Good Binding Research Articles

Network Pharmacology and Molecular Docking: Exploring the Mechanism of Peppermint in Mastitis Prevention and Treatment in Dairy Cows

In order to elucidate the active ingredients, potential targets, and mechanisms of action of peppermint in treating bovine mastitis, this study utilized network pharmacology analysis and molecular docking to conduct an exploratory, prospective investigation. Using the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database, all compounds and targets of peppermint were retrieved. After removing duplicates, a total of 133 compounds and 272 targets were obtained. Targets were then standardized to gene names using the UniProt database to construct a drug–component-target network. A total of 183 disease targets related to bovine mastitis were retrieved from the GeneCards database. We obtained 28 cross targets of peppermint targets and bovine mastitis targets, and constructed a protein–protein interaction (PPI) network using the STRING database. A visual network was built using Cytoscape 3.10.0 software, and seven core targets were analyzed and obtained. GO and KEGG pathway enrichment analysis was performed using the Metascape database. Molecular docking was conducted using AutoDockTools–1.5.6 software on some small–molecule compounds and the seven targets to evaluate the stability of binding between peppermint and core targets. Apigenin, luteolin, and ursolic acid are the three main components in peppermint. Core targets (TNF, IL–6, STAT–3, IL–1β, FGF–2, IFNG, and ESR–1) were selected based on the PPI network. The enrichment analysis suggested that the major signaling pathways in network pharmacology may include AGEs–RAGE, IL–17, NF–κB, TLRs, HIF–1, TGF–β, PI3K–Akt, and MAPK. The molecular docking results showed that one of the main components of mint, ursolic acid, exhibited good binding activity with all core targets of bovine mastitis. Other constituents also produced favorable binding with some core targets. This study elucidates the mechanisms of mint in treating bovine mastitis, providing data to support the potential development of new therapies for bovine mastitis using mint and its constituents.

Comparative study of essential oil extraction from lime peel (Citrus amblycarpa Hassk. Ochse): Impact on chemical composition and bioactivities of anti-tyrosinase, anti-elastase, and antimicrobial properties

Essential oil is a natural substance widely utilized in the cosmetics industry. However, there has been no study on the cosmetic bioactivity of essential oil from the peel of Citrus amblycarpa Hassk. Ochse (CAEO). This study aimed to investigate the potential of CAEO, obtained through NaCl hydrodistillation (HD) and microwave-assisted hydrodistillation (MAHD), as an anti-tyrosinase, anti-elastase, and antibacterial agent, along with analyzing its chemical constituents. GC-MS analysis identified β-pinene (28.3%) and limonene (27.1%) as the main compounds in CAEO (HD), while limonene (13.75%) and citronellal (7.49%) predominated in CAEO (MAHD). The CAEO obtained from HD with 25% NaCl as the solvent exhibited significantly higher (p < 0.05) anti-tyrosinase (IC50 = 235.94 ± 1.34 μg/mL) and anti-elastase (IC50 = 41.59 ± 0.47 μg/mL) activities compared to the CAEO from the MAHD method. This result was consistent with the in silico pharmacological study, which revealed that limonene, the major component of CAEO, showed good binding affinity to the active sites of elastase and tyrosinase. The CAEO obtained from both methods exhibited strong and selective antibacterial activity in a disc diffusion assay against two bacteria associated with skin acne: Propionibacterium acnes ATCC 11827 (CAEO HD = 24.08 ± 5.85 mm) and Staphylococcus aureus ATCC 25923 (CAEO MAHD = 20.5 ± 0.27 mm). These results pave the way for further studies on the bioassay-guided isolation of CAEO to enhance its effectiveness as a cosmetic agent. Moreover, this study provides the first report indicating the potential of CAEO for development as a cosmetic agent.

Chitosan functionalized two-dimensional covalent organic framework nanosheets with high hydrophilicity for efficient glycopeptide enrichment.

Glycopeptides are an important biomarker, which play a crucial role in various biological processes. Due to their low abundance and the presence of interfering macromolecular proteins, enrichment of glycopeptides is necessary before testing. However, most materials for enriching glycopeptides have high site resistance, relatively low surface area, and limited recognition sites. Herein, a highly hydrophilic two-dimensional (2-D) covalent organic framework (NUS-10) loaded with chitosan (CS) (denoted as NUS-10@CS) had been synthesized. After enrichment with NUS-10@CS, a total of 34 glycopeptides from horseradish peroxidase (HRP) tryptic digests were detected, demonstrating a high enrichment efficiency for glycopeptides from model glycoprotein digestion. Meanwhile, the material exhibited ultra-high adsorption capacity (1 fmol/μL HRP), excellent selectivity (HRP tryptic digest/bovine serum albumin (BSA) tryptic digest=1:2000), macromolecular protein anti-interference ability (HRP tryptic digest/BSA = 1:2000) and good binding capacity (200mg/g). Additionally, 712 glycopeptides corresponding to 200 glycoproteins were identified from 3µL human serum. NUS-10@CS was promising for glycopeptide analysis, helping to identify potential disease biomarkers more efficiently, and leading to easier and more accurate diagnosis of diseases, which was essential for early intervention and treatment.

Efficacy and mechanism of Qianjinweijing Decoction for asthma: Integrating systematic review with meta-analysis and network pharmacology.

Asthma seriously affects people's survival and quality of life, causing a huge economic burden on society. Modern clinical use of Qianjinweijing Decoction (QJWJ) for the treatment of asthma has achieved good results. However, there is still a lack of research on its efficacy and mechanism of action. Therefore, the purpose of this study is to evaluate the efficacy of QJWJ in the treatment of asthma by systematic review and meta-analysis, and to explore its potential mechanism by network pharmacology. The meta-analysis was performed to search for studies published before May 2023 in 7 databases, and Revman 5.4 and R language softwares were used for analysis. Network pharmacology was based on open databases and softwares such as Cytoscape, Perl, Autoduck Vina, and R language. A total of 14 studies were included, involving 1200 patients. The results of the meta-analysis showed that QJWJ could significantly improve the clinical efficacy of asthma patients compared with routine pharmacotherapy (risk ratio = 1.22, 95% CI [1.16, 1.28], P < .00001), enhance lung function, such as FEV1/FVC (mean difference [MD] = 5.63, 95% CI [1.45, 9.81], P = .008), FEV1% (MD = 5.03, 95% CI [4.32, 5.74], P < .00001), PEF (standardized mean difference = 1.37, 95% CI [1.03, 1.71], P < .00001), and increase traditional Chinese medicine syndrome score (MD = -2.50, 95% CI [-4.81, -0.19], P = .03). The results of network pharmacology suggested that the 4 traditional Chinese medicines in QJWJ included 35 active ingredients and 34 potential targets for the treatment of asthma. The core ingredients involved were stigmasterol, β-sitosterol, hederagenin, and gibberellin 7. The core targets were PTGS2, BCL2, and CASP3. The interaction pathway between QJWJ and asthma was mainly enriched in p53, cyclic guanosine monophosphate-protein kinase G, IL-17, and advanced glycation end products-receptor for advanced glycation end products signaling pathways. Molecular docking showed that the core ingredients had good binding activity with the core targets. QJWJ is effective in the treatment of asthma, and the therapeutic mechanism may be related to its regulation of inflammation, immunity, and apoptosis.

Synthesis, antibacterial and anticancer activity of azo-based and aminomethylene derivatives of cyclic β-keto sulfones.

Herein we describe efficient and cost-effective synthesis of azo-based and aminomethylene derivatives of cyclic β-keto sulfones, specifically, 4,4-disubstituted (E)-2-(2-phenylhydrazineylidene)dihydrothiophen-3(2H)-one 1,1-dioxides and 4,4-disubstituted (E)-2-((methylamino)methylene)dihydrothiophen-3(2H)-one 1,1-dioxides. The azo-based derivatives were prepared through the azo coupling of cyclic β-keto sulfones with aromatic diazonium acetates, in situ prepared by diazotization of appropriate substituted amino benzenes. The aminomethylene derivatives were synthesized through the condensation of cyclic β-keto sulfones with DMF-DMA followed by a transamination reaction with primary amines. The target products were obtained in good yields and their structure was unambiguously established based on NMR spectra data, X-ray diffraction study, and DFT calculations. These compounds were designed as cost-effective and multitarget biologically active compounds. Further in vitro study showed antibacterial activity against Staphylococcus aureus and cytotoxicity against the MDA-MB-231 breast cancer cell line without affecting non-malignant cells MAEC. The molecular docking study confirmed good binding affinity of the studied compounds for DHPS, crucial for the bacterial life cycle and for CLK4 and IDO1, which are the therapeutic targets in cancer treatment.

Phytotoxic impact of di-butyl phthalate (DBP) on physiological, biochemical, and oxidative stress parameters of rice (Oryza sativa).

Phthalates are synthetic compounds, well-known plasticizers, with numerous applications and reported to have adverse effects on all living organisms residing in terrestrial and aquatic environments. In this study, the rice (Oryza sativa) seedlings were exposed to di-butyl phthalate (DBP) exogenously for 7days, with varying concentrations of 0, 200, 400, 800, and 1600mg/L, to explore the toxicological, physiological, and biochemical consequences by measuring various parameters such as pigment, lipid, and H2O2 (hydrogen peroxide) contents. The biochemical analysis of seedlings showed that the pigments, lipids, and H2O2 concentrations were altered abnormally. After 7days of exposure, the maximum amount of DBP was accumulated and translocated in both the shoot and root of the grown seedlings, and all morphological parameters (i.e., length and weight of both shoot and root) and pigment content (such as total carotenoid, chlorophyll a and b) were declined significantly. Superoxide dismutase (SOD), H2O2, and thiobarbituric acid reactive substance (TBARS) levels in seedlings increase as the stress increases due to the higher exposure dose of DBP. Cell viability was observed under a confocal microscope confirming the damage of the plasma membrane. Additionally, molecular docking studies indicated that DBP has a good binding affinity with key antioxidant enzymes of Oryza sativa, interacting via hydrogen bonds with specific amino acids. This suggests a potential mechanistic pathway for the observed biochemical changes in Oryza sativa.

Mechanisms of wogonoside in the treatment of atherosclerosis based on network pharmacology, molecular docking, and experimental validation

BackgroundAtherosclerosis serves as the fundamental pathology for a variety of cardiovascular disorders, with its pathogenesis being closely tied to the complex interplay among lipid metabolism, oxidative stress, and inflammation. Wogonoside is a natural flavonoid extracted from Scutellaria baicalensis with a variety of biological activities, including anti-inflammatory, hypolipidemic, and cardiac function improvement properties. Despite these known effects, the specific role of wogonoside in the context of atherosclerosis remains to be elucidated.PurposeTo validate the efficacy of wogonoside in the treatment of atherosclerosis and to investigate its possible therapeutic mechanisms.MethodsNetwork pharmacology was used to obtain the core targets and signaling pathways that may be efficacious in the treatment of atherosclerosis with wogonoside, which were validated using molecular docking and molecular dynamics simulations. To further validate the core targets in the signaling pathway, we performed in vivo experiments using apolipoprotein E (ApoE)-/- mice. This included pathological morphology and lipid deposition analysis of mouse aorta, serum lipid level analysis, Elisa analysis, oxidative stress analysis, reactive oxygen species (ROS) fluorescence assay, immunohistochemical analysis and protein blot analysis.ResultsPredictions were obtained that wogonoside treatment of atherosclerosis has 31 core targets, which are mainly focused on pathways such as Toll-like receptor (TLR) signaling pathway and NF-kappa B (NF-κB ) signaling pathway. Molecular docking and molecular dynamics simulations showed that wogonoside has good binding properties to the core targets. In vivo experimental results showed that wogonoside significantly inhibited aortic inflammatory response and lipid deposition, significantly reduced the release levels of total cholesterol (TC), triglycerides (TG), low-density-lipoprotein cholesterol (LDL-C), oxidized low density (ox-LDL) and free fatty acid (FFA), and significantly inhibited the release of inflammatory factors TNF-α, IL-1β, IL-6 and oxidative stress in ApoE-/- mice. Further molecular mechanism studies showed that wogonoside significantly inhibited the activation of TLR4/NF-κB signaling pathway in ApoE-/- mice.ConclusionWogonoside may be an effective drug monomer for the treatment of atherosclerosis, and its mechanism of action is closely related to the inhibition of the activation of the TLR4/NF-κB signaling pathway.Graphical

Investigating the Inhibitory Effects of Paliperidone on RAGEs: Docking, DFT, MD Simulations, MMPBSA, MTT, Apoptosis, and Immunoblotting Studies

Chronic diseases such as diabetes and cancer are the leading causes of mortality worldwide. Receptors for Advanced Glycation End products (RAGEs) are ubiquitous factors that catalyse Advanced Glycation End products (AGEs), proteins, and lipids that become glycated from sugar ingestion. RAGEs are cell surface receptor proteins and play a broad role in mediating the effects of AGEs on cells, contributing to modifying biological macromolecules like proteins and lipids, which can cause Reactive Oxygen Species (ROS) generation, inflammation, and cancer. We targeted RAGE inhibition analysis and screening of United States Food and Drug Administration (FDA) libraries through molecular docking studies that identified the four most suitable FDA compounds: Zytiga, Paliperidone, Targretin, and Irinotecan. We compared them with the control substrate, Carboxymethyllysine, which showed good binding interaction through hydrogen bonding, hydrophobic interactions, and π-stacking at active site residues of the target protein. Following a 100 ns simulation run, the docked complex revealed that the Root Mean Square Deviation (RMSD) values of two drugs, Irinotecan (1.3 ± 0.2 nm) and Paliperidone (1.2 ± 0.3 nm), were relatively stable. Subsequently, the Molecular Mechanics Poisson–Boltzmann Surface Area (MMPBSA) determined that the Paliperidone molecule had a high negative energy of −13.49 kcal/mol, and the Absorption, Distribution, Metabolism, and Excretion (ADME) properties were in control for use in the mentioned cases. We extended this with many in vitro studies, including an immunoblotting assay, which revealed that RAGEs with High Mobility Group Box 1 (HMGB1) showed higher expression, while RAGEs with Paliperidone showed lower expressions. Furthermore, cell proliferation assay and Apoptosis assay (Annexin-V/PI staining) results revealed that Paliperidone was an effective anti-glycation and anti-apoptotic drug—however, more extensive in vivo studies are needed before its use.

Synthesis and Anticancer Activity Evaluation of N-(3,4,5- Trimethoxycinnamoyl)anthranilic Acid Derivatives

There is an urgent need to search for more efficient, better-tolerated anticancer drugs. Tranilast, a synthetic analog of tryptophan metabolite, has been shown to inhibit the growth of various cancer cells. In our previous study, the tranilast analog TRA01 [N-(3,4,5- trimethoxycinnamoyl)anthranilic acid] was identified as a promising agent against the proliferation of MCF-7 and HepG2 cell lines along with good binding affinity on the transforming growth factor beta 1 (TGFβ1) target. To further explore the structure-activity relationship, a series of N-(3,4,5- trimethoxycinnamoyl)anthranilic acid derivatives (5a–i) were successfully synthesized via a two-step synthetic procedure based on N-acylation and Knoevenagel-Doebner reactions and their structures were determined using 1 H-NMR, 13C-NMR, and MS spectra. The derivatives (5a–i) were evaluated for in vitro activity using MTT assay and in silico docking on TGFβ1 target by AutoDockTools–1.5.6 software. The bioactivity results of 5a–i showed 12.30–27.04% and 19.13–46.23% inhibition on proliferation of MCF-7 and HepG2 cell lines at 100 µM concentration, respectively, with the fluorinated derivatives (5a and 5e) possessing the strongest inhibitory activities. The molecular docking also found the best binding affinity of 5a (-8.70 Kcal/mol) and 5e (-8.71 Kcal/mol) on the TGFβ1 target. The SAR result revealed that substituents on the benzene ring of anthranilic acid were not favored for anticancer activity. Although these derivatives exhibited weak anticancer properties, the good binding affinity to the TGFβ1 target suggested certain potential of this scaffold for further study on this target.

Management of intraocular pressure and inflammation using febuxostat film: in vitro - in vivo correlation

Background and purpose: Urate crystal accumulation may lead to the condition of ocular tophaceous gout, causing ocular inflammation and increased intraocular pressure (IOP) due to the triggering of several inflammatory receptors like NLRP3, A2A, and TLR4. The study has been undertaken to manage intraocular pressure and inflammation using febuxostat (FBX) film formulation for sustained and improved activity, particularly for long-term tophaceous gout patients. Experimental approach: Hydroxypropyl methyl-cellulose K100 matrix-based hydrogel film of FBX has been fabricated in the presence of plasticizers like triethanolamine, dimethyl-sulphoxide (DMSO), or polyethylene glycol 600 using casting and evaporation technique. Carrageenan was injected into the upper palpebral region to induce ocular inflammation, and a normotensive rabbit eye model was used for monitoring IOP. Key results: Amorphization of the drug was observed from the differential scanning calorimetry and X-ray diffraction results. In vitro release study revealed an improved and diffusion-controlled sustained drug release for more than 5 h (62.69 to 84.76 %). Compared to its absence, decreased IOP was extended up to 5 h using film (with DMSO). Disappearance of ocular inflammation was also observed in the test animals after 2.5 h of film application, whereas acute inflammation was continued in the group without treatment for more than 4 h. Docking study revealed good binding interaction of drug and NLRP3, A2A, and TLR4 receptor. Conclusion: Febuxostat-loaded hydrogel-forming plasticized film could be utilized to better manage and control ocular inflammation and associated IOP, particularly in ocular tophaceous gout patients.

Anti-TMV activity based flavonol derivatives containing piperazine sulfonyl: Design, synthesis and mechanism study.

A series of flavonoid derivatives containing piperazine sulfonate were designed and synthesized. The results of antiviral experiments in vivo showed that some target compounds had good inhibitory effect on tobacco mosaic virus (TMV). The EC50 values of S15 and S19 curative activity were 174.5 and 110.4μg/mL, respectively, which were better than 253.7μg/mL of Ningnanmycin (NNM). The EC50 values of S4 and S19 protection activity were 140.3 and 116.1μg/mL, respectively, better than that of NNM (247.1μg/mL). Microscale thermophoresis (MST) and molecular docking experiments showed that S19 had a good molecular binding force with TMV. Transmission electron microscopy (TEM) results show that S19 can fracture TMV particles and affect self-assembly. S19 treatment had almost no effect on the growth of seeds and seedlings, and can change the content of chlorophyll malondialdehyde (MDA) and superoxide dismutase (SOD) in tobacco to a certain extent, and improve the disease resistance of tobacco.

Ginkgolide B binds to GPX4 and FSP1 to alleviate cerebral ischemia/reperfusion injury in rats.

Ischemia/reperfusion (I/R) injury can increase the anomalous permeability of the blood-brain barrier and the risk of hemorrhagic conversion. Ginkgolide B (Gin B) has been recognized for its neuroprotective properties in stroke treatment. This study aimed to analyze the association of Gin B with GPX4 and FSP1 in cerebral I/R injury treatment. HT22 cells were induced by oxygen-glucose deprivation/reoxygenation (OGD/R) and treated with a series of Gin B (10, 20, 40 μM) for 24 h. It found that the Gin B treatment declined the OGD/R-induced cellular ROS and lipid ROS with increasing concentrations. Moreover, the Gin B treatment improved the OGD/R-induced ferroptotic cell death by activating the GPX4-GSH and FSP1-CoQ10-NADH pathways with increasing concentrations. Molecular docking showed there is a good binding activity of Gin B to GPX4 (score = -6.4 kcal/mol) and FSP1 (score = -6.7 kcal/mol), and the microscale thermophoresis (MST) assay confirmed that Gin B can directly bind to GPX4 and FSP1. In vivo, rats were induced by middle cerebral artery occlusion (MCAO)/R and treated with 20 mg/kg of Gin B to analyze its effects on the GPX4-GSH and FSP1-CoQ10-NADH pathways. The GPX4 inhibitor (RSL3) and the FSP1 inhibitor (iFSP1) were used to confirm the mechanism of Gin B in the MCAO/R-treated rats. It showed that the Gin B treatment alleviated the MACO/R-induced brain injury by activating the GPX4-GSH and FSP1-CoQ10-NADH pathways. This study showed that Gin B improved cerebral I/R-induced ferroptotic cell death by activating the GPX4-GSH and FSP1-CoQ10-NADH pathways, providing a new mechanism of Gin B for cerebral I/R treatment.

Mechanisms of the Compound of Magnoliae Flos and Xanthii Fructus Essential Oils for the Treatment of Allergic Rhinitis based on the Integration of Network Pharmacology, Molecular Docking, and Animal Experiment.

Magnoliae Flos (Chinese name: Xin-Yi) and Xanthii Fructus (Chinese name: Cang-Er-Zi) are Chinese herbal medicines and have been used to treat allergic rhinitis (AR). However, the therapeutic effect, active ingredients, and probable processes of a compound of Magnoliae Flos and Xanthii Fructus in the form of essential oils (CMFXFEO) in treating AR have not been reported. This study aims to determine the efficacy of the CMFXFEO on ovalbumin (OVA)-induced AR in a rat model and to use network pharmacology and molecular docking to reveal the hub genes, biological functions, and signaling pathways of CMFXFEO against AR. Animal experiments were applied to validate the role of CMFXFEO in the treatment of AR. 20 rats were randomly divided into four groups: control group (CON, n=5), positive control group (AR, n=5), CMFXFEO-treated group (AR+CMFXFEO, n=5), and budesonide-treated group (AR+Budesonide, n=5). Rats were stimulated with OVA to induce AR. Symptom scores assessment and histo-pathomorphological evaluation was performed. The serum level of OVA-specific immunoglobulin (Ig) E was measured. Gas Chromatograph-Mass Spectrometer analysis (GC-MS) was used to identify the monomer chemical composition of CMFXFEO. The target genes of CMFXFEO were obtained by using PubChem and SwissTargetPrediction databases. The target genes of AR were screened using GeneCards, DisGeNET, and OMIM databases. The target genes were intersected using the venny2.1 website to obtain the potential therapeutic targets of CMFXFEO for treating AR and to construct the PPI network. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were used to reveal associated signaling pathways. The Sybyl tool was used to dock the CMFXFEO with key therapeutic targets molecularly. Intranasal CMFXFEO administration significantly suppressed the allergic symptoms, reduced the inflammatory cell infiltration, and the serum level of OVA-specific immunoglobulin (Ig) E. The main components of CMFXFEO obtained through the GC-MS analysis, listed as γ-terpinene (9.4908%), limonene (7.2693%), menthol (7.1821%), β-pinene (7.1190%), β-caryophyllene (7.0396%), eucalyptol (6.1367%), linalool(5.9686%), eugenol (5.0776%). A total of 398 CMFXFEO targets and 488 AR-related targets were screened, of which 42 were common targets. The GO and KEGG pathway analyses unveiled that CMFXFEO were strongly associated with several signaling pathways, including the AGE-RAGE signaling pathway, TNF signaling pathway, and Chemokine signaling pathway. PPI network construction screened six hub genes as therapeutic targets, including STAT3, IL1B, TLR4, PTGS2, ICAM1, and VCAM1. The molecular docking verification indicated that CMFXFEO have good binding activity with therapeutic targets, and β-Pinene's docking ability with TLR4 is particularly prominent. The anti-inflammatory and anti-allergic effects of CMFXFEO are to inhibit the infiltration of inflammatory cells in the OVA-induced AR rat model. The results of the network pharmacology and molecular docking deduced that the CMFXFEO may have the potential to treat AR by multiple pathways through relieving inflammatory, anti-oxidative stress response, and modulating the immune system.

Evaluating the potential of Kalanchoe pinnata, Piper amalago amalago, and other botanicals as economical insecticidal synergists against Anopheles gambiae

BackgroundSynergists reduce insecticide metabolism in mosquitoes by competing with insecticides for the active sites of metabolic enzymes, such as cytochrome P450s (CYPs). This increases the availability of the insecticide at its specific target site. The combination of both insecticides and synergists increases the toxicity of the mixture. Given the demonstrated resistance to the classical insecticides in numerous Anopheles spp., the use of synergists is becoming increasingly pertinent. Tropical plants synthesize diverse phytochemicals, presenting a repository of potential synergists.MethodsExtracts prepared from medicinal plants found in Jamaica were screened against recombinant Anopheles gambiae CYP6M2 and CYP6P3, and Anopheles funestus CYP6P9a, CYPs associated with anopheline resistance to pyrethroids and several other insecticide classes. The toxicity of these extracts alone or as synergists, was evaluated using bottle bioassays with the insecticide permethrin. RNA sequencing and in silico modelling were used to determine the mode of action of the extracts.ResultsAqueous extracts of Piper amalago var. amalago inhibited CYP6P9a, CYP6M2, and CYP6P3 with IC50s of 2.61 ± 0.17, 4.3 ± 0.42, and 5.84 ± 0.42 μg/ml, respectively, while extracts of Kalanchoe pinnata, inhibited CYP6M2 with an IC50 of 3.52 ± 0.68 μg/ml. Ethanol extracts of P. amalago var. amalago and K. pinnata displayed dose-dependent insecticidal activity against An. gambiae, with LD50s of 368.42 and 282.37 ng/mosquito, respectively. Additionally, An. gambiae pretreated with K. pinnata (dose: 1.43 μg/mosquito) demonstrated increased susceptibility (83.19 ± 6.14%) to permethrin in a bottle bioassay at 30 min compared to the permethrin only treatment (0% mortality). RNA sequencing demonstrated gene modulation for CYP genes in anopheline mosquitoes exposed to 715 ng of ethanolic plant extract at 24 h. In silico modelling showed good binding affinity between CYPs and the plants’ secondary metabolites.ConclusionThis study demonstrates that extracts from P. amalago var. amalago and K. pinnata, with inhibitory properties, IC50 < 6.95 μg/ml, against recombinant anopheline CYPs may be developed as natural synergists against anopheline mosquitoes. Novel synergists can help to overcome metabolic resistance to insecticides, which is increasingly reported in malaria vectors.Graphical

Genetic susceptibility and potential therapeutic targets of unruptured intracranial aneurysms: A genome-wide study based on Mendelian randomization.

At present, although some studies have offered certain insights into the genetic factors related to unruptured intracranial aneurysms (uIAs), the potential genetic targets associated with uIAs remain largely unknown. Thus, this research adopted Mendelian randomization (MR) analysis to study two genome-wide association studies on uIAs, aiming to determine the reliable genetic susceptibility and potential therapeutic targets for uIAs. This study summarizes the data of expression quantitative trait loci (eQTL) as exposure data. The outcome data of uIAs were derived from the study by Bakker et al. and the FinnGen Biobank (version R10). The reliable genetic susceptibility and potential therapeutic targets of uIAs were identified by means of Mendelian randomization (MR) methods, with the inverse variance weighting (IVW) method as the primary analytical approach. Simultaneously, sensitivity and pleiotropy analyses were carried out, and the results were visualized. Subsequently, drug predictions and molecular docking were conducted for the potential gene targets to verify their reliability. The MR analysis of the training cohort identified 100 targets related to uIAs. Then, these 100 gene targets and eQTL data were verified by MR Analysis again with the testing cohort. Finally, 7 gene targets were selected, namely MTMR3, SERINC1, CITED2, NKX3-1, ATOX1, MYADM and SLC20A1-DT.GO/KEGG enrichment analysis confirmed that the 7 gene targets mainly participate in the process Biological functions and pathways such as art development, cellular response to hypoxia, male Gonad development, RNA polymerase II specific DNA binding transcription factor binding, DNA binding transcription factor binding, Mineral absorption, Inositol phase metabolism, Photoshatidylinositol signaling system, etc.The protein-protein interaction（PPI） network describes the interactions between seven gene targets and related proteins.The molecular docking diagram shows good binding between candidate drugs and proteins related to gene targets. The study identified 7 reliable gene susceptibility and potential therapeutic targets associated with uIAs, offering new insights for clinical diagnosis and treatment of uIAs, and suggesting novel research directions for understanding the etiology and molecular mechanisms of uIAs.

Duhuo Jisheng Mixture attenuates neuropathic pain by inhibiting S1PR1/P2Y1R pathway after Chronic Constriction Injury in mice.

The pathogenesis of neuropathic pain is complex and lacks effective clinical treatment strategies. Medical plants and herbal extracts from traditional Chinese medicine with multi-target comprehensive effects have attracted great attention from scientists. To investigate the pharmacological active components and mechanism underlying the anti-neuralgia effect of classic analgesic formulas Duhuo Jisheng Mixture (DJM). Chronic Constriction Injury (CCI) surgery was used to assess the efficiency of DJM in treating neuropathic pain. Forty-two C57BL/6 mice were evenly divided into seven groups: sham-operated, sham treatment with DJM (20 ml/kg), CCI treatment with DJM (0, 10, 20, or 40 ml/kg) and CCI treatment with Morphine (3 mg/kg). DJM irrigation stomach was implemented from post-operative day (POD) 0 to POD 21. Paw withdrawal threshold and Thermal latency were conducted on POD 0, 1, 3, 5, 7, 14 and 21. On POD 14, eEPSP in spinal dorsal horn neurons was recorded by patch-clamp. mRNA transcription in spinal cord was also monitored to screen molecular targets and signaling pathways on POD 14. To further validate the influence of DJM on the S1PR1 signaling pathway, 1 mg/kg W146 was infused into mice intrathecally and delivered to the cerebral spinal fluid through one 30 G needle with a whole 20 μl intervertebral between the L5 and L6. Furthermore, the active ingredients of DJM were identified through LC/MS equipment and predicted through Molecular Docking. We demonstrated that DJM treatment reversed CCI-induced pain sensitivity and the excitability of neurons, decreased up-regulation of astrocyte S1PR1, inhibited astrocyte activation, and further inhibited the expression of Purinergic P2Y1R and its downstream molecule p-JNK in mouse spinal cord, as well as the release of inflammatory factors. Interestingly, due to the regulatory role of astrocytes on neurons, the effects of DJM on astrocytes ultimately manifest in pain-effector neurons, resulting in decreased p-ERK, p-CREB, and pain-marking protein c-fos in neurons. S1PR1 Antagonist W146 possessed an equivalent analgesic effect as DJM and inhibited S1PR1 and c-fos expression. According to LC/MS analysis results, a total of 33 active ingredients were screened, of which 20 active ingredients had good binding activity with S1PR1 and were considered to be the main active ingredients for analgesia. This study is the first to clarify the effect and molecular mechanism of DJM for anti-neuralgia in particular, which confirms the clinical value of DJM in relieving neuropathic pain. Furthermore, this study innovatively identifies the potential pharmacological components of DJM through LC/MS and Bioinformatics technology, which forms a framework for people's understanding of DJM treatment for neuropathic pain and provides a sufficient theoretical basis of DJM for clinical application.

Dapagliflozin attenuates skeletal muscle atrophy in diabetic nephropathy mice through suppressing Gasdermin D-mediated pyroptosis.

Skeletal muscle atrophy is a clinical concern in diabetic nephropathy, and without effective therapeutic approaches. Massive evidence has demonstrated that dapagliflozin, a sodium-glucose co-transporter 2 inhibitor can relieve diabetic nephropathy by inhibiting glucose re-absorption or podocyte pyroptosis. Nevertheless, whether dapagliflozin could treat skeletal muscle atrophy or the potential protection mechanism in diabetic nephropathy mice is unclear. The variety of approaches were used to assess the particular histology-associated characteristics, mRNA, and protein expression. These included examing the morphology of renal and skeletal muscle tissues through H&E staining, detecting mRNA and protein expression through real-time PCR and Western blot analysis, and monitoring fasting blood glucose levels by using Blood Glucose Monitor Test Kits. Dapagliflozin mitigated renal tissue injury with podocyte protein-nephrin and skeletal muscle atrophy effectively with mitochondrial-related proteins. Meanwhile, our research revealed that Casp3 was the target gene and dapagliflozin could decrease the expressions. Subsequently, we verified that dapagliflozin can effectively decrease canonical pyroptosis pathway proteins, which include Gasdermin D, NLRP3, Casp1, and ASC. Meanwhile, Palmitic acid (PA) induced Gasdermin E-N fragment (non-canonical pyroptosis protein) in C2C12 cells, and then released the inflammatory molecules such as IL-1β, IL-18, and NF-kappaB, which were suppressed by dapagliflozin treatment. Aside from that, dapagliflozin demonstrated a good binding affinity to the Casp3 and Gasdermin D protein, whereas it had a less binding affinity with NLRP3, Casp1, ASC, and Gasdermin E. At last, the Gasdermin D inhibitor can reverse the therapeutic effect of dapagliflozin. Dapagliflozin alleviates skeletal muscle atrophy in diabetic nephropathy mice, which is through the Gasdermin D-mediated canonical pyroptosis pathway.

Novel Protective Role for Gut Microbiota-derived Metabolite PAGln in Doxorubicin-induced Cardiotoxicity.

Doxorubicin (Dox) is a classic anthracycline chemotherapy drug with cause cumulative and dose-dependent cardiotoxicity. This study aimed to investigate the potential role and molecular mechanism of phenylacetylglutamine (PAGln), a novel gut microbiota metabolite, in Dox-induced cardiotoxicity (DIC). DIC models were established in vivo and in vitro, and a series of experiments were performed to verify the cardioprotective effect of PAGln. RNA sequencing (RNA-seq) was employed to explore the mechanism of PAGln in DIC. Subsequently, the differentially expressed genes (DEGs) were subjected to comprehensive analysis using diverse public databases, and RT-PCR was used to confirm the expression levels of the candidate genes. Finally, molecular docking techniques were used for validation. PAGln effectively prevented both in vivo and in vitro Dox-induced myocardial injury and cell apoptosis. RNA-seq results showed that 40 genes were up-regulated and 54 down-regulated in the Dox group compared to the Con group, displaying opposite changes in the Dox + PAGln group. Enrichment analysis highlighted several mechanisms by which PAGln alleviated Dox-induced cardiotoxicity, including the lipid metabolic process, calcium-mediated signaling, positive regulation of store-operated calcium channel activity, and hypertrophic cardiomyopathy. In vitro and in vivo experiments confirmed that PAGln treatment could reverse the changes in the expression levels of Klb, Ece2, Nmnat2, Casq1, Pak1, and Apob in Dox. Molecular docking results showed that these genes had good binding activity with PAGln. PAGln shows potential in alleviating Dox-induced cardiotoxicity, with Ece2 identified as key regulatory molecules related to endothelial dysfunction.

Discovery of key genes and plant extracts in Sjögren’s syndrome with yin deficiency syndrome based on female patients clinical data &amp; molecular docking

Background: Primary Sjögren’s Syndrome (pSS) is an autoimmune disease, for which Traditional Chinese Medicine has shown significant therapeutic effects based on different syndrome types of Sjögren’s Syndrome. However, the research findings between it and related plant extracts are limited. Objective: To explore the symptoms of patients with yin deficiency syndrome and non-yin deficiency syndrome of Sjögren’s Syndrome and their relationship with plant extracts under the background of Traditional Chinese Medicine. Methods: Whole blood samples were collected from SS patients with yin deficiency syndrome (5), SS patients without yin deficiency syndrome (5), and healthy volunteers (5) for proteomic testing. Using healthy volunteers as the control group, the differential protein expression of SS with yin deficiency syndrome was screened for enrichment analysis and functional annotation; combining previous effective clinical cases and prescription drugs for data mining and network pharmacological analysis, potential effective plant extracts were screened and enriched. Finally, molecular docking was used to validate the results of network analysis, providing a highly probable new idea for the correlation between potential drug targets for SS patients with yin deficiency syndrome and potential effective plant extracts. Results: 1. By analyzing the proteomic data of clinical samples, the differential genes of healthy individuals versus Sjögren’s Syndrome with yin deficiency versus Sjögren’s Syndrome without yin deficiency were discovered. By overlapping the above data, it was found that the key target genes for treating Sjögren’s Syndrome are HBA1, MST1, PCSK9, and HBB. 2. Enrichment analysis of plant extracts (PE) through clinical data mining of effective cases revealed the tremendous potential of four Chinese botanical drug - β-sitosterol, quercetin, berberine, and kaempferol in treating Sjögren’s Syndrome patients with yin deficiency syndrome. 3. The pathogenesis and regulation of Sjögren’s Syndrome with yin deficiency syndrome may be related to interleukin-related pathways, DNA synthesis, and oxidative stress. 4. Molecular docking suggests that berberine has good binding activity with MST1. Conclusion: We believe that berberine and MST1 may have significant research value in the treatment of Sjögren’s Syndrome with Female Patients. This study provides valuable reference points for future research on Sjögren’s Syndrome with yin deficiency syndrome.

Multipotent Effect of Clozapine on Lipopolysaccharide-Induced Acetylcholinesterase, Cyclooxygenase-2,5-Lipoxygenase, and Caspase-3: In Vivo and Molecular Modeling Studies.

Dual inhibition of cyclooxygenase-2 (COX-2) and lipoxygenase (LOX) is a recognized strategy for enhanced anti-inflammatory effects in small molecules, offering potential therapeutic benefits for individuals at risk of dementia, particularly those with neurodegenerative diseases, common cancers, and diabetes type. Alzheimer's disease (AD) is the most common cause of dementia, and the inhibition of acetylcholinesterase (AChE) is a key approach in treating AD. Meanwhile, Caspase-3 catalyzes early events in apoptosis, contributing to neurodegeneration and subsequently AD. Structure-based virtual screening of US-FDA-approved molecules from the ZINC15 database identified clozapine (CLOZ) as the dual inhibitor of COX-2 and AChE, with significant binding affinity. Further molecular docking of CLOZ in the active site of LOX and Caspase-3 also showed significant binding potential. Further, the results from molecular docking were validated using molecular dynamics simulation (MDS) studies, confirming the results from molecular docking. The results from MDS showed good binding potential and interactions with key residues. The CLOZ was further assessed using lipopolysaccharide (LPS)-challenged rats treated for thirty days at doses of 5 and 10 mg/kg, p.o. The results demonstrated modulation of COX-2, 5-LOX, AChE, Caspase-3, and MDA in LPS-induced brains. Additionally, the expression level of IL-10 was also measured. Our results showed a significant decrease in the levels of COX-2, 5-LOX, AChE, Caspase-3, and MDA. Our results also showed a significant decrement in the pro-inflammatory markers NF-κB, TNF-α, and IL-6 and an improvement in the levels of anti-inflammatory markers IL-10 and TGF-β1. Overall, the findings indicate that CLOZ has potential for neuroprotective effects against LPS-treated rats and can be explored.