Network Pharmacology Research Articles

Erxian decoction ameliorates myocardial tissue damage through activating PI3K/AKT signaling pathway in ovariectomized rats

Background Erxian decoction (EXD) is an empirical formula for treating cardiovascular disease, our previous work has shown that EXD could improve the cardiovascular structure and function in ovariectomized (OVX) rats, but its pharmacological mechanism is still unclear. Materials and Methods Network pharmacology was utilized to assess the key active components and central targets of EXD in treating postmenopausal cardiovascular disease. Then, an OVX rat model was established, HE staining and transmission electron microscope were utilized to observe myocardial tissue morphology, TUNEL staining was utilized to detect cardiomyocyte apoptosis, western blot, and ELISA were used to confirm efficacy and pathway of EXD. Results The network pharmacology prediction results showed that 129 common targets were identified by intersecting EXD targets and postmenopausal cardiovascular disease targets, including AKT1, TNF, IL-6, IL-1β, PTGS2 and other core targets, apoptosis, PI3K/AKT, and other signaling pathways may be closely related to postmenopausal cardiovascular disease. After ovariectomy, the myocardial tissue of rats was damaged, the expression level of PI3K/AKT pathway-related molecules in the myocardial tissue were decreased, the apoptosis index of cardiomyocytes was increased, and the levels of inflammatory factors (TNF-α, IL-6, and IL-1β) were enhanced. EXD intervention could improve myocardial tissue injury, EXD could up-regulate the protein expression of PI3K and p-AKT in myocardial tissue, and thereby prevent myocardial cell apoptosis. At the same time, EXD downregulated the levels of inflammatory factors in serum of ovariectomized rats. Conclusion EXD may prevent myocardial tissue damage through induction of the PI3K/AKT signaling pathway, thereby reducing cardiomyocyte apoptosis and inflammation. EXD may be a potential drug for the treatment of postmenopausal cardiovascular disease.

Investigation of the regulatory mechanisms of Guiqi Yimu Powder on dairy cow fatty liver cells using a multi-omics approach.

Fatty liver disease in dairy cows is a metabolic disorder that significantly affects their health and productivity, imposing a notable economic burden on the global dairy industry. Traditional Chinese medicine (TCM), characterized by its multi-component and multi-target features, has shown unique advantages in the prevention and treatment of various diseases. Guiqi Yimu Powder, a traditional TCM formula, enhances growth, boosts production efficiency, and strengthens immune function in livestock by regulating antioxidant along with anti-inflammatory pathways. However, its specific regulatory mechanisms on fatty liver in dairy cows remain unclear. This study aims to investigate the molecular-level effects and potential regulatory mechanisms of Guiqi Yimu Powder in a Trimethylamine N-oxide (TMAO) induced fatty liver cell model of dairy cows. We employed a comprehensive analysis integrating transcriptomics, proteomics, metabolomics, and network pharmacology. An in vitro dairy cow fatty liver cell model was established using TMAO to induce lipid accumulation. Cells were treated with the optimal TMAO concentration identified through preliminary experiments, and further divided into a lipid accumulation group and Guiqi Yimu Powder treatment groups. The treatment groups received varying concentrations of Guiqi Yimu Powder (10, 20, 30, 40, or 50 g/L). High-throughput omics sequencing technologies were utilized to perform a comprehensive analysis of the treated cells. Bioinformatics methods were applied to explore the regulatory effects, aiming to elucidate the specific impacts of Guiqi Yimu Powder on lipid metabolism, liver function, and related signaling pathways, thereby providing scientific evidence for its potential application in the prevention and treatment of fatty liver in dairy cows. Guiqi Yimu Powder treatment significantly affected 1,536 genes, 152 proteins, and 259 metabolites. KEGG enrichment analysis revealed that the significantly altered molecules are involved in multiple pathways related to the pathology of fatty liver, including metabolic pathways, glutathione metabolism, hepatitis B, and AMPK signaling pathway (p < 0.05). Notably, joint analysis highlighted the regulatory mechanisms of Guiqi Yimu Powder on glutathione cycling, with L-5-Oxoproline identified as an important metabolic compound. These findings indicate its impact on oxidative stress, energy metabolism, and liver function, suggesting potential therapeutic applications for fatty liver in dairy cows. This study elucidated the regulatory mechanisms of Guiqi Yimu Powder on fatty liver cells in dairy cows, providing new scientific evidence for its potential application in the prevention and treatment of fatty liver disease.

Astaxanthin Alleviates Chronic Prostatitis via the ERK1/2 Signaling Pathway: Evidence from Network Pharmacology and Experimental Validation.

Astaxanthin (AST) has been widely recognized for its therapeutic potential in chronic inflammatory ailments. This study investigates the therapeutic efficacy and underlying mechanisms of AST in the management of chronic prostatitis (CP). Male Sprague-Dawley (SD) rats were randomly divided into control, complete Freund's adjuvant (CFA), and CFA + AST groups. CFA was used to induce the CP model, and saline was used for the control group. Inflammation of the prostate was detected 28 days after oral administration of AST. qRT-PCR and ELISA were used to detect pro-inflammatory factors in RWPE-1 and WPMY-1 cells. Potential targets of AST for CP were explored by network pharmacology, and related proteins were detected by Western blotting. Oral administration of AST alleviated the increase in prostate stroma and reduced inflammatory cell infiltration in CP rats. The IC50 of AST-treated RWPE-1 and WPMY-1 cells for 48 h were 171 and 212.1 μM, respectively. AST pretreatment reduced IL-6 and IL-8 expression in these cells. PPI network, GO, and KEGG enrichment analyses suggested that the antiinflammatory effect of AST was associated with the ERK1/2 pathway. Western blotting showed that AST inhibited ERK1/2 phosphorylation. In addition, AST and ERK1/2 pathway inhibitors (U0126) synergistically inhibited LPS-induced inflammation in prostate cells. Our study identified the potential of AST in the treatment of CP. However, subsequent randomized controlled trials are needed to validate its clinical efficacy.

Network pharmacology combined with experimental validation show that apigenin as the active ingredient of Campsis grandiflora flower against Parkinson's disease by inhibiting the PI3K/AKT/NF-κB pathway.

The exploration of novel natural products for Parkinson's disease (PD) is a focus of current research, as there are no definitive drugs to cure or stop the disease. Campsis grandiflora (Thunb.) K. Schum (Lingxiaohua) is a traditional Chinese medicine (TCM), and the exact active constituents and putative mechanisms for treating PD are unknown. Through data mining and network pharmacology, apigenin (APi) was identified as the main active ingredient of Lingxiaohua, and key targets (TNF, AKT1, INS, TP53, CASP3, JUN, BCL2, MMP9, FOS, and HIF1A) of Lingxiaohua for the treatment of PD were discovered. The primary routes implicated were identified as PI3K/AKT, Apoptosis, TNF, and NF-κB pathways. Subsequently, therapeutic potential of APi in PD and its underlying mechanism were experimentally evaluated. APi suppressed the release of mediators of inflammation and initiation of NF-κB pathways in MES23.5 cells induced by MPP+. APi suppressed caspase-3 activity and apoptosis and elevated p-AKT levels in MES23.5 cells. Pretreatment with LY294002, a PI3K inhibitor, resulted in APi treatment blocking the activation of NF-κB pathway and expression of inflammatory factors in MES23.5 cells by activating the PI3K/AKT pathway. In conclusion, APi protects dopaminergic neurons by controlling the PI3K/AKT/NF-κB pathway, giving novel insights into the pharmacological mechanism of Lingxiaohua in treating PD.

O-Allyloxy chalcone derivatives: design, synthesis, anticancer activity, network pharmacology and molecular docking

O-Allyloxy chalcone derivatives: design, synthesis, anticancer activity, network pharmacology and molecular docking

Taiwan Chingguan Yihau may improve post-COVID-19 respiratory complications through PI3K/AKT, HIF-1, and TNF signaling pathways revealed by network pharmacology analysis.

The emergence of new SARS-CoV-2 variants with a higher contagious capability and faster transmissible speed has imposed an incessant menace on global health and the economy. The SARS-CoV-2 infection might reoccur and last much longer than expected. Thence, there is a high possibility that COVID-19 can cause long-term health problems. This condition needs to be investigated thoroughly, especially the post-COVID-19 complications. Respiratory tract disorders are common and typical complications after recovery. Until now, there has been a lack of data on specialized therapeutic medicine for post-COVID-19 complications. The clinical efficacy of NRICM101 has been demonstrated in hospitalized COVID-19 patients. This herbal medicine may also be a promising therapy for post-COVID-19 complications, thanks to its phytochemical constituents. The potential pharmacological mechanisms of NRICM101 in treating post-COVID-19 respiratory complications were investigated using network pharmacology combined with molecular docking, and the results revealed that NRICM101 may exert a beneficial effect through the three primary pathways: PI3K/AKT, HIF-1, and TNF signaling pathways. Flavonoids (especially quercetin) have a predominant role and synergize with other active compounds to produce therapeutic effectiveness. Most of the main active compounds exist in three chief herbal ingredients, including Liquorice root (Glycyrrhiza glabra), Scutellaria root (Scutellaria baicalensis), and Mulberry leaf (Morus alba). To our knowledge, this is the first study of the NRICM101 effect on post-COVID-19 respiratory complications. Our findings may provide a better understanding of the potential mechanisms of NRICM101 in treating SARS-CoV-2 infection and regulating the immunoinflammatory response to improve post-COVID-19 respiratory complications.

Exploring the mechanisms of Cornus officinalis in the treatment of non-alcoholic steatohepatitis using network pharmacology, molecular docking and molecular dynamics simulations

Non-alcoholic steatohepatitis (NASH) is a predominant metabolic liver disease, typically characterized by hepatic steatosis, oxidative stress, and inflammation. The traditional Chinese medicine Cornus officinalis possesses anti-inflammatory and hepatoprotective pharmacological properties and has shown ameliorative effects on NASH. however, its mechanism of action remains unclear. This study aims to elucidate the mechanisms by which C. officinalis ameliorates NASH. The active components of C. officinalis were analyzed using the Traditional Chinese Medicine Systems Pharmacology Database (TCMSP), and the corresponding targets were predicted. Subsequently, the DisGeNET, GeneCards, and GEO databases were employed to identify NASH-related targets. Venn diagrams were used to intersect the C. officinalis targets with the NASH targets. Protein–protein interaction (PPI) networks were constructed using the STRING database, and PPI network analysis was performed using Cytoscape. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted using the Database for Annotation, Visualization, and Integrated Discovery (DAVID), followed by molecular docking validation. Cornus officinalis was found to contain 20 major active ingredients corresponding to 672 potential targets, 61 of which overlapped with NASH targets. PPI network, GO, and KEGG pathway analyses identified four targets with the highest correlation, and molecular docking results indicated that the active ingredients of C. officinalis exhibited strong binding affinities to NASH targets. The treatment of NASH with C. officinalis is characterized by multiple active ingredients and multiple targets, underscoring the major advantage of traditional Chinese medicine in treating NASH.

Exploring the protective effect and molecular mechanism of betulin in Alzheimer's disease based on network pharmacology, molecular docking and experimental validation.

Alzheimer's disease (AD) is a neurodegenerative disorder that impairs learning and memory, with high rates of mortality. Birch bark has been traditionally used in the treatment of various skin ailments. Betulin (BT) is a key compound of birch bark that exhibits diverse pharmacological benefits and therapeutic potential in AD. However, the therapeutic effects and molecular mechanisms of BT in AD remain unclear. The present study aimed to predict the potential therapeutic targets of BT in the treatment of AD, and to determine the specific underlying molecular mechanisms through network pharmacology analysis and experimental validation. PharmMapper was used to predict the target genes of BT, and four disease databases were searched to screen for AD targets. The intersection targets were identified using the jveen website. Drug‑disease target protein‑protein interaction networks and hub genes were obtained and visualized using the Search Tool for the Retrieval of Interacting Genes/Proteins database and Cytoscape. The Database for Annotation, Visualization and Integrated Discovery was used for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, and AutoDock was used for molecular docking analysis of BT and hub genes. Subsequently, the network‑predicted mechanisms of BT in AD were verified in vitro. A total of 495 BT and 1,386 AD targets were identified, and 120 were identified as potential targets of BT in the treatment of AD. The results of the molecular docking analysis revealed a strong binding affinity between BT and the hub genes. In addition, enrichment analyses of GO and KEGG pathways indicated that the neuroprotective effects of BT mainly involved the 'PI3K‑Akt signaling pathway'. The results of in vitro experiments demonstrated that pretreatment with BT for 2 h may ameliorate formaldehyde (FA)‑induced cytotoxicity and morphological changes in HT22 cells, and decrease FA‑induced Tau hyperphosphorylation and reactive oxygen species levels. Furthermore, the PI3K/AKT signaling pathway was activated and the expression levels of downstream proteins, namely GSK3β, Bcl‑2 and Bax, were modified following pre‑treatment with BT. Overall, the results of network pharmacology and in vitro analyses revealed that BT may reduce FA‑induced AD‑like pathology by modulating the PI3K/AKT signaling pathway, highlighting it as a potential multi‑target drug for the treatment of AD.

System biology-based assessment of the molecular mechanism of IMPHY000797 in Parkinson’s disease: a network pharmacology and in-silico evaluation

IMPHY000797 derivatives have been well known for their efficacy in various diseases. Moreover, IMPHY000797 derivatives have been found to modulate such genes involved in multiple neurological disorders. Hence, this study seeks to identify such genes and the probable molecular mechanism that could be involved in the pathogenesis of Parkinson’s disease. The study utilized various biological tools such as DisGeNET, STRING, Swiss target predictor, Cytoscape, AutoDock 4.2, Schrodinger suite, ClueGo, and GUSAR. All the reported genes were obtained using DisGeNET, and further, the common genes were incorporated into the STRING to get the KEGG pathway, and all the data was converted to a protein/pathway network via Cytoscape. The clustering of the genes was performed for the gene-enriched data using two-sided hypergeometrics (p-value). The binding affinity of the IMPHY000797 was verified with the highest regulated 25 proteins via utilizing the “Monte Carlo iterated search technique” and the “Emodel and Glide score” function. Three thousand five hundred eighty-three genes were identified for Parkinson’s disease and 31 genes for IMPHY000797 compound, among which 25 common genes were identified. Further, the “FOXO-signaling pathway” was identified to be a modulated pathway. Among the 25 proteins, the highest modulated genes and highest binding affinity were exhibited by SIRT3, FOXO1, and PPARGC1A with the compound IMPHY000797. Further, rat toxicity analysis provided the efficacy and safety of the compound. The study was required to identify the probable molecular mechanism, which needs more confirmation from other studies, which is still a significant hit-back.

Simultaneous recycling of flavonone and polymethoxyflavonoids from Chenpi processing residue based on their complementary action on glycolipid lowering activity: Innovative combination use of yeast and aqueous two-phase system

This study aimed at recycling flavonoids with glycolipid lowering activity from Chenpi processing residue produced by essential oil extraction. The crude Chenpi flavonoids obtained by ethanol reflux mainly contained flavonone (hesperidin) and polymethoxyflavonoids (nobiletin, tangeretin, sinensetin, and isosinensetin), which complementally exerted strong glycolipid lowering activity as revealed by digestive enzyme inhibitory activity, molecular docking and network pharmacology. The reducing sugar and water-soluble impurities were efficiently removed by yeast fermentation and aqueous two-phase system (ATPS) extraction, respectively. The optimal combination use of yeast (2 % w/v, CN36) and ATPS formed by 22 % (w/w) (NH4)2SO4 and 25 % (w/w) ethanol improved flavonoid purity to 60.25 %, enriched flavonone and polymethoxyflavonoids by 6.69-fold, and enhanced digestive enzyme inhibitory activity, which was superior to the use of individual yeast or ATPS, unoptimized combination, macroporous resin (XAD-16), and ethyl acetate. This study was the first to develop a recyclable biophysical method for simultaneous enrichment of flavonone and polymethoxyflavonoids.

Deciphering farnesol's anti-arthritic and immunomodulatory potential by targeting multiple pathways: a combination of network pharmacology guided exploration and experimental verification.

Farnesol (FAR), a sesquiterpene alcohol, has documented FAR's anti-inflammatory and antioxidant activities. Current study was undertaken to assess the efficacy and mechanism of FAR in arthritis by employing network pharmacology and experimental models. Two experimental models comprising formaldehyde- and complete Freund's adjuvant (CFA)-induced arthritis evaluated the efficacy of FAR in treating arthritis. Various parameters were assessed. Then, a network pharmacology approach was applied to gain further insight into the potential mechanism and signaling pathways. FAR significantly reduced paw volume and the arthritic score and improved the hematological and biochemical changes. Radiographic and histological examination showed the anti-arthritic efficacy of FAR, which was associated with down-regulation of pro-inflammatory mediators and upregulation of anti-inflammatory mediators. Network pharmacology analysis revealed that FAR may exert its anti-arthritic effects by targeting specific genes associated with arthritis. Pathway analysis revealed the involvement of three key signaling pathways (IL-17 signaling, TNF signaling, and toll-like receptor signaling) in the development and progression of arthritis. The results pointed out the protective attributes of farnesol against formaldehyde and CFA-induced arthritis via modulation of multiple targets. This study provides a valuable reference for the development of a new treatment or complementary therapy for arthritis.

From Life's Essential 8 to metabolic syndrome: insights from NHANES database and network pharmacology analysis of quercetin.

Metabolic syndrome (MetS), or syndrome X, is a collection of metabolic illnesses that affect the body's health, particularly insulin resistance and obesity. The prevalence of MetS is on the rise, particularly among younger individuals. Quercetin, a natural flavonoid found in many traditional Chinese medicines, can impact various pathways to disrupt the pathological advancement of MetS with few negative effects. The American Heart Association recently introduced a cardiovascular health assessment termed Life's Essential 8 (LE8), which might impact the treatment of MetS. Quercetin targets and their functions in MetS pathways were identified using a network pharmacology method and molecular docking techniques. The study examined quercetin's direct and indirect interactions with proteins linked to the pathogenic processes of MetS. Data were collected regarding the American Heart Association's LE8 cardiovascular health indicators, which include health behaviors (diet, physical activity, nicotine exposure, and sleep) and health factors (body mass index, non-high-density lipoprotein cholesterol, blood glucose, and blood pressure). The study assessed the connection between LE8 and the occurrence of MetS, taking into account dietary quercetin consumption as a variable of interest. The negative correlation between MetS and LE8 indicates that individuals with higher LE8 scores are less likely to develop MetS. Individuals in the fully adjusted highest group (LE8 ≥ 80) demonstrated a 79% lower likelihood of developing MetS than those in the lowest group (OR = 0.21; 95% CI, 0.17-0.26, p < 0.0001). Network pharmacology and molecular docking results show that quercetin may exert its therapeutic effects by modulating various biological response processes, including those related to xenobiotic stimuli, bacterial molecules, lipopolysaccharides, and oxidative stimuli. These processes involve key pathways associated with diabetic complications, such as the AGE-RAGE signaling pathway, pathways related to diabetic complications, and pathways involved in lipids and atherosclerosis. Therefore, quercetin may reduce cardiovascular risk, improve glucose-lipid metabolism, and alleviate insulin resistance and other biological processes by influencing multiple aspects of the lipid profile, blood glucose, and insulin resistance, ultimately impacting the links between LE8 score and MetS. This study discovered that an optimal LE8 score is a marker of adopting a lifestyle of wellness and is connected with a reduced likelihood of developing MetS. Quercetin acts on core targets such as IL6, BCL2, TP53, IL1B, MAPK1, and CCL2, and then plays a therapeutic role in regulating lipid metabolism, anti-inflammation, immunomodulation, autophagy, etc., through the pathways of diabetic complications, lipids, atherosclerosis, etc., and has the characteristics of multi-targets, multi-pathways, and multi-functions in regulating interventions for MetS.

Bioinformatics-based drug repositioning and prediction of the main active ingredients and potential mechanisms of action for the efficacy of Dan-Lou tablet

Drug repositioning is gaining attention as a method for developing new drugs due to its low cost, short cycle time, and high success rate. One important approach is to explore new uses for already marketed drugs. In this study, we utilized the strategy of drug repositioning, focusing on the Dan-Lou tablet. We predicted the efficacy of Dan-Lou tablet against non-small cell lung cancer based on gene expression similarity and verified it by in vitro experiments. Next, we performed further analysis and validation using network pharmacology, molecular docking and molecular dynamics. Based on the results, it was concluded that Dan-Lou tablet mainly acted through nine compounds, Quercetin, Luteolin, Scoparone, Isorhamnetin, Eugenol, Genistein, Coumestrol, Hederagenin, Succinic Acid, and mainly targeted CCL2, FEN1, TPI1, RMI2 by six pathways. This discovery not only provides a new idea for the development of Dan-Lou tablet but also provides useful predictive information for clinical treatment. The method we adopted has great development prospects as a way to predict the efficacy of new drugs and their main mechanisms of action, and it has a positive impact on the research and development of new drugs using drug repositioning and the modernization of traditional Chinese medicine.

A network pharmacology approach to elucidate the anti-inflammatory and antioxidant effects of bitter leaf (Vernonia amygdalina Del.)

The therapeutic potential of bitter leaf (Vernonia amygdalina Del.) has been established both empirically and in various scientific investigations. However, the molecular pathways related to its possible anti-inflammatory and antioxidant properties remain unclear. Therefore, the aim of this study was to elucidate the molecular interactions between bitter leaf's bioactive compounds and cellular targets involved in these activities. The compounds in bitter leaf were identified using gas chromatography-mass spectrometry (GC-MS) analysis, and subsequently, a network pharmacology approach was employed together with molecular docking and dynamics simulations. Acetonitrile (4.5%) and dimethylamine (4.972%) were the most prevalent compounds among the 38 identified by the GC-MS analysis of bitter leaf extract. The proto-oncogene tyrosine-protein kinase (SRC) demonstrated significant connectivity within the antioxidant network, highlighting its pivotal role in facilitating inter-protein communication. It also exhibited strategic positioning in anti-inflammatory mechanisms based on closeness centrality (0.385). The enrichment analysis suggested multifaceted mechanisms of bitter leaf compounds, including transcriptional regulation and diverse cellular targeting, indicating broad antioxidant and anti-inflammatory effects. Eicosapentaenoyl ethanolamide (EPEA) displayed strong interactions with multiple proteins, including SRC (-7.17 kcal/mol) and CYP3A4 (-6.88 kcal/mol). Moreover, EPEA demonstrated to form a stable interaction with SRC during a 100 ns simulation. In conclusion, the computational simulations revealed that the hypothetical antioxidant and anti-inflammatory actions of bitter leaf compounds were achieved by specifically targeting SRC. However, confirmation using either in vitro or in vivo techniques is necessary.

Network Pharmacology Revealing the Therapeutic Potential of Bioactive Components of Triphala and Their Molecular Mechanisms against Obesity.

Obesity, characterized by the excessive accumulation of fat, is a prevalent metabolic disorder that poses a significant global health concern. Triphala, an herbal combination consisting of Phyllanthus emblica Linn, Terminalia chebula Retz, and Terminalia bellerica (Gaertn) Roxb, has emerged as a potential solution for addressing concerns related to obesity. This study aimed to investigate the network pharmacology and molecular docking of Triphala to identify its bioactive ingredients and their interactions with pathways associated with obesity. The bioactive compounds present in Triphala and genes linked to obesity were identified, followed by an analysis of the protein-protein interaction networks. Enrichment analysis, including Gene Ontology analysis and Kyoto Encyclopedia of Genes and Genomes pathway analysis, was conducted. Prominent genes and compounds were selected for further investigation through molecular docking studies. The study revealed a close correlation between obesity and the AKT1 and PPARG genes. The observed binding energy between beta-sitosterol, 7-dehydrosigmasterol, peraksine, α-amyrin, luteolin, quercetin, kaempferol, ellagic acid, and phyllanthin with AKT1 and PPARG indicated a favorable binding affinity. In conclusion, nine compounds showed promise in regulating these genes for obesity prevention and management. Further research is required to validate their specific effects.

Screening the effective components of Lysionotus pauciflorus Maxim. on the treatment of LPS induced acute lung injury mice by integrated UHPLC-Q-TOF-MS/MS and network pharmacology

Ethnopharmacological relevanceAcute lung injury (ALI) is an inflammatory reaction produced through various injury-causing factors acting on the lungs in a direct or indirect way, with a high morbidity and mortality rate. A review of clinical experience has revealed that Lysionotus pauciflorus Maxim (LP) has a significant therapeutic effect on ALI. However, the comprehensive effective components of LP are uncertain, and the mechanisms, especially the potential therapeutic target for anti-ALI, are still unknown. Aims of the studyIn vitro and in vivo validation of the pharmacodynamics of LP in the treatment of ALI and exploration of its potential mechanism of action based on network pharmacology, molecular docking and experimental validation. Materials and methodsUltra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) was employed to identify the ingredients of LP extracts. The potential bioactive ingredients, key targets and signalling pathways were identified by network pharmacology, based on the results of the mass spectrometry analysis. Subsequently, molecular docking was performed on the screened core components and key targets to calculate their molecular binding energies and binding potentials, and to explore the mutual binding modes of small-molecule ligands and large-molecule proteins. Finally, lipopolysaccharide (LPS)-induced RAW264.7 cell model and ALI mice model were used to validate the therapeutic effects and potential mechanism of LP extract towards ALI. ResultsFrom the mass spectrometry results of LP extracts, a total of 89 chemical components were identified, including 46 phenylethanol glycosides, 26 flavonoids, 9 organic acids and their derivatives and 8 other compounds. And furthermore 39 core active components were screened by network pharmacology. The top 10 core components (4 phenylethanol glycosides, 6 flavonoids) have been screened in the composition -target-disease network, and 37 core targets related to LP efficacy were obtained by fitting PPI network analysis. 10 signalling pathways and their targets associated with LP treatment of ALI were obtained by GO/KEGG analysis, indicating that LP could regulate TLR4 and NF-κB signalling pathways through 4 key targets, namely NFKB1, RELA, TLR4 and TNF. The results of the molecular docking procedure indicated a strong affinity, with the binding energies between each component and the target site being less than −6 kcal/mol. The binding modes included Hydrogen Bonds, Pi-Pi interaction, Hydrophobic Interactions, Salt Bridges, Pi-cation interactions. These observations were subsequently validated in vitro and in vivo experiments. The outcomes of in vitro and in vivo experiments demonstrated that LP was effective in reducing the infiltration of inflammatory bacteria in lung tissues and attenuated the expression of pro-inflammatory cytokines in LPS-stimulated mice bronchoalveolar lavage fluid (BALF) and RAW264.7 cells. Furthermore, LP inhibited the expression and phosphorylation of TLR4 protein and NF-κB protein, thus playing a role in the prevention of ALI. ConclusionsIn this study, mass spectrometry analysis was combined with biomolecular networks to initially elucidate the potential of LP to treat ALI by modulating the TLR4/NF-κB pathway. This offers a definitive experimental basis for the development of new LP drugs.

Integrative study to determine the anti-tumor role and mechanism of Chouchunpi San in colorectal cancer

IntroductionThe herbal formula Chouchunpi San (CCPS) is a traditional formula that has been widely used and proven effective in various clinical cancer treatments. However, the mechanism of its anticancer effect remains unclear. This study aims to determine the anti-tumor effect of CCPS on colorectal cancer (CRC) in vivo and in vitro and to explore the underlying molecular mechanisms. MethodsCell counting kit-8 assays, colony-forming assays, and flow cytometry for cell cycle and apoptosis assays were employed to determine the anti-tumor roles of CCPS. Liquid chromatography-mass spectrometry (LC-MS), network pharmacology, molecular docking, analysis of real-world clinical datasets of CRC, and western blotting were conducted to explore the molecular mechanism of CCPS on CRC. CRC xenografted mouse model, western blotting, and public CRC data analysis were conducted to evaluate the anti-tumor efficacy and mechanisms of CCPS on CRC. ResultsCCPS suppresses the growth of CRC cells and increases the number of apoptotic cells dose-dependently. CCPS targets and significantly downregulates RPA1 and enhances the phosphorylation of its downstream effectors, ATR and CHK1, which are critical for CRC progression. Additionally, CCPS shows a comparable anti-tumor effect in CRC xenografted mouse models compared to Capecitabine, a chemotherapy drug commonly used in clinics. DiscussionOur findings demonstrate the potential use of CCPS in cancer treatment by suppressing cancer cell growth and modulating the RPA1/ATR/CHK1 signaling pathway in CRC. Further investigations on the application of CCPS in cancer therapies could be extended to evaluate the potential in vivo toxicity and adverse events, as well as the synergistic effect of CCPS in combination with other chemotherapeutic agents.

Study on the Mechanism of Guizhi Fuling Decoction in Treating Hyperlipidemia Based on Network Pharmacology and Experimental Methods

Background Hyperlipidemia, caused by abnormal lipid metabolism, has become a significant health concern, especially in younger populations. While statins are commonly used for treatment, they often cause severe side effects with long-term use, leading to increased interest in finding natural alternatives. Purpose This study aimed to investigate how Guizhi Fuling Decoction (GZFL) treats hyperlipidemia using network pharmacology and experimental validation. Materials and Methods Network pharmacology analyzed GZFL’s active components, targets, and treatment mechanisms for hyperlipidemia. Hyperlipidemia rat and high-fat cell models were established. Experimental validation included enzyme-linked immunosorbent assay, Western blot, quantitative polymerase chain reaction, Oil Red O staining, and other methods. Statistical analysis was performed using one-way analysis of variance followed by Tukey’s post hoc test for multiple comparisons. Pearson correlation analysis was used to identify correlations between key protein expressions and lipid levels. Results GZFL comprises 85 active components and targets 218 proteins. Hyperlipidemia involves 3,852 targets, with 175 overlapping targets. Key targets included estrogen receptor 1, prostaglandin-endoperoxide synthase 2, interleukin-6 (IL-6), protein kinase B (AKT) 1, tumor necrosis factor, interleukin-1 beta, peroxisome proliferator-activated receptor gamma (PPARG), tumor protein p53 (TP53), and fructo oligosaccharide (FOS). Major active components were quercetin, phytosterols, kaempferol, and baicalein. Enrichment analysis highlighted processes like lipopolysaccharide response and lipid signaling pathways. Animal studies showed GZFL significantly reduced serum total cholesterol, triglycerides, and low-density lipoprotein cholesterol levels, and increased apolipoprotein A1 ( p &lt; 0.05). Western blot indicated that GZFL influenced AKT, TP53, IL-6, PPARγ expression, and mRNA levels in the phosphatidylinositol 3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) pathway ( p &lt; 0.01). Serum pharmacology experiments demonstrated GZFL-medicated serum reduced lipid droplet formation in high-fat cell models. Conclusion GZFL contains multiple pharmacologically active components, such as quercetin, phytosterols, and kaempferol, which can influence lipid metabolism. It may exert its effects by regulating proteins like AKT, TP53, IL-6, and PPARγ, participating in lipid and atherosclerosis signaling pathways and the PI3K/AKT/mTOR pathway.

Shenmai injection improves lipid metabolism in post-myocardial infarction heart failure based on network pharmacology and experimental validation

BackgroundShenmai injection (SMI), a traditional Chinese medicine formulation derived from the herbal decoction Shenmai Yin, is widely used in treating cardiovascular disorders. This study extensively investigated the effects and mechanisms of action of SMI on lipid metabolism in post-myocardial infarction heart failure (pMIHF). MethodsNetwork pharmacology was employed to predict the key targets and associated pathways involved in lipid metabolism for potential SMI treatments in post-myocardial infarction heart failure (pMIHF). Subsequently, a pMIHF mouse model and an ischemia/reperfusion (I/R) cell model were established to delve deeper into and validate the underlying mechanism of action. ResultsWe performed network pharmacology analysis, which identified 48 active components in SMI and 201 common gene targets. Subsequent screening using the protein–protein interaction network identified 26 core targets, including interleukin (IL)-6, tumor necrosis factor (TNF)-α, peroxisome proliferator-activated receptor alpha (PPARα), and sirtuin 1 (SIRT1). Based on Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses, we predicted that SMI might act on lipid metabolism in pMIHF through the PPARα pathway, a hypothesis supported by the strong binding affinity between this receptor and the active components of SMI, as confirmed via molecular docking. In a left anterior descending artery-ligation mouse model, SMI significantly improved cardiac function, reduced serum free fatty acid levels, decreased inflammatory cell infiltration and myocardial fibrosis, and maintained myocardial mitochondrial morphology. In ischemia-reperfusion (I/R) cells, SMI reduced cell apoptosis, improved mitochondrial membrane potential, and decreased mRNA expression levels of IL-6 and TNF-α, while increasing protein levels of PPARα, SIRT1, and PPARα co-activator-1 alpha (PGC1α). ConclusionCollectively, our findings suggest that SMI enhances myocardial lipid metabolism and ameliorates pMIHF by upregulating the PPARα/SIRT1/PGC1α pathway.

Protective effects and mechanisms of HuDiChangRong capsule on TNBS-induced ulcerative colitis in mice

Ethnopharmacological relevanceUC, characterized by chronic inflammation primarily affecting the colon and rectum, follows a protracted relapsing course marked by inflammation and an abundance of free radicals at the onset. Hudichangrong Capsule (HDCRC), a traditional Chinese medicinal formula, has long been employed in the treatment of UC and chronic bacillary dysentery, exhibiting positive therapeutic outcomes and a high rate of cure in clinical practice. Aim of the studyThe precise mechanism underlying its efficacy for UC remains elusive. Our objective was to investigate the anti-inflammatory effect and underlying mechanisms of HDCRC on TNBS-induced UC. Materials and methodsHere, we introduced HDCRC and induced UC using TNBS. SPF BALB/c mice were divided into 6 groups as follows: control group, colitis model group, colitis treated with sulfasalazine (400 mg/kg) group, and colitis treated with HDCRC (156, 312, and 624 mg/kg) groups. To assess the effects of HDCRC on colitis, we measured body weight loss, disease activity index (DAI), colon length, tissue damage, degree of inflammation, immune capacity, and oxidative stress. Additionally, we evaluated the TLR-4/MyD88 pathway and its downstream signaling using immunohistochemistry, real-time qPCR, and Western blot. Network pharmacology was used for main target prediction. 16s rRNA was employed for gut microbiota detechtion and UPLC-QTOF-MS was used for its and its metabonomics. ResultsHDCRC significantly slowed weight loss, ameliorated DAI, restored colon length, alleviated TNBS-induced tissue damage. It exerted the therapeutic effects via reducing oxidative stress, restoring immune balance, normalizing the inflammatory mediator levels and restoring intestinal barrier integrity. Furthermore, HDCRC mainly alleviate UC via suppressing the TLR-4/MyD88 pathway and its downstream signaling. The key components of the downstream pathway, including TLR-4, MyD88, NF-κB p65, ERK, p-JNK, p38, p-JAK1, JAK1, p-STAT3, and STAT3, were improved, thereby ameliorating the TNBS-induced injury. In addition, HDCRC could regulate gut microbiota (eg. Erysipelaloclostridium,etc.) and its metabonomics (eg. Vitamin B6 metabolism) in UC mice. ConclusionsIn conclusion, HDCRC exerts a protective effect against TNBS-induced UC in mice by inhibiting the TLR-4/MyD88 pathway and its downstream signaling, and partially JAK1/STAT3, suppressing oxidative stress, regulating immunity, restoring intestinal barrier integrity, and regulating gut microbiota and its metabonomics.