Search Results (43)

Chronic atrophic gastritis (CAG) is a precancerous gastric condition with limited therapeutic interventions, and the mechanisms underlying the benefits of Coptis chinensis Franch. (CCF) remain insufficiently defined. This study employed an integrated computational strategy to clarify the molecular basis of CCF activity against CAG. Network pharmacology was used to identify potential targets of the major CCF constituents berberine, coptisine, and palmatine, followed by molecular docking, machine learning-based IC₅₀ prediction, and molecular dynamics simulations. Fifty-eight overlapping targets between CCF compounds and CAG-related genes were identified, highlighting SRC, STAT3, MAPK1, and NFKB1 as central nodes enriched in inflammatory and immune pathways, including TNF and MAPK signaling. Docking analyses revealed strong interactions between all three compounds and SRC kinase, and machine learning models predicted IC₅₀ values in the low micromolar range (1.38–1.82 μM). Molecular dynamics simulations further suggest that berberine may stabilize the crucial regulatory regions of SRC, specifically the activation loop. It is hypothesized that this stabilization maintains the inactive conformation of the kinase domain and potentially shields Tyr416 from phosphorylation, thus potentially influencing kinase activation. These findings suggest that CCF may modulate key inflammatory and immune pathways implicated in CAG progression, with SRC emerging as a central node for further investigation. Full article

Background: Using Huanglian Jiedu decoction (HJD) as a model, this study systematically compares the traditionally prepared combined decoction with mixtures of separately decocted components, focusing on differences in chemical composition, material properties, and transcriptomic responses. Methods: To maintain consistency, both the combined HJD decoction and the single-herb mixture were prepared using a standardized method, and their chemical profiles were analyzed by HPLC and UPLC-MS/MS to identify constituent differences. Physical properties were examined through key parameter measurements and phase behavior analysis, and the integration of chemical and physical data identified the components driving the observed material changes. Transcriptome sequencing compared the two decoction types, highlighting differentially expressed genes and the major regulatory pathways involved. Results: HPLC analysis showed a clear redistribution of components between the two decoction methods, with the combined decoction containing higher levels of alkaloids such as coptisine chloride, epiberberine, palmatine chloride, jatrorrhizine hydrochloride, and phellodendrine chloride, while the single decoction mixture had higher levels of berberine hydrochloride and baicalin. In the combined decoction, the berberine hydrochloride content was 37.04 mg/g, and the baicalin content was 15.57 mg/g; in the single decoction, the berberine hydrochloride content was 41.15 mg/g; in the combined decoction, the baicalin content was 40.07 mg/g. UPLC-MS/MS analysis confirmed clear differences between the two decoctions, mainly in flavonoid and alkaloid compositions. The combined decoction contains 110 flavonoid compounds and 67 alkaloid compounds, while the single decoction contains 100 flavonoid compounds and 80 alkaloid compounds. Physical measurements showed that the combined decoction had higher total dissolved solids, conductivity, and salinity, while the single decoction had higher resistivity. The combined decoction had a TDS of 2480 mg/L, σ of 4.95 ms/cm, S of 0.26%, and ρ of 202 Ω·cm; the single decoction had a TDS of 1190 mg/L, σ of 2.37 ms/cm, S of 0.12%, and ρ of 419 Ω·cm. Phase separation analysis indicated that the combined decoction formed a stable nanoscale phase structure, whereas the single decoction remained unstable. Transcriptome sequencing at various concentrations revealed marked differences in gene expression between the two preparations, reflecting their distinct biological activities. Conclusions: Analysis showed clear differences in chemical composition, physical properties, and gene expression in the combined decoction. Taking Huanglian Jiedu decoction as a representative example, we systematically compared the specific differences between combined and single decoction methods, providing a reference basis for subsequent pharmacodynamic evaluation and clinical application. Full article

Coptis trifolia (threeleaf goldthread) offers a valuable comparative system for investigating the evolution and regulation of benzylisoquinoline alkaloid (BIA) synthesis. In this study, we analyzed the leaf and root transcriptomes of C. trifolia using both long-read and short-read RNA-Sequencing. We assembled 41,926 unigenes (≥500 bp) and identified 37 genes related to BIA biosynthesis, including two transcription factors, bHLH1 and WRKY1. The number of BIA genes identified in C. trifolia was comparable to that in other Coptis species. Transcriptome analysis revealed that most of these genes were more highly expressed in roots than leaves. Consistent with previous studies, C. trifolia contained a single (S)-stylopine synthase (SPS) gene homolog, potentially multifunctional for (S)-canadine synthase (CAS), (S)-cheilanthifoline synthase (CFS), and SPS. Transcriptome and untargeted metabolomic data indicated greater variation in root samples than leaf samples, although slightly more differentially expressed transcripts and metabolites were observed in leaves. Targeted metabolite profiling showed higher BIA accumulation in roots, with epiberberine being the most abundant, followed by coptisine, berberine, and columbamine. These results provide essential genomic resources for comparative analysis of the BIA pathway across Ranunculaceae, targeted gene function studies for metabolic bioengineering, and conservation strategies for C. trifolia, a member of an early-diverging clade within the genus with limited genetic resources. Full article

Excessive osteoclast activity in bone remodeling can lead to an imbalance between bone resorption and formation, a common occurrence in abnormal bone metabolic diseases. This research investigates the effect of Coptidis rhizoma water extract (CRW) on osteoclastogenesis provoked by RANKL in vitro and bone destruction mediated by ovariectomy (OVX) in vivo. CRW, prepared from dried Coptidis rhizoma (CR), was analyzed for its active compounds—coptisine and berberine—using HPLC analysis. CRW markedly decreased the size and number of TRAP-positive multinucleated cells (TRAP⁺ MNCs), suppressed F-actin ring formation, and diminished bone resorption in RANKL-treated cultures. In the early phase of differentiation, CRW suppressed the phosphorylation of MAPKs p38, JNK, and ERK, as well as NF-κB p65, Iκ-Bα, and Akt. CRW also down-regulated RANKL-mediated induction of c-Fos and NFATc1 and attenuated the activation of NFATc1- dependent genes, such as OSCAR, ATP6V0D2, ACP5 (TRAP), OC-STAMP, DC-STAMP, CTSK (cathepsin K), CALCR (calcitonin receptor), and MMP-9. In ovariectomized rats, micro-CT and histological analyses showed that CRW alleviated femoral bone destruction. These findings indicate that CRW restrains osteoclast differentiation and function and may have therapeutic potential for disorders driven by excessive osteoclast activity. Full article

Various metabolic modulators have been widely used in recent years to increase the accumulation of desired secondary metabolites in medicinal plants, although most studies to date have focused on in vitro systems. Although simpler and cheaper, their potential application in vivo is still limited. Therefore, the aim of this study was to compare the effect of three chemically different elicitors (150 mg/L chitosan lactate—ChL; 10 mg/L selenium as selenite—Se; 100 mg/L salicylic acid—SA) applied to the soil substrate on some aspects of the secondary metabolism and physiological responses of Chelidonium majus L. Using HPLC-DAD, six isoquinoline alkaloids were identified and quantified in shoot extracts. LC-ESI-TOF-MS analysis confirmed the molecular identity of all target alkaloids, supporting the identification. The strongest stimulatory effect on the accumulation of protopine, berberine, and allocryptopine was observed with the Se and SA treatment, whereas ChL was less effective. In turn, the dominant alkaloids (coptisine and chelidonine) remained unaffected. There was also an increase in total phenolic compounds, but not in soluble flavonols. The elicitor treatments caused an increase in the antioxidant activity of the plant extracts obtained. Regardless of the metabolic modulator type, the strongest effect was generally observed on days 7 and 10 after application. No visual signs of toxicity and no effect on shoot biomass were found, although some elicitor-induced changes in the oxidative status (increased H₂O₂ accumulation and enhanced lipid peroxidation) and free proline levels in leaves were observed. We suggest that Se or SA can be applied to C. majus grown in a controlled pot culture to obtain high-quality raw material and extracts with increased contents of valuable specialized metabolites and enhanced antioxidant capacity. Full article

Berberine, palmatine, and coptisine are bioactive alkaloids commonly found in medicinal plants, including Coptis chinensis Franch. (Ranunculaceae). To address the limitations of conventional volatile organic solvents, this study employed eco-friendly solvents—aqueous solutions of surfactants, carboxylic acids, and deep eutectic solvents—to extract these alkaloids. Among the solvents tested, lactic acid, malic acid, and pyruvic acid exhibited the highest extraction efficiencies. Optimal extraction conditions for ultrasound-assisted extraction were determined via response surface methodology. For lactic acid, optimal conditions included a concentration of 96% (w/w), a liquid-to-solid ratio of 30.0 mL/g, and a temperature of 60.0 °C, yielding 139.6 ± 0.2 mg/g of total alkaloids. Malic acid at 40.0% (w/w), 30.0 mL/g, and 80.0 °C produced 133.0 ± 0.5 mg/g, while pyruvic acid at 88.0% (w/w), 30.0 mL/g, and 75.0 °C resulted in 146.3 ± 0.4 mg/g. The recovery efficiencies of these alkaloids were further enhanced using macroporous resins. The XAD-8 and AB-8 resins achieved recovery rates of 80.11 ± 0.78% and 79.00 ± 1.06%, respectively, for lactic acid extracts. The LSA-40 resin yielded efficiencies of 95.58 ± 1.40% and 89.86 ± 0.90% for malic and pyruvic acid extracts, respectively. Notably, the combination of malic acid as an extraction solvent and the HPD-400 resin achieved an impressive alkaloid recovery yield of 79.52% from C. chinensis. This work represents the first reported application of this approach and highlights the potential of green solvents and macroporous resins for sustainable and efficient alkaloid extraction from C. chinensis. Full article

Irritable bowel syndrome (IBS) is a chronic disorder of the gastrointestinal tract. Its pathogenesis involves multiple factors, including visceral hypersensitivity and immune activation. NLRP3 inflammasome is part of the nucleotide-binding oligomerization domain-like receptor (NLR) family, a crucial component of the innate immune system. Preclinical studies have demonstrated that inhibiting NLRP3 reduces visceral sensitivity and IBS symptoms, like abdominal pain, and diarrhea, suggesting that targeting the NLRP3 might represent a novel therapeutic approach for IBS. This review aims to assess the NLRP3 inhibitors (tranilast, β-hydroxybutyrate, Chang-Kang-fang, paeoniflorin, coptisine, BAY 11-7082, and Bifidobacterium longum), highlighting the signaling pathways, and their potential role in IBS symptoms management was assessed. Although premature, knowledge of the action of synthetic small molecules, phytochemicals, organic compounds, and probiotics might make NLRP3 a new therapeutic target in the quiver of physicians’ therapeutic choices for IBS symptoms management. Full article

Coptisine (Cop) exerts a neuroprotective effect on central nervous system disease, particularly ischemic stroke. However, its protective mechanism is still unclear. This study aimed to investigate the protective effect of Cop on cerebral ischemia–reperfusion (IR) rats with a middle cerebral artery occlusion model by integrating a gas chromatography–mass spectrometry (GC–MS)-based metabolomics approach with biochemical assessment. Our results showed that Cop could improve neurobehavioral function and decrease the ischemia size in IR rats. In addition, Cop was found to decrease inflammatory mediators (e.g., prostaglandin D2 (PGD2) and tumor necrosis factor-α (TNF-α) and attenuate oxidative stress response (e.g., increase the superoxide dismutase (SOD) expression and decrease 8-iso-PGF2α level). Furthermore, the GC-MS-based cerebrospinal fluid (CSF) metabolomics analysis indicated that Cop influenced the level of glycine, 2,3,4-trihydroxybutyric acid, oleic acid, glycerol, and ribose during IR injury. Cop exhibited a good neuroprotective effect against cerebral IR injury and metabolic alterations, which might be mediated through its antioxidant and anti-inflammatory properties. Full article

The activation of innate antiviral immunity is a promising approach for combatting viral infections. In this study, we screened Chinese herbs that activated human immunity and identified coptisine as a potent inhibitor of the influenza virus with an EC₅₀ of 10.7 μM in MDCK cells. The time of an addition assay revealed that pre-treatment with coptisine was more effective at reducing viral replication than co-treatment or post-treatment. Our bulk RNA-sequencing data showed that coptisine upregulated the p21 signaling pathway in MDCK cells, which was responsible for its antiviral effects. Specifically, coptisine increased the expression of p21 and FOXO1 in a dose-dependent manner while leaving the MELK expression unchanged. Docking analysis revealed that coptisine likely inhibited MELK activity directly by forming hydrogen bonds with ASP-150 and GLU-87 in the catalytic pocket. These findings suggest that coptisine may be a promising antiviral agent that regulates the p21 signaling pathway to inhibit viral replication. Full article

Cancer therapy with clinically established anticancer drugs is frequently hampered by the development of drug resistance of tumors and severe side effects in normal organs and tissues. The demand for powerful, but less toxic, drugs is high. Phytochemicals represent an important reservoir for drug development and frequently exert less toxicity than synthetic drugs. Bioinformatics can accelerate and simplify the highly complex, time-consuming, and expensive drug development process. Here, we analyzed 375 phytochemicals using virtual screenings, molecular docking, and in silico toxicity predictions. Based on these in silico studies, six candidate compounds were further investigated in vitro. Resazurin assays were performed to determine the growth-inhibitory effects towards wild-type CCRF-CEM leukemia cells and their multidrug-resistant, P-glycoprotein (P-gp)-overexpressing subline, CEM/ADR5000. Flow cytometry was used to measure the potential to measure P-gp-mediated doxorubicin transport. Bidwillon A, neobavaisoflavone, coptisine, and z-guggulsterone all showed growth-inhibitory effects and moderate P-gp inhibition, whereas miltirone and chamazulene strongly inhibited tumor cell growth and strongly increased intracellular doxorubicin uptake. Bidwillon A and miltirone were selected for molecular docking to wildtype and mutated P-gp forms in closed and open conformations. The P-gp homology models harbored clinically relevant mutations, i.e., six single missense mutations (F336Y, A718C, Q725A, F728A, M949C, Y953C), three double mutations (Y310A-F728A; F343C-V982C; Y953A-F978A), or one quadruple mutation (Y307C-F728A-Y953A-F978A). The mutants did not show major differences in binding energies compared to wildtypes. Closed P-gp forms generally showed higher binding affinities than open ones. Closed conformations might stabilize the binding, thereby leading to higher binding affinities, while open conformations may favor the release of compounds into the extracellular space. In conclusion, this study described the capability of selected phytochemicals to overcome multidrug resistance. Full article

In cell-based regenerative medicine, induced pluripotent stem cells (iPSCs) generated from reprogrammed adult somatic cells have emerged as a useful cell source due to the lack of ethical concerns and the low risk of immune rejection. To address the risk of teratoma formation, which is a safety issue in iPSC-based cell therapy, it is essential to selectively remove undifferentiated iPSCs remaining in the iPSC-derived differentiated cell product prior to in vivo transplantation. In this study, we explored whether an ethanol extract of coptidis rhizoma (ECR) exhibited anti-teratoma activity and identified the active components involved in the selective elimination of undifferentiated iPSCs. Transcriptome analysis of iPSCs confirmed that cell death-related pathways were significantly altered by ECR treatment. Our results demonstrate that ECR effectively induced apoptotic cell death and DNA damage in iPSCs, and that reactive oxygen species generation, mitochondrial damage, caspase activation, and p53 activation were involved in ECR-mediated iPSC death. However, in iPSC-derived differentiated cells (iPSC-Diff), reduced cell viability and the DNA damage response were not observed after ECR treatment. We co-cultured iPSCs and iPSC-Diff and found that ECR treatment selectively removed iPSCs, whereas iPSC-Diff remained intact. Prior to in ovo implantation, ECR treatment of a mixed cell culture of iPSCs and iPSC-Diff significantly suppressed iPSC-derived teratoma formation. Among the main components of the ECR, berberine and coptisine showed selective cytotoxicity to iPSCs but not to iPSC-Diff. Together, these results indicate the usefulness of ECRs in preparing safe and effective iPSC-based therapeutic cell products with no risk of teratoma formation. Full article

The Scutellaria baicalensis—Coptis chinensis pair is an herbal combination used for the treatment of various heat-related diseases. During the extraction process, two herbs can mutually influence the extraction efficiency of the chemical constituents contained in each herb. The concentrations of five flavonoids from S. baicalensis and seven alkaloids from C. chinensis were compared in paired or single hot-water extracts at different temperatures (80, 90, and 100 °C) and extraction times (60, 90, and 120 min). Temperature- and time-dependent increases in marker compound concentrations were observed in both paired and single extracts, with the exception of baicalin, berberine, and coptisine in the paired extracts at 100 °C. However, the extractions of the compounds in the paired and single extracts were affected differently by the extraction conditions. Furthermore, the concentrations of most marker compounds in single extracts were 1.09–44.13 times those in paired extracts. The contents of baicalin, wogonoside, coptisine, and berberine, known to be easily aggregated by the flavonoid–alkaloid complex, were changed by 0.024–0.764-fold in the paired extract. The effect of extraction temperature and time on the formation of the flavonoid–alkaloid complex was not significant. The extraction efficiency of the flavonoids and alkaloids can be affected by the pair of S. baicalensis—C. chinensis, which is a primary factor in the chemical modification of two herb-containing herbal extracts. Full article

Scutellaria baicalensis is often used to treat breast cancer, but the molecular mechanism behind the action is unclear. In this study, network pharmacology, molecular docking, and molecular dynamics simulation are combined to reveal the most active compound in Scutellaria baicalensis and to explore the interaction between the compound molecule and the target protein in the treatment of breast cancer. In total, 25 active compounds and 91 targets were screened out, mainly enriched in lipids in atherosclerosis, the AGE–RAGE signal pathway of diabetes complications, human cytomegalovirus infection, Kaposi-sarcoma-associated herpesvirus infection, the IL-17 signaling pathway, small-cell lung cancer, measles, proteoglycans in cancer, human immunodeficiency virus 1 infection, and hepatitis B. Molecular docking shows that the two most active compounds, i.e., stigmasterol and coptisine, could bind well to the target AKT1. According to the MD simulations, the coptisine–AKT1 complex shows higher conformational stability and lower interaction energy than the stigmasterol–AKT1 complex. On the one hand, our study demonstrates that Scutellaria baicalensis has the characteristics of multicomponent and multitarget synergistic effects in the treatment of breast cancer. On the other hand, we suggest that the best effective compound is coptisine targeting AKT1, which can provide a theoretical basis for the further study of the drug-like active compounds and offer molecular mechanisms behind their roles in the treatment of breast cancer. Full article

As part of a project on fused medium-sized ring systems as potential drugs, we have previously demonstrated the usefulness of Density Functional Theory (DFT) to evaluate amine nitrogen-based transannular interactions across the central 10-membered ring in the bioactive dibenzazecine alkaloid, protopine. A range of related hypothetical systems have been investigated, together with transannular interactions involving ring-embedded imino or azo group nitrogens and atoms or groups (Y) across the ring. Electrostatic potential energies mapped onto electron density surfaces in the different ring conformations were evaluated in order to characterise these conformations. Unexpectedly, the presence of sp² hybridised nitrogen atoms in the medium-sized rings did not influence the conformations appreciably. The strength and type of the N^…Y interactions are determined primarily by the nature of Y. This is also the case when the substituent on the interacting nitrogen is varied from CH₃ (protopine) to H or OH. With Y = BOH, very strong interactions were observed in protopine analogues, as well as in rings incorporating imino or azo groups. Strong to moderate interactions were observed with Y = CS, CO and SO in all ring systems. Weaker interactions were observed with Y = S, O and weaker ones again with an sp³ hybridised carbon (Y = CH₂). The transannular interactions can influence conformational preferencing and shape and change electron distributions at key sites, which theoretically could modify properties of the molecules while providing new or enhanced sites for biological target interactions, such as the H or OH substituent. The prediction of new strong transannular interaction types such as with Y = BOH and CS should be helpful in informing priorities for synthesis and other experimental studies. Full article

Gandouling (GDL) Pill is a novel Traditional Chinese medicinal drug to treat Wilson’s disease in clinics. It is composed of six separate herbal medicines, including Rhei Radix ET Rhizoma, Coptidis Rhizoma, Salviae Miltiorrhizae Radix ET Rhizoma, Spatholobi Caulis, Curcumae Rhizoma, and Curcumae Longae Rhizoma. In this study, a strategy was proposed to investigate the chemical constituents and to quantify the potential bioactive components in GDL Pill. Firstly, the mass fragmentation behaviors of representative compounds were investigated, and, in total, 69 compounds were characterized in GDL Pill using full scan/dd-MS² scan mode by ultra-high-performance liquid chromatography (UPLC)/Q-Orbitrap mass spectrometry (MS). These compounds included 18 alkaloids, 18 ketones, 16 phenolic compounds, 11 organic acids, and 6 tanshinones. Seventeen of the compounds were unambiguously identified by comparison with reference standards. Secondly, the absorption components of GDL Pill in rat plasma were investigated by using target-Selected Ion Monitoring (t-SIM) scan mode built in Q-Orbitrap MS. A total of 18 components were detected, which were considered as potential bioactive components of GDL Pill. Thirdly, 10 major absorption components were simultaneously determined in six batches of samples by UPLC/diode array detector (DAD). The method was fully validated with respect to linearity, precision, repeatability, stability, and recovery. Alkaloids from Coptidis Rhizoma, such as coptisine (8), berberine (18), palmatine (19), were the most abundant bioactive compounds for GDL Pill that possess the potential be used as quality markers. The proposed strategy is practical and efficient for revealing the material basis of GDL Pill, and also provides a simple and accurate method for quality control. Full article