders cause the occurrence and development of CN. Therefore, this study used Mendelian randomization (MR) analysis to evaluate the relationship between CN and psychiatric disorders.

We used genome-wide association studies (GWAS) to assess the relationship between constipation (N = 411, 623) and four psychiatric disorders, including SP (N = 77, 096), BD (N = 51, 710), SD (N = 210, 962), PD (N = 482, 730), using bidirectional MR analysis. Inverse variance weighting (IVW), MR Egger (ME) and Weighted median (WM) were used as causal analysis methods. Cochran's Q test, funnel plot, MR Egger intercept test and Leave‐one‐out analysis were used to detect sensitivity. Confounding factors were analyzed and eliminated by LDtrait to avoid influencing the final MR Analysis result.

The results of positive MR analysis indicated that there was no evidence of influence of constipation on SP (OR 1.043, 95%CI 0.946 - 1.149, P value = 0.398), BD (OR 1.114, 95%CI 0.995 - 1.248, P value = 0.062), SD (OR 0.934, 95%CI 0.674 - 1.294, P value = 0.682) and PD (OR 1.118, 95%CI 0.918 - 1.361, P value = 0.269) under gene prediction. Reverse MR analysis suggested that SP (OR 1.030, 95% CI 1.001-1.060, P value = 0.042) had a causal relationship with constipation. BD (OR 0.993, 95% CI 0.962-1.025, P value = 0.664), SD (OR 1.021, 95% CI 0.984-1.059, P value = 0.265) and PD (OR 1.004, 95% CI 0.974-1.035, P value = 0.790) were not associated with CN.

There was a positive association between SP and CN. CN may have no exact causal relationship with BD, SD and PD, and the interaction mechanism between these diseases needs to be further explored.

Bladder cancer (BC) is one of the most common urological malignancies, ranking as the eleventh most common cause of cancer-related deaths worldwide. The lack of specific and sensitive prognostic biomarkers presents a significant challenge in the early diagnosis and treatment of BC.

This study utilizes the Gene Expression Omnibus (GEO) dataset GSE13507 and the Cancer Genome Atlas (TCGA) database to screen differentially expressed genes related to BC. By using Weighted Gene Co-expression Network Analysis (WGCNA), two modules associated with BC were investigated in GSE13507 and TCGA. Hub genes were identified through Protein-Protein Interaction (PPI) network analysis, and their functions were validated through multiple approaches, including Gene Expression Profiling Interactive Analysis (GEPIA), Western Blotting (WB) assay, Human Protein Atlas (HPA), Oncomine analysis, and quantitative Real-Time PCR (qRT-PCR) analysis. Additionally, miRNAs associated with hub gene expression were identified using various databases to predict the progression and prognosis of BC.

WGCNA and a differential gene expression analysis identified 171 common genes as target genes. Ten genes (MYH11, ACTA2, TPM2, ACTG2, CALD1, MYL9, TPM1, MYLK, SORBS1, and LMOD1) were identified using the PPI tool. The CALD1 and MYLK genes showed a significant prognostic value for overall survival and disease-free survival in patients with BC. CALD1 and MYLK expression levels were significantly lower in BC tissues than in normal tissues. Furthermore, miR-155 showed a significant positive correlation with MYLK.

This study established MYLK as a direct target gene of miR-155, functioning as an actionable survival-related gene correlated with BC development.

This study aims to clarify the role of kelch like family member 17 (KLHL17) in cervical cancer (CESC) is unclear.

To clarify this uncertainty, our research employed bioinformatics analysis coupled with experimental corroboration.

We utilized the Cancer Genome Atlas (TCGA) database to assess the expression of KLHL17 in various cancers, specifically CESC, and to explore its association with clinical characteristics, diagnostic utility, and prognostic significance in CESC. The current investigation delved into the potential regulatory pathways related to KLHL17, examining its connection with the infiltration of immune cells, the expression of immune checkpoint genes, the status of microsatellite instability (MSI), and the efficacy of diverse therapeutic agents in CESC. The research analyzed KLHL17 expression patterns using single-cell sequencing data from CESC samples and investigated the genetic variations of KLHL17 within this context. KLHL17 expression was validated using GSE145372. The presence and levels of KLHL17 in different cell lines were validated through quantitative real-time PCR (qRT-PCR) assays.

KLHL17 exhibited irregular expression profiles across various cancer types, including CESC. Furthermore, increased KLHL17 levels in CESC patients were significantly associated with a lower progression-free survival (PFS) rate (hazard ratio: 1.62; 95% confidence interval: 1.01–2.60, p = 0.044). Moreover, KLHL17 expression emerged as a distinct prognostic indicator for CESC patients (p = 0.031). It has been associated with various biological pathways, such as cytokine-cytokine receptor interaction, primary immunodeficiency, cell adhesion molecules (CAMs), chemokine signaling pathway, steroid hormone biosynthesis, and others. The expression levels of KLHL17 were found to correlate with the presence of immune cells, the expression of immune checkpoint genes, and the status of MSI within CESC. Furthermore, KLHL17 expression exhibited a significant and inverse correlation with XMD15-27, rTRAIL, Paclitaxel, tp4ek, and tp4ek-k6. Furthermore, KLHL17 was found to be significantly positively regulated in CESC cell lines.

The findings suggest that KLHL17 is involved in the progression of CESC and may serve as a potential prognostic marker and therapeutic target. KLHL17's association with immune cell infiltration and immune checkpoint genes indicates a role in immuneevasion. Future research should focus on validating these findings through independent datasets and experimental studies to elucidate the molecular mechanisms underlying KLHL17's role in CESC progression and immune regulation.

KLHL17 is a promising prognostic marker and potential therapeutic target in CESC.

For clarifying the “multi genes and multi targets” characteristic of the treatment of Erxia Decoction (EXD), the aim of this study was to employ network pharmacology technology to perform cluster analysis on selected EXD targets.

EXD, a famous Chinese herbal prescription, consisting of Pinelliae Rhizoma (PR) and Prunellae Spica (PS), was mainly used to treat sleep disorder (SLD).

Using network pharmacology combined with proteomics to find out the main active components and core targets of EXD in the treatment of SLD.

By constructing the network of drug–component–target, the key protein targets of EXD for the treatment of SLD were screened. Then the interaction of the main active components of EXD and predicted candidate targets were verified. Then the proteomic analysis was used to screen the core targets in BV2 cells treated with EXD or the chemical ingredients, and the expression level was validated by Western blotting. Finally, molecular docking was used to further evaluate the mechanism of the action of the main ingredients and the core targets.

The 24 components of EXD mainly participate in the SLD treatment process by acting on 15 important key genes, and the core signal pathways were identified in the process of the action of EXD in treating SLD. Four key ingredients and five core targets were revealed from the results of network pharmacological analysis combination with proteomics, and then the AKT1 protein as a key target was validated by PCR and Western blotting.

This study preliminarily revealed EXD, morin (MOR) and quercetin (QUE) mainly inhibited the AKT1 core targets for the treatment of SLD using the network pharmacological analysis, proteomics, Western blotting and molecular docking.

The results elucidated partly the molecular mechanism and provided clues and a basis for further research.

Immunoglobulin A (IgA) Nephropathy (IgAN) is characterized by pIgA1 dep-osition in the glomerular mesangium, resulting in podocyte Epithelial-to-Mesenchymal Transi-tion (EMT). Traditional Chinese Medicines (TCMs) have been used for the treatment of IgAN for several years and have demonstrated positive efficacy. The present study aimed to establish a High-Content Screening (HCS) method for identifying wind-dampness dispelling TCM extracts that can mitigate podocyte EMT induced by pIgA1.

IgA1 from IgAN patients was used to establish a podocyte EMT model. The expression of EMT markers, including desmin, podocalyxin, and podocin, was assessed by Immunocytofluorescence (IF). An image-based HCS method was established to identify wind- dampness dispelling TCMs with the anti-EMT activity of podocyte EMT by automatic acquisi- tion and processing of dual-fluorescent labeled images.

A total of 21 wind-dampness-dispelling TCM extracts were screened using the HCS system, leading to the identification of eight wind-dampness-dispelling TCM extracts exhibiting anti-EMT activity. These eight extracts inhibited the pIgA-induced expression of desmin while upregulating the expression of podocalyxin and podocin.

By quantifying the changes in the expression of EMT markers during pIgA1- induced EMT, this study successfully identified wind-dampness dispelling TCM extracts with anti-EMT properties using an HCS system. In addition, the proposed approach presents a novel avenue for the identification of wind-dampness-dispelling TCMs for the treatment of pIgA1- induced EMT.

Allergic asthma is an inflammatory disease of the airways that causes great distress to the patient's normal life. Astragalus Polysaccharide (APS) is the main active ingredient in the traditional Chinese medicine Astragalus mongholicus Bunge, which has the effect of regulating immune function.

This study aimed to evaluate the effect of APS on allergic asthma and investigate its potential mechanism of action.

This study utilized network pharmacology to predict the relevant targets and signaling pathways of APS treatment for allergic asthma. Subsequently, an animal model was established using Ovalbumin (OVA) induction. The efficacy of APS was verified using histopathologic staining and Airway Hyperresponsiveness (AHR) assay. Signaling pathways were examined using Western Blot (WB). Finally, bioinformatics analysis was utilized to explore the correlation between the progression of allergic asthma and signaling pathways.

Network pharmacology analysis identified 15 intersection targets significantly enriched in the PI3K/AKT signaling pathway. The results of molecular docking showed that small molecule drugs have a strong binding ability to target proteins. The experiments confirmed APS successfully suppressed the pathological symptoms in allergic asthma model mice. Subsequently, WB provided evidence supporting that APS has potential therapeutic effects mediated through the PI3K/AKT signaling pathway. The bioinformatics results confirmed that disease progression in allergic asthma patients does correlate with the PI3K/AKT signaling pathway.

Our study suggests that APS may treat allergic asthma by targeting the PI3K/AKT signaling pathway. This provides a basis for preliminary research on the clinical application of APS for treating allergic asthma.

Our objective is to assess the therapeutic impact of HEC on OAB rats and investigate potential mechanisms.

Overactive bladder (OAB) is a syndrome of urinary storage symptoms characterized by “urinary urgency with or without urinary acute incontinence, usually accompanied by increased daytime and nocturnal urination”, which impacts patients’ quality of life. We found the potential therapeutic impact of HuangE capsules (HEC) on OAB patients through clinical practice. However, the exact effect and mechanism of action remain unclear.

We developed a “drugs- active ingredients- targets- diseases” network and employed the pathway enrichment analysis to identify the potential mechanisms of HEC on OAB. Bladder outlet obstruction (BOO) models and sham-operated ones were established in healthy male Wistar rats through surgical procedures. Following 28 days of continuous gavage administration of HEC, saturated copper sulfate test paper was utilized to quantify the frequency of urination over a 24-hour period. Subsequently, cystostomy was conducted to perform cystometry, and Masson staining was applied to a portion of the bladder tissue. Finally, we investigated the Rho/Rho-kinase pathway's expression and assessed the oxidative stress and inflammatory factor levels in the rat bladder through western blotting and ELISA techniques.

Through network pharmacological analysis, we identified RhoA/Rho-kinase pathway and cytokine including TNF-α, IL-6, SOD and MDA as potential mechanisms of HEC on OAB. The rats in the 2× HuangE group exhibited significantly enhanced urodynamic outcomes and decreased 24-hour urination frequency compared to the model group. Masson staining indicated a decrease in the proportion of collagenous tissue and an improvement in histomorphology. We observed a decrease expression of RhoA, ROCK1, and ROCK2 protein in the bladder tissue of 2× HuangE group rats, along with elevated SOD levels and decreased levels of TNF-α, IL-6, and MDA.

HEC could improve bladder function and morphology in BOO-induced OAB rats by reducing the expression of RhoA, ROCK1, and ROCK2 and lowering levels of oxidative stress and inflammation.

Shenhuang Liuwei powder (SHLWP) is frequently used to treat diabetic ulcers (DUs), but its mechanism of action remains poorly understood. This study aimed to identify the active compounds and mechanisms by which SHLWP alleviates DUs.

The chemical components of SHLWP were analyzed using high-resolution mass spectrometry (HRMS). Network pharmacology based on HRMS data identified SHLWP-associated targets and signaling pathways. Its antibacterial activity was assessed using Kirby-Bauer disc diffusion and minimum inhibitory concentration (MIC) tests. Its in vivo pharmacological effects were evaluated in a streptozotocin-induced diabetic ulcer model in Sprague-Dawley (SD) rats.

Seventy-three components were identified in SHLWP, with key constituents including caffeic acid (13.11 ± 0.14 μg/g), ferulic acid (20.40 ± 0.24 μg/g), quercetin (8.49 ± 0.18 μg/g), luteolin (36.63 ± 0.19 μg/g), apigenin (82.14 ± 1.60 μg/g), and linoleic acid (507.59 ± 1.46 μg/g). SHLWP exhibited strong antibacterial activity against Staphylococcus aureus (MIC = 7.8125 μg/mL), Streptococcus pyogenes (MIC < 3.90625 μg/mL), and Streptococcus epidermidis (MIC < 3.90625 μg/mL). Network pharmacology revealed significant enrichment of the AGE/RAGE, HIF-1, and PI3K-Akt pathways, which was validated in vivo using qPCR, immunohistochemistry, and Western blot.

SHLWP alleviated streptozotocin-induced diabetic ulcers by inhibiting the AGE/RAGE pathway and promoting antibacterial activity and angiogenesis via the PI3K/Akt/eNOS/HIF-1α pathway, providing a biological basis for its therapeutic effects.

Xuebijing (XBJ) injection, a Traditional Chinese medicine (TCM) widely used in China for treating sepsis and multiple organ dysfunction, has shown neuroprotective effects in traumatic brain injury (TBI). However, the mechanisms underlying these effects remain unclear. This study aims to elucidate the neuroprotective and pharmacological molecular mechanisms of XBJ and its active monomer, Hydroxy-safflor yellow A (HSYA), in treating TBI through network pharmacology and experimental validation.

Potential therapeutic targets for TBI were collected from TCMSP, TTD, OMIM, and GeneCards databases. Active compounds and targets of XBJ injection were obtained from TCMSP. The STRING database and Cytoscape software constructed a protein-protein interaction (PPI) network. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed using the DAVID database and visualized with Bioinformatics tools. Neuroprotective effects of XBJ were verified in vitro using BV2 and primary microglia cells stimulated by Lipopolysaccharide (LPS). Additionally, a TBI mice model was used to identify microglial activation in vivo.

A total of 161 common targets related to TBI were identified. Network pharmacological analysis suggested that XBJ targets proteins involved in inflammation. In vitro results showed that XBJ and HSYA inhibited LPS-induced microglial activation via the NF-κB pathway. Furthermore, XBJ was found to inhibit microglial activation in TBI mice.

These findings indicate that XBJ and HSYA may treat TBI by repressing microglial activation through the NF-κB pathway. Our study provides valuable evidence supporting XBJ as an effective therapy for TBI.

Limited treatments for silicosis necessitate further study of pneumoconiosis characteristics and pathophysiology. This study employs metabolomics to investigate metabolite changes and identify biomarkers for understanding pneumoconiosis pathogenesis.

18 healthy SPF male SD rats were divided into three groups: control, coal dust, and silica. Rats were exposed to coal dust, silica, or sterile saline for 8 weeks, after which blood, lung tissue, and feces were collected. Lung pathology was assessed, and inflammatory factors (IL-6, IL-11) were measured. 16S rDNA sequencing and UHPLC-QTOFMS metabolomics were used to analyze intestinal flora and fecal metabolites.

After 8 weeks of dust exposure, silica-exposed rats showed significantly reduced weight and elevated serum IL-6 and IL-11 levels compared to controls (P < 0.05). Lung tissue pathology revealed silica group rats exhibited lung damage, intensified inflammation, and silicon nodule formation. Coal dust group rats showed lung tissue changes with fibroblast aggregation. ? diversity analysis showed decreased Shannon index and increased Simpson index in the coal dust group, and a decreased Simpson index in the silica group. ? diversity analysis confirmed significant differences in gut microbiota between dust-exposed groups and controls. Metabolomics identified 11 differential metabolites in rat feces, meeting criteria of Fold change > 2, VIP > 1, and P < 0.05.

Dust exposure disrupts intestinal flora and metabolic state, with potential metabolic markers identified in both coal dust and silica groups, implicating fructose and mannose metabolism in coal dust exposure and sphingolipid metabolism in silica exposure.

Diminished ovarian reserve (DOR) is accompanied by abnormal initiation and development of primordial follicles. Reporting that curcumin can protect the ovarian reserve, we used rats as a model to explore the regulatory mechanism of curcumin on primordial follicle priming.

Curcumin restores the ovarian microenvironment of DOR model rats by AMPK/SIRT 1 signaling pathway, thus regulating the initiation of primordial follicles.

The study used the ovaries of 3-day-old female rats, after replicating the DOR model by triptolide (TP), then used curcumin intervention for 3 days. Histomorphological analysis was counted by H & E staining; ELISA test was used to count ovarian hormone [follicle stimulating hormone (FSH) / luteinizing hormone (LH) ratio and estradiol (E2)] concentration in the culture supernatant. Spectrophotometric measurement was used to count of superoxide dismutase (SOD) and the malondialdehyde (MDA). The protein and mRNA expression of the pathway and key indicators for follicle initiation were determined by Western Blot and Q-PCR (AMPK, SIRT 1, PTEN, PGC-1 α, and AMH).

After curcumin treatment, the number of growing follicles increased (P < 0.05). FSH/LH ratio decreased but the content and expression of E2 and AMH increased (P < 0.05). The protein and mRNA expression of characteristic indicators of inhibiting primordial follicle initiation (PTEN) was decreased (P < 0.05). Oxidation-reduction-related SOD activity increased and the content of MDA decreased (P < 0.05), while the protein and mRNA expression of PGC-1α increased (P < 0.05). The protein and mRNA expression of the pathway (AMPK, SIRT 1) were increased (P < 0.05).

Curcumin restored the ovarian local oxidant-antioxidant balance and promoted primordial follicle priming through AMPK/SIRT 1 signaling pathway in the DOR model rats.

Chronic hyperglycemia in diabetes is a significant contributor to endothelial injury through the induction of oxidative stress. Paeonol is anticipated to address oxidative stress with the aim of ameliorating endothelial injury. Our study delved into the effects of paeonol on endothelial damage induced by diabetes and elucidated the underlying mechanisms.

This research presented a novel endothelial injury model employing advanced glycation end products (AGEs) in human umbilical vein endothelial cells (HUVECs). Additionally, a network analysis was carried out to pinpoint the targets influenced by paeonol, with pivotal targets substantiated via polymerase chain reaction (PCR), western blot analysis, and immunofluorescence staining. Ultimately, the introduction of small interfering RNA transfection validated the involvement of SIRT1 in AGEs-induced HUVECs injury.

Twelve metabolites of paeonol were conclusively detected in vivo. Paeonol demonstrated substantial efficacy in ameliorating and diminishing levels of various cytokines and biochemical indicators, including AGEs, Col IV, ET-1, E-selectin, FN, hs-CRP, ICAM-1, MMP2, and sVCAM-1. Notably, network analysis accentuated the pivotal role of the MAPK signaling pathway. Furthermore, paeonol exhibited significantly elevated mRNA and protein levels of SIRT1 and ERK across varying dosage regimens compared to the model group while displaying relatively decreased mRNA expression levels of p38MAPK.

This research revealed that paeonol inhibited the activation of p38 and ERK within the MAPK signaling pathway. Moreover, the regulatory influence of paeonol over p38 and ERK was compromised subsequent to the silencing of SIRT1, indicating a SIRT1-dependent suppressive action of paeonol on the MAPK pathway. The potential therapeutic utility of SIRT1 in mitigating diabetic endothelial impairment and its concomitant cardiovascular ramifications is underscored by these findings.

Renal interstitial fibrosis (RIF) is the primary pathological progression in chronic kidney disease (CKD). Given the constraints related to cost and adverse effects of current treatments, it is crucial to explore novel and efficacious therapeutic strategies. The purpose of this study was to elucidate the potential of Jiawei Danggui Buxue Decoction (JDBD) to reduce apoptosis and epithelial-mesenchymal transition (EMT) in RIF by regulating the Janus kinase 2 (JAK2)/Signal Transducer and Activator of Transcription 3 (STAT3) pathway.

An angiotensin II (Ang II)-induced HK-2 cells model and a unilateral ureteral obstruction (UUO) animal model were employed to replicate the RIF model. A total of 48 male Wistar rats (weighing 200-220g) were acclimated for 1 week and then randomly divided into 6 groups (sham operation, UUO, Losartan potassium tablets, and three JDBD dosage groups: high, medium, and low, n=8). After the acclimatization period, UUO models were established in 40 rats through surgery, excluding the sham operation group. Each group received the corresponding drug via gavage for 2 weeks. After 2 weeks, rats were anesthetized, and tissues were collected for subsequent analysis. Renal function tests and histological stains were used to evaluate renal damage and histopathological alterations in rats. Cell viability was examined using the CCK-8 assay. Apoptosis was identified through the utilization of flow cytometry and assessment of mitochondrial membrane potential, along with other techniques. We identified and examined the expression of EMT and extracellular matrix (ECM)-related factors, as well as the JAK2/STAT3 pathway.

In vivo experiments indicated that JDBD effectively reduced renal dysfunction in UUO rats, ameliorated pathological changes in renal tissues, and significantly modulated the JAK2/STAT3 signaling pathway to inhibit EMT and apoptosis, thereby reducing ECM deposition. Furthermore, JDBD markedly increased the survival rate of Ang II-treated HK-2 cells and reduced apoptosis. The in vitro experimental results further confirmed that JDBD ameliorates RIF by regulating the JAK2/STAT3 pathway.

JDBD exhibits anti-apoptotic and EMT-inhibiting functions in RIF, potentially mediated by targeting and inhibiting JAK2/STAT3 signaling transduction.