Combinatorial Chemistry & High Throughput Screening - Online First

The intricate relationship between the gut microbiome and myopia is increasingly recognized, underscoring the need to explore its causal dynamics. Despite emerging evidence, the influence of Gut Microbiota (GM) on ocular development remains underexplored.

This study utilized Mendelian Randomization (MR) to investigate the causal impact of GM on the development of myopia. Instrumental variables (IVs) were identified from Genome-Wide Association Studies (GWAS), focusing on genetic variants significantly associated with microbiome composition. A comprehensive array of MR techniques was applied to ensure a robust estimation of causal effects and to adjust for potential confounders and pleiotropy.

The Inverse-Variance Weighted (IVW) method was used to identify significant associations between GM and myopia. Increased risk of myopia was linked to the class Betaproteobacteria (OR=1.01, 95% CI 1.004-1.017, P=0.003), the order Burkholderiales (OR=1.009, 95% CI 1.001-1.016, P=0.02), the family Oxalobacteraceae (OR=1.005, 95% CI 1.001-1.01, P=0.023), and several genera including Eubacterium xylanophilum group (OR=1.007, 95% CI 1.001-1.013, P=0.033), and Bifidobacterium (OR=1.005, 95% CI 1-1.01, P=0.038). Protective effects were noted for the order Mollicutes RF9 (OR=0.994, 95% CI 0.99-0.999, P=0.014), the genus Allisonella (OR=0.996, 95% CI 0.993-0.999, P=0.019), the genus Lachnospiraceae UCG001 (OR=0.994, 95% CI 0.989-1, P=0.045), and the family Enterobacteraceae (OR=0.991, 95% CI 0.982-1, P=0.047) and order Enterobacteriales (OR=0.991, 95% CI 0.982-1, P=0.047). Sensitivity analyses further confirmed the robustness of these findings.

This study provides causal evidence for the “Microbiome-Gut-Eye Axis” in myopia development, identifying specific gut microbiota that influence myopia risk. These findings suggest potential for microbiota-targeted interventions, warranting further research in diverse populations.

The findings support the “Microbiome-Gut-Eye Axis” as a potential factor in myopia pathogenesis and highlight microbiota-targeted interventions as novel therapeutic strategies for managing myopia. This study lays the groundwork for further research on how modifying GM can influence eye health and offers new perspectives on preventive health strategies.

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.

The specific role of necroptosis in the pathogenesis of cataracts remains unclear. This study aimed to identify and validate the genes related to necroptosis in the development of cataracts through bioinformatics analysis.

We utilized RNA sequencing data (GSE161701) from the Gene Expression Omnibus (GEO) database and employed R software to perform differential expression analysis of necroptosis-related genes (NRGs) in lens epithelial cells (LECs) under oxidative stress. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were conducted to evaluate the functions of necroptosis-related differentially expressed genes (NRDEGs) and their associated pathways. Additionally, a diagnostic model was established using LASSO regression to select hub genes, and protein-protein interaction (PPI) networks, mRNA-miRNA, and mRNA-drug regulatory networks were constructed. Immune infiltration analysis was performed using the xCell and CIBERSORT algorithms, and the differential expression of hub genes was validated in a UVB-induced rat cataract model using RT-qPCR and immunohistochemistry.

The results indicated that oxidative stress promoted necroptosis in LECs, involving 86 NRDEGs and nine hub genes. GO and KEGG analyses revealed significant enrichment in necroptosis-associated pathways. Furthermore, we identified 58 mRNA-miRNA interactions and 131 potential molecular compounds or drugs. The immune infiltration analysis showed that certain immune cells exhibited significantly elevated expression in the cataract group, with notable correlations between some immune cells and hub genes. RT-qPCR and immunohistochemistry confirmed the expression of 9 hub genes and 3 key necroptosis genes. BAX, CXCL1, EPAS1, JUN, LRP1, RBM14, SERTAD1, and TNFAIP3 were highlighted as potential diagnostic and therapeutic targets.

This study identified key NRDEGs involved in the pathogenesis of cataracts under oxidative stress through bioinformatics analyses, potentially providing new targets and research directions for future cataract prevention and treatment.

Chenopodium album Linn. is a nutritionally and pharmacologically significant herb that generally grows in the winter season along with other crops. It is rich in fibers, protein, minerals (Mg, Ca, Fe, K, P, and others), vitamins (ascorbic acids, thiamine, riboflavin, and others), and several other biologically active chemical components like flavonoids, saponins, steroids and many more. In this article, the authors briefly describe and assess the chemistry and pharmacology of this nutritionally significant plant.

This study is based on several literature searches conducted via Google Scholar, Research Gate, PubMed, and many other online sources.

Due to its richness with bioactive phytochemicals, it has become a valuable functional food. C. album has several medicinal properties like antioxidant, antimicrobial, anti-arthritic, anti-diabetic, anti-infection, anti-ulcer, and many others. Even after its rich nutritional values, chemical compositions, and a broad spectrum of pharmacological properties, this is a highly ignored herb worldwide. Therefore, extensive research and awareness regarding the functional role of this herb is needed.

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.

Porcine epidemic diarrhea virus (PEDV), a member of the Coronaviridae, is responsible for acute diarrhea, vomiting, and dehydration, which can lead to high mortality in neonatal piglets. Previous research has indicated the antiviral potential of forsythia essential oil (FEO); however, its active components and mechanisms of action remain inadequately defined. This study aims to investigate the antiviral effects of FEO and elucidate its potential mechanisms for treating PEDV.

The primary components of FEO were identified using gas chromatography-mass spectrometry (GC/MS) in conjunction with the National Institute of Standards and Technology Standard Spectrum (NIST) Database. Network pharmacology and weighting coefficients were employed to determine the key signaling pathways associated with PEDV-related diseases. Molecular docking simulations were conducted to explore the interactions between the active ingredients and their corresponding targets. The safety profile of FEO was assessed through cell viability assays utilizing the CCK8 method. Subsequently, immunofluorescence assays (IFA) and reverse transcription-quantitative polymerase chain reaction (RT-Q-PCR) were performed to provide evidence of the anti-PEDV effects. Additionally, the viral replication cycle was analyzed to identify the stages at which FEO exerts its antiviral effects. Finally, key targets were validated through RT-Q-PCR to further investigate the anti-PEDV mechanisms of FEO.

The IL-17 signaling pathway was identified as a critical pathway for the treatment of PEDV with FEO based on network pharmacology and weighting coefficient analyses. Furthermore, results from RT-Q-PCR and IFA demonstrated that FEO influenced the replication of PEDV during the attachment and internalization phases. Specifically, during the viral attachment phase, FEO significantly upregulated the expression of HSP90AA1 while downregulating MAPK14 expression, leading to a reduction in associated inflammatory factors. At the high dose of FEO, the expression of HSP90AA1 was higher than that of the model group by about 5-fold, and the expression of MAPK14 was lower than that of the model group by about 2-fold. Cell viability assay showed no significant cytotoxicity of FEO at 0.63 µL/mL, thus confirming its safety.

The findings of this study suggest that FEO possesses potential antiviral properties against PEDV. Its novel mechanisms of action warrant further investigation, which may contribute to the development of effective therapeutic strategies for managing PEDV infections.