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Search Results (814)

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Keywords = ethyl acetate fraction

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20 pages, 2822 KiB  
Article
Nanoparticle Formulation Generated from DDGS and Its Anthraquinone Synthesis Elicitation in Rubia tinctorum Hairy Roots
by Gonzalo Galaburri, Yazmín R. Kalapuj, María Perassolo, Julián Rodríguez Talou, Patricio G. Márquez, Romina J. Glisoni, Antonia Infantes-Molina, Enrique Rodríguez-Castellón and Juan M. Lázaro-Martínez
Polymers 2025, 17(15), 2021; https://doi.org/10.3390/polym17152021 - 24 Jul 2025
Viewed by 299
Abstract
A nanoparticle formulation was generated from distiller dried grains with solubles (DDGS), and its effect on the production of anthraquinones (AQs) was evaluated on Rubia tinctorum hairy roots. The DDGS material was washed with water and ethyl acetate to remove mainly the soluble [...] Read more.
A nanoparticle formulation was generated from distiller dried grains with solubles (DDGS), and its effect on the production of anthraquinones (AQs) was evaluated on Rubia tinctorum hairy roots. The DDGS material was washed with water and ethyl acetate to remove mainly the soluble organic/inorganic molecules and reduce the fat content, respectively, followed by an alkaline treatment to remove the polysaccharides. The resulting alkaline solutions were then lyophilized and redispersed in deionized water to generate a monodispersed nanoparticulate formulation (DDGS-NP) with a hydrodynamic diameter and zeta potential of 227 ± 42 nm and −53 ± 7 mV, respectively. The formulation demonstrated good colloidal stability over time, and sterilized DDGS-NPs maintained comparable physicochemical properties. The nanoparticles were enriched in protein fractions, unsaturated fatty acids, and orthophosphate anion components from DDGS, as determined by solid-state Nuclear Magnetic Resonance (NMR), X-ray photoelectron spectroscopy (XPS), organic elemental analysis (OEA), and inductively coupled plasma optical emission spectrometry (ICP-OES) techniques. The DDGS-NPs were tested at different concentrations on Rubia tinctorum hairy roots, in comparison to or in combination with methyl jasmonate (MeJ), for their capacity to induce the production of AQs. All DDGS-NP concentrations increased the production of specific AQs to 7.7 (100 mg L−1), 7.8 (200 mg L−1), and 9.3 µmol/gFW (500 mg L−1), with an extracellular AQ accumulation of 18 µM for the highest DDGS-NP concentration, in comparison with the control hairy roots (~2 µM AQ). The plant growth was not affected at any of the tested nanoparticle concentrations. Interestingly, the combination of DDGS-NPs and MeJ resulted in the highest extracellular AQ accumulation in R. tinctorum root cultures. Full article
(This article belongs to the Section Polymer Composites and Nanocomposites)
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21 pages, 17488 KiB  
Article
Mechanistic Study on the Inhibitory Effect of Dandelion Extract on Breast Cancer Cell Proliferation and Its Induction of Apoptosis
by Weifeng Mou, Ping Zhang, Yu Cui, Doudou Yang, Guanjie Zhao, Haijun Xu, Dandan Zhang and Yinku Liang
Biology 2025, 14(8), 910; https://doi.org/10.3390/biology14080910 - 22 Jul 2025
Viewed by 771
Abstract
This study aimed to investigate the underlying mechanisms by which dandelion extract inhibits the proliferation of breast cancer MDA-MB-231 cells. Dandelion root and leaf extracts were prepared using a heat reflux method and subjected to solvent gradient extraction to obtain fractions with different [...] Read more.
This study aimed to investigate the underlying mechanisms by which dandelion extract inhibits the proliferation of breast cancer MDA-MB-231 cells. Dandelion root and leaf extracts were prepared using a heat reflux method and subjected to solvent gradient extraction to obtain fractions with different polarities. MTT assays revealed that the ethyl acetate fraction exhibited the strongest inhibitory effect on cell proliferation. LC-MS analysis identified 12 potential active compounds, including sesquiterpenes such as Isoalantolactone and Artemisinin, which showed significantly lower toxicity toward normal mammary epithelial MCF-10A cells compared to tumor cells (p < 0.01). Mechanistic studies demonstrated that the extract induced apoptosis in a dose-dependent manner, with an apoptosis rate as high as 85.04%, and significantly arrested the cell cycle at the S and G2/M phases. Label-free quantitative proteomics identified 137 differentially expressed proteins (|FC| > 2, p < 0.05). GO enrichment analysis indicated that these proteins were mainly involved in cell cycle regulation and apoptosis. KEGG pathway analysis revealed that the antitumor effects were primarily mediated through the regulation of PI3K-Akt (hsa04151), JAK-STAT (hsa04630), and PPAR (hsa03320) signaling pathways. Moreover, differential proteins such as PI3K, AKT1S1, SIRT6, JAK1, SCD, STAT3, CASP8, STAT2, STAT6, and PAK1 showed strong correlation with the core components of the EA-2 fraction of dandelion. Molecular docking results demonstrated that these active compounds exhibited strong binding affinities with key target proteins such as PI3K and JAK1 (binding energy < −5.0 kcal/mol). This study elucidates the multi-target, multi-pathway synergistic mechanisms by which dandelion extract inhibits breast cancer, providing a theoretical basis for the development of novel antitumor agents. Full article
(This article belongs to the Section Cell Biology)
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23 pages, 8387 KiB  
Article
Solvent Fractionation of Polygonum cuspidatum Sieb. et Zucc. for Antioxidant, Biological Activity, and Chromatographic Characterization
by Yuchen Cheng, Yuri Kang and Woonjung Kim
Int. J. Mol. Sci. 2025, 26(14), 7011; https://doi.org/10.3390/ijms26147011 - 21 Jul 2025
Viewed by 316
Abstract
This study investigated the natural bioactive compounds in Polygonum cuspidatum Sieb. et Zucc. (P. cuspidatum) by fractionating a 70% ethanol extract using n-hexane, chloroform, ethyl acetate, n-butanol, and water. The total polyphenol and flavonoid contents of each fraction were [...] Read more.
This study investigated the natural bioactive compounds in Polygonum cuspidatum Sieb. et Zucc. (P. cuspidatum) by fractionating a 70% ethanol extract using n-hexane, chloroform, ethyl acetate, n-butanol, and water. The total polyphenol and flavonoid contents of each fraction were determined, and their antioxidant activities were evaluated using DPPH, ABTS, and FRAP assays. Additionally, the anti-diabetic potential was assessed via α-glucosidase inhibitory activity, while anti-obesity activity was evaluated using lipase inhibitory activity. The fractions were also tested for tyrosinase and elastase inhibitory activities to assess their skin-whitening and anti-wrinkle potential, and their antibacterial activity against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa was determined using the agar diffusion method. Finally, bioactive compounds were identified and quantified using HPLC and GC–MSD. The results showed that the ethyl acetate fraction possessed the highest total polyphenol content (0.53 ± 0.01 g GAE/g) and total flavonoid content (0.19 ± 0.02 g QE/g). It also exhibited strong antioxidant activity, with the lowest DPPH radical scavenging IC50 (0.01 ± 0.00 mg/mL), ABTS radical scavenging IC50 (0.06 ± 0.00 mg/mL), and the highest FRAP value (6.02 ± 0.30 mM Fe2+/mg). Moreover, it demonstrated potent enzyme inhibitory activities, including tyrosinase inhibitory activity (67.78 ± 2.50%), elastase inhibitory activity (83.84 ± 1.64%), α-glucosidase inhibitory activity (65.14 ± 10.29%), and lipase inhibitory activity (85.79 ± 1.04%). In the antibacterial activity, the ethyl acetate fraction produced a clear inhibitory zone of 19.50 mm against Staphylococcus aureus, indicating notable antibacterial activity. HPLC-PDA and GC–MSD analyses identified tannic acid and emodin as the major bioactive constituents. These findings suggest that the ethyl acetate fraction of P. cuspidatum extract, rich in polyphenol and flavonoid compounds, is a promising natural source of bioactive ingredients for applications in the food, pharmaceutical, and cosmetic industries. Further research is needed to explore its mechanisms and therapeutic applications. Full article
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16 pages, 1978 KiB  
Article
Comparative Analysis of Anti-Inflammatory Flavones in Chrysanthemum indicum Capitula Using Primary Cultured Rat Hepatocytes
by Keita Minamisaka, Airi Fujii, Cheng Li, Yuto Nishidono, Saki Shirako, Teruhisa Kawamura, Yukinobu Ikeya and Mikio Nishizawa
Molecules 2025, 30(14), 2996; https://doi.org/10.3390/molecules30142996 - 16 Jul 2025
Viewed by 377
Abstract
The capitula of Chrysanthemum indicum Linné or C. morifolium Ramatuelle (Kikuka in Japanese) are included in several formulae of Kampo medicines (traditional Japanese medicines), such as Chotosan, which is used for headache and dizziness. Luteolin, the principal constituent of C. indicum [...] Read more.
The capitula of Chrysanthemum indicum Linné or C. morifolium Ramatuelle (Kikuka in Japanese) are included in several formulae of Kampo medicines (traditional Japanese medicines), such as Chotosan, which is used for headache and dizziness. Luteolin, the principal constituent of C. indicum, has antioxidant and anti-inflammatory activities. However, the effects of other flavonoids on this crude drug have not yet been thoroughly investigated. To evaluate and compare anti-inflammatory effects, we used primary cultured rat hepatocytes, which produce proinflammatory mediators, such as nitric oxide (NO) and proinflammatory cytokines, in response to interleukin (IL)-1β. Eight derivatives of 5,7-dihydroxyflavone were purified and identified in the ethyl acetate-soluble fraction of a C. indicum capitulum extract: luteolin (Compound 1), apigenin (2), diosmetin (3), 5,7-dihydroxy-3′,4′,5′-trimethoxyflavone (4), acacetin (5), eupatilin (6), jaceosidin (7), and 6-methoxytricin (8). Luteolin is the most abundant compound in this fraction. All compounds significantly suppressed NO production in hepatocytes, with apigenin and acacetin showing the greatest efficacy. The comparison of the IC50 values of the inhibition of NO production suggests that substitutions by hydroxyl and methoxy groups at the C-3′ and C-4′ positions of 5,7-dihydroxyflavone may be at least essential for the suppression of NO production. In hepatocytes, acacetin and luteolin decreased the levels of mRNAs encoding inducible nitric oxide synthase (iNOS), proinflammatory cytokines, including tumor necrosis factor, IL-6, and type 1 IL-1 receptor, which regulates inflammatory responses. Based on the comparison of the IC50 values and the content, luteolin, jaceosidin, and diosmetin may be responsible for the anti-inflammatory effects of C. indicum capitula. Full article
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16 pages, 644 KiB  
Article
Isolation and Identification of Secondary Metabolites in Rheum tataricum L.fil. Growing in Kazakhstan and Surveying of Its Anticancer Potential
by Aiman A. Turgunbayeva, Nurgul A. Sultanova, Mohammad Saleh Hamad, Victor A. Savelyev, Elena I. Chernyak, Irina Yu. Bagryanskaya, Mikhail A. Pokrovsky, Andrey G. Pokrovsky, Nadezhda G. Gemejiyeva and Elvira E. Shults
Molecules 2025, 30(14), 2978; https://doi.org/10.3390/molecules30142978 - 15 Jul 2025
Viewed by 409
Abstract
Rheum tataricum L.fil., known for its high tolerance to drought, salinity, and nutritional deficiency, is the least studied species of wild rhubarb. Extract of roots and rhizomes of R. tataricum has been traditionally used for the treatment of different diseases such as liver, [...] Read more.
Rheum tataricum L.fil., known for its high tolerance to drought, salinity, and nutritional deficiency, is the least studied species of wild rhubarb. Extract of roots and rhizomes of R. tataricum has been traditionally used for the treatment of different diseases such as liver, kidney, womb, and bladder diseases and also relapsing fever. An ethanol extract of the roots of R. tataricum was prepared and further successively fractionated by extraction with tert-butyl methyl ether (TBME) and ethyl acetate (EtOAc). The obtained extract fractions were subjected to a series of chromatographic separations on silica gel for the isolation of its individual compounds. A total of 12 individual compounds, 2-O-β-D-glucopyranoside of R-(4-hydroxyphenyl)-2-butanol (rhododendrin) 1, gallic acid 2, 2-O-β-D-glucopyranoside of S-4-(4-hydroxyphenyl)-2-butanol (epi-rhododendrin) 3, their aglycones (-)-(2R)-rhododendrol 4 and (+)-(2S)-rhododendrol 5, gallotannin β-glucogallin 6, chlorogenic acids (3,5-di-O-caffeoylquinic acid 7 and 5-O-caffeoyl-3-O-(p-coumaroyl) quinic acid 8), 4-(4-hydroxyphenyl)-2-butanon (raspberry ketone) 9 and three stilbenes (rhaponticin 10, desoxyrhaponticin 11 and resveratroloside 12), were isolated and characterized. The structure of desoxyrhaponticin 11 was confirmed by X-ray diffraction analyses. The results of in vitro biological assays (the MTT test) showed that ethanol extract Rheum tataricum was non-toxic against the normal epithelial VERO cells. The isolated compounds 1, 4, 11 and 12 exhibited cytotoxicity against a cervical cancer cell line (CaSki), breast adenocarcinoma (MCF7) and glioblastoma cell line (SNB-19) at low micromolar concentrations. Polyhydroxystilbenes 11 and 12 showed the best potency against adenocarcinoma cells (GI50 = 7–8 μM). The inhibition activity towards cancer cells was comparable to those of the standard drug doxorubicin. The available from R. tataricum secondary metabolites may serve as new leads for the discovery of anticancer drugs. Full article
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14 pages, 1114 KiB  
Article
Deciphering Important Odorants in a Spirulina (Arthrospira platensis) Dietary Supplement by Aroma Extract Dilution Analysis Using Offline and Online Fractionation Approaches
by Aikaterina Paraskevopoulou, Veronika Mall, Theodoros M. Triantis, Triantafyllos Kaloudis, Anastasia Hiskia, Dimitra Dimotikali and Martin Steinhaus
Int. J. Mol. Sci. 2025, 26(14), 6767; https://doi.org/10.3390/ijms26146767 - 15 Jul 2025
Viewed by 635
Abstract
Investigating the volatiles isolated from a commercial spirulina (Arthrospira platensis) dietary supplement by gas chromatography–olfactometry (GC–O) in combination with an aroma extract dilution analysis (AEDA) resulted in 29 odor events with flavor dilution (FD) factors between 8 and 2048. Identification experiments, [...] Read more.
Investigating the volatiles isolated from a commercial spirulina (Arthrospira platensis) dietary supplement by gas chromatography–olfactometry (GC–O) in combination with an aroma extract dilution analysis (AEDA) resulted in 29 odor events with flavor dilution (FD) factors between 8 and 2048. Identification experiments, including various offline and online fractionation approaches, led to the structure assignment of 30 odorants, among which the most potent were sweaty 2- and 3-methylbutanoic acid (FD 2048), roasty, earthy, shrimp-like 2-ethyl-3,5-dimethylpyrazine (FD 2048), vinegar-like acetic acid (FD 1024), and floral, violet-like β-ionone (FD 1024). Static headspace dilution analysis revealed sulfuric, cabbage-like methanethiol (FD factor ≥ 32) as an additional potent odorant. In summary, 31 important spirulina odorants were identified in this study, and 14 were reported for the first time as spirulina constituents. Our data will provide a basis for future odor optimization of spirulina-based food products. Full article
(This article belongs to the Special Issue Recent Research of Natural Products from Microalgae and Cyanobacteria)
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17 pages, 4202 KiB  
Article
The Dichloromethane Fraction of Sanguisorba tenuifolia Inhibits Inflammation in Cells Through Modulation of the p38/ERK/MAPK and NF-κB Signaling Pathway
by Yue Wang, Yiming Lu, Fuao Niu, Siqi Fa, Li Nan and Hyeon Hwa Nam
Int. J. Mol. Sci. 2025, 26(14), 6732; https://doi.org/10.3390/ijms26146732 - 14 Jul 2025
Viewed by 213
Abstract
Sanguisorba tenuifolia is a wild plant of the genus Sanguisorba officinalis. This study aimed to investigate the regulatory effect of the dichloromethane fraction of Sanguisorba tenuifolia on LPS-induced inflammatory responses in RAW264.7 cells, thereby providing a new scientific basis for the medicinal [...] Read more.
Sanguisorba tenuifolia is a wild plant of the genus Sanguisorba officinalis. This study aimed to investigate the regulatory effect of the dichloromethane fraction of Sanguisorba tenuifolia on LPS-induced inflammatory responses in RAW264.7 cells, thereby providing a new scientific basis for the medicinal development of Sanguisorba tenuifolia. Initially, we used 75% ethanol to crudely extract the roots of Sanguisorba tenuifolia, followed by fractional extraction using dichloromethane (CH2Cl2), ethyl acetate (EtOAc), butanol (BuOH), and distilled water (DW) as solvents. By measuring the inhibitory effects of each fractionated extract on NO production, we determined that the SCE (Dichloromethane fraction of Sanguisorba tenuifolia) exhibited the most potent anti-inflammatory activity, leading to its progression to the next experimental stage. Subsequently, we evaluated the effects of SCE on cell viability and LPS-induced inflammatory cytokine secretion in RAW264.7 cells. A rat model of reflux esophagitis was also used to validate the in vivo anti-inflammatory effects of SCE. Additionally, we utilized UPLC/MS-MS to identify and analyze the active components of SCE. The results indicated that SCE could effectively inhibit LPS-induced cellular inflammation by modulating the p38/ERK/MAPK and NF-κB signaling pathways, and also reduced the damage of the esophageal mucosa in rats with reflux esophagitis. UPLC/MS-MS analysis of SCE identified 423 compounds, including 12 active ingredients such as triterpenoids, phenols, and steroids. This discovery not only provides scientific support for the potential of Sanguisorba tenuifolia as an anti-inflammatory agent but also lays the groundwork for the development of new therapeutics for the treatment of inflammatory diseases. Full article
(This article belongs to the Section Molecular Pharmacology)
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27 pages, 6079 KiB  
Article
Bioactive Cyclopeptide Alkaloids and Ceanothane Triterpenoids from Ziziphus mauritiana Roots: Antiplasmodial Activity, UHPLC-MS/MS Molecular Networking, ADMET Profiling, and Target Prediction
by Sylvestre Saidou Tsila, Mc Jesus Kinyok, Joseph Eric Mbasso Tameko, Bel Youssouf G. Mountessou, Kevine Johanne Jumeta Dongmo, Jean Koffi Garba, Noella Molisa Efange, Lawrence Ayong, Yannick Stéphane Fotsing Fongang, Jean Jules Kezetas Bankeu, Norbert Sewald and Bruno Ndjakou Lenta
Molecules 2025, 30(14), 2958; https://doi.org/10.3390/molecules30142958 - 14 Jul 2025
Viewed by 395
Abstract
Malaria continues to pose a significant global health burden, driving the search for novel antimalarial agents to address emerging drug resistance. This study evaluated the antiplasmodial potential of Ziziphus mauritiana Lam. (Rhamnaceae) roots through an integrated phytochemical and pharmacological approach. The ethanol extract, [...] Read more.
Malaria continues to pose a significant global health burden, driving the search for novel antimalarial agents to address emerging drug resistance. This study evaluated the antiplasmodial potential of Ziziphus mauritiana Lam. (Rhamnaceae) roots through an integrated phytochemical and pharmacological approach. The ethanol extract, along with its derived fractions, demonstrated potent in vitro activity against the chloroquine-sensitive Plasmodium falciparum strain 3D7 (Pf3D7), with the ethyl acetate-soluble (IC50 = 11.35 µg/mL) and alkaloid-rich (IC50 = 4.75 µg/mL) fractions showing particularly strong inhibition. UHPLC-DAD-ESI-QTOF-MS/MS-based molecular networking enabled the identification of thirty-two secondary metabolites (132), comprising twenty-five cyclopeptide alkaloids (CPAs), five of which had not yet been described (11, 20, 22, 23, 25), and seven known triterpenoids. Bioactivity-guided isolation yielded thirteen purified compounds (5, 6, 14, 2630, 3236), with betulinic acid (30; IC50 = 19.0 µM) and zizyberenalic acid (32; IC50 = 20.45 µM) exhibiting the most potent antiplasmodial effects. Computational ADMET analysis identified mauritine F (4), hemisine A (10), and nummularine R (21) as particularly promising lead compounds, demonstrating favourable pharmacokinetic properties, low toxicity profiles, and predicted activity against both family A G protein-coupled receptors and evolutionarily distinct Plasmodium protein kinases. Quantitative analysis revealed exceptionally high concentrations of key bioactive constituents, notably zizyberenalic acid (24.3 mg/g) in the root extracts. These findings provide robust scientific validation for the traditional use of Z. mauritiana in malaria treatment while identifying specific cyclopeptide alkaloids and triterpenoids as valuable scaffolds for antimalarial drug development. The study highlights the effectiveness of combining advanced metabolomics, bioassay-guided fractionation, and computational pharmacology in natural product-based drug discovery against resistant malaria strains. Full article
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17 pages, 5655 KiB  
Article
Trans-p-Coumaryl Alcohol as a Bioactive Compound and Anti-Inflammatory Agent in Wannachawee Recipe for Psoriasis
by Supreeya Tantipat, Kongkiat Trisuwan, Phraepakaporn Kunnaja, Seewaboon Sireeratawong, Surapol Natakankitkul, Surasak Imiam and Sunee Chansakaow
Pharmaceutics 2025, 17(7), 864; https://doi.org/10.3390/pharmaceutics17070864 - 30 Jun 2025
Viewed by 367
Abstract
Background/Objectives: Wannachawee recipe (WCR) has been listed in the Hospital Traditional Medicine Formulary and has been used as a Thai medicine to treat psoriasis in the Thai Traditional Medicine Clinic of Prapokklao Hospital since 2006. Previous reports have found that WCR demonstrates [...] Read more.
Background/Objectives: Wannachawee recipe (WCR) has been listed in the Hospital Traditional Medicine Formulary and has been used as a Thai medicine to treat psoriasis in the Thai Traditional Medicine Clinic of Prapokklao Hospital since 2006. Previous reports have found that WCR demonstrates good results for the treatment of patients with psoriasis. Among 136 Thai psoriasis patients who received WCR, 92.80% responded well. Although WCR is effective, there is still a lack of scientific data, especially relating to the bioactive compound in WCR. Therefore, this study aims to evaluate the phytochemicals in WCR via bioassay-guided isolation. Methods: In this study, the WCR was extracted via decoction with water, in a process based on traditional Thai medicine. The water extract was concentrated and dried using a spray dryer. The crude water extract was isolated using the partition technique with organic solvents, namely petroleum ether and ethyl acetate. These fractions were then separated and tested for anti-inflammatory activity using the bioassay-guided fractionation method. Results: Two particular types of pro-inflammatory cytokines are involved in inflammation and are among the factors that cause psoriasis—TNF-α and IL-6. Thus, we evaluated the isolated samples in terms of anti-inflammatory activity. The isolation resulted in two pure compounds—p-coumaryl aldehyde and trans-p-coumaryl alcohol. In the efficacy test of the isolated compounds, compared to the standard indomethacin at the same concentration of 12.5 ug/mL, trans-p-coumaryl alcohol was found to have the best efficacy, inhibiting TNF-α by 29.28% and IL-6 by 36.75%, with the standard compound showing inhibitions rates of 15.80% for TNF-α and 27.44% for IL-6. Conclusions: This study is the first report to identify the bioactive compound of WCR as trans-p-coumaryl alcohol or 4-hydroxycinnamyl alcohol. Full article
(This article belongs to the Special Issue Natural Pharmaceuticals Focused on Anti-inflammatory Activities)
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18 pages, 1769 KiB  
Article
Antioxidant and Pancreatic Lipase Inhibitory Activities of Panax japonicus (T. Nees) C.A. Meyer
by Jinfeng Yang, Wenxuan Jiang, Ju Hee Park, Eun Soo Seong, Yong Soo Kwon and Myong Jo Kim
Plants 2025, 14(13), 2003; https://doi.org/10.3390/plants14132003 - 30 Jun 2025
Viewed by 314
Abstract
Obesity and its associated complications, including oxidative stress, pose significant global health challenges. Natural products offer a promising avenue for developing novel therapeutic strategies. In this study, we investigated the potential of Panax japonicus (T. Nees) C.A. Meyer, a traditional medicinal plant known [...] Read more.
Obesity and its associated complications, including oxidative stress, pose significant global health challenges. Natural products offer a promising avenue for developing novel therapeutic strategies. In this study, we investigated the potential of Panax japonicus (T. Nees) C.A. Meyer, a traditional medicinal plant known for its antioxidant and anti-obesity properties. A methanol extract of Panax japonicus and its fractions were evaluated for their in vitro antioxidant activities (tested using DPPH and reducing power assays), pancreatic lipase (PL) inhibitory capacities, and underlying mechanisms of action. The results indicated that the ethyl acetate fraction of P. japonicus (PJEA) exhibited the greatest potency, demonstrating strong antioxidant activity and significantly inhibiting digestive enzyme activity (pancreatic lipase). Mechanistic studies revealed that the PL inhibition was of a mixed type, combining both competitive and non-competitive mechanisms. Furthermore, PJEA demonstrated the ability to inhibit the differentiation of preadipocytes, primarily exerting its anti-adipogenic effects by downregulating the mRNA expression of PPARγ and the gene expression of C/EBPα. In addition, the extract suppressed the gene expression of FAS and ACC in adipose tissue. Isolation of the bioactive compounds from PJEA identified kaempferol 3-O-α-L-rhamnoside and catechin, which potentially contribute to the observed anti-obesity effects. Overall, this study highlights P. japonicus as a promising natural ingredient for scavenging free radicals and managing obesity, suggesting its potential for development into functional foods or therapeutic agents. Full article
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18 pages, 17059 KiB  
Article
Unraveling Potential Compounds of Uncaria gambir (W.Hunter) Roxb. as Antikeloid Agent: In Silico, In Vitro and Ex Vivo Experimental Validation
by Sri Suciati Ningsih, Sri Widia A. Jusman, Rahimi Syaidah, Muhamad Arif Budiman, Alfi Khatib and Fadilah Fadilah
Biologics 2025, 5(3), 18; https://doi.org/10.3390/biologics5030018 - 27 Jun 2025
Viewed by 427
Abstract
Background/Objectives: Keloid treatment remains challenging due to limited effectiveness and patient dissatisfaction. Herbal-based therapy offers promising alternatives that require further investigation. Uncaria gambir (W.Hunter) Roxb., an original plant from Indonesia, possesses an antifibrotic effect. However, its potential as an antifibrotic agent in keloid [...] Read more.
Background/Objectives: Keloid treatment remains challenging due to limited effectiveness and patient dissatisfaction. Herbal-based therapy offers promising alternatives that require further investigation. Uncaria gambir (W.Hunter) Roxb., an original plant from Indonesia, possesses an antifibrotic effect. However, its potential as an antifibrotic agent in keloid management remains unclear. This study aims to bridge this gap by evaluating the bioactive compound from gambir and its effects on keloid fibroblast primary culture. Methods: The bioactive compounds of gambir extract and fractions (ethanol, hexane, and ethyl acetate fractions) were identified by using liquid chromatography–mass spectrometry (LCMS/MS) analysis. The mechanism of gambir bioactive compounds for keloid was predicted using the compound–protein interaction network and enrichment analysis, and validated using molecular docking and dynamic simulation. The experimental study results, including cytotoxic and bioactivity effects, were represented as IC50 and selectivity index (SI) values, and the ex vivo analysis of keloid tissue explants. Results: Uncariagambiriine was identified as the most potent compound with the lowest binding energy and high stability to the core protein targets: AKT1 and TGFB1. The ethanol fraction was determined to have the highest abundance of gambir’s typical bioactive compounds, with the lowest IC50 (128.76 ± 0.24 µg/mL) and the highest SI (6.32) value. Furthermore, the results of the ex vivo analysis indicated the significant inhibition of keloid fibroblast proliferation and migration by the gambir ethanolic fraction. Conclusions: This study underlines the potential of the gambir ethanolic fraction as an antifibrotic agent in keloid, warranting further investigation and development for clinical applications. Full article
(This article belongs to the Section Natural Products)
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12 pages, 1434 KiB  
Article
Protective Effects of the Ethyl Acetate Fraction of Distylium racemosum Against Metabolic Dysfunction-Associated Steatohepatitis
by Young-Hyeon Lee, Min-Ho Yeo, Kyung-Soo Chang, Weon-Jong Yoon, Hye-Sook Kim, Jongwan Kim and Hye-Ran Kim
Appl. Sci. 2025, 15(13), 7238; https://doi.org/10.3390/app15137238 - 27 Jun 2025
Viewed by 309
Abstract
Metabolic dysfunction-associated steatohepatitis (MASH), previously referred to as non-alcoholic steatohepatitis (NASH), which is a progressive non-alcoholic fatty liver disease, is accompanied by hepatic steatosis, inflammation, and fibrosis. Despite its increasing prevalence, available treatment options for MASH are limited. Here, we investigated the protective [...] Read more.
Metabolic dysfunction-associated steatohepatitis (MASH), previously referred to as non-alcoholic steatohepatitis (NASH), which is a progressive non-alcoholic fatty liver disease, is accompanied by hepatic steatosis, inflammation, and fibrosis. Despite its increasing prevalence, available treatment options for MASH are limited. Here, we investigated the protective effects of the Distylium racemosum ethyl acetate fraction (DRE) using MASH models and explored its key physiologically active components. Palmitic acid (PA)-induced AML12 hepatocytes and high-fat methionine- and choline-deficient-fed C57BL/6 mice were used as MASH models. Lipid accumulation was evaluated via triglyceride measurement, oil red O staining, and histological analysis. Lipid accumulation, inflammation, and fibrosis-associated gene expression were evaluated via real-time polymerase chain reaction. The physiologically active components of DRE were identified via high-performance liquid chromatography. Lipid accumulation and triglyceride levels were significantly reduced in PA-treated AML12 cells following DRE treatment. Additionally, DRE inhibited the expression of genes involved in lipogenesis (FAS and SREBP1c), inflammation (CD68, IL-6, and MCP-1), and fibrosis (COL1A1, COL1A2, and TIMP1). DRE reduced the liver weight, liver-to-body weight ratio, and hepatic steatosis in MASH model mice. It increased carnitine palmitoyltransferase-1 levels and decreased CD36 and transforming growth factor-β levels in the MASH mouse liver. High-performance liquid chromatography revealed that the extract contained rutin flavonoid family members. Overall, DRE was involved in lipid metabolism, inflammation, and fibrosis regulation, exerting potent hepatoprotective effects partly attributed to rutin and serving as a potential preventive candidate for MASH. Full article
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14 pages, 857 KiB  
Article
Rapid and Effective Recovery of Oleanolic and Maslinic Acids from Olive Leaves Using SFE and pH-Zone Centrifugal Partition Chromatography
by Lemonia Antoniadi, Apostolis Angelis, Theodora Nikou, Dimitris Michailidis and Leandros A. Skaltsounis
Molecules 2025, 30(13), 2709; https://doi.org/10.3390/molecules30132709 - 24 Jun 2025
Viewed by 354
Abstract
Olive leaves, the main byproducts of olive cultivation, are characterized by a plethora of bioactive metabolites with significant nutritional value. Their main pentacyclic triterpenes, Oleanolic Acid (OA) and Maslinic Acid (MA), are two high added-value compounds with remarkable activities. This study aimed to [...] Read more.
Olive leaves, the main byproducts of olive cultivation, are characterized by a plethora of bioactive metabolites with significant nutritional value. Their main pentacyclic triterpenes, Oleanolic Acid (OA) and Maslinic Acid (MA), are two high added-value compounds with remarkable activities. This study aimed to develop an efficient methodology for extracting and purifying OA and MA, utilizing Supercritical Fluid Extraction (SFE) and Centrifugal Partition Chromatography (CPC)—two modern, scalable, and green techniques. A total of 21 g of olive leaves were subjected to SFE using supercritical CO2 and ethanol as co-solvent. The extraction employed a step gradient mode, starting with 100% CO2 and incrementally increasing ethanol (0–10% w/w) every 20 min. Fractions rich in OA and MA (500 mg) were further purified via CPC, utilizing pH zone refining to exploit the protonation and deprotonation properties of acidic triterpenes. The biphasic solvent system consisted of n-hexane, ethyl acetate, ethanol, and water (8:2:5:5 v/v/v/v), with trifluoroacetic acid added to the stationary phase and triethylamine added to the mobile phase. This two-step process yielded 89.5 mg of OA and 28.5 mg of MA with over 95% purity, as confirmed by HPLC-ELSD and 1H-NMR. Moreover, purified compounds and SFE fractions exhibited promising elastase and collagenase inhibition, highlighting them as dermocosmetic agents. Full article
(This article belongs to the Special Issue Supercritical Fluid Extraction of Natural Bioactive Compounds)
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15 pages, 2149 KiB  
Article
Biofilm-Forming Lactic Acid Bacteria in Sausages: Isolation, Characterization, and Inhibition Using Eisenia bicyclis-Based Nanoparticles
by Do Kyung Oh, Du-Min Jo, Minji Kim, Jeong-Bin Jo, Ji-Hwan Choi, Jeong Mi Choi, Geum-Jae Jeong, Se Yun Jeong, Fazlurrahman Khan and Young-Mog Kim
Antibiotics 2025, 14(7), 637; https://doi.org/10.3390/antibiotics14070637 - 22 Jun 2025
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Abstract
Background/Objectives: Lactic acid bacteria produce biofilms in meat products that contribute to the products’ deterioration, reduction in quality, and shortened shelf life. Although LAB are generally considered benign, certain strains create slime and cause significant drops in pH. The study’s goal was to [...] Read more.
Background/Objectives: Lactic acid bacteria produce biofilms in meat products that contribute to the products’ deterioration, reduction in quality, and shortened shelf life. Although LAB are generally considered benign, certain strains create slime and cause significant drops in pH. The study’s goal was to identify and characterize LAB strains from sausage products that are capable of biofilm formation, and to evaluate the inhibitory effects of E. bicyclis methanol extract, its ethyl acetate fraction, and phloroglucinol, as well as to synthesize AuNPs, and assess their efficacy in controlling biofilm formation. Methods: Slime or biofilm-producing LAB bacteria were isolated from commercial sausages and identified using 16S rRNA gene sequencing. Lactobacillus sakei S10, which can tolerate high salt concentrations and cold temperatures, was chosen as a representative strain. The isolates were subsequently tested for hemolytic activity, salt and temperature tolerance, and carbohydrate consumption patterns. To evaluate antibiofilm potential, marine-derived compounds from Eisenia bicyclis, such as phloroglucinol (PG), crude methanolic extracts, ethyl acetate fractions, and gold nanoparticle (AuNP) formulations, were tested in situ on sausage surfaces against L. sakei S10 and common pathogens (Pseudomonas aeruginosa and Staphylococcus aureus). The biofilm-inhibitory effects of the extracts, PG, and PG-AuNPs were estimated using the colony-counting method. Results: The PG-AuNPs had an average particle size of 98.74 nm and a zeta potential of −29.82 mV, indicating nanoscale dimensions and considerable colloidal stability. Structural analysis confirmed their spherical form and crystalline structure, as well as the presence of phenolic groups in both reduction and stabilization processes. Among the studied treatments, the PG and PG-AuNPs had the strongest antibiofilm activities, dramatically lowering biofilm biomass, particularly for P. aeruginosa and L. sakei S10. However, the inhibitory effects were less prominent in in situ conditions than in in vitro testing, highlighting the complexity of real food matrices. Conclusions: The results of this study indicate that polyphenolic compounds obtained from marine sources, particularly in nano-formulated forms, have a great deal of potential as natural antibiofilm products. Enhancing the microbiological safety of processed meat products and extending their shelf life could be accomplished through the application of these polyphenolic compounds in food packaging or surface treatments. Full article
(This article belongs to the Section Antibiofilm Strategies)
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27 pages, 7392 KiB  
Article
Skin-Whitening, Antiwrinkle, and Moisturizing Effects of Astilboides tabularis (Hemsl.) Engl. Root Extracts in Cell-Based Assays and Three-Dimensional Artificial Skin Models
by Nam Ho Yoo, Hyun Sook Lee, Sung Min Park, Young Sun Baek and Myong Jo Kim
Int. J. Mol. Sci. 2025, 26(12), 5725; https://doi.org/10.3390/ijms26125725 - 15 Jun 2025
Viewed by 539
Abstract
This study investigated the potential cosmetic properties of the ethyl acetate (EtOAc) fraction obtained from the roots of Astilboides tabularis (Hemsl.) Engl., focusing on skin-whitening, antiwrinkle, and moisturizing effects using cell-based assays and three-dimensional (3D) artificial skin models (Neoderm-ED and Neoderm-ME). The EtOAc [...] Read more.
This study investigated the potential cosmetic properties of the ethyl acetate (EtOAc) fraction obtained from the roots of Astilboides tabularis (Hemsl.) Engl., focusing on skin-whitening, antiwrinkle, and moisturizing effects using cell-based assays and three-dimensional (3D) artificial skin models (Neoderm-ED and Neoderm-ME). The EtOAc fraction showed significant dose-dependent inhibitory activity against tyrosinase (TYR) (72.0% inhibition at 50 µg/mL), comparable to that of kojic acid. In α-melanocyte-stimulating hormone (α-MSH)-stimulated Neoderm-ME artificial skin containing melanocytes, the EtOAc fraction reduced melanin synthesis at concentrations of 50 and 75 µg/mL and decreased melanogenesis-related gene expression, including TYR, microphthalmia-associated transcription factor (MITF), tyrosinase-related protein-1 (TRP-1) and TRP-2. In the antiwrinkle assays, the EtOAc fraction effectively inhibited elastase activity (41.5% inhibition at 10 µg/mL), exceeding the efficacy of ursolic acid. In the Neoderm-ED artificial skin model, the EtOAc fraction reversed structural damage induced by particulate matter (PM10), restoring epidermal thickness and dermal density. This improvement was supported by the increased expression of skin barrier and antiwrinkle genes, including filaggrin, hyaluronic acid synthase-1 (HAS-1), HAS-2, aquaporin-3 (AQP-3), collagen type I alpha 1 chain (COL1A1), elastin, tissue inhibitor of metalloproteinases-1 (TIMP-1), and TIMP-2, as well as decreased expression of matrix metalloproteinases (MMP-1, MMP-3, and MMP-9). Our results indicate that the EtOAc fraction from A. tabularis root has considerable potential as a multifunctional cosmetic. Full article
(This article belongs to the Section Bioactives and Nutraceuticals)
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