Search Results (41)

Background: Acquired resistance to the third-generation EGFR tyrosine kinase inhibitor osimertinib, often mediated by EGFR triple mutations, poses a major clinical challenge in non-small cell lung cancer (NSCLC) treatment. Among these, some rare mutations, such as L858R/T790M/L792H and L858R/T790M/G796R, create steric hindrance that directly interferes with osimertinib binding, yet effective targeted therapeutic strategies for these specific mutations remain lacking. Cudratricusxanthone A (CTXA), a natural xanthone derivative isolated from Cudrania tricuspidata Bur., has demonstrated various pharmacological activities, but its effects against EGFR triple-mutant NSCLC have not been systematically investigated. Methods: Stable Ba/F3 and NIH/3T3 cell lines expressing EGFR L858R/T790M/L792H or L858R/T790M/G796R triple mutations were generated via electroporation. The antiproliferative effects of CTXA were evaluated by MTT/MTS assays, colony formation, and wound healing assays. Cell cycle distribution and apoptosis were analyzed by flow cytometry. Protein expression of EGFR signaling pathway components (p-EGFR, p-ERK, p-AKT, p-STAT3) and cell cycle regulators (Cyclin D1, CDK4) were examined by Western blotting. Molecular docking and 200 ns molecular dynamics simulations were performed to investigate the stability and binding modes of CTXA to the mutant EGFR kinase domains. Results: The successfully established triple-mutant cell lines exhibited high EGFR expression, IL-3-independent growth, and significant resistance to osimertinib. CTXA inhibited the proliferation of all triple-mutant cell lines in a time- and concentration-dependent manner, with 48 h IC₅₀ values ranging from 0.362 to 2.488 μM. Mechanistically, CTXA suppressed EGFR autophosphorylation and downregulated downstream p-ERK, p-AKT, and p-STAT3. CTXA induced G1 phase cell cycle arrest by downregulating Cyclin D1 and CDK4, significantly promoted apoptosis, and inhibited cell migration. Molecular docking revealed that while osimertinib binding was blocked by steric hindrance from His-792 or Arg-796, CTXA adapted to the mutated ATP-binding pockets through multiple hydrogen bonds and extensive hydrophobic interactions. Molecular dynamics simulations confirmed the stable binding of CTXA to both mutant EGFR proteins over the 200 ns simulations. Conclusions: This study demonstrates for the first time that the natural compound CTXA possesses antitumor efficacy against EGFR L858R/T790M/L792H and L858R/T790M/G796R mutants by regulating EGFR-ERK/AKT/STAT3 signaling. Our findings position CTXA as a promising lead compound for tackling this challenging form of acquired resistance and highlight the value of natural products in multi-target antitumor drug discovery. Full article

This study explored the efficacy, residue dynamics, and dietary risks of sulfoxaflor and flupyradifurone in Cudrania tricuspidata. Following two applications, residue levels of sulfoxaflor and flupyradifurone decreased from 0.254 to 0.012 mg/kg and 0.732 to 0.016 mg/kg, respectively, over a period of 22 days. The half-lives (t_1/2) in fruits and leaves ranged from 7.0 to 13.6 days. LC-MS/MS analysis showed recovery rates of 79.8–94.9% and RSD < 8.5%. Both pesticides effectively controlled hemipteran pests, reducing aphid and spotted lanternfly populations by >90%. Acute and chronic dietary risk assessments indicated acute hazard index (aHI) and chronic hazard quotient (HQ) values remarkably < 1, suggesting a negligible health risk. According to these results, sulfoxaflor and flupyradifurone have recently been registered as pesticides for C. tricuspidata against hemipteran pests, with a recommended pre-harvest interval of 7 days, as projected residue levels (0.078–5.213 mg/kg) were below established maximum residue limits (MRLs). These findings indicate a low dietary risk associated with sulfoxaflor and flupyradifurone in C. tricuspidata when applied according to the evaluated application rates and pre-harvest interval. Full article

A high-performance liquid chromatography (HPLC) method using a diode array detector (DAD) was developed and validated for the simultaneous quantification of chlorogenic acid, rutin, and isoquercitrin, which are key bioactive compounds in Cudrania tricuspidata leaves. The method demonstrated excellent specificity, precision, and accuracy in accordance with the guidelines of the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH). Calibration curves showed outstanding linearity (r² > 0.99), with recovery rates of 101.63%, 101.81%, and 102.18% for chlorogenic acid, rutin, and isoquercitrin, respectively. The limits of detection (LOD) were 0.286, 0.411, and 0.201 μg/mL, and the limits of quantification (LOQ) were 1.246, 0.866, and 0.608 μg/mL for chlorogenic acid, rutin, and isoquercitrin, respectively. Additionally, response surface methodology (RSM) based on a Box–Behnken design was employed to optimize the extraction conditions of the three marker compounds. The second-order regression models showed high coefficients of determination (r²) and significant ANOVA results (p < 0.05). RSM analysis revealed that extraction temperature and ethanol concentration exerted the most significant effects on the extraction yields, while extraction time played a supportive role. The optimal conditions (70 °C, 40% ethanol, 120 min) significantly enhanced compound recovery while reducing solvent and energy consumption, thereby contributing to the development of efficient and sustainable extraction processes. Collectively, the validated HPLC-DAD method and the optimized extraction strategy developed in this study provide a reliable framework for the quality standardization and industrial application of C. tricuspidata leaf extracts in functional food, cosmetic, and pharmaceutical products. Full article

This study evaluated the therapeutic potential of 70% ethanol extracts from six medicinal plants (Chenopodium album, Cassia tora, Cudrania tricuspidata, Dioscorea polystachya, Lonicera japonica, Solidago virgaurea subsp. gigantea) through their antibacterial, antioxidant, cytotoxic, and immunomodulatory activities, targeting applications in aquaculture. All extracts exhibited potent antibacterial activity (MIC ≤ 10 μg/mL) against Aeromonas spp. and Photobacterium damselae subsp. damselae, but limited efficacy against Streptococcus parauberis. C. tricuspidata (CTR) and C. tora (CTO) demonstrated superior antioxidant activity (IC₅₀ = 1292 μg/mL and IC₅₀ = 227 μg/mL, respectively), correlating with high polyphenol content (1498 and 1409 mg GAE/g). CTR displayed significant concentration-dependent cytotoxicity (IC₅₀ = 904.2 μg/mL), while C. album (CA) promoted cell proliferation (132.3% viability). In LPS-stimulated CHSE-214 cells, D. polystachya (DP) induced the highest IL-8 expression (207-fold), followed by Chenopodium album (CA) (194-fold IL-8, 49-fold TNF-α) and CTR (245-fold RIPK2), activating NF-κB, MAPK, and NOD-like receptor pathways critical for teleost immunity. Lonicera japonica (LJ) suppressed TNF-α (0.4-fold) and IRF1 (0.3-fold), indicating anti-inflammatory potential, while S. virgaurea subsp. gigantea (SV) showed biphasic TNF-α modulation (79-fold at 10 μg/mL, 5-fold at 100 μg/mL). These diverse bioactivities, particularly the robust immunomodulatory effects, highlight the promise of these extracts as natural therapeutic agents for fish health management in aquaculture. Full article

The purpose of this study was to evaluate the gene expression pattern of the caries-inhibiting effect of Cudrania tricuspidata (C. tricuspidata) extract on cariogenic bacteria Streptococcus mutans (S. mutans). We examined bacterial growth, tooth surface attachment, biofilm formation, acid production, free calcium release, and toxicity gene expression. The major components of the extract were investigated by UPLC-Q-TOF-MS analysis. Exposure to C. tricuspidata inhibited bacterial growth and attachment at concentrations of ≥15 μg/mL. Inhibition effects on biofilm formation, acid production, and free calcium release due to acid production were observed at concentrations ≥ 30 μg/mL. S. mutans virulence gene expression analysis showed that it inhibited the expression of gbpB and spaP, which mediate bacterial attachment to the tooth surface, and that of genes contributing to biofilm formation (gtfB, gtfC, and gtfD) and acid resistance (brpA and relA), and regulation (vicR). Analysis using UPLC–Q–TOF–MS/MS showed that the main component was phenylpropanoids. These results suggest that C. tricuspidata may inhibit the cariogenic properties associated with the expression of caries-related genes in S. mutans. Full article

Microglia play a crucial role as immune cells responsible for the brain’s defense mechanisms. Similar to the actions of macrophages in the body, microglial cells elicit an inflammatory immune response in the brain. Recent papers highlight activated microglial cells as pivotal contributors to inflammatory responses in the brain, leading to damage to nerve tissue and the onset of Alzheimer’s disease (AD). In the brains of AD patients, elevated levels of inflammatory cytokines such as interleukin-6 (IL-6) and oxidized cholesterol metabolites (oxysterols) are observed. These factors are closely associated with inflammatory diseases in the brain. 4′-O-Methylalpinum isoflavone (mAI), derived from Cudrania tricuspidata fruit, possesses antioxidant, neuroprotective, and anti-inflammatory properties. Consequently, this study examined the effect of mAI on the expression of IL-6, a major inflammatory cytokine. The HMC3 microglial cell line was treated with oxysterols to assess the effectiveness of mAI in mitigating this inflammatory response. The results indicated that mAI inhibited the gene expression and protein secretion of IL-6 induced by 25-hydroxycholesterol (25OHChol) and 27-hydroxycholesterol (27OHChol). Furthermore, the expression of MHC class II, a marker for microglial activation, was reduced to baseline levels. These findings suggest that mAI may serve as a viable option for suppressing and treating brain inflammatory diseases induced by cholesterol oxidation products. This is achieved by curtailing the expression of the inflammatory cytokine resulting from the activation of microglial cells by immuno-oxysterol. Full article

We investigated the antioxidant activity and neuroprotective effects of an enzymatic hydrolyzate from silkworms fed the leaves of Cudrania tricuspidata (ESLC) produced by enzymatic treatment. The proteins in silkworms fed the leaves of Cudrania tricuspidata (SLC) were effectively hydrolyzed using enzymatic treatment. The total polyphenols of ESLC were significantly higher, and the superoxide dismutase-like activity and 2,2′-azino-bis (3-thylbenzothiazoline-6-sulfonicacid) diammonium salt radical scavenging capacity of ESLC were significantly increased compared to the SLC group. We evaluated the properties of ESLC to protect SH-SY5Y cells from H₂O₂-induced oxidative stress. The viability rate of SH-SY5Y neuroblastoma cells was significantly restored when treated with ESLC at a concentration of 100 μg/mL or 250 μg/mL. Furthermore, the mitogen-activated protein kinase (MAPK) pathway was investigated, and ESLC significantly inhibited the phosphorylation of c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p38. Therefore, ESLC can potentially be used as an antioxidant. The polyphenol content increases owing to hydrolysis through enzymatic treatment, which increases the antioxidant effect and inhibits the phosphorylation of JNK, ERK, and p38 by activating the MAPK pathway, which inhibits cell death from oxidative stress and exerts cytoprotective effects. Full article

This study aimed to isolate the proteolytic fraction from the silkworm thorn fruit (Cudrania tricuspidata) through ethanol precipitation at different ratios, and to determine its proteolytic activity and optimal activity conditions. Furthermore, the hydrolysis characteristics and antioxidant activity of soy protein isolate (SPI) and whey protein concentrate (WPC) hydrolyzates obtained through the enzymatic hydrolysis of freeze-dried silkworm thorn fruit powder (SF) were evaluated. For isolation and partial purification of proteolytic fraction, the water-solubilized fraction of the silkworm thorn fruit was purified through ethanol precipitation at four different ratios of 1:1, 1:2, 1:4, and 1:6 (v/v). The protein recovery rate, caseinolytic activity, protein pattern, and optimal activity (pH, temperature, and inhibitors) of fractional ethanol precipitate obtained from the silkworm thorn fruit (ESF) were evaluated. The proteolytic fraction obtained from silkworm thorn fruit exhibited a major protein band around 65–70 kDa and showed the highest proteolytic activity at a 1:4 ratio of ethanol precipitation (p < 0.05). The optimal activity of the measured enzyme fraction was determined to be at pH 9.0 and 50 °C, and the proteolytic activity of ESF was almost inhibited by phenyl methyl sulphonyl fluoride (PMSF, 2 mM), a serine protease inhibitor. Compared to Alcalase and papain, extensively used as commercial enzymes, the silkworm thorn fruit powder was less effective in hydrolyzing SPI and WPC. Nevertheless, SPI and WPC hydrolyzates mediated with silkworm thorn fruit powder showed even better antioxidant activities than those mediated with Alcalase and papain. Thus, our results show the potential application of silkworm thorn fruit as a novel source of plant protease for producing human-grade protein hydrolyzates. Full article

Neuroinflammation activated by microglia affects inflammatory pain development. This study aimed to explore the anti-inflammatory properties and mechanisms of 1,6,7-trihydroxy-2-(1,1-dimethyl-2-propenyl)-3-methoxyxanthone (THMX) from Cudrania tricuspidata in microglia activation-mediated inflammatory pain. In RAW 264.7 and BV2 cells, THMX has been shown to reduce lipopolysaccharide (LPS)-induced inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and pro-inflammatory mediators and cytokines, including nitric oxide (NO), prostaglandin (PG) E2, interleukin (IL)-6, and tumor necrosis factor alpha (TNF-α). THMX also decreased LPS-induced phosphorylation of mitogen-activated protein kinase (MAPK) and the activation of p65 nuclear factor kappa B (NF-κB). Interestingly, THMX also activated heme oxygenase (HO)-1 expression. These findings suggest that THMX is a promising biologically active compound against inflammation through preventing MAPKs and NF-ĸB and activating HO-1 signaling pathways. Full article

This study investigated the memory-improving effects and mechanisms of action of hydrolysate from silkworm fed Cudrania tricuspidata leaves (HSCT) in rats with scopolamine-induced memory impairment. Thirty-two rats were categorized into 4 groups and the experiment was conducted for 6 weeks. The experimental diet groups are as follows: control, control diet; scopolamine, control diet + scopolamine (0.7 mg/kg); HSCT, HSCT diet + scopolamine (0.7 mg/kg); and donepezil, control diet + scopolamine (0.7 mg/kg) + donepezil (1.0 mg/kg). Except for the control group, the experimental groups received scopolamine 0.7 mg/kg intraperitoneally to induce decline in memory. Animal behavioral tests such as the Barnes maze, novel object recognition test, and Y-maze were performed to investigate the effects of HSCT on memory improvement. In addition, brain acetylcholine (ACh) concentration and acetylcholinesterase (AChE) activity were assessed to investigate the mechanism of memory improvement. Results of the animal behavior test suggested that the rats administered HSCT displayed improved memory behavior compared to those administered scopolamine (p < 0.05). The concentration of Ach was increased in the HSCT group compared to the scopolamine group (p < 0.05), and the activity of AChE was significantly reduced (p < 0.05). These results suggest that HSCT effectively improves memory by reducing AChE activity and increasing ACh content in the brains of rats with scopolamine-induced memory impairment. As a natural compound, HSCT has the potential to be used as a material to improve memory. Full article

The study investigated the antioxidant effects of roasted Cudrania tricuspidata (C. tricuspidata) fruits by comparing them with unroasted C. tricuspidata fruits. The results showed that the roasted C. tricuspidata fruits (150 °C, 120 min) exhibited significantly higher antioxidant activity, especially in terms of anti-inflammatory effects, than the unroasted fruits. Interestingly, there is a high correlation between the color of the roasted fruit and the antioxidant activity. Heating disrupts cells and deactivates endogenous oxidative enzymes, leading to an increase in flavonoid content. Moreover, heat treatment may also interfere with plant metabolism, thereby influencing flavonoid content. Moreover, an HPLC analysis of roasted fruits in our study showed that the increase in antioxidant activity was attributed to the increase in flavan-3-ols and phenolic acids in the roasted C. tricuspidata fruits. To the best of our knowledge, this is the first time the antioxidant activity and anti-inflammation of roasted C. tricuspidata fruits was studied. The study concluded that roasted C. tricuspidata fruits could be a valuable natural source of antioxidants for various food and medicinal applications. Full article

Cudrania tricuspidata is a traditional medicinal herb in East Asia. The compounds of plants vary depending on environmental factors, such as soil, temperature, drainage, and so on. However, few to no studies have been done on the correlation among environment, growth, and compounds in C. tricuspidata. Thus, we aimed to investigate their relationship. Samples of C. tricuspidata fruit and cultivation soil were collected from 28 cultivation sites in October 2021. Six growth characteristics, eleven soil physicochemical properties, seven meteorological data points, and three active compounds were investigated in this study. We developed and validated an optimized method for quantifying active compounds using UPLC and performed correlation analysis of the environment, growth characteristics, and active compounds. The UPLC-UV method for determining active compounds was validated by measuring the linearity, LOD, LOQ, precision, and accuracy using UPLC. The LOD and LOQ were 0.01–0.03 µg/mL and 0.04–0.09 µg/mL, respectively. The precision was acceptable with RSD% values less than 2%. The recoveries ranged from 97.25 to 104.98% with RSD values <2%, within the acceptable limits. The active compounds were negatively correlated with the size of the fruit, and the growth characteristics were negatively correlated with some environmental factors. The results of this study can be used as basic data for the standard cultural practices and quality control of C. tricuspidata fruits. Full article

Alpinumisoflavone is a natural prenylated isoflavonoid extracted from the raw fruit of Cudrania tricuspidata. Several studies have reported the beneficial characteristics of alpinumisoflavone, such as its antioxidant, anti-inflammation, anti-bacterial, osteoprotective, and neuroprotective effects. Alpinumisoflavone also has anti-cancer effects on thyroid, renal, and ovarian cancers, but its therapeutic effects on hepatocellular carcinoma (HCC) have not yet been demonstrated. We investigated the anti-cancer effects of alpinumisoflavone on HCC using human liver cancer cell lines, Hep3B and Huh7. Our results confirmed that alpinumisoflavone inhibited viability and regulated the MAPK/PI3K pathway in Hep3B and Huh7 cells. We also verified that alpinumisoflavone can depolarize the mitochondrial membrane potential and suppress the mitochondrial respiration in HCC cells. Moreover, we confirmed the dysregulation of the mitochondrial complexes I, III, and V involving mitochondrial oxidative phosphorylation at the mRNA level and the accumulation of calcium ions in the mitochondrial matrix. Lastly, we demonstrated that alpinumisoflavone induced mitochondria-mediated apoptosis via regulation of the Bcl-xL and BAK proteins. This study elucidates the anti-cancer effects of alpinumisoflavone on HCC. Full article

6,8-Diprenylorobol is a flavonoid compound extracted from Cudrania tricuspidata. It has various biological functions, such as inhibiting melanin synthesis and inducting cell death in cancerous cells. In addition, Cudrania tricuspidata is known to be effective in female diseases, and previous studies have shown anticancer effects in cervical cancer, a female reproductive disease. Outside of that, Cudrania tricuspidata has various physiological effects. However, the effect of 6,8-diprenylorobol is not well known in other benign and chronic diseases, even in endometriosis, which commonly arises in the female reproductive tract. In the present study, we determined the inhibitory effects of 6,8-diprenylorobol on the growth of endometriosis VK2/E6E7 and End1/E6E7 cells. Results indicated that 6,8-diprenylorobol suppressed cellular proliferation and increased the disruption of the cell cycle, mitochondrial membrane potential (MMP), generation of reactive oxygen species, and Ca²⁺ homeostasis in both endometriosis cells. However, the proliferation of normal stromal cells isolated from endometrial tissue was not altered by 6,8-diprenylorobol. The change in Ca²⁺ levels was estimated in fluo-4- or rhod-2-stained VK2/E6E7 and End1/E6E7 cells after the treatment of the intracellular calcium regulators 2-aminoethoxydiphenyl borate (2-APB) and ruthenium red (RUR) with 6,8-diprenylorobol. A combination of 6,8-diprenylorobol with each regulator decreased the calcium accumulation in endometriosis cells. Furthermore, Western blot analysis indicated that 6,8-diprenylorobol inactivated AKT pathways, whereas it activated P38 MAPK pathways. In addition, 6,8-diprenylorobol decreased mitochondrial respiration, leading to the reduction in ATP production in VK2/E6E7 and End1/E6E7 cells. Collectively, our results suggested that 6,8-diprenylorobol might be a potential therapeutic agent or adjuvant therapy for the management of endometriosis. Full article

Monoamine serotonin is a major neurotransmitter that acts on a wide range of central nervous system and peripheral nervous system functions and is known to have a role in various processes. Recently, it has been found that 5-HT is involved in cognitive and memory functions through interaction with cholinergic pathways. The natural flavonoid kaempferol (KAE) extracted from Cudrania tricuspidata is a secondary metabolite of the plant. Recently studies have confirmed that KAE possesses a neuroprotective effect because of its strong antioxidant activity. It has been confirmed that KAE is involved in the serotonergic pathway through an in vivo test. However, these results need to be confirmed at the molecular level, because the exact mechanism that is involved in such effects of KAE has not yet been elucidated. Therefore, the objective of this study is to confirm the interaction of KAE with 5-HT_3A through electrophysiological studies at the molecular level using KAE extracted from Cudrania tricuspidata. This study confirmed the interaction between 5-HT_3A and KAE at the molecular level. KAE inhibited 5-HT_3A receptors in a concentration-dependent and voltage-independent manner. Site-directed mutagenesis and molecular-docking studies confirmed that the binding sites D177 and F199 are the major binding sites of human 5-HT_3A receptors of KAE. Full article