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20 pages, 3045 KiB  
Article
Using In Vitro and In Silico Analysis to Investigate the Chemical Profile and Biological Properties of Polygonum istanbulicum Extracts
by Giancarlo Angeles Flores, Gaia Cusumano, Gokhan Zengin, Mehmet Veysi Cetiz, Abdullahi Ibrahim Uba, Ismail Senkardes, Ismail Koyuncu, Ozgur Yuksekdag, Alina Kalyniukova, Carla Emiliani, Roberto Venanzoni and Paola Angelini
Plants 2024, 13(23), 3421; https://doi.org/10.3390/plants13233421 (registering DOI) - 5 Dec 2024
Abstract
The present study investigates the chemical profile and biological activities of Polygonum istanbulicum M. Keskin, a species endemic to Turkey, aiming to explore its potential applications in pharmacology. We assessed its phenolic and flavonoid content by employing ethyl acetate, methanol, and water as [...] Read more.
The present study investigates the chemical profile and biological activities of Polygonum istanbulicum M. Keskin, a species endemic to Turkey, aiming to explore its potential applications in pharmacology. We assessed its phenolic and flavonoid content by employing ethyl acetate, methanol, and water as extraction solvents. The methanol extract demonstrated the highest concentrations of these compounds, with liquid chromatography–quadrupole time-of-flight tandem mass spectrometry (LC-MS-qTOF) analysis identifying a wide range of bioactive substances, such as derivatives of quercetin and myricetin. Antioxidant capacity was evaluated using 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), cupric-reducing antioxidant capacity (CUPRAC), ferric-reducing antioxidant power (FRAP), and phosphomolybdenum assays, with the methanol extract showing the most potent activity (DPPH: 892.22 mg Trolox equivalent (TE)/g; ABTS: 916.21 mg TE/g; CUPRAC: 1082.69 mg TE/g; FRAP: 915.05 mg TE/g). Enzyme inhibition assays highlighted the efficacy of P. istanbulicum extracts against key enzymes, with potential implications for managing Alzheimer’s disease, hyperpigmentation, and type 2 diabetes. Cytotoxicity tests against various cancer cell lines showed notable activity, particularly with the aqueous extract on the HGC-27 cell line (IC50: 29.21 µg/mL), indicating potential for targeted anti-cancer therapy. Molecular docking and molecular dynamics simulations further supported the binding affinities of quercetin and myricetin derivatives to cancer-related proteins, suggesting significant therapeutic potential. This study underscores the value of P. istanbulicum as a source of bioactive compounds with applications in antioxidant, anti-cancer, and enzyme-inhibitory treatments. Full article
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<p>(<b>a</b>) PCA scores of <span class="html-italic">Polygonum istanbulicum</span> extracts in negative ionization mode. (<b>b</b>) PCA scores of <span class="html-italic">Polygonum istanbulicum</span> extracts in positive ionization mode. (<b>c</b>) PLSDA analysis of <span class="html-italic">Polygonum istanbulicum</span> extracts in negative ionization mode. (<b>d</b>) PLSDA analysis of <span class="html-italic">Polygonum istanbulicum</span> extracts in positive ionization mode.</p>
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<p>Heatmap of <span class="html-italic">Polygonum istanbulicum</span> extracts in negative (<b>a</b>) and positive (<b>b</b>) ionization mode.</p>
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<p>Enzyme and protein active sites with compounds showing the best binding energy. (<b>a</b>) Interaction between AChE and quercetin 3-O-rhamnoside. (<b>b</b>) Interaction between AChE and quercetin 4-O-glucoside. (<b>c</b>) Interaction between AChE and quercetin 3-O-galactoside. (<b>d</b>) Interaction between BChE and myricetin 3-O-rutinoside.</p>
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<p>MM/PBSA binding free energy analysis. (<b>a</b>) ARO_quercetin 3-O-rutinoside complex. (<b>b</b>) CDK4_quercetin 3-O-arabinoside complex. (<b>c</b>) EGFR_myricetin 3-O-rhamnoside complex.</p>
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<p>Presentation of molecular dynamics simulations in graphical form; (<b>a</b>) RMSD of CDK4_quercetin 3-O-arabinoside. EGFR_myricetin 3-O-rhamnoside and ARO_quercetin 3-O-rutinoside complexes. (<b>b</b>) RMSF of CDK4_quercetin 3-O-arabinoside. EGFR_myricetin 3-O-rhamnoside and ARO_quercetin 3-O-rutinoside complexes. (<b>c</b>) Solvent accessibility of AChE_quercetin 3-O-glucoside. BChE_quercetin 3-O-rhamnoside-7-O-glucoside. glucosidase-kaempferol 3,6-acetylglucoside-7-rhamnoside, and tyrosinase-quercetin 3-caffeoylrobinobioside complexes. (<b>d</b>) Minimum distance of CDK4_quercetin 3-O-arabinoside. EGFR_myricetin 3-O-rhamnoside and ARO_quercetin 3-O-rutinoside complexes.</p>
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<p>Hydrogen bonds in complexes. (<b>a</b>) Hydrogen bonds in CDK4_quercetin 3-O-arabinoside. EGFR_myricetin 3-O-rhamnoside and ARO_quercetin 3-O-rutinoside complexes. (<b>b</b>) Hydrogen bonds in CDK4_quercetin 3-O-arabinoside. EGFR_myricetin 3-O-rhamnoside and ARO_quercetin 3-O-rutinoside complexes. (<b>c</b>) Hydrogen bonds in CDK4_quercetin 3-O-arabinoside. EGFR_myricetin 3-O-rhamnoside and ARO_quercetin 3-O-rutinoside complexes.</p>
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<p><span class="html-italic">Polygonum istanbulicum</span>. (<b>a</b>) Plant in natural habitat; (<b>b</b>) herbarium specimen.</p>
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15 pages, 4848 KiB  
Article
Therapeutic Efficacy and Underlying Mechanisms of a Mannoglucan from Hirsutella sinensis Mycelium on Dextran Sulfate Sodium-Induced Inflammatory Bowel Disease in Mice: Modulation of the Intestinal Barrier, Oxidative Stress and Gut Microbiota
by Weihua Ni, Yu Li, Jingyue Feng, Boxuan Liu, Hongyan Yuan, Guixiang Tai and Hongtao Bi
Int. J. Mol. Sci. 2024, 25(23), 13100; https://doi.org/10.3390/ijms252313100 (registering DOI) - 5 Dec 2024
Abstract
Hirsutella sinensis (H. sinensis), a non-sexual form of the valuable Chinese medicinal herb, demonstrates various biological activities, such as immune modulation and antioxidative capabilities. Nonetheless, the effects of bioactive polysaccharides derived from H. sinensis on colitis have yet to be investigated. In [...] Read more.
Hirsutella sinensis (H. sinensis), a non-sexual form of the valuable Chinese medicinal herb, demonstrates various biological activities, such as immune modulation and antioxidative capabilities. Nonetheless, the effects of bioactive polysaccharides derived from H. sinensis on colitis have yet to be investigated. In our prior research, we extracted a mannoglucan (HSWP-1d) from H. sinensis and found that it attenuates TGF-β1-induced epithelial-mesenchymal transition. The present study investigated the protective effects of HSWP-1d against colitis induced by dextran sulfate sodium (DSS) in mice. The results demonstrate that HSWP-1d effectively ameliorates symptoms of colitis and preserves the intestinal barrier’s stability by enhancing the expression of tight junction proteins. The administration of HSWP-1d results in a reduction in oxidative stress through the augmentation of antioxidative enzyme activities, concomitant with the suppression of oxidative product generation. Simultaneously, HSWP-1d reduced the levels of pro-inflammatory cytokines while elevating the levels of anti-inflammatory cytokines, effectively mitigating the inflammatory response. Furthermore, HSWP-1d influences and alters short-chain-fatty-acid (SCFA) levels, thereby enhancing the intestinal microenvironment. In conclusion, HSWP-1d contributes to intestinal well-being and holds potential as both a therapeutic choice and a supplier of essential nutrients for the amelioration of colitis. Full article
(This article belongs to the Special Issue Health Promoting Benefits of Natural Products and Functional Foods)
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<p>HSWP-1d ameliorated colitis symptoms in mice with DSS-induced colitis. (<b>A</b>) Experimental design diagram: mice were categorized into 5 groups (<span class="html-italic">n</span> = 6) receiving either plain water or water containing 3% DSS for a duration of 7 days while concurrently being administered HSWP-1d (5, 10, and 20 mg/kg) via intragastrical administration. (<b>B</b>) Changes of body weight. (<b>C</b>) Scores for the DAI. (<b>D</b>) Morphological assessment of the mouse colon. (<b>E</b>) Length measurements of the mouse colon. (<b>F</b>–<b>I</b>) Serum concentrations of ALT, AST, BUN, and Cr. Results are shown as means ± SDs, ns, not significant, *, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>HSWP-1d ameliorated histopathologic damage and intestinal barrier damage in mice with DSS-induced colitis. (<b>A</b>) H&amp;E staining of colon tissue. (<b>B</b>–<b>E</b>) Immunohistochemical assay and average integrated optical density of claudin 1, occludin, and ZO-1. Scale bar represents 100 µm. Results are shown as means ± SDs. *, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>HSWP-1d mitigated oxidative stress in mice with DSS-induced colitis. (<b>A</b>–<b>H</b>) The activities of SOD, CAT, and GSH-Px, along with the MDA level, in both serum (<b>A</b>–<b>D</b>) and colon tissue (<b>E</b>–<b>H</b>) from colitis mice were measured. Results are shown as means ± SDs. *, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>HSWP-1d regulated inflammatory cytokines in the serum and colon in mice with DSS-induced colitis. (<b>A</b>–<b>H</b>) The IL-1β, IL-6, TNF-α, and IL-10 concentrations in the serum (<b>A</b>–<b>D</b>) and colon tissue (<b>E</b>–<b>H</b>) of mice with colitis were measured. Results are shown as means ± SDs. *, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>HSWP-1d influenced the intestinal microbiota composition in mice with DSS-induced colitis. (<b>A</b>) Venn diagram. (<b>B</b>) Principal-coordinate analysis (PCoA). (<b>C</b>,<b>D</b>) The distribution of gut microbiota at the phylum classification. (<b>E</b>,<b>F</b>) The breakdown of gut microbiota at the genus classification. (<b>G</b>) LDA scores for taxa with significant abundance differences (LDA &gt; 4.5). Results are shown as means ± SDs, ns, not significant, **, <span class="html-italic">p</span>  &lt; 0.01.</p>
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<p>HSWP-1d influenced the structure and metabolites of intestinal flora in mice. (<b>A</b>) Heatmap of differentially abundant metabolite clustering among different samples (data were not normalized). (<b>B</b>) Correlation plot of the intestinal flora with metabolites between the DSS and HSWP-1d (10 mg/kg) groups. *, <span class="html-italic">p</span>  &lt; 0.05; **, <span class="html-italic">p</span>  &lt; 0.01; ***, <span class="html-italic">p</span>  &lt; 0.001.</p>
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32 pages, 2628 KiB  
Review
Flavonoids and Other Polyphenols: Bioactive Molecules from Traditional Medicine Recipes/Medicinal Plants and Their Potential for Phytopharmaceutical and Medical Application
by Aekkhaluck Intharuksa, Sompop Kuljarusnont, Yohei Sasaki and Duangjai Tungmunnithum
Molecules 2024, 29(23), 5760; https://doi.org/10.3390/molecules29235760 (registering DOI) - 5 Dec 2024
Abstract
Currently, natural bioactive ingredients and/or raw materials are of significant interest to scientists around the world. Flavonoids and other polyphenols are a major group of phytochemicals that have been researched and noted as bioactive molecules. They offer several pharmacological and medical benefits. This [...] Read more.
Currently, natural bioactive ingredients and/or raw materials are of significant interest to scientists around the world. Flavonoids and other polyphenols are a major group of phytochemicals that have been researched and noted as bioactive molecules. They offer several pharmacological and medical benefits. This current review aims to (1) illustrate their benefits for human health, such as antioxidant, anti-aging, anti-cancer, anti-inflammatory, anti-microbial, cardioprotective, neuroprotective, and UV-protective effects, and also (2) to perform a quality evaluation of traditional medicines for future application. Consequently, keywords were searched on Scopus, Google Scholar, and PubMed so as to search for related publications. Then, those publications were carefully checked in order to find current and non-redundant studies that matched the objective of this review. According to this review, researchers worldwide are very interested in discovering the potential of flavonoids and other polyphenols, used in traditional medicines and taken from medicinal plants, in relation to medical and pharmaceutical applications. Many studies focus on the health benefits of flavonoids and other polyphenols have been tested using in silico, in vitro, and in vivo models. However, few studies have been carried out using clinical trials that have trustworthy subject sizes and are in accordance with clinical practice guidelines. Additionally, interesting research directions and perspectives for future studies are highlighted in this work. Full article
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<p>General structures of flavonoids (<b>A</b>) and other polyphenols (non-flavonoids, i.e., coumarins (<b>B</b>), phenolic acids (<b>C</b>), and stilbenes (<b>D</b>)).</p>
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<p><span class="html-italic">Nelumbo nucifera</span> Gaertn., an example of a medicinal plant and traditional medicinal recipe: (<b>A</b>) <span class="html-italic">N. nucifera</span> medicinal plants in their aquatic natural habitat; (<b>B</b>) Thai traditional medicinal recipe made from the dried stamens of <span class="html-italic">N. nucifera</span>. All photos are taken by Assoc. Prof. Dr. Duangjai Tungmunnithum.</p>
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<p>The summary on flavonoids and other polyphenols from traditional medicines/medicinal plants and their phytopharmaceutical and medical potentials. Created with BioRender.com/j25g033 by Aekkhaluck Intharuksa.</p>
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14 pages, 952 KiB  
Article
Identification of Viral Diseases and Influences on Yield and Quality of Angelica sinensis
by Jinjuan Li and Ting Li
Horticulturae 2024, 10(12), 1300; https://doi.org/10.3390/horticulturae10121300 (registering DOI) - 5 Dec 2024
Abstract
Angelica sinensis is a perennial herbaceous species mainly cultivated in the Gansu, Yunnan, and Qinghai provinces of China, and its dried roots have been widely used for nourishing blood and harmonizing vital energy, largely relying on its bioactive compounds (e.g., alkylphthalides, polysaccharides, and [...] Read more.
Angelica sinensis is a perennial herbaceous species mainly cultivated in the Gansu, Yunnan, and Qinghai provinces of China, and its dried roots have been widely used for nourishing blood and harmonizing vital energy, largely relying on its bioactive compounds (e.g., alkylphthalides, polysaccharides, and flavonoids). In recent years, viral diseases have been suspected to be present in A. sinensis in field cultivation. In order to reveal the infection status and causes, a survey and the identification of viral diseases and their influence on the yield and quality of A. sinensis were conducted in four different counties of Gansu province. The results showed viral disease rates of ca. 21% to 37% for potato virus Y (PVY) and tomato mosaic virus (ToMV), as well as ca. 2.8- to 8.9-fold decreases in root yield on a unit-area basis; meanwhile, the contents of the main bioactive compounds (i.e., ferulic acid, ligustilide, and polysaccharides) were significantly lower in the virus-infected plants (VIPs) compared with the virus-free plants (VFPs); there were significant positive relationships of the viral disease rate with planting density and expression levels of the PVY-coat protein (CP) and ToMV-CP genes (p < 0.01). The above-mentioned observations indicate that it is necessary and urgent to take measures (e.g., controlling plant density, rational rotation, and using virus-free seedlings) to prevent the spread of plant viruses. Full article
(This article belongs to the Special Issue Breeding, Cultivation, and Metabolic Regulation of Medicinal Plants)
13 pages, 5662 KiB  
Article
Bioengineered Extracellular Vesicle Hydrogel Modulating Inflammatory Microenvironment for Wound Management
by Yunfei Mu, Liwen Ma, Jia Yao, Dan Luo and Xianguang Ding
Int. J. Mol. Sci. 2024, 25(23), 13093; https://doi.org/10.3390/ijms252313093 - 5 Dec 2024
Abstract
Chronic wounds, frequently arising from conditions like diabetes, trauma, or chronic inflammation, represent a significant medical challenge due to persistent inflammation, heightened infection risk, and limited treatment solutions. This study presents a novel bioengineered approach to promote tissue repair and improve the healing [...] Read more.
Chronic wounds, frequently arising from conditions like diabetes, trauma, or chronic inflammation, represent a significant medical challenge due to persistent inflammation, heightened infection risk, and limited treatment solutions. This study presents a novel bioengineered approach to promote tissue repair and improve the healing environment. We developed a bioactive hydrogel patch, encapsulated zeolitic imidazolate framework-8 (ZIF-8) into extracellular vesicles (EVs) derived from anti-inflammatory M2 macrophages, and synthesized ZIF@EV, then embedded it in the sodium alginate matrix. This hydrogel structure enables the controlled release of therapeutic agents directly into the wound site, where it stimulates endothelial cell proliferation and promotes new blood vessel formation. These processes are key components of effective tissue regeneration. Crucially, the EV-infused patch influences the immune response by polarizing macrophages towards an M2 phenotype, shifting the wound environment from inflammation toward regenerative healing. When applied in a murine model of chronic wounds, the EV hydrogel patch demonstrated notable improvements in healing speed, quality, and tissue integration compared to traditional approaches such as growth factor therapies and foam dressings. These promising findings suggest that this bioactive hydrogel patch could serve as a versatile, practical solution for chronic wound management, providing an adaptable platform that addresses both the biological and logistical needs of wound care in clinical settings. Full article
(This article belongs to the Special Issue Recent Research of Nanomaterials in Molecular Science)
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<p>Synthesis of ZIF@EV–Gel ink. (<b>A</b>) Schematic illustration of the synthetic process of ZIF@EV nanoparticles and corresponding TEM images of the prepared ZIF@EV. Scale bar: 100 nm. (<b>B</b>) NTA of the prepared ZIF@EVs demonstrating the average size of around 180 nm. (<b>C</b>) Zeta potential of EV and ZIF@EV. (<b>D</b>) The gelation of ZIF@EV–Gel ink. (<b>E</b>) SEM image of the macrostructure of ZIF@EV–Gel ink. Scale bar: 3 μm. (<b>F</b>) The degradation curve of ZIF@EV forms the Gel. (<b>G</b>) The continuous release profile of ZIF@EV from Gel. ns denotes non-significant difference.</p>
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<p>Angiogenesis ability of ZIF@EV. (<b>A</b>) Representative images of the transwell migration assay of HUVECs under the treatment of ZIF@EV. Scale bar, 200 μm. (<b>B</b>) Quantification of the migratory capacity of HUVECs. (<b>C</b>) Images of HUVEC migration under ZIF@EV treatment at different time points. Scale bar, 200 μm. (<b>D</b>) Corresponding quantification of HUVEC migration at different time points. (<b>E</b>) Formation of tubes by HUVECs with various treatments. (<b>F</b>) percentage area of vessels as a representation of tube formation capability in various groups (n = 3). ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, and **** <span class="html-italic">p</span> &lt; 0.0001.</p>
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<p>ZIF@EV induced macrophage polarization into M2 phenotype. (<b>A</b>) Representative image of macrophages under different treatments. Scale bar, 50 μm. (<b>B</b>) Flow cytometry analysis of CD206+ macrophages. (<b>C</b>) Flow cytometry analysis of CD86+ macrophages. (<b>D</b>) Quantification of CD206+ macrophages (n = 3). (<b>E</b>) Histogram analysis of CD206+ macrophages. (<b>F</b>) Quantification of the ratio of M1 and M2 macrophages (n = 3). (<b>G</b>) Relative protein expression of M1 and M2 macrophage markers after macrophages incubated with various treatments (n = 3). The color contour from blue to red denotes the intensified signal. **** <span class="html-italic">p</span> &lt; 0.0001.</p>
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<p>In vivo wound healing efficacy of ZIF@EV-Gel. (<b>A</b>) Representative image of wound size on different days. (<b>B</b>) Fluorescence images of cutaneous wounds extracted at 6 and 12 h after different treatments. The color contour from blue to red denotes the intensified signal. (<b>C</b>) Hematoxylin and eosin (HE) and Masson stain of the wounded skin on day 12. Scale bar = 100 μm. (<b>D</b>) Quantification of wound size in different groups. (<b>E</b>) Quantitative analysis of epidermal thickness. (<b>F</b>) Quantitative analysis of collagen deposition (n = 3). * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, and **** <span class="html-italic">p</span> &lt; 0.0001.</p>
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<p>Therapeutic mechanism of ZIF@EV-Gel ink. ZIF@EV-Gel accelerated wound healing in vivo. (<b>A</b>) Image of immunofluorescence staining to assess the formation of new blood vessels and the inflammatory status of the wounds. The blue color indicate nucleus and red color indicate the presence of CD31 and CD206 separately. (<b>B</b>) bactericidal effects of ZIF@EV-Gel. (<b>C</b>) Quantification of blood vessels with various treatments. (<b>D</b>) Quantitative analysis of activated M2 macrophages in epidermal tissues. **** <span class="html-italic">p</span> &lt; 0.0001.</p>
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13 pages, 3062 KiB  
Article
Isolation and Antioxidant Mechanism of Polyphenols from Sanghuangporous vaninii
by Peng Liu, Yuyang Wang, Daoyou Chen, Zhengpeng Li, Di Wu, Zhong Zhang, Wanchao Chen, Wen Li and Yan Yang
Antioxidants 2024, 13(12), 1487; https://doi.org/10.3390/antiox13121487 - 5 Dec 2024
Abstract
Sanghuangporous vaninii, as an edible and medicinal macrofungus, represents a high source of polyphenols with considerable antioxidant activities. However, due to the significant differences in polyphenol content and bioactivity caused by different cultivation substrates, its antioxidant mechanism has not been fully determined. [...] Read more.
Sanghuangporous vaninii, as an edible and medicinal macrofungus, represents a high source of polyphenols with considerable antioxidant activities. However, due to the significant differences in polyphenol content and bioactivity caused by different cultivation substrates, its antioxidant mechanism has not been fully determined. In this paper, five groups of S. vaninii fruiting bodies were collected from cultivation substrates from different areas. The ethanol extracts of mulberry sawdust from Haining City (HNMS) had the highest polyphenol content, as well as excellent antioxidant activity. HNMS3, a polyphenol component with promising antioxidant capacity, was further isolated through optimization with different extractants, silica gel column chromatography, and thin layer chromatography analysis. UPLC-Q-TOF-MS analysis showed that HNMS3 was composed of 33 compounds, corresponding to 257 targets of oxidative stress by network pharmacology analysis, which were strongly associated with mental health and neurodegenerative diseases. Protein–protein interaction and molecular docking analysis indicated that eight hub genes (PPARG, IL-6, STAT3, PTGS2, SRC, MTOR, ERS1, and EGFR) are attributed to the regulation of the key compounds hispidin, inoscavin A, inoscavin_C, and phellibaumin B. Consequently, this study obtains S. vaninii polyphenolic component HNMS3 with excellent antioxidant capacity, simultaneously revealing its potential antioxidant mechanisms, providing new insights into the application of S. vaninii. Full article
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<p><span class="html-italic">S. vaninii</span> fruiting bodies from different regions or cultivation substrates exhibited different composition components. (<b>A</b>) Five groups of <span class="html-italic">S. vaninii</span> fruiting bodies were collected in China. (<b>B</b>) HPLC fingerprint of the ethanol extracts from these five groups of <span class="html-italic">S. vaninii</span> fruiting bodies. (<b>C</b>) Analysis of key component contents, the values are presented as means (<span class="html-italic">n</span> = 3).</p>
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<p>Analysis of antioxidant capacity in vitro of alcohol extracts from <span class="html-italic">S. vaninii</span> fruiting bodies. (<b>A</b>) DPPH radical scavenging assay and (<b>B</b>) ABST radical scavenging assay were employed. All the values are presented as means ± standard error of mean (SEM) (<span class="html-italic">n</span> = 3).</p>
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<p>Optimization of extractants. (<b>A</b>) Schematic diagram of liquid–liquid extraction. (<b>B</b>) HPLC fingerprint and (<b>C</b>) polyphenol content of extracts from different extractants. All the values are presented as means ± standard error of mean (SEM) (<span class="html-italic">n</span> = 3). Different letters above the bars represent results that were significantly different, <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Isolation and purification of HNMS purified by liquid-liquid extraction were further implemented. HPLC fingerprint of ethyl acetate extract and six recombinant samples obtained by silica gel column chromatography analysis.</p>
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<p>Analysis of antioxidant capacity in vitro of HNMS1 ~HNMS5. (<b>A</b>) DPPH radical scavenging assay and (<b>B</b>) ABST radical scavenging assay were employed. All the values are presented as means ± standard error of mean (SEM) (<span class="html-italic">n</span> = 3).</p>
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<p>Network pharmacology analysis. (<b>A</b>) Venn diagram of the targets of the active ingredients and oxidative stress-related targets. (<b>B</b>) The network of the 257 common targets and 20 compounds from HNMS3. (<b>C</b>) KEGG and (<b>D</b>) Disease–gene Associations (DISEASES) enrichment analysis of the common targets. (<b>E</b>) PPI network of the common and core targets, the deepened node color was proportional to the degree of interaction.</p>
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<p>Molecular docking. (<b>A</b>) Molecular docking analysis, the deepened node color of active compounds was proportional to the degree of interaction. The heat map of binding energy (kcal/mol) of key targets and compounds of HNMS3. (<b>B</b>) Visualized display of molecular docking between representative active compounds and core targets.</p>
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23 pages, 9260 KiB  
Article
Neuroprotective and Anti-Inflammatory Effects of Dimethyl Fumarate, Monomethyl Fumarate, and Cannabidiol in Neurons and Microglia
by Alicia Sánchez-Sanz, María José Coronado-Albi, Rafael Muñoz-Viana, Antonio García-Merino and Antonio J. Sánchez-López
Int. J. Mol. Sci. 2024, 25(23), 13082; https://doi.org/10.3390/ijms252313082 - 5 Dec 2024
Abstract
Dimethyl fumarate (DMF) is an immunomodulatory treatment for multiple sclerosis (MS) that can cross the blood–brain barrier, presenting neuroprotective potential. Its mechanism of action is not fully understood, and there is a need to characterize whether DMF or its bioactive metabolite monomethyl fumarate [...] Read more.
Dimethyl fumarate (DMF) is an immunomodulatory treatment for multiple sclerosis (MS) that can cross the blood–brain barrier, presenting neuroprotective potential. Its mechanism of action is not fully understood, and there is a need to characterize whether DMF or its bioactive metabolite monomethyl fumarate (MMF) exerts neuroprotective properties. Moreover, the combination of adjuvant agents such as cannabidiol (CBD) could be relevant to enhance neuroprotection. The aim of this study was to compare the neuroprotective and immunomodulatory effects of DMF, MMF, and CBD in neurons and microglia in vitro. We found that DMF and CBD, but not MMF, activated the Nrf2 antioxidant pathway in neurons. Similarly, only DMF and CBD, but not MMF, prevented the LPS-induced activation of the inflammatory pathway NF-kB in microglia. Additionally, the three drugs inhibited the production of nitric oxide in microglia and protected neurons against apoptosis. Transcriptomically, DMF modulated a greater number of inflammatory and Nrf2-related genes compared to MMF and CBD in both neurons and microglia. Our results show that DMF and MMF, despite being structurally related, present differences in their mechanisms of action that could be relevant for the achievement of neuroprotection in MS patients. Additionally, CBD shows potential as a neuroprotective agent. Full article
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<p>Cell viability of the neurons and microglia cells treated with dimethyl fumarate (DMF), monomethyl fumarate (MMF), or cannabidiol (CBD). (<b>A</b>) Cell viability of neurons treated with DMF. (<b>B</b>) Cell viability of microglia cells treated with DMF. (<b>C</b>) Cell viability of neurons treated with MMF. (<b>D</b>) Cell viability of microglia cells treated with MMF. (<b>E</b>) Cell viability of neurons treated with CBD. (<b>F</b>) Cell viability of microglia cells treated with CBD. (<b>A</b>–<b>F</b>) Dose–response relationships of each drug at 24, 48, and 72 h. The mean with standard deviation is represented of the percentage of cell viability with respect to the logarithm of the tested concentrations (10, 30, 50, 100, and 200 µM for DMF and MMF; 1, 3, 5, 10, and 30 µM for CBD). The percentage of cell viability was calculated relative to the vehicle-treated cells. The half-maximal inhibitory concentration (IC<sub>50</sub>) and the coefficient of determination (R<sup>2</sup>) values are indicated below each graph.</p>
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<p>Effect of dimethyl fumarate (DMF), monomethyl fumarate (MMF), and cannabidiol (CBD) on the activation of Nrf2 in neurons and microglia. (<b>A</b>) Effect of DMF on Nrf2 activation in neurons and microglia. (<b>B</b>) Effect of MMF on Nrf2 activation in neurons and microglia. (<b>C</b>) Effect of CBD on Nrf2 activation in neurons and microglia. (<b>D</b>) Representative immunofluorescence images from neurons showing the Nrf2 protein location (green). TO-PRO (blue) was used as a nuclear counterstain. Scale bar: 20 µm, shown in the bottom-right corner of each image. (<b>E</b>) Representative immunofluorescence images from microglia showing Nrf2 protein location (green). TO-PRO (blue) was used as a nuclear counterstain. Scale bar: 20 µm, shown in the bottom-right corner of each image. (<b>A</b>–<b>C</b>) White bars represent neurons and black dotted lines represent microglia. (<b>A</b>–<b>E</b>) Neurons and microglia were treated for 4 h with either the vehicle (VEH) or drug. Bars represent the fold change of the mean nuclear fluorescence intensity of Nrf2 in the drug-treated cells compared to their respective vehicle. Data (mean ± standard deviation) are representative of three different experiments. The Kruskal–Wallis test was used to compare the different concentrations of each drug against their respective vehicle. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001 in neurons; ### <span class="html-italic">p</span> &lt; 0.001, #### <span class="html-italic">p</span> &lt; 0.0001 in microglia.</p>
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<p>Immunomodulatory effects of dimethyl fumarate (DMF), monomethyl fumarate (MMF), and cannabidiol (CBD) in microglia. (<b>A</b>–<b>C</b>) Effect of DMF (<b>A</b>), MMF (<b>B</b>), and CBD (<b>C</b>) on the activation of NF-kB p65. Microglia cells were treated for 30 min with either vehicle (VEH), lipopolysaccharide (LPS), or LPS in combination with DMF, MMF, or CBD. Bars represent the fold change of the mean nuclear fluorescence intensity of NF-kB p65 in the drug-treated cells compared to the vehicle (represented by black dotted lines). (<b>D</b>–<b>F</b>) Effect of DMF (<b>D</b>), MMF (<b>E</b>), and CBD (<b>F</b>) on the production of nitric oxide (NO). Microglia cells were treated for 24, 48, or 72 h with either the VEH, LPS, or LPS in combination with DMF, MMF, or CBD. Bars represent the concentration (µM) of NO. (<b>G</b>–<b>I</b>) Effect of DMF (<b>G</b>), MMF (<b>H</b>), and CBD (<b>I</b>) on neuronal apoptosis. Neurons were cultured for 4 h with microglia-conditioned medium. Microglia had been previously treated with the VEH, LPS, or LPS in combination with DMF, MMF, or CBD for 48 h. Bars represent the fold change of the number of apoptotic cells (mean ± standard deviation) in the drug-treated cells compared to their respective vehicle. (<b>A</b>–<b>I</b>) Data are representative of three different experiments. # <span class="html-italic">p</span> &lt; 0.05, ## <span class="html-italic">p</span> &lt; 0.01, ### <span class="html-italic">p</span> &lt; 0.001, #### <span class="html-italic">p</span> &lt; 0.0001 LPS compared to the vehicle (Mann–Whitney test); * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001 LPS + drug compared to LPS (Kruskal–Wallis test).</p>
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<p>Transcriptomic profile of neurons treated with dimethyl fumarate (DMF), monomethyl fumarate (MMF), or cannabidiol (CBD). (<b>A</b>) Principal component analysis of the gene expression profile of neurons treated with 30 µM DMF, 30 µM MMF, 6 µM CBD, or their respective vehicles (DMSO or EtOH) for 4 and 24 h. Each condition presents experimental triplicates. The x-axis represents the first principal component (PC1), and the y-axis represents the second principal component (PC2). The percentages indicated in parentheses represent the percentages of variation explained by the principal components. (<b>B</b>) Venn diagram showing the number and overlap of the differentially expressed genes (DEGs) in neurons treated with DMF or CBD for 4 and 24 h. (<b>C</b>) Heat map representing the expression levels of genes from the Nrf2 signaling pathway. Gene expression levels are represented with the log2 of the fold change (FC), indicating the ratio of the gene expression in the drug-treated cells compared to their respective vehicle. * <span class="html-italic">p</span>-adj &lt; 0.05.</p>
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<p>Transcriptomic profile of the lipopolysaccharide (LPS)-activated microglia cells treated with dimethyl fumarate (DMF), monomethyl fumarate (MMF), or cannabidiol (CBD). (<b>A</b>) Heat map representing the expression levels of genes related to inflammation. Gene expression levels are represented with the log2 of the fold change (FC), indicating the ratio of the gene expression in the LPS-treated cells compared to the vehicle or drug-treated cells compared to LPS. * <span class="html-italic">p</span>-adj &lt; 0.05. (<b>B</b>,<b>C</b>) Principal component analysis of the gene expression profile of the LPS-activated microglia cells treated with 30 µM DMF, 30 µM MMF, 6 µM CBD, or their respective vehicles (DMSO or EtOH) for 4 h (<b>B</b>) and 24 h (<b>C</b>). Each condition presents experimental triplicates. The x-axis represents the first principal component (PC1), and the y-axis represents the second principal component (PC2). The percentages indicated in parentheses represent the percentages of variation explained by the principal components.</p>
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<p>Transcriptomic modulation of the NF-kB and Nrf2 pathways in the lipopolysaccharide (LPS)-activated microglia cells treated with dimethyl fumarate (DMF), monomethyl fumarate (MMF), or cannabidiol (CBD). (<b>A</b>) Heat map representing the expression levels of genes related to the NF-kB pathway. (<b>B</b>) Heat map representing the expression levels of genes related to the Nrf2 pathway. (<b>A</b>,<b>B</b>) LPS-activated microglia cells were treated with 30 µM DMF, 30 µM MMF, 6 µM CBD, or their respective vehicles (DMSO or EtOH) for 4 and 24 h. The gene expression levels are represented with the log2 of the fold change (FC), indicating the ratio of the gene expression in the LPS-treated cells compared to the vehicle or drug-treated cells compared to LPS. * <span class="html-italic">p</span>-adj &lt; 0.05.</p>
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23 pages, 869 KiB  
Article
Synthesis of Enantiostructured Triacylglycerols Possessing a Saturated Fatty Acid, a Polyunsaturated Fatty Acid and an Active Drug Intended as Novel Prodrugs
by Lena Rós Jónsdóttir and Gudmundur G. Haraldsson
Molecules 2024, 29(23), 5745; https://doi.org/10.3390/molecules29235745 - 5 Dec 2024
Abstract
This report describes the asymmetric synthesis of a focused library of enantiopure structured triacylglycerols (TAGs) comprised of a single saturated fatty acid (C6, C8, C10, C12, C14 or C16), a pure bioactive n-3 polyunsaturated fatty acid (EPA or DHA) and a potent drug [...] Read more.
This report describes the asymmetric synthesis of a focused library of enantiopure structured triacylglycerols (TAGs) comprised of a single saturated fatty acid (C6, C8, C10, C12, C14 or C16), a pure bioactive n-3 polyunsaturated fatty acid (EPA or DHA) and a potent drug (ibuprofen or naproxen) intended as a novel type of prodrug. One of the terminal sn-1 or sn-3 positions of the glycerol backbone is occupied with a saturated fatty, the remaining one with a PUFA, and the drug entity is present in the sn-2 position. This was accomplished by a six-step chemoenzymatic approach starting from enantiopure (R)- and (S)-solketals. The highly regioselective immobilized Candida antarctica lipase (CAL-B) played a crucial role in the regiocontrol of the synthesis. All combinations, a total of 48 such prodrug TAGs, were prepared, isolated and fully characterized, along with 60 acylglycerol intermediates, obtained in very high to excellent yields. Full article
(This article belongs to the Section Organic Chemistry)
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Graphical abstract

Graphical abstract
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<p>The structure of TAG prodrugs <b>1</b> and <b>2</b> belong to the first category prodrugs, and TAG products <b>3</b> and <b>4</b> belong to the second category prodrugs.</p>
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<p>Chemoenzymatic synthesis of the first category TAG prodrug diastereomer series (<span class="html-italic">S</span>,<span class="html-italic">S′</span>)-<b>10a</b>–<b>f</b>–<b>13a</b>–<b>f</b>, starting from 1-<span class="html-italic">O</span>-benzyl-<span class="html-italic">sn</span>-glycerol. In the scheme, SFA-CO-, PUFA-CO- and Drug-CO- refer to the corresponding saturated fatty acyl, polyunsaturated fatty acyl and drug acyl group substituents, respectively. In box: (<span class="html-italic">S′</span>)-ibuprofen and (<span class="html-italic">S′</span>)-naproxen attached as esters to acylglycerols (AG).</p>
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<p>Chemoenzymatic synthesis of the first category TAG prodrug diastereomer series (<span class="html-italic">R</span>,<span class="html-italic">S′</span>)-<b>10a</b>–<b>f</b>–<b>13a</b>–<b>f</b>, starting from 3-<span class="html-italic">O</span>-benzyl-<span class="html-italic">sn</span>-glycerol. In the scheme, SFA-CO-, PUFA-CO- and Drug-CO- refer to the corresponding saturated fatty acyl, polyunsaturated fatty acyl and drug acyl group substituents, respectively. In box: (<span class="html-italic">S′</span>)-ibuprofen and (<span class="html-italic">S′</span>)-naproxen attached as esters to acylglycerols (AG).</p>
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14 pages, 5116 KiB  
Article
Enhanced Bioactivity of Quercetin–Tetrahydroisoquinoline Derivatives: Effect on Lipophilicity, Enzymes Inhibition, Antioxidant Potential, and Cytotoxicity
by Marija Vučkovski, Ana Filipović, Milka Jadranin, Lela Korićanac, Jelena Žakula, Bojan P. Bondžić and Aleksandra M. Bondžić
Int. J. Mol. Sci. 2024, 25(23), 13076; https://doi.org/10.3390/ijms252313076 - 5 Dec 2024
Viewed by 16
Abstract
Quercetin, a well-known flavonoid with significant medicinal potential, was derivatized at the C8 position with a tetrahydroisoquinoline (THIQ) moiety, and physicochemical and pharmacological properties, inhibition potential, antioxidant activity, and cytotoxicity of new compounds were evaluated. Physicochemical and pharmacological properties, including lipophilicity, membrane permeability, [...] Read more.
Quercetin, a well-known flavonoid with significant medicinal potential, was derivatized at the C8 position with a tetrahydroisoquinoline (THIQ) moiety, and physicochemical and pharmacological properties, inhibition potential, antioxidant activity, and cytotoxicity of new compounds were evaluated. Physicochemical and pharmacological properties, including lipophilicity, membrane permeability, and P-glycoprotein substrate affinity, were assessed theoretically using the SwissADME software. The metal-chelating ability of the new compounds was evaluated on metal ions Fe2+, Zn2+, and Cu2+, whose homeostasis disruption is linked to the development of Alzheimer’s disease. Inhibition potential was tested on the cholinergic enzymes acetylcholinesterase and butyrylcholinesterase, as well as Na+, K+-ATPase, an enzyme commonly overexpressed in tumours. Antioxidant potential was assessed using the DPPH assay. Cytotoxicity studies were conducted on healthy MRC-5 cells and three cancer cell lines: HeLa, MDA-231, and MDA-468. The results indicated that derivatization of quercetin with THIQ yielded compounds with lower toxicity, preserved chelating ability, improved antioxidant potential, increased selectivity toward the cholinergic enzyme butyrylcholinesterase, and enhanced inhibition potential toward Na+, K+-ATPase and butyrylcholinesterase compared to quercetin alone. Therefore, the synthesized derivatives represent compounds with an improved profile and could be promising candidates for further optimization in developing drugs for neurodegenerative and cancer diseases. Full article
(This article belongs to the Section Bioactives and Nutraceuticals)
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<p>Antioxidative activity (%) of quercetin, Q; its tetrahydroisoquinoline derivates, <b>2a</b> and <b>2b</b>; ascorbic acid; 1,2,3,4-tetrahydroisoquinoline, <b>1a</b>; and 6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline, <b>1b</b>, toward DPPH radical at 0.01, 0.1, and 1mM concentration after 30 min of incubation.</p>
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<p>UV/Vis spectra of 5 × 10<sup>−</sup><sup>5</sup> M (<b>a</b>) Q, (<b>b</b>) <b>2a</b>, and (<b>c</b>) <b>2b</b> in the presence of different metal ions: Cu<sup>2+</sup> (red), Zn<sup>2+</sup> (blue), and Fe<sup>2+</sup> (pink) at concentration 1 × 10<sup>−</sup><sup>4</sup> M, recorded 30 min after the addition of metal ions (t = 30 min).</p>
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<p><math display="inline"><semantics> <mrow> <mo>∆</mo> <mrow> <mi mathvariant="normal">A</mi> <mo>=</mo> <mi mathvariant="normal">f</mi> </mrow> <mfenced> <mrow> <msub> <mi mathvariant="normal">c</mi> <mrow> <msup> <mi mathvariant="normal">M</mi> <mrow> <mrow> <mn>2</mn> <mo>+</mo> </mrow> </mrow> </msup> </mrow> </msub> <msub> <mrow> <mrow> <mo>/</mo> <mi mathvariant="normal">c</mi> </mrow> </mrow> <mrow> <mi>compound</mi> </mrow> </msub> </mrow> </mfenced> </mrow> </semantics></math> at absorption maxima of complexes for different metal ions: (<b>a</b>) Cu<sup>2+</sup>, (<b>b</b>) Zn<sup>2+</sup>, and (<b>c</b>) Fe<sup>2+</sup>; <b>2a</b>—black squares; <b>2b</b>—red circles.</p>
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<p>Viability of HeLa cells obtained by SRB assay 24, 48, and 72 h after treatment with quercetin (<b>a</b>), compound <b>2a</b> (<b>b</b>), and compound <b>2b</b> (<b>c</b>). Applied concentrations were 1 × 10<sup>−6</sup>, 1 × 10<sup>−5</sup>, and 1 × 10<sup>−4</sup> M. Data obtained from four experiments are presented as mean ± S.D. Asterisks indicate statistical significance compared to the untreated control: * 0.01 &lt; <span class="html-italic">p</span> &lt; 0.05; ** 0.001 &lt; <span class="html-italic">p</span> &lt; 0.01; *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Synthesis of quercetin derivatives <b>2a</b> and <b>2b</b>.</p>
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15 pages, 4917 KiB  
Article
Optimization of Ultraviolet-B Treatment for Enrichment of Total Flavonoids in Buckwheat Sprouts Using Response Surface Methodology and Study on Its Metabolic Mechanism
by Jiyuan Xue, Meixia Hu, Jia Yang, Weiming Fang and Yongqi Yin
Foods 2024, 13(23), 3928; https://doi.org/10.3390/foods13233928 - 5 Dec 2024
Viewed by 50
Abstract
Buckwheat possesses significant nutritional content and contains different bioactive compounds, such as total flavonoids, which enhance its appeal to consumers. This study employed single-factor experiments and the response surface methodology to identify the optimal germination conditions for enhancing the total flavonoid content in [...] Read more.
Buckwheat possesses significant nutritional content and contains different bioactive compounds, such as total flavonoids, which enhance its appeal to consumers. This study employed single-factor experiments and the response surface methodology to identify the optimal germination conditions for enhancing the total flavonoid content in buckwheat sprouts through ultraviolet-B treatment. The research showed that buckwheat sprouts germinated for 3 days at a temperature of 28.7 °C while being exposed to ultraviolet-B radiation at an intensity of 30.0 μmol·m−2·s−1 for 7.6 h per day during the germination period resulted in the highest total flavonoid content of 1872.84 μg/g fresh weight. Under these specified conditions, ultraviolet-B treatment significantly elevated the activity and gene expression levels of enzymes related to the phenylpropanoid metabolic pathway, including phenylalanine ammonia-lyase, cinnamic acid 4-hydroxylase, 4-coumarate coenzyme A ligase, and chalcone isomerase. Ultraviolet-B treatment caused oxidative damage to buckwheat sprouts and inhibited their growth, but ultraviolet-B treatment also enhanced the activity of key enzymes in the antioxidant system, such as catalase, peroxidase, superoxide dismutase, and ascorbate peroxidase. This research provided a technical reference and theoretical support for enhancing the isoflavone content in buckwheat sprouts through ultraviolet-B treatment. Full article
(This article belongs to the Section Food Engineering and Technology)
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<p>Effects of germination time (<b>a</b>), UV-B treatment time (<b>b</b>), germination temperature (<b>c</b>), and UV-B intensity (<b>d</b>) on total flavonoid content. Different lowercase letters represent significant differences between treatment groups.</p>
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<p>Plots of the interaction of variables on total flavonoid enrichment. Response surface plot of UV−B treatment time and Germination time (<b>a</b>), contour map of UV−B treatment time and Germination time (<b>b</b>), response surface plot of Germination temperature and Germination time (<b>c</b>), contour map of Germination temperature and Germination time (<b>d</b>), response surface plot of UV−B intensity and Germination time (<b>e</b>), contour map of UV−B intensity and Germination time (<b>f</b>), response surface plot of Germination temperature and UV−B treatment time (<b>g</b>), contour map of Germination temperature and UV−B treatment time (<b>h</b>), response surface plot of UV−B intensity and UV−B treatment time (<b>i</b>), contour map of UV−B intensity and UV−B treatment time (<b>j</b>), response surface plot of UV−B intensity and Germination temperature (<b>k</b>) and contour map of UV−B intensity and Germination temperature (<b>l</b>).</p>
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<p>Effects of UV-B treatment on the content of total flavonoid content (<b>a</b>), total phenolic (<b>b</b>), and growth performance (<b>c</b>) of buckwheat sprouts. * Indicates significant differences in indicators among treatments according to ANOVA and Tukey’s test (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effect of UV-B treatment on the content of MDA (<b>a</b>), <math display="inline"><semantics> <mrow> <msubsup> <mrow> <mi>O</mi> </mrow> <mrow> <mn>2</mn> </mrow> <mrow> <mo>−</mo> </mrow> </msubsup> </mrow> </semantics></math>. (<b>b</b>), and H<sub>2</sub>O<sub>2</sub> (<b>c</b>), ABTS free-radical scavenging rate (<b>d</b>), DPPH scavenging capacity (<b>e</b>), and FRAP (<b>f</b>). * Indicates significant differences in indicators among treatments according to ANOVA and Tukey’s test (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effects of UV-B treatment on the activity of APX (<b>a</b>), SOD (<b>b</b>), POD (<b>c</b>), and CAT (<b>d</b>), and the relative expression of <span class="html-italic">FtAPX</span> (<b>e</b>), <span class="html-italic">FtSOD</span> (<b>f</b>), <span class="html-italic">FtPOD</span> (<b>g</b>), and <span class="html-italic">FtCAT</span> (<b>h</b>). * Indicates significant differences in indicators among treatments according to ANOVA and Tukey’s test (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effects of UV-B treatment on the activity of 4CL (<b>a</b>), PAL (<b>b</b>), C4H (<b>c</b>), and CHI (<b>d</b>), and the relative expression of <span class="html-italic">Ft4CL</span> (<b>e</b>), <span class="html-italic">FtPAL</span> (<b>f</b>), <span class="html-italic">FtC4H</span> (<b>g</b>), <span class="html-italic">FtCHI</span> (<b>h</b>), <span class="html-italic">FtCHS</span> (<b>i</b>) and <span class="html-italic">FtF3H</span> (<b>j</b>). * Indicates significant differences in indicators among treatments according to ANOVA and Tukey’s test (<span class="html-italic">p</span> &lt; 0.05).</p>
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16 pages, 3250 KiB  
Article
Enhancing Lettuce (Lactuca sativa) Productivity: Foliar Sprayed Fe-Alg-CaCO3 MPs as Fertilizers for Aquaponics Cultivation
by Davide Frassine, Roberto Braglia, Francesco Scuderi, Enrico Luigi Redi, Federica Valentini, Michela Relucenti, Irene Angela Colasanti, Andrea Macchia, Ivo Allegrini, Angelo Gismondi, Gabriele Di Marco and Antonella Canini
Plants 2024, 13(23), 3416; https://doi.org/10.3390/plants13233416 - 5 Dec 2024
Viewed by 58
Abstract
Aquaponics is an innovative agricultural method combining aquaculture and hydroponics. However, this balance can lead to the gradual depletion of essential micronutrients, particularly iron. Over time, decreasing iron levels can negatively impact plant health and productivity, making the monitoring and management of iron [...] Read more.
Aquaponics is an innovative agricultural method combining aquaculture and hydroponics. However, this balance can lead to the gradual depletion of essential micronutrients, particularly iron. Over time, decreasing iron levels can negatively impact plant health and productivity, making the monitoring and management of iron in aquaponic systems vital. This study investigates the use of Fe-Alg-CaCO3 microparticles (MPs) as foliar fertilizer on lettuce plants in an aquaponic system. The research investigated Lactuca sativa L. cv. Foglia di Quercia Verde plants as the experimental cultivar. Three iron concentrations (10, 50, and 250 ppm) were tested, with 15 plants per treatment group, plus a control group receiving only sterile double-distilled water. The Fe-Alg-CaCO3 MPs and ultrapure water were applied directly to the leaves using a specialized nebulizer. Foliar nebulization was chosen for its precision and minimal resource use, aligning with the sustainability goals of aquaponic cultivation. The research evaluated rosette diameter, root length, fresh weight, soluble solids concentration, levels of photosynthetic pigments, and phenolic and flavonoid content. The 250 ppm treatment produced the most notable enhancements in both biomass yield and quality, highlighting the potential of precision fertilizers to boost sustainability and efficiency in aquaponic systems. In fact, the most significant increases involved biomass production, particularly in the edible portions, along with photosynthetic pigment levels. Additionally, the analysis of secondary metabolite content, such as phenols and flavonoids, revealed no reduction compared to the control group, meaning that the proposed fertilizer did not negatively impact the biosynthetic pathways of these bioactive compounds. This study opens new possibilities in aquaponics cultivation, highlighting the potential of precision fertilizers to enhance sustainability and productivity in soilless agriculture. Full article
(This article belongs to the Section Horticultural Science and Ornamental Plants)
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<p>(<b>A</b>) SEM magnification 10 K: a microsphere is illustrated, it has a rough surface, due to incomplete fusion of constituent subunits. (<b>B</b>) SEM magnification 10 K: this image shows the microsphere inner cavity; the surface is roughest than (<b>A</b>), and constituent subunits are well visible; they have a minimum diameter of 100 nm, inset. (<b>C</b>) Region of interest (ROI) for EDX analysis. (<b>D</b>) EDX analysis element graph shows the presence of calcium, oxygen, and a small amount of Fe. Platinum, copper, and silver peaks are due to the platinum coating, the copper grid where the sample is placed, and the aluminum supporting stub.</p>
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<p>Plant samples collected on the 55th day from sowing at the end of each Fe-Alg-CaCO<sub>3</sub> MPs treatment (CT, 10, 50, and 250 ppm).</p>
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<p>Morpho-biometrical parameters. In detail, (<b>A</b>) rosette diameter; (<b>B</b>) root length; (<b>C</b>) rosette fresh weight; (<b>D</b>) root fresh weight. The <span class="html-italic">x</span>-axis denotes the treatments, while the <span class="html-italic">y</span>-axis represents the units of measurement. The significance resulting from the comparisons between the various treatments is indicated by asterisks: * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.005.</p>
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<p>Qualitative and quantitative data from spectrophotometric assays. In detail, (<b>A</b>) chlorophyll <span class="html-italic">a</span>; (<b>B</b>) chlorophyll <span class="html-italic">b</span>; (<b>C</b>) total chlorophyll; (<b>D</b>) carotenoids; (<b>E</b>) total phenolic content; (<b>F</b>) total flavonoid content. The <span class="html-italic">x</span>-axis denotes the treatments, and the <span class="html-italic">y</span>-axis represents units of measurement. The significance resulting from the comparisons between the various treatments is indicated by asterisks: * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.005.</p>
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<p>Representative flow chart for the biomineralization synthetic approach able to produce CaCO<sub>3</sub> NPs (i.e., the chemical precursor) for the second step to obtain functionalized Fe-Alg-CaCO<sub>3</sub> MPs, which can be able to act as micro-carriers for plant nutrients. Created with <a href="http://BioRender.com" target="_blank">BioRender.com</a>.</p>
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19 pages, 3649 KiB  
Review
Unlocking the Therapeutic Potential of Adipose-Derived Stem Cell Secretome in Oral and Maxillofacial Medicine: A Composition-Based Perspective
by Chiara Giannasi, Francesca Cadelano, Elena Della Morte, Camilla Baserga, Camilla Mazzucato, Stefania Niada and Alessandro Baj
Biology 2024, 13(12), 1016; https://doi.org/10.3390/biology13121016 - 5 Dec 2024
Viewed by 85
Abstract
The adipose-derived stem cell (ADSC) secretome is widely studied for its immunomodulatory and regenerative properties, yet its potential in maxillofacial medicine remains largely underexplored. This review takes a composition-driven approach, beginning with a list of chemokines, cytokines, receptors, and inflammatory and growth factors [...] Read more.
The adipose-derived stem cell (ADSC) secretome is widely studied for its immunomodulatory and regenerative properties, yet its potential in maxillofacial medicine remains largely underexplored. This review takes a composition-driven approach, beginning with a list of chemokines, cytokines, receptors, and inflammatory and growth factors quantified in the ADSC secretome to infer its potential applications in this medical field. First, a review of the literature confirmed the presence of 107 bioactive factors in the secretome of ADSCs or other types of mesenchymal stem cells. This list was then analyzed using the Search Tool for Retrieval of Interacting Genes/Proteins (STRING) software, revealing 844 enriched biological processes. From these, key processes were categorized into three major clinical application areas: immunoregulation (73 factors), bone regeneration (13 factors), and wound healing and soft tissue regeneration (27 factors), with several factors relevant to more than one area. The most relevant molecules were discussed in the context of existing literature to explore their therapeutic potential based on available evidence. Among these, TGFB1, IL10, and CSF2 have been shown to modulate immune and inflammatory responses, while OPG, IL6, HGF, and TIMP1 contribute to bone regeneration and tissue repair. Although the ADSC secretome holds great promise in oral and maxillofacial medicine, further research is needed to optimize its application and validate its clinical efficacy. Full article
(This article belongs to the Special Issue Advances in Biological Research of Adipose-Derived Stem Cells)
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<p>(<b>a</b>) Overview of the protein−protein interaction network of the 107 factors quantified in the ADSC secretome, generated using STRING software (version 12.0) with the interaction score threshold set to 0.900 for the highest confidence level. (<b>b</b>) Histogram showing the fold enrichment and false discovery rate (FDR) for seven selected pathways within the enriched biological processes highlighted by STRING analysis (<a href="#app1-biology-13-01016" class="html-app">Supplementary S3</a>). Fold enrichment was calculated as follows: (number of observed proteins/number of proteins in the list)/(background gene count/number of protein-coding genes).</p>
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<p>(<b>a</b>) Overview of the protein−protein interaction network of the 107 factors quantified in the ADSC secretome, generated using STRING software (version 12.0) with the interaction score threshold set to 0.900 for the highest confidence level. (<b>b</b>) Histogram showing the fold enrichment and false discovery rate (FDR) for seven selected pathways within the enriched biological processes highlighted by STRING analysis (<a href="#app1-biology-13-01016" class="html-app">Supplementary S3</a>). Fold enrichment was calculated as follows: (number of observed proteins/number of proteins in the list)/(background gene count/number of protein-coding genes).</p>
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<p>Venn diagram showing the unique and shared factors across the three major fields of ADSC secretome application in oral and maxillofacial medicine: immunomodulation, bone regeneration, wound healing, and soft tissue regeneration.</p>
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<p>Protein–protein interaction network of the 73 factors associated with the pathways GO:0006955 (immune response) and GO:0006954 (inflammatory response). The network visualization was generated using STRING software (version 12.0, minimum required interaction score set to 0.900).</p>
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<p>Protein–protein interaction network of the 13 factors associated with the pathways GO:0045667 (regulation of osteoblast differentiation) and GO:0045670 (regulation of osteoclast differentiation). The network visualization was generated using STRING software (version 12.0, minimum required interaction score set to 0.900).</p>
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<p>Protein–protein interaction network of the 27 factors associated with the pathways GO:0042060 (wound healing), GO:0031099 (regeneration), and GO:0050678 (regulation of epithelial cell proliferation). The network visualization was generated using STRING software (version 12.0, minimum required interaction score set to 0.900).</p>
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19 pages, 1053 KiB  
Article
Effect of the Storage Conditions on the Microbiological Quality and Selected Bioactive Compound Content in Fruit Mousses for Infants and Young Children
by Aleksandra Purkiewicz, Patryk Wiśniewski, Małgorzata Tańska, Gulden Goksen and Renata Pietrzak-Fiećko
Appl. Sci. 2024, 14(23), 11347; https://doi.org/10.3390/app142311347 - 5 Dec 2024
Viewed by 126
Abstract
Fruit mousses, as low-processed products, are highly susceptible to external conditions, and storage leads to the degradation of bioactive compounds, particularly phenolic compounds and vitamins, as well as promoting the growth of yeasts and molds. This study investigated the impact of storage conditions [...] Read more.
Fruit mousses, as low-processed products, are highly susceptible to external conditions, and storage leads to the degradation of bioactive compounds, particularly phenolic compounds and vitamins, as well as promoting the growth of yeasts and molds. This study investigated the impact of storage conditions on the microbiological quality and degradation of selected bioactive compounds in fruit mousses from various producers (from apples, pears, and multi-components). Total phenolic (TPC) and total flavonoid (TFC) contents, vitamin C level, antioxidant capacity (AC, measured by the DPPH assay), and concentrations of macro- and microminerals were evaluated in fresh mousses and those stored for 48 h at 23 °C and 4 °C. Changes in total aerobic mesophilic bacteria (TAMB), yeast and mold counts, and selected microbial groups were also checked. It was found that the analyzed compounds varied depending on the components of the mousses. Multi-component mousses contained the highest levels of TPC, TFC, and vitamin C, and had 2–5 times higher AC values compared to apple and pear mousses. Storage at room temperature resulted in TFC lowering of up to 25% in apple mousses and vitamin C reductions of up to 22% in multi-component mousses. During refrigerated storage, the highest losses were observed in pear mousses, with TPC decreasing by up to 13% and vitamin C by up to 11%. Among the minerals, magnesium and zinc levels decreased most significantly in apple mousses stored at 23 °C (up to 33% and up to 29%, respectively). Microbiological analysis revealed variability in TAMB, yeast, and mold counts, with refrigeration (4 °C) generally limiting microbial growth compared to room temperature (23 °C). Notably, no pathogenic bacteria were detected under any storage conditions, and the mousses retained a high microbiological quality even after room-temperature storage. Full article
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<p>Antioxidant capacity (DPPH assay values) of the studied fruit mousses (fresh and stored at room temperature and refrigerated temperature). Different lowercase letters (a,b,c,d) placed above the bars for mousses of the same type, separate for fresh, stored at 23 °C, and stored at 4 °C, indicate significant differences (<span class="html-italic">p</span> ≤ 0.05), whereas different uppercase letters (A,B,C) placed above the bars for various storage conditions, separate for each mousse sample, indicate significant differences (<span class="html-italic">p</span> ≤ 0.05).</p>
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<p>Principal component analysis (PCA) results presented as score plots of the analyzed fresh fruit mousses (<b>a</b>,<b>c</b>,<b>e</b>) and loading plots of the analyzed parameters (TPC, TFC, DPPH, sum of macrominerals, and sum of microminerals) in the analyzed fresh fruit mousses (<b>b</b>,<b>d</b>,<b>f</b>). Explanations: A1, A2, A3, A4—apple mousses; P1, P2, P3, P4—pear mousses; M1, M2, M3, M4—multi-component mousses; F—fresh mousses; CT—mousses stored at refrigerated temperature (48 h/4 °C); RT—mousses stored at room temperature (48 h/23 °C).</p>
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21 pages, 3538 KiB  
Article
Hemostatic Antimicrobial Hydrogels Based on Silicon, Iron, Zinc, and Boron Glycerolates for Wound Healing Applications
by Tat’yana Khonina, Semyon Alekseenko, Elena Shadrina, Il’ya Ganebnykh, Alexander Mekhaev, Leonid Larionov, Maria Dobrinskaya, Nadezhda Izmozherova, Irina Antropova, Maxim Karabanalov, Muza Kokhan, Natali’ya Evstigneeva and Oleg Chupakhin
Gels 2024, 10(12), 795; https://doi.org/10.3390/gels10120795 - 5 Dec 2024
Viewed by 126
Abstract
The use of glycerolates of biogenic elements as biocompatible precursors in sol–gel synthesis is an innovative direction and opens up new scientific and practical prospects in chemistry and technology of producing practically important biomedical materials, including hemostatic, antimicrobial, and wound healing materials. Using [...] Read more.
The use of glycerolates of biogenic elements as biocompatible precursors in sol–gel synthesis is an innovative direction and opens up new scientific and practical prospects in chemistry and technology of producing practically important biomedical materials, including hemostatic, antimicrobial, and wound healing materials. Using biocompatible precursors, silicon, zinc, boron, and iron glycerolates, new bioactive nanocomposite hydrogels were obtained by the sol–gel method. The composition and structural features of the hydrogels were studied using a complex of modern analytical techniques, including TEM, XRD, AES, and ESI MS. Hemostatic activity of the hydrogels was studied in the in vivo experiments; using the example of silicon-iron-zinc-boron glycerolates hydrogel, primary toxicological studies were carried out. Antimicrobial properties of hydrogels were studied using the agar diffusion method. The structural features of hydrogels and their relationship to medical and biological properties were revealed. It was shown that glycerolates hydrogels are non-toxic, and exhibit pronounced hemostatic activity, generally comparable to the commercial hemostatic drug Capramine. Antimicrobial activity is more pronounced for silicon-iron-zinc-boron and silicon-iron-boron glycerolates gel. The results obtained indicate that these glycerolates hydrogels are potential hemostatic and antibiotic-independent antimicrobial agents for topical wound healing applications in medical and veterinary practice. Full article
(This article belongs to the Special Issue Designing Gels for Antibacterial and Antiviral Agents)
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Graphical abstract

Graphical abstract
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<p>The biocompatible precursors used in the sol–gel synthesis of the glycerolates hydrogels: (<b>a</b>) silicon tetraglycerolate, (<b>b</b>) boron bisglycerolates, (<b>c</b>) zinc monoglycerolate, and (<b>d</b>) iron(III) monoglycerolate.</p>
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<p>IR spectra of (<b>a</b>) Si-Fe– (for comparison), (<b>b</b>) Si-Fe-Zn–, (<b>c</b>) Si-Fe-B–, and (<b>d</b>) Si-Fe-Zn-B–gel.</p>
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<p>TEM micrographs of dried suspension: (<b>a</b>) Si-Fe–(for comparison), (<b>b</b>) Si-Fe-Zn–, (<b>c</b>) Si-Fe-B–, (<b>d</b>) Si-Fe-Zn-B–gel in ethanol. (<b>a</b>–<b>d</b>) High-resolution TEM image, inserts show electron diffraction area.</p>
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<p>Thermal analysis data for (<b>a</b>) Si-Fe–, (<b>b</b>) Si-Fe-Zn–, (<b>c</b>) Si-Fe-B–, (<b>d</b>) Si-Fe-Zn-B–gel.</p>
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<p>ESI mass spectrum in negative mode of Si-Fe-Zn-B–gel liquid medium (* averaged for scan number from 60 to 80).</p>
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<p>XRD patterns of extracted solid phase of (<b>a</b>) Si-Fe– (for comparison), (<b>b</b>) Si-Fe-Zn–gel, (<b>c</b>) Si-Fe-B–gel, and (<b>d</b>) Si-Fe-Zn-B–gel.</p>
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<p>Histological analysis of Si-Fe-Zn-B–gel treated group 14 days after administration, hematoxylin-eosin, magnification ×100.</p>
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<p>Comparative assessment of bleeding time in mice with incised liver wounds.</p>
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<p>Strain growth inhibition zones: (<b>1</b>) <span class="html-italic">E. coli</span> ATCC 8739; (<b>2</b>) <span class="html-italic">P. aeruginosa</span> ATCC 9027; (<b>3</b>) clinical strain <span class="html-italic">S. aureus</span> (MRSA); (<b>4</b>) <span class="html-italic">S. pyogenes</span> ATCC 19615. (<b>5</b>) <span class="html-italic">C. albicans</span> RCPF <sub>Y</sub>-401/NCTC-885-653: (<b>a</b>) Si-Fe-Zn-B-gel; (<b>b</b>) positive control; (<b>c</b>) silicon glycerolates gel (negative control).</p>
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18 pages, 3157 KiB  
Article
Bioactivated Glucoraphanin Improves Cell Survival, Upregulating Phospho-AKT, and Modulates Genes Involved in DNA Repair in an In Vitro Alzheimer’s Disease Model: A Network-Transcriptomic Analysis
by Aurelio Minuti, Emanuela Mazzon, Renato Iori, Luigi Chiricosta and Osvaldo Artimagnella
Nutrients 2024, 16(23), 4202; https://doi.org/10.3390/nu16234202 - 5 Dec 2024
Viewed by 177
Abstract
Background/Objectives: Alzheimer’s disease (AD) is one of the most common neurodegenerative diseases, for which a definitive cure is still missing. Recently, natural compounds have been investigated for their possible neuroprotective role, including the bioactivated product of glucoraphanin (GRA), the sulforaphane (SFN), which is [...] Read more.
Background/Objectives: Alzheimer’s disease (AD) is one of the most common neurodegenerative diseases, for which a definitive cure is still missing. Recently, natural compounds have been investigated for their possible neuroprotective role, including the bioactivated product of glucoraphanin (GRA), the sulforaphane (SFN), which is highly rich in cruciferous vegetables. It is known that SFN alleviates neuronal dysfunction, apoptosis, and oxidative stress in the brain. In the light of this evidence, the aim of this study was to investigate the molecular effects of SFN pre-treatment in differentiated SH-SY5Y neurons exposed to β-amyloid (Aβ). Methods: To this end, we first evaluated first cell viability via the Thiazolyl Blue Tetrazolium Bromide (MTT) assay, and then we analyzed the transcriptomic profiles by next-generation sequencing (NGS). Finally, we used a network analysis in order to understand which biological processes are affected, validating them by Western blot assay. Results: SFN pre-treatment counteracted Aβ-induced loss of cell viability. The network-transcriptomic analysis revealed that SFN upregulates genes associated with DNA repair, such as ABRAXAS1, BRCA1, BRCA2, CDKN1A, FANCA, FANCD2, FANCE, NBN, and XPC. Finally, SFN also increased the phosphorylation of AKT, which is associated with DNA repair and cell survival. Conclusions: These data suggest that SFN is a natural compound that could be suitable in the prevention of AD, thanks to its neuroprotective role in increasing cell survival, potentially restoring DNA damage induced by Aβ exposure. Full article
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<p>Chemical structures of glucoraphanin (GRA) and sulforaphane (SFN), along with a graphical representation of SFN’s biochemistry. Glucoraphanin is converted into sulforaphane through the action of the enzyme myrosinase. The chemical structures of GRA and SFN were obtained using PubChem compound records (accessed on 28 November 2024). Information about the molecule’s properties is available at the following links: <a href="https://pubchem.ncbi.nlm.nih.gov/compound/9548634" target="_blank">https://pubchem.ncbi.nlm.nih.gov/compound/9548634</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/compound/Sulforaphane" target="_blank">https://pubchem.ncbi.nlm.nih.gov/compound/Sulforaphane</a> (both accessed on 28 November 2024).</p>
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<p>(<b>A</b>) Cell viability after SFN treatment. SFN at the concentrations tested did not affect cell viability. (<b>B</b>) Exposure with Aβ 10 µM decreased the cell viability of RA-SH-SY5Y-differentiated neurons. Interestingly, SFN at a concentration of 5 µM was able to increase cell viability. <span class="html-italic">N</span> = 6 independent biological replicates. Data are expressed as mean ± Standard Error of the Mean (SEM). ** <span class="html-italic">p</span> &lt; 0.01; **** <span class="html-italic">p</span> &lt; 0.0001. The complete primary data are reported in <a href="#app1-nutrients-16-04202" class="html-app">Table S1</a>.</p>
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<p>Cluster of over-represented biological terms obtained by ShinyGO. The terms are sorted by fold enrichment after FDR. The color palette shows the log FDR. The size of the bubble represents the number of DEGs in the cluster.</p>
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<p>Final subnetworks of DEGs included in the network with the highest diameter excluding ribosomal proteins. The color of the nodes represents the deregulation of DEGs; green nodes are downregulated DEGs while red nodes are upregulated.</p>
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<p>Degree and betweenness score values for DEGs included in the network with the highest diameter excluding ribosomal proteins. The color of the nodes represents the deregulation of DEGs so that green nodes are downregulated while red nodes are upregulated DEGs. Labels are shown only for nodes with betweenness higher than 0.5 and degree higher than 2 for higher readability.</p>
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<p>KEGG pathway terms obtained by ShinyGO. The terms are sorted by FDR. The color palette shows the fold enrichment. The size of the bubble represents the number of DEGs in the cluster.</p>
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<p>Western blot analysis for p-AKT and cleaved-CASP3. (<b>Left</b>) Aβ treatment induced a light decrease in p-AKT, while SFN increased p-AKT levels. (<b>Right</b>) In addition, SFN treatment reduced levels of cleaved-CASP3 compared to Aβ. <span class="html-italic">N</span> = 3 independent biological replicates. The results are indicated by mean ± Standard Error of the Mean (SEM). * <span class="html-italic">p</span> &lt; 0.05. The complete primary data are reported in <a href="#app1-nutrients-16-04202" class="html-app">Table S1</a>.</p>
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<p>Schematic representation of putative molecular mechanism of how SFN could be involved in DNA repair process and cell survival in AD model. In red we indicated all genes/proteins upregulated. The arrow in red indicates the upregulation of DNA repair and cell survival process. β-amyloid structure (9CZP) was retrieved by PDB. Figure was drawn using vector image bank of Servier Medical Art by Servier (<a href="http://smart.servier.com" target="_blank">smart.servier.com</a>) (accessed on 3 November 2024). Licensed under Creative Commons Attribution 3.0 Unported License (<a href="http://creativecommons.org/licenses/by/3.0/" target="_blank">creativecommons.org/licenses/by/3.0/</a>) (accessed on 3 November 2024).</p>
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