Anti‐inflammatory components of the essential oil from the leaves of Citrus × aurantium L. based on network pharmacology study

Context: Breast cancer therapy currently presents several uncomfortable side effects in patients, including effects on non-malignant tissues, recurrence, and resistance, which restrict their utilization. Consequently, researchers have directed their attention toward studying plant-derived anticancer compounds that exhibit high efficacy and safety profiles. Eugenol, a major component found in clove plants, demonstrates promising potential as a therapeutic agent for both estrogen receptor-positive and estrogen receptor-negative breast cancer. Aims: To predict the target of eugenol in estrogen receptor–positive breast cancer using network pharmacology and molecular docking analyses. Methods: Network pharmacology analysis was performed using the Chemical Toxigenomic Database, STITCH, GeneCards, Cytoscape, Enrichr, and Stringdb. Subsequently, molecular docking was performed using protein targets obtained from the RCSB-PDB and analyzed using AutoDock software. Results: Network pharmacology study and molecular docking revealed the anticancer effect of eugenol against breast cancer estrogen receptor–positive, especially in cancer and apoptotic pathways, by acting on caspase-3 (CASP3), epidermal growth factor receptor (EGFR), and poly [ADP-ribose] polymerase 1 (PARP1) signaling pathways. The docking results between the protein targets and eugenol showed that eugenol has the strongest binding with CASP3 (ligand binding energy: -5.78 kcal/mol), followed by eugenol binding with EGFR (ligand binding energy: -5.58 kcal/mol), and eugenol binding with PARP1 (ligand binding energy: -5.58 kcal/mol). Conclusions: Eugenol is a potential candidate for breast cancer therapy, especially for apoptosis mediated by CASP3 in breast cancer luminal A.

BackgroundRosa rugosa cv. Plena, a cultivar of Rosa rugosa, has a history of more than 1300 years of application in both medicine and food in China. The essential oil of Rosa rugosa cv. Plena (PREO) is one of the most frequently used additives in food, cosmetics and aromatherapy. PREO exhibits some anti-inflammation, antioxidant and nerve alleviating effects. However, the mechanisms behind these effects are still unclear.MethodsThe composition of PREO was determined by GC‒MS. Network pharmacology was performed to predict the possible compound-target network and analyze the possible targets against inflammation and oxidative stress. An inflammatory immune cell model was constructed by exposing RAW 264.7 cells to LPS. A series of experiments, including biochemical assays, RT‒PCR, and western blotting, were conducted to investigate the anti-inflammatory and antioxidative effects of PREO.ResultsPREO treatment significantly (p < 0.05) alleviated inflammatory and oxidative biomarkers such as NO, ROS, and MDA and preserved SOD and CAT activities. GC‒MS analysis revealed that PREO consists of 57 compounds, mainly monoterpenoids. Network pharmacology revealed that citronellol, farnesol, ethyl octanoate, geranyl acetate, and methyl eugenol were active components interacting with several inflammatory pathway proteins. By measuring the gene and protein expression of possible targets by qRT‒PCR and western blotting, PREO anti-inflammatory responses in LPS-treated RAW 264.7 cells might be associated with the regulation of NF-κB signaling. Molecular docking showed that PREO components can interact with different proteins involved in the NF-κB pathway.ConclusionThe integrated study of molecular analysis and network pharmacology suggested that PREO might be a potential anti-inflammatory agent to treat inflammation and oxidative stress.

Network pharmacology and in-silico studies for molecular mechanisms of analgesic, anti-inflammatory, and anti-arthritic effects of phytoconstituents of Withania somnifera (L.) Dunal

Epilepsy is a common neurological disorder, and the exploration of potential therapeutic drugs for its treatment is still ongoing. Vitamin D has emerged as a promising treatment due to its potential neuroprotective effects and anti-epileptic properties. This study aimed to investigate the effects of vitamin D on epilepsy and neuroinflammation in juvenile mice using network pharmacology and molecular docking, with a focus on the mammalian target of rapamycin (mTOR) signaling pathway. Experimental mouse models of epilepsy were established through intraperitoneal injection of pilocarpine, and in vitro injury models of hippocampal neurons were induced by glutamate (Glu) stimulation. The anti-epileptic effects of vitamin D were evaluated both in vivo and in vitro. Network pharmacology and molecular docking analysis were used to identify potential targets and regulatory pathways of vitamin D in epilepsy. The involvement of the mTOR signaling pathway in the regulation of mouse epilepsy by vitamin D was validated using rapamycin (RAPA). The levels of inflammatory cytokines (TNF-α, IL-1β, and IL-6) were assessed by enzyme-linked immunosorbent assay (ELISA). Gene and protein expressions were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, respectively. The terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL) staining was used to analyze the apoptosis of hippocampal neurons. In in vivo experiments, vitamin D reduced the Racine scores of epileptic mice, prolonged the latency of epilepsy, and inhibited the production of TNF-α, IL-1β, and IL-6 in the hippocampus. Furthermore, network pharmacology analysis identified RAF1 as a potential target of vitamin D in epilepsy, which was further confirmed by molecular docking analysis. Additionally, the mTOR signaling pathway was found to be involved in the regulation of mouse epilepsy by vitamin D. In in vitro experiments, Glu stimulation upregulated the expressions of RAF1 and LC3II/LC3I, inhibited mTOR phosphorylation, and induced neuronal apoptosis. Mechanistically, vitamin D activated the mTOR signaling pathway and alleviated mouse epilepsy via RAF1, while the use of the pathway inhibitor RAPA reversed this effect. Vitamin D alleviated epilepsy symptoms and neuroinflammation in juvenile mice by activating the mTOR signaling pathway via RAF1. These findings provided new insights into the molecular mechanisms underlying the anti-epileptic effects of vitamin D and further supported its use as an adjunctive therapy for existing anti-epileptic drugs.

Banxia Xiexin decoction (BXD) is a traditional prescription widely used to treat gastrointestinal conditions, including gastric cancer. Through network pharmacology, bioinformatics, and molecular docking analysis, this study aimed to investigate the potential mechanism of the antigastric cancer effect of BXD and pave the way for future research. The network pharmacology analysis used BXD index components to improve reliability and validity. Prognosis-related genes identified through Lasso and Cox regression analysis were considered potential BXD core targets for gastric cancer. Functional enrichment analysis was conducted to uncover the potential mechanism of action of BXD in gastric cancer. In addition, molecular docking of the index components of BXD and the core targets was used to validate the results. The present study obtained six index components of BXD and 155 corresponding antigastric cancer targets. ANXA5, CYP19A1, FGF1, and F2 in the prognostic signature model were identified as core targets of the index components of BXD. Protein-protein interaction networks and functional enrichment analysis indicated that proteoglycans in cancer, PI3K-Akt, and other pathways were involved. According to molecular docking results, six index components showed good-to-strong binding affinities to the core targets. The results indicated that the index components of BXD act on multiple pathways and targets of gastric cancer. Our study paved the way for further investigation of the antigastric cancer activity and mechanisms of BXD.

This study investigated the chemical composition and anti-inflammatory effects of essential oils extracted from Citrus aurantium flower, Citrus sinensis, Brazilian Citrus sinensis, Citrus limon, Citrus bergamia, and Citri Reticulatae Pericarpium using steam distillation and gas chromatography-mass spectrometry (GC-MS). Their anti-inflammatory activities were assessed in LPS-stimulated RAW 264.7 cells. Among them, Citri Reticulatae Pericarpium essential oil (CRPEO) exhibited the most potent anti-inflammatory effects, with D-Limonene (76.51%), α-Pinene (2.68%), and Linalool (2.11%) as its primary constituents. The CCK-8 assay showed that the essential oil exhibited no cytotoxicity on HaCaT cells at a concentration of 50 μg/mL. CRPEO significantly preserved cell viability and reduced the production of pro-inflammatory mediators, including tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-1β, and nitric oxide (NO). Gene expression analysis via RT-qPCR further confirmed the downregulation of TNF-α, IL-6, IL-1β, and inducible nitric oxide synthase (iNOS) at the mRNA level. Network pharmacology and molecular docking studies were employed to identify α-Bulnesene as a key bioactive component of CRPEO and revealed that its principal target is the NLR Family Pyrin Domain-Containing 3 (NLRP3) inflammasome. These findings highlight the strong anti-inflammatory potential of CRPEO and suggest its promising therapeutic application for inflammation-related conditions.

Essential oil from Ligusticum chuanxiong Hort. Alleviates lipopolysaccharide-induced neuroinflammation: Integrating network pharmacology and molecular mechanism evaluation

BackgroundHepatic ischemia-reperfusion injury (HIRI) is a major complication in clinical situations such as liver transplant, hypovolemic shock, and severe trauma, critically affecting patient prognosis. Therapeutic interventions targeting HIRI are limited....

Effects of Jowiseungki-tang on high fat diet-induced obesity in mice and functional analysis on network pharmacology and metabolomics analysis

BackgroundThis study aimed to undertake a network pharmacology analysis to identify the active compounds of the herbal extract Christina Loosestrife, or Lysimachia Christinae (Jin Qian Cao), in the treatment of nephrolithiasis.Material/MethodsThe active components of Christina Loosestrife were identified from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database and analysis platform and the online Taiwan TCM database. The potentially active compounds were screened based on their parenteral bioavailability identified from the TCMSP database. The PharmMapper integrated pharmacophore matching platform was used for target identification of active compounds in nephrolithiasis. The identified active compounds were validated by molecular docking using the systemsDock network pharmacology website. Biological functions and pathway outcomes of effective targets were analyzed using the Metascape gene annotation resource. The results were used to construct the pharmacological networks, which were visualized and integrated using Cytoscape software.ResultsThere were 16 active compounds of Christina Loosestrife and 11 nephrolithiasis-associated targets that were obtained. Functional enrichment analysis showed that Christina Loosestrife might exert its therapeutic effects by regulating pathways that included purine salvage, interleukin-4 (IL-4) and IL-13 signaling, and neutrophil degranulation.ConclusionsNetwork pharmacology analysis of the herbal extract, Christina Loosestrife, identified multiple active compounds, targets, and pathways involved in the effects on nephrolithiasis.

Altered metabolic profiles and targets relevant to the protective effect of acteoside on diabetic nephropathy in db/db mice based on metabolomics and network pharmacology studies

This work aims to combine network pharmacology and metabolomics to explore the mechanism of action of dioscin on hyperuricemia (HUA). The preventative impact of dioscin on HUA and its putative mechanism were examined using network pharmacological analysis and metabonomics. Network pharmacology study further pointed out the potential targets of dioscin after a review of the relevant biomarker pathways discovered by metabolomic analysis. Molecular docking was then used to examine how the active chemicals interacted with the target proteins. The therapeutic effect of dioscin on HUA was shown to be mediated by 13 potentially important metabolites as a result of metabonomic research. Most of these metabolites are regulated after dioscin therapy to help patients recover. Based on network pharmacology, we identified 10 central genes, which is partly in agreement with metabolomics data. Using metabolomics and network pharmacology, this study investigated the primary targets and mechanisms of dioscin in the treatment of HUA. It is advantageous that dioscin has been developed as an additional drug for the treatment of HUA.

Research indicated that berberine (BBR) plays a protective role in modulating Alzheimer’s disease (AD). This study aimed to explore the target genes of BBR associated with AD therapy using a network pharmacology study. Through network pharmacology analysis, two main potential target genes, β-amyloid precursor protein (APP) and peroxisome proliferator-activated receptor gamma (PPARG), of BBR for AD therapy were screened out. Further experiments demonstrated that BV2 and C8-D1A treated with BBR were decreased in the mRNA and protein expression of APP and presenilin 1 while PPARG was increased with a reduction in the NF-κB pathway. A similar result was shown in vivo. Through a network pharmacology study, this study supported that BBR played a protective role in the AD mice model via blocking APP processing and amyloid plaque formation. It also promotes PPARG expression to blockage of NF-κB pathway-mediated inflammatory response and neuroinflammation.

Quality markers of Baizhu dispensing granules based on multi-component qualitative and quantitative analysis combined with network pharmacology and chemometric analysis

The aim of this study is to elucidate the pharmacological mechanism underlying the effects of Ginseng Radix et Rhizoma (ginseng) in heart failure (HF), providing a theoretical foundation for its clinical application. The potential mechanism of ginseng in the context of HF was investigated using systems pharmacology that combined network pharmacology, Gene Expression Omnibus (GEO) analysis, molecular docking, and experimental verification. Network pharmacology was employed to identify drug-disease targets. Core gene targets were subsequently subjected to enrichment analysis by integrating network pharmacology with GEO. Molecular docking was utilized to predict the binding affinities between identified targets and ginseng compounds. Furthermore, the therapeutic efficacy of ginseng was validated in an isoproterenol (ISO)-induced rat model of HF. The modulation of key signaling pathways by ginseng was confirmed through Western blot analysis. A total of 154 potential targets of ginseng in the treatment of HF were identified through network pharmacology analysis. The analysis of GSE71613 revealed that the PI3K-Akt pathway, reactive oxygen species, oxidative phosphorylation, MAPK signaling, and Ras signaling pathways are predominantly associated with patients with HF. By integrating the findings from network pharmacology and GEO analysis, ginsenoside Rg1 and ginsenoside Rb3 were identified as the potential components in ginseng, while FN1 and PRKAA2 were recognized as key targets involved in the PI3K-AKT and AMPK pathways, respectively. Molecular docking analysis revealed a strong affinity between the potential components and the identified core targets. In vivo experiments indicated that the extract of ginseng (EPG) significantly ameliorated ISO-induced cardiac dysfunction by improving cardiac parameters such as cardiac left ventricular internal systolic diameter, left ventricular end-diastolic volume, left ventricular end systolic volume, and left ventricular ejection fraction, while also reducing malondialdehyde production. In addition, EPG was found to enhance superoxide dismutase activity and ATP levels, while concurrently reducing the levels of interleukin (IL)-1β, IL-6, and TNF-α. The extract also reduced myocardial oxygen consumption, inflammatory cell infiltration, and the number of damaged myocardial fibers. Moreover, EPG was observed to upregulate the expression of p-PI3K, p-AKT, p-AMPK, and Bcl-2, while downregulating the expression of p-NFκB, TGF-β, and Bax. The therapeutic effects of ginseng on HF are primarily mediated through the PI3K-Akt and AMPK pathways. Ginsenoside Rg1 and ginsenoside Rb3 have been identified as potential therapeutic agents for HF.