Pharmacological Analysis Research Articles

Antidepressant effects of fisetin: Identifying molecular mechanisms by network pharmacology and molecular docking

Major depressive disorder (MDD) is a heterogeneous condition influenced by a complex interplay of social, psychological, and biological factors. Fisetin (FT), a flavonoid polyphenol found in various plants, has demonstrated neuroprotective properties that may be beneficial in treating MDD. This research aims to evaluate the potential molecular mechanisms of FT in treating MDD using network pharmacology analysis, with validation through molecular docking methods. We assessed the drug-like properties of FT using the TCMSP and SwissADME platforms. Potential drug targets for FT were identified through SuperPred and SwissTargetPrediction. We compiled MDD-associated targets from established databases and identified common genes shared between FT and MDD. The common targets were analyzed for protein-protein interactions using the STRING database to identify essential targets. Consequently, these key targets were further investigated through Kyoto Encyclopedia of Genes and Genomes and Gene Ontology (GO) enrichment analyses with the help of ShinyGO software. The results indicated that FT targets are linked to specific pathways involved in the pathogenesis of MDD, with the IL-17 signaling pathway emerging as a significant pathway of interest. Strong binding affinities were found between FT and key proteins, including glycogen synthase kinase 3 beta, monoamine oxidase A, acetylcholinesterase, matrix metalloproteinase 9, and myeloperoxidase, suggesting that FT may serve as a promising therapeutic agent for MDD by targeting components of the IL-17 pathway. In conclusion, this research successfully employed computational methods to elucidate the potential effectiveness of FT in managing MDD. It offered important perspectives on the regulatory mechanisms involved and emphasized the IL-17 signaling pathway as a possible target for MDD therapy.

Mechanism of Qilong Capsule against Myocardial Ischemia-Reperfusion Injury Based on Network Pharmacology and Experimental Validation.

Qilong capsule (QC) has been used clinically to treat ischemic stroke in China. This study evaluated the therapeutic effects of QC on myocardial ischemia-reperfusion injury (MIRI) and its potential mechanisms. The components and candidate targets of QC against MIRI were predicted by network pharmacology via relevant databases such as TCMSP, BATMAN-TCM, GeneCards. The potential mechanisms were predicted by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses and verified by enzyme-linked immunosorbent assay (ELISA) and Western blot. Network pharmacology analysis indicated that the cardioprotective effect of QC against MIRI was associated with inflammatory pathways. We further confirmed that QC effectively decreased the levels of inflammatory factors, including hs-CRP and MCP-1, and suppressed the expression of TNF-α and the phosphorylation of STAT3. This study provides evidence for further clinical applications of QC for MIRI therapy.

Integrating network pharmacology and component analysis to investigate the potential mechanisms of Sheng-Hui-Yi-Zhi decoction in the treatment of Alzheimer's disease.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by cognitive impairment. To elucidate the potential mechanisms of Sheng-Hui-Yi-Zhi (SHYZ) for the treatment of AD and explore the effective substances of SHYZ. Liquid chromatography-mass spectrometry (LC-MS) was used to identify the active components of SHYZ. Network pharmacology was employed to predict the potential targets and pathways of SHYZ in the treatment of AD. SAMP8 mice were used as a model for AD and were treated with SHYZ. The Morris water maze was utilized to assess the learning and memory capabilities of mice. Additionally, the levels of TNF-α, IL-1β, and IL-6 in the brain hippocampus of mice were quantified using ELISA. The protein expression of PI3 K/p-PI3 K, AKT/p-AKT, MAPK38/p-MAPK38, and NFκB p65/p-NFκB p65 in the hippocampus was analyzed using Western blotting. Additionally, qRT-PCR was employed to assess the gene expressions of TNF-α, IL-1β, and IL-6 in the hippocampus. The network pharmacological prediction results showed that the treatment of AD with SHYZ was closely related to the inhibition of inflammatory response. Behavioral experiments revealed that SHYZ significantly reduced the time taken to escape, increased the number of times the platform was crossed, and prolonged the residence time in the target quadrant. Meanwhile, SHYZ treatment suppressed the expression of Aβ1-42 protein and inflammatory factors. SHYZ significantly inhibited the expression of proteins of PI3 K, AKT, MAPK p38, and NF-κB p65. SHYZ has been shown to effectively ameliorate learning and memory impairment in SAMP8 AD mice by inhibiting the expression of Aβ1-42 and reducing the increase of inflammatory factors.

Network pharmacological analysis and in vitro testing of the rutin effects on triple-negative breast cancer.

This study aims to assess the potential mechanism of rutin to treat triple-negative breast cancer (TNBC) based on network pharmacology followed by in vitro experiments. The potential rutin targets were predicted, and the DisGeNET database was used to obtain the disease targets. The intersection targets were identified with Venny 2.1 software, with the String database subsequently used as input to produce the "drug-target-disease" visual network employing Cytoscape 3.7.2. Gene ontology. Kyoto Encyclopaedia of Genes and Genomes analyses were performed for intersection targets, while AutoDock Vina was used for molecular docking and visualization. Cell viability was assessed using the Colorimetric CCK-8 test, and apoptosis was analyzed using PI/Annexin V. The predicted core targets were confirmed by qPCR and western blotting assays. EGFR, IL6, TNF, and INS were found as the primary targets. The molecular docking analysis revealed the rutin interaction with the core targets. The in vitro results confirmed that rutin inhibited the growth of the MDA-MB-231 cell line. Rutin also induced cell death and decreased the expressions of IL6, TNF, INS, and EGFR. Rutin's multi-target effects and molecular mechanism for treating TNBC were confirmed through preliminary results. The results provide a theoretical base for rutin's possible function in breast cancer treatment.

Integrated Network Pharmacology and Transcriptomics Analysis to Elucidate the Mechanism of Huoxue Tongluo Qiwei Decoction in the Treatment of Erectile Dysfunction in Spontaneously Hypertensive Rats through Angii-Activated Pkcε Pathway.

As a classical formula to invigorate blood circulation, Huoxue Tongluo Qiwei Decoction (HTQD) can effectively treat hypertensive erectile dysfunction (ED), but its exact mechanism of action is not yet clear. The goal of this research was to explore the potential mechanism of HTQD in improving hypertensive erectile dysfunction in rats through transcriptomics, network pharmacology, and associated animal experimentations. The HTQD chemical constituents were screened using high-performance liquid chromatography- tandem mass spectrometry (HPLC-MS/MS). Furthermore, transcriptomics analysis was performed via mRNA sequencing to identify significantly differentially expressed proteins. Moreover, the key target proteins of HTQD in the treatment of hypertensive ED were screened by network pharmacology and transcriptomics. In addition, the endothelial cells of the corpus cavernosum were assessed using hematoxylin-eosin staining. The transcript and protein expressions were evaluated via western blotting and Real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The network pharmacology and transcriptome mRNA sequencing revealed that KCNE1 may be the target protein of HTQD in improving hypertensive ED. After HTQD treatment, the systolic and diastolic blood pressure (BP) of hypertensive rats decreased, the number of erections increased, and the pathological structure of the penis was improved. Moreover, HTQD downregulated the protein and mRNA expression of AngII, AT1R, DAG, and PKCε, whereas it upregulated the transcript and protein expression of KCNE1. HTQD may activate the PKCε pathway through AngII, inhibit the expression of KCNE1 protein, relax vascular smooth muscles, and improve erectile function.

The Anti-Inflammatory Effects of Formononetin, an Active Constituent of Pueraria montana Var. Lobata, via Modulation of Macrophage Autophagy and Polarization.

Pueraria montana var. lobata (Willd.) Maesen & S.M.Almeida ex Sanjappa & Predeep (P. lobata) is a medicinal herb widely used in the food and pharmaceutical industries, and studies have shown that P. lobata possesses significant anti-inflammatory pharmacological activities. In this paper, a total of 16 compounds were isolated and identified from P. lobata, among which compounds 1-3, 7, 14, and 16 were isolated from P. lobata for the first time. The results of an in vitro anti-inflammatory activity screening assay showed that compounds 1, 4, 6, 8, and 15 were able to significantly reduce the levels of pro-inflammatory cytokines IL-6 and IL-1β in LPS-induced RAW264.7 macrophages, with the most obvious effect produced by compound 6 (formononetin), while formononetin was able to significantly reduce the number of macrophages at the site of inflammation in transgenic zebrafish. In addition, network pharmacological analysis revealed that the anti-inflammatory activity of formononetin is closely related to autophagy and polarization targets such as TNF, EGFR, PTGS2, and ESR1. In vitro validation experiments showed that formononetin could enhance the expression of LCII/LCI and reduce the expression of P62 protein, reduce the expression of CD86, and enhance the expression of CD206, which further indicated that formononetin could reduce inflammation by regulating macrophage autophagy and polarization processes.

A Network Pharmacology Study and Experimental Validation to Identify the Potential Mechanism of Heparan Sulfate on Alzheimer's Disease-Related Neuroinflammation.

Heparan sulfate (HS) is a polysaccharide that is found on the surface of cells and has various biological functions in the body. The purpose of this study was to predict the pharmacological effects and molecular mechanisms of HS on Alzheimer's disease (AD) and neuroinflammation (NI) through a network pharmacology analysis and to experimentally verify them. We performed functional enrichment analysis of common genes between HS target genes and AD-NI gene sets and obtained items such as the "Cytokine-Mediated Signaling Pathway", "Positive Regulation Of MAPK Cascade", and "MAPK signaling pathway". To confirm the predicted results, the anti-inflammatory effect of HS was investigated using lipopolysaccharide (LPS)-stimulated BV2 microglia cells. HS inhibited the production of nittic oxide, interleukin (IL)-6, and tumor necrosis factor-α in LPS-stimulated BV2 cells, but not IL-1β. In addition, HS inactivated P38 in the MAPK signaling pathway. These findings suggest the potential for HS to become a new treatment for AD and NI.

Using network pharmacology analysis and molecular docking of quercetin compound for treatment of Alzheimer

Using network pharmacology analysis and molecular docking of quercetin compound for treatment of Alzheimer

The multi-target mechanism of action of Selaginella doederleinii Hieron in the treatment of nasopharyngeal carcinoma: a network pharmacology and multi-omics analysis

Nasopharyngeal carcinoma (NPC) presents significant treatment challenges due to its complex etiology and late-stage diagnosis. The traditional Chinese medicine Selaginella doederleinii Hieron (S. doederleinii) has shown potentiality in NPC treatment due to its multi-target, multi-pathway anti-cancer mechanisms. First, we identified NPC related target genes from databases like GeneCards, OMIM, and DisGeNET, and performed WGCNA analysis on the GSE53819 dataset to identify several important gene modules related to NPC. Active components and their targets in S. doederleinii were screened from the TCMSP and other databases, identifying 32 overlapping genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that these genes are primarily involved in critical biological processes like protein phosphorylation and cell cycle regulation. A protein–protein interaction network was constructed, and cytoHubba identified six key genes (BCL2, MAPK14, ABCB1, PLK1, ATM, HMOX1). Kaplan–Meier analysis and immune infiltration analysis further showed that these key genes are closely related to the prognosis and immune microenvironment of NPC patients. Single-cell RNA sequencing analysis revealed the expression distribution of key genes across different immune cell types and explored their roles in the differentiation process of malignant cells through pseudotime trajectory analysis. Molecular docking and dynamics simulation results indicated that the Berberine-MAPK14 and Matairesinol-PLK1 complexes have high binding affinity and stability. Binding free energy calculations confirmed the stability of these complexes. Based on our comprehensive multi-level analysis, the active components of S. doederleinii may play a significant role in the treatment of NPC through multi-pathway and multi-target synergistic effects.

Shenghui decoction inhibits neuronal cell apoptosis to improve Alzheimer's disease through the PDE4B/cAMP/CREB signaling pathway.

Shenghui Decoction (SHD) is a frequently utilized traditional Chinese medicine formula in clinical settings for addressing cognitive impairment in elderly individuals. Nevertheless, the precise mechanism by which SHD exerts its effects on the most prevalent form of dementia, Alzheimer's disease (AD), remains to be elucidated. Temperature-induced transgenic C. elegans assess Aβ deposition and toxicity. Behavioral experiments are utilized to assess learning and memory capabilities as well as cognitive impairment in APP/PS1 mice. Immunofluorescence and immunohistochemistry are employed to identify Aβ deposits, while UHPLCOE/MS combine network pharmacology is utilized to characterize chemical composition, predict target and analyze the biological processes and signaling pathways modulated by SHD. Molecular biology methodologies confirm the functionality of regulatory pathways. Molecular docking, molecular dynamic simulations (MD) and ultrafiltration-liquid chromatography/mass spectrometry (LC/MS) are employed for the assessment of the binding interactions between active ingredients of SHD and target proteins. SHD effectively reduced the deposition of Aβ in the head of C. elegans and mitigated its toxicity, as well as improved the learning deficits and cognitive impairment in APP/PS1 mice. Network pharmacology analyses revealed that G protein-coupled receptors (GPCRs) and cell apoptosis are the primary biological processes modulated by SHD, with KEEG results indicating that SHD regulated the cAMP signaling pathway. Subsequent experimental investigations demonstrated that SHD attenuated the loss of neurons in APP/PS1 mice, upregulated the expression of anti-apoptotic protein Bcl-2 and downregulated the expression of pro-apoptotic proteins like cleave-Caspase-3 both in vivo and in vitro. Additionally, SHD decreased intracellular AMP levels while increasing cAMP levels, leading to the phosphorylation of PKA to activate CREB. This process ultimately regulated the expression of Bcl-2, Bdnf, among others, to prevent cell apoptosis and safeguard neurons. Molecular docking, MD, and ultrafiltration-LC/MS revealed that the active constituents of SHD formed stable interactions with the cAMP hydrolysis enzyme phosphodiesterase 4B (PDE4B). SHD regulated the cAMP/CREB signaling pathway to inhibit neuronal cell apoptosis and improve AD. Furthermore, it is worth noting that this mechanism may be associated with the specific and consistent binding of SHD active ingredients to PDE4B, potentially offering promising candidates for drug development aimed at addressing AD.

Danggui Liuhuang Decoction Ameliorates Endothelial Dysfunction by Inhibiting the JAK2/STAT3 Mediated Inflammation

Ethnopharmacological relevanceVascular endothelial dysfunction (VED) is recognized as a key triggering diabetic vascular complications. Danggui Liuhuang Decoction (DGLHD) has shown potential in mitigating these complications. However, the clinical efficacy of DGLHD in enhancing endothelial function, as well as the molecular mechanisms underlying its alleviation of Type 2 Diabetes-Related Vascular Endothelial Dysfunction (T2DM-VED), remains insufficiently understood. Aim of the studyThis study aims to validate the therapeutic efficacy of DGLHD in ameliorating T2DM-VED through clinical research. Furthermore it seeks to analyze the pharmacodynamic basis and molecular mechanisms of DGLHD, elucidating the biological processes through which DGLHD alleviates VED in type 2 diabetes mellitus (T2DM). Materials and methodsPatients diagnosed with "Yin deficiency with hyperactive fire syndrome", who are at a high risk for atherosclerotic cardiovascular disease (ASCVD) associated with T2DM, were recruited for this study. The effect of DGLHD on vascular endothelial function in T2DM was assessed by measuring the levels of pro-inflammatory factors through enzyme-linked immunosorbent assay (ELISA) and flow-mediated dilation (FMD). The primary components of DGLHD were analyzed using the UHPLC-Q-Exactive Orbitrap system. Potential therapeutic targets of DGLHD were predicted using network pharmacology and molecular docking analysis. To validate the mechanism of DGLHD on T2DM-VED, endothelial injury and inflammation cell models were established using human umbilical vein endothelial cells (HUVECs). A mouse model of diabetic endothelial injury was also developed to observe the effects of DGLHD on pro-inflammatory factors and vascular endothelial factors were observed through immunohistochemistry. Additionally, the effects on the JAK2/STAT3 signaling pathway were observed through Western blot experiments. ResultsDGLHD was found to contain 201 active components. Network pharmacology analysis indicated that the treatment of T2DM-VED with DGLHD is associated with modulation of the JAK2/STAT3 signaling pathway. Molecular docking analysis demonstrated that small molecules in DGLHD interact with JAK2 and STAT3. Our clinical study demonstrated that DGLHD significantly reduces the levels of pro-inflammatory factors and improves FMD readings in diabetic patients, thereby alleviating T2DM-VED. DGLHD was shown to inhibit the phosphorylation of JAK2 and STAT3, which blocks the JAK2/STAT3 signaling pathway transmission, reducing the release of pro-inflammatory and vascular endothelial growth factors, and preventing the inflammatory response in vivo and in vitro. ConclusionThis study demonstrates the potential efficacy of DGLHD in improving endothelial function in T2DM patients at high risk for ASCVD. By inhibiting the JAK2/STAT3 signaling pathway, DGLHD effectively reduces the release of pro-inflammatory factors and vascular endothelial growth factors, alleviating VED.

Integrating lipidomics, 16S rRNA sequencing, and network pharmacology to explore the mechanism of Qikui granule in treating diabetic kidney disease mice.

Qikui granule (QKG), a hospital preparation of traditional Chinese medicine, has been widely used for diabetic kidney disease (DKD) in clinical practice. However, its holistic therapeutic effects and the underlying therapeutic mechanisms remain unclear. In the present study, the integrated analysis of network pharmacology, 16S rRNA sequencing, and non-targeted lipidomics was performed to explore the anti-DKD effects of QKG and the underlying mechanisms in db/db mouse DKD model. The results of the network pharmacology analysis identified the PI3K-AKT, EGFR, MAPK, JAK-STAT, FoxO, and AGE-RAGE signaling pathways as the potential molecular mechanisms responsible for the efficacy of QKG. Importantly, these signaling pathways were found to be closely related to lipid metabolism and gut microbiota. The therapeutic effectiveness of QKG against DKD was manifested by reducing body weight, alleviating oxidative stress, improving kidney function indicators, promoting the recovery of renal histopathological damage, and regulating the lipid metabolic profile of serum and kidney in db/db mice. A total of 26 lipid metabolites were identified as potential pharmacological biomarkers (PPBs) of QKG for the treatment of DKD, which were mainly involved in glycerophospholipid metabolism. Meanwhile, QKG could alleviate DKD-induced gut microbiota dysbiosis primarily by enriching Candidatus_Arthromitus, which showed a negative correlation with all 26 lipid PPBs as well as 5 biochemical parameters, including 2 oxidative stress factors and 3 kidney function indices. In conclusion, our findings suggest that QKG may upregulate the gut level of Candidatus_Arthromitus to suppress the abnormal activation of PI3K-AKT related signaling pathway, thereby reducing the levels of PC and LPC in the glycerophospholipid metabolism, to finally ameliorate the progression of DKD in db/db mice.

The effect and mechanism of sanguinarine against PCV2 based on the analysis of network pharmacology and TMT quantitative proteomics.

Porcine circovirus type 2 (PCV2) is highly prevalent in nature and serves as the primary pathogen responsible for porcine circovirus-associated disease (PCVD/PCVAD), posing a significant threat to pig production. Currently, vaccination alone could not provide the complete protection for PCV2 infection. The active ingredients of traditional Chinese medicine have shown a positive effect in combating viral infections. This study employed tandem mass tag (TMT) labeled proteomic analyses and network pharmacology to examine the effect and mechanism of sanguinarine against PCV2. IFIH1, IFITM1, p38α, and JNK were identified as the key targets of sanguinarine against PCV2 based on proteomics and network pharmacology. Using PCV2-infected PK-15 cells, it was discovered that sanguinarine inhibited the expression of the PCV2 CAP gene by upregulating IFIH1, thereby promoting STAT1 phosphorylation and activating MAVS expression. This, in turn, facilitated IRF3 phosphorylation, leading to increase IFITM1 expression. Simultaneously, sanguinarine suppressed the expression of the PCV2 CAP gene by inhibiting the expression of p38α, JNK, and p-JNK protein. In conclusion, the results of this study suggest that sanguinarine exerts anti-PCV2 effects through different targets and pathways, which lays the foundation for the subsequent development of new anti-PCV2 veterinary drugs.

Comprehensive Pharmacological Network Analysis of Kaempferol Targeting Proteins in Breast Cancer Pathways for Receptive Therapy

Comprehensive Pharmacological Network Analysis of Kaempferol Targeting Proteins in Breast Cancer Pathways for Receptive Therapy

Study on the protective mechanism of Xuemaitong Capsule against acute myocardial ischemia rat based on network pharmacology and metabolomics

BackgroundXuemaitong Capsule (XMT) is a widely recognized traditional Miao medicine extensively utilized in Chinese clinical settings. Previous studies have demonstrated XMT protective effects against acute myocardial ischemia (AMI). However, the mechanism by which XMT provides protection to AMI rats is yet to be fully understood. Aim of the studyThe purpose of this study was to investigate the protective mechanism of XMT on AMI rats through network pharmacology, traditional pharmacodynamics and metabolomics. Material and methodsThe components and potential targets of XMT were identified through the application of traditional Chinese medicine system pharmacology and traditional Chinese medicine molecular mechanism bioinformatics analysis tools. We constructed herb-composition-target networks and analyzed protein–protein interaction (PPI) networks. The potential mechanism was explored by pathway enrichment analysis. Subsequently, the AMI model was constructed by ligation of the anterior descending branch of the left coronary artery, and XMT protective effects on AMI rats were evaluated by analyzing the myocardial enzyme profiles, electrocardiograms(ECG), Triphenyltetrazolium chloride(TTC) staining, and Hematoxylin-Eosin (HE) staining in AMI rats. Metabolomics based on UHPLC-Q-Exactive Orbitrap MS was used to observe the protective effect of XMT on the serum metabolic profile of AMI, and multivariate statistical analysis further revealed the differential patterns of metabolites after XMT treatment. Finally, integrated pathway analysis was carried out to reveal the biological metabolic mechanism. ResultsA total of 392 active components of XMT acted with 624 targets for treating AMI. Pathway enrichment analysis revealed that XMT could treat AMI through TNF, MAPK and PI3K-Akt signaling pathways. Further, XMT could effectively prevent ST-segment elevation in the ECG, reduce the size of myocardial infarction, decrease cardiac weight index and cardiac enzyme levels, and mitigate histological damage in the hearts of AMI rats. In addition, XMT callback 117 metabolites and four metabolic pathways, including taurine and hypotaurine metabolism, phenylalanine metabolism, pyrimidine metabolism and retinol metabolism. Through integrating network pharmacology and metabolomics, we explored the biological mechanism by which XMT treats AMI. It was speculated that the mechanism of XMT is to regulate TNF signaling, PI3K-Akt pathway and MAPK signaling pathway, and participate in cell apoptosis, oxidative stress, immune and inflammatory reaction and other biological processes. ConclusionXMT plays a protective role in AMI rats by regulating multiple metabolic biomarkers, multiple targets and pathways. Therefore, XMT may provide a potential strategy for the treatment of AMI.

Screening of SIRT2 inhibitors from natural product databases using computer-aided drug design and molecular dynamics simulation

Abstract This study is committed to searching for inhibitors of deacetylase SIRT2 within the natural product database via computer-aided drug design techniques. A comprehensive computer-aided drug design platform has been successfully established by integrating various techniques such as drug-likeness screening, pharmacokinetic prediction, molecular docking, and molecular dynamics simulation. The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (YaTCM) has been thoroughly explored to identify SIRT2 inhibitors, and the discovered compounds have been validated using molecular dynamics simulation. Through the computer-aided drug design method, five compounds capable of binding to SIRT2 have been successfully screened out from 47,696 natural product compounds derived from 6,220 herbs in the YaTCM database. Molecular dynamics simulation reveals that Artonin E and Paleatin B can form stable receptor-ligand complexes in the active pocket of SIRT2 inhibitors. Based on computer-aided drug design and virtual screening and verification techniques, Artonin E and Paleatin B have been identified as inhibitors of SIRT2, which is a key therapeutic target for the treatment of neuroblastoma. Applying computer aided drug design techniques to identify potential drug molecules from natural products holds profound significance for drug research.

Integration of transcriptomics and metabolomics reveals the mechanism of Glycyrrhizae Radix Et Rhizoma extract inhibiting CCL5 in the treatment of acute pharyngitis.

Acute pharyngitis (AP) is a common condition marked by inflammation of the oropharynx, which can lead to severe throat swelling, breathing difficulties, and even suffocation, significantly impacting quality of life. Despite the beneficial anti-inflammatory activity of Glycyrrhizae Radix Et Rhizoma (GRER) and Isoliquiritigenin (ISL), their pharmacological mechanisms against AP remain unclear. This study explores the mechanisms by which GRER treats AP, utilizing both transcriptomics and metabolomics approaches. We identified the chemical components of GRER and those that enter the bloodstream using UPLC-MS/MS. Based on15 % ammonia-induced AP model, this study integrates transcriptomics and metabolomics to investigate the mechanism of GRER and ISL in the AP treatment. The results indicated that GRER has significantly protective and anti-inflammatory effects against AP. Our analysis identified 144 components of GRER in vitro and 17 components in vivo. Network pharmacology and quantitative analysis highlighted ISL as a key active ingredient responsible for GRER's anti-AP effects. Transcriptomics and metabolomics results indicate that GRER and its active ingredient ISL exert therapeutic effects on AP by inhibiting the expression of CCL5 in pharyngeal tissue, thereby downregulating the levels of pro-inflammatory metabolites malic acid and fumaric acid in the tricarboxylic acid (TCA) cycle pathway. The data in this article demonstrated that GRER and ISL has significantly anti-inflammatory effects and protective effects for AP by regulating CCL5 expression to reduce the levels of pro-inflammatory metabolites within TCA cycle pathway. It provides a scientific basis for prevention and treatment of AP.

Molecular insights into the therapeutic mechanisms of Bushen-Qiangdu-Zhilv decoction for ankylosing spondylitis.

Ankylosing spondylitis (AS) is a chronic rheumatic immune disease characterized by high disability rates, significantly affecting patients' quality of life. BuShen-QiangDu-ZhiLv Decoction (BQZD), developed by the renowned traditional Chinese medicine practitioner Jiao Shude, has been traditionally used for AS treatment. However, the bioactive components and the precise mechanisms underlying BQZD's therapeutic effects remain largely unexplored. To investigate the protective effects and elucidate the molecular mechanisms of BQZD in treating ankylosing spondylitis. Ultra Performance Liquid Chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) was used to identify active compounds in BQZD. Bulk RNA-seq and Gene Set Variation Analysis (GSVA) were conducted to assess changes in pathway activity in AS patients before and after three months of BQZD treatment. We also utilized network pharmacology and molecular docking analyses to predict potential mechanisms, identifying key target genes such as IL-6, NFATc1, and c-FOS. Animal experiments were performed to validate these findings. UPLC-MS/MS identified 28 active compounds in BQZD capable of entering the bloodstream, with potential anti-inflammatory, immunomodulatory, and bone metabolism-regulating effects. BQZD treatment led to a significant reduction in ESR, CRP, and ASDAS-CRP scores, indicating clinical improvement in AS patients. RNA-seq analysis showed decreased GSVA scores for the ossification pathway, with moderate reductions in inflammatory response and RANKL signaling pathways. Positive correlations were found between pathway activity and clinical indicators. Network pharmacology and transcriptomic analysis identified IL-6, NFATc1, and c-FOS as key targets. In vivo experiments confirmed that BQZD reduced TNF-α and IL-1β levels, inhibited ectopic ossification, and modulated the expression of DKK-1, MMP-9, and OPN in the CAIA model. BQZD exerts therapeutic effects in AS by regulating inflammation and abnormal ossification through multi-pathway, multi-target mechanisms. The identification of key target genes such as IL-6, NFATc1, and c-FOS provides a solid foundation for future research and clinical applications of BQZD in AS management.

Antiviral activity of HuaganJiedu decoction (HGJDD) against hepatitis B virus (HBV) through FOXO4/ERK/HNF4α signal pathway.

Chronic hepatitis B virus (HBV) infection is still a widespread global health issue. HuaganJiedu Decoction (HGJDD) is a common prescription for treating HBV in China, which has the effect of enhancing antiviral efficacy and improving clinical efficacy. However, its precise mechanism of action remains unclear, warranting further investigation to elucidate its therapeutic potential and integration into standard medical practices. This study aims to explore the therapeutic mechanism of HuaganJiedu Decoction (HGJDD) in HBV. We investigated the therapeutic potential of HGJDD, and LC-MS analysis characterized the chemical profile of HGJDD. In vitro, we utilized HepG2.2.15cell line to assess cytotoxicity and treatment efficacy of HGJDD compared to Entecavir controls. In vivo, assessments included monitoring HBV-related biomarkers and viral load. Network pharmacology and RNA-seq analyses identified molecular pathways and targets influenced by HGJDD treatment. Immunofluorescence and Western blotting provided further insights into the therapeutic mechanisms underlying HGJDD for HBV. HGJDD showed no toxicity on HepG2.2.15cells at 10%, 20%, 40%, and 80% serum concentrations. In vitro, HGJDD reduced HBsAg, HBeAg, and HBV DNA levels by dose-dependently and time-dependently. HGJDD can decrease the levels of HBsAg, HBeAg, and HBV DNA in serum and liver levels, meanwhile the therapeutic effect of high-dose HGJDD approach to EVT's in HBV Tg mice. According to intersection of network pharmacology and transcriptome, FOXO signal pathway was highlighted as potential targets and Immunofluorescence find that FOXO4D protein expression lever was increased in three HGJDD group, especially in high-dose HGJDD group. Western blotting confirmed increased level of FOXO4, ERK, and p-ERK and decreased levels of HNF4α, which reflected that the therapeutic effect was closely to FOXO4/ERK/HNF4α signal pathway. Traditional Chinese medicine (TCM) offers diverse herbal treatments for HBV, with HGJDD showing efficacy in reducing HBsAg, HBeAg, and HBV DNA levels at cellular and animal levels. This study identified that FOXO4/ERK/HNF4α signal pathway played an important role in HGJDD's therapeutic effects. These findings support HGJDD's potential in HBV treatment, providing a scientific basis for clinical use.

Anti-Candida albicans effect and mechanism of Pachysandra axillaris Franch.

Pachysandra axillaris Franch., a traditional herbal medicine in Yunnan, has been used to treat traumatic injuries and stomach ailments, some of which were related to microbial infections in conventional applications, but, to the best of our knowledge, the antifungal bioactivity of this plant and its main antifungal components have not been previously reported. To identify the antifungal compounds of P. axillaris against fluconazole-resistant C. albicans in vitro and in vivo, and then elucidate the underlying mechanism of action. The antifungal compounds were obtained by bioguided isolation, and then they were investigated in vitro by MIC, growth curves, time-kill assay, and drug resistance induction. The antifungal mechanism was explored using combined network pharmacology and metabolomic analysis, and further supported by analyzing sterol composition using LC-MS/MS, scanning and transmission electron microscopy observation of fungal cell morphology, examining its effects on cell membranes using the fluorescent probes and RT-qPCR. Additionally, the antifungal effect in vivo was evaluated by a murine C. albicans skin infection model. Three bioactive compounds from P. axillaris efficiently inhibited fluconazole-resistant C. albicans (MIC=4μg/mL), in which the major compound, pachysamine M, affected the ergosterol biosynthesis pathway by inhibiting ERG genes (ERG1, ERG4, ERG7, ERG9, and ERG24), leading to the accumulation of squalene, lanosterol, and zymosterol. So, pachysamine M targeted cell membranes in vitro by reducing the ergosterol level, to avoid drug resistance. In addition, it promoted wound healing, reduced fungal load, and alleviated inflammation in vivo. Pachysamine M, an antifungal compound without reported before, inhibited fluconazole-resistant C. albicans efficiently in vitro and in vivo, and its mechanism targeted cell membranes, reducing the risk of drug resistance, which validated the traditional use of P. axillaris for the treatment of fungal skin infections.