Search Results (17)

Background/Objectives: Ampelopsis grossedentata, a novel food resource (vine tea), is a therapeutically valuable species wherein all parts exhibit bioactive potential. The therapeutic value and health benefits of A. grossedentata are rooted in its metabolomic profile, yet the metabolites in its various parts remain incompletely characterized. In this study, the tissue-specific biochemical properties and metabolic profiles of A. grossedentata were systematically characterized to identify the optimal tissue type for harvesting, with the goal of maximizing the production of bioactive compounds. Methods: The biochemical properties of various A. grossedentata tissues were assessed using a spectrophotometer, and their metabolic profiles were characterized through LC-MS/MS-based untargeted metabolomics. Results: The results demonstrate that, compared with the stems and roots, the tender leaves and floral tissues contained a higher TFC and TPC, correlating with superior antioxidant activity (DPPH, ABTS, and FRAP). A metabolic analysis revealed that the tender leaves accumulated multiple bioactive flavonoids beyond dihydromyricetin, including naringenin 7-O-beta-D-glucoside, (-)-epigallocatechin, tricetin, and cyanidin 3-O-sophoroside. The floral tissues displayed a comparable antioxidant capacity and dihydromyricetin level to the leaves, as well as unique compounds, such as leucodelphinidin, naringenin, epicatechin, dihydroquercetin, astilbin, and myricitrin. A metabolomic analysis further demonstrated a high accumulation of L-glutamine and L-glutamic acid in the floral tissues, potentially contributing to the characteristic umami flavor profile of vine tea made from A. grossedentata flowers. Conclusions:A. grossedentata flowers could be considered a promising raw material for developing novel functional foods and premium herbal tea products, as they possess superior antioxidant activity and abundant flavonoids and amino acids. Full article

Lung cancer, particularly non-small-cell lung cancer (NSCLC), remains a leading cause of cancer-related mortality, with cisplatin-based chemotherapy being a standard treatment. However, the development of chemoresistance significantly limits its efficacy, necessitating alternative therapeutic approaches. Here, we demonstrate the anticancer effects of the extracts of Allium pseudojaponicum Makino (APE), a salt-tolerant plant, in cisplatin-resistant NSCLC. Metabolite profiling using UPLC-Q-TOF-MS^E identified 13 major compounds, predominantly alkaloids (71.65%) and flavonoids (8.81%), with key bioactive constituents such as lycorine (29.81%), tazettine (17.22%), and tricetin (8.19%). APE significantly inhibited cell viability in A549 and H460 cells, reducing viability to 38.6% (A549-Ctr), 37.2% (A549-CR), 28.4% (H460-Ctr), and 30.4% (H460-CR) at 40 µg/mL after 48 h. APE also suppressed colony formation by over 90% in both 2D and soft agar assays, while showing no cytotoxicity in normal human keratinocytes up to 80 µg/mL. Flow cytometry analysis revealed APE-induced G1 phase arrest, with the G1 population increasing from 50.4% to 56.6% (A549-Ctr) and 47.5% to 58.4% (A549-CR), accompanied by reduced S phase populations. This effect was associated with the downregulation of G1/S transition regulators, including cyclin D1, CDK4, cyclin E, and CDK2. Furthermore, proteomic analysis identified STAT3 signaling as a major target of APE; APE decreased phosphorylated STAT3 and c-Myc expression, and STAT3 knockdown phenocopied the effects of APE. These findings highlight the potential of APE as a natural product-based therapeutic strategy for overcoming cisplatin resistance in NSCLC. Full article

Background: Scientific and rational fertilizer management can not only improve the yield and quality of hazelnut (Corylus heterophylla × Corylus avellana) but also reduce the negative impact on the environment. Methods: Liquid Chromatography–tandem Mass Spectrometry (LC-MS/MS) technology was used to reveal the contents of various metabolites in hazelnut seedlings, and differential metabolites were screened by principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA). Results: The results showed that a total of 178 up-regulated differential metabolites (Fold change > 1) and 175 down-regulated differential metabolites (Fold change < 1) were detected in 6 comparison groups (DWF0 vs. DWF4, DWF0 vs. DWF5, DWF0 vs. DWF6, DWF4 vs. DWF5, DWF4 vs. DWF6, DWF5 vs. DWF6). Interestingly, the flavonoid metabolic pathway was dramatically enriched, and it was involved in each fertilization combination. The metabolites of the flavonoid pathway in different fertilized and unfertilized groups were compared and analyzed, which displayed that metabolites tricetin, eriodictyol, garbanzol, apigenin, and biochanin A were significantly up-regulated, while garbanzol and astraglin were significantly down-regulated. More interestingly, the determination of flavonoid content and real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) displayed that the application of trace element water-soluble fertilizer could significantly enhance the flavonoid content and the expression of genes related to the flavonoid biosynthesis pathway, such as phenylalanine ammonia-lyase (PAL) and cinnamate 4-hydroxylase (C4H), with the DWF4 treatment displaying the most significant values. Conclusions: Overall, the application of trace element water-soluble fertilizer (especially the DWF4 treatment) markedly affected the changes in key metabolites of the flavonoid pathway and the expression levels of key genes, thus promoting the growth and development of the hazelnut, which offers an important starting point for future analysis through genetic engineering. Full article

Cereal crops are important staple foods, and their defense metabolites hold significant research importance. In this study, we employed LC-MS-based untargeted and widely-targeted metabolomics to profile the leaf metabolome of nine cereal species, including rice, wheat, maize, barley, sorghum, common oat, foxtail millet, broomcorn millet, and adlay. A total of 9869 features were detected, among them, 1131 were annotated, encompassing 18 classes such as flavonoids, lipids, and alkaloids. Results revealed that 531 metabolites were detected in all species, while each cereal crop possessed 4 to 12 unique metabolites. Focusing on defense metabolites, we identified eight benzoxazinoids uniquely present in maize, wheat, and adlay. Hierarchical clustering based on metabolite abundance divided all metabolites into nine clusters, and subsequent pathway enrichment analysis revealed that the stress-related flavonoid biosynthesis pathway was enriched in multiple clusters. Further analysis showed that four downstream compounds of HBOA (2-hydroxy-1,4-benzoxazin-3-one) in the benzoxazinoid biosynthesis pathway were enriched in maize. Wheat uniquely accumulated the 4′-methylated product of tricin, trimethoxytricetin, whereas adlay accumulated the tricin precursor tricetin in the flavonoid biosynthesis pathway. In summary, this study elucidates the metabolic diversity in defense metabolites among various cereal crops, providing valuable background information for the improvement of stress resistance in cereal crops. Full article

The color of Zanthoxylum bungeanum Maxim. (Z. bungeanum) is a key quality indicator and a factor limiting the development of its industry. However, the underlying mechanisms governing color formation remain largely unexplored. In this study, an integrative analysis of transcriptome and metabolome profiles was conducted across four developmental stages to elucidate the color formation mechanism in Z. bungeanum. A total of 137 flavonoids were identified as the fruits ripened, with high levels of differentially accumulated metabolites (DAMs), including tricetin and (-)-epigallocatechin, which were strongly associated with color formation. This suggests their significant contribution to the pigmentation process. Nine differentially expressed genes (DEGs) were identified as candidate genes involved in color development. Additionally, 15 transcription factors (TFs) (12 MYB and 3 bHLH) exhibited expression patterns similar to those of structural genes in the flavonoid biosynthetic pathway, indicating their role in regulating flavonoid synthesis. The bioinformatics analysis of three key flavonoid synthesis genes—ZbCHI, ZbFLS, and ZbANR—revealed that all three proteins exhibit hydrophobic structures without transmembrane domains. Among them, ZbANR possesses signal peptide regions, whereas ZbCHI and ZbFLS do not. Subcellular localization predictions suggest that ZbCHI is most likely localized in the chloroplast, ZbFLS in the cytoplasm, and ZbANR in the membrane. Functional analyses revealed that their transient expression in Nicotiana benthamiana (N. benthamiana) increased the flavonoid content, with ZbANR overexpression producing a distinct white phenotype in the plants. This study enriches transcriptomic data and provides a comprehensive understanding of flavonoid metabolism and the molecular basis of color formation in Z. bungeanum, offering a valuable theoretical foundation for future breeding programs. Full article

Raisins are an important source of polyphenolic compounds in plant foods, and polyphenols are associated with antioxidant and anti-aging activity. In this work, 628 polyphenols in raisin extracts were characterized using UPLC-MS/MS, mainly including tricetin 3′-glucuronide, diisobutyl phthalate, butyl isobutyl phthalate, isoquercitrin and 6-hydroxykaempferol-7-O-glucoside. The oxidative stress in H₂O₂-induced HepG2 cells and D-gal-induced aging mice was alleviated by raisin polyphenols (RPs) via increases in the cellular levels of superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH), along with decreases in malonaldehyde (MDA), reactive oxygen species (ROS) and advanced glycosylation end-products (AGEs) levels. In addition, it was observed that RPs enhanced Sirt1 and Sirt3 expression, initiating the Keap1-Nrf2 signaling pathway, by upregulating the levels of nuclear Nrf2, facilitating the expressions of the antioxidant proteins NQO1 and HO-1, and downregulating Keap1 and cytoplasmic Nrf2 protein levels in H₂O₂-induced HepG2 cells and D-gal-induced aging mice. In summary, RP exerted antioxidant and anti-aging effects via regulating the Sirt1–Nrf2 signaling pathway in vitro and in vivo. Full article

Lentil is a leguminous crop with a high content of health-beneficial polyphenols. Lentil sprouts are popularly consumed in fresh vegetable markets, although their phytochemical qualities are not well understood. In this study, we investigated the accumulation of phenolics in lentil sprouts in response to photosynthetic and stress light qualities, including fluorescent light (FL), red LED (RL), blue LED (BL), ultraviolet A (UV-A), and ultraviolet B (UV-B). Three lentil cultivars, Lentil Green (LG), French Green (FG), and Lentil Red (LR), were used to evaluate sprouts grown under each light condition. The adequate light intensities for enhancing the antioxidant activity of lentil sprouts were found to be 11 W/m² under photosynthetic lights (FL, RL, BL), and 1 W/m² under stress lights (UV-A, UV-B). Subsequently, HPLC-ESI/Q-TOF MS analysis was conducted for the quantitative analysis of the individual phenolics that were accumulated in response to light quality. Four main phenolic compounds were identified: ferulic acid, tricetin, luteolin, and kaempferol. Notably, tricetin accumulation was significantly enhanced under BL across all three lentil cultivars examined. Furthermore, the study revealed that the other phenolic compounds were highly dependent on FL, BL, or UV-B exposure, exhibiting cultivar-specific variations. Additionally, the antioxidant activities of lentil extracts indicated that BL was most effective for LG and FG cultivars, whereas FL was most effective for enhancing antioxidant activity of LR cultivars as the sprouts grew. Full article

The phytohormone gibberellic acids (GAs) play a crucial role in the processes of growth, organ development, and secondary metabolism. However, the mechanism of exogenous GA₃ regulating the growth and flavonoid synthesis in Phellodendron chinense Schneid (P. chinense Schneid) seedlings remains unclear. In this study, the physicochemical properties, gene expression level, and secondary metabolite of P. chinense Schneid seedlings under GA₃ treatment were investigated. The results showed that GA₃ significantly improved the plant height, ground diameter, fresh weight, chlorophyll content, soluble substance content, superoxide dismutase, and peroxidase activities. This was accompanied by elevated relative expression levels of Pc(S)-GA2ox, Pc(S)-DELLA, Pc(S)-SAUR50, Pc(S)-PsaD, Pc(S)-Psb 27, Pc(S)-PGK, Pc(S)-CER3, and Pc(S)-FBA unigenes. Conversely, a notable reduction was observed in the carotenoid content, catalase activity and the relative expression abundances of Pc(S)-KAO, Pc(S)-GID1/2, and Pc(S)-GH 3.6 unigenes in leaves of P. chinense Schneid seedlings (p < 0.05). Furthermore, GA₃ evidently decreased the contents of pinocembrin, pinobanksin, isosakuranetin, naringin, naringenin, (−)-epicatechin, tricetin, luteolin, and vitexin belonged to flavonoid in stem bark of P. chinense Schneid seedlings (p < 0.05). These results indicated that exogenous GA₃ promoted growth through improving chlorophyll content and gene expression in photosynthesis and phytohormone signal pathway and inhibited flavonoid synthesis in P. chinense Schneid seedlings. Full article

The aim of this study was to compile a detailed phytochemical profile and assess the antioxidant properties of bee-collected pollen (PBP) obtained from corn poppy (Papaver rhoeas L.) plants. To achieve this, a lipid fraction was prepared for quantifying fatty acids using GC-FID. Extractable and alkaline-hydrolysable PBP fractions (obtained from a defatted sample) were used to determine the qualitative and quantitative profiles of phenolic compounds, phenylamides and alkaloids using UHPLC/Q-ToF-MS. Additionally, various spectrophotometric assays (TAC, FRP, CUPRAC, DPPH^⦁) were conducted to evaluate the antioxidant properties. Phenolic compounds were more present in the extractable fraction than in the alkaline-hydrolysable fraction. Luteolin was the predominant compound in the extractable fraction, followed by tricetin and various derivatives of kaempferol. This study presents one of the first reports on the quantification of tricetin aglycone outside the Myrtaceae plant family. The alkaline-hydrolysable fraction exhibited a different phenolic profile, with a significantly lower amount of phenolics. Kaempferol/derivatives, specific compounds like ferulic and 5-carboxyvanillic acids, and (epi)catechin 3-O-gallate were the predominant compounds in this fraction. Regarding phenylamides, the extractable fraction demonstrated a diverse range of these bioactive compounds, with a notable abundance of different spermine derivatives. In contrast, the hydrolysable fraction contained six spermine derivatives and one spermidine derivative. The examined fractions also revealed the presence of seventeen different alkaloids, belonging to the benzylisoquinoline, berberine and isoquinoline classes. The fatty-acid profile confirmed the prevalence of unsaturated fatty acids. Furthermore, both fractions exhibited significant antioxidant activity, with the extractable fraction showing particularly high activity. Among the assays conducted, the CUPRAC assay highlighted the exceptional ability of PBP’s bioactive compounds to reduce cupric ions. Full article

Acute pancreatitis (AP) poses a worldwide challenge due to the growing incidence and its potentially life-threatening course and complications. Specific targeted therapies are not available, prompting the identification of new pathways and novel therapeutic approaches. Flavonoids comprise several groups of biologically active compounds with wide-ranging effects. The flavone compound, tricetin (TCT), has not yet been investigated in detail but sporadic reports indicate diverse biological activities. In the current study, we evaluated the potential protective effects of TCT in AP. TCT (30 μM) protected isolated primary murine acinar cells from the cytotoxic effects of cerulein, a cholecystokinin analog peptide. The protective effects of TCT were observed in a general viability assay (calcein ester hydrolysis), in an apoptosis assay (caspase activity), and in necrosis assays (propidium iodide uptake and lactate dehydrogenase release). The effects of TCT were not related to its potential antioxidant effects, as TCT did not protect against H₂O₂-induced acinar cell death despite possessing radical scavenging activity. Cerulein-induced expression of IL1β, IL6, and matrix metalloproteinase 2 and activation of nuclear factor-κB (NFκB) were reduced by 30 μM TCT. In vivo experiments confirmed the protective effect of TCT in a mouse model of cerulein-induced AP. TCT suppressed edema formation and apoptosis in the pancreas and reduced lipase and amylase levels in the serum. Moreover, TCT inhibited interleukin-1β (IL1β), interleukin-6 (IL6), and tumor necrosis factor-α (TNFα) expression in the pancreas and reduced the activation of the oxidative DNA damage sensor enzyme poly(ADP-ribose) polymerase-1 (PARP-1). Our data indicate that TCT can be a potential treatment option for AP. Full article

Tricetin (5,7,3′,4′,5′-pentahydroxyflavone) is a dietary flavone from flowers of Myrtales plants with demonstrated functions in promoting human health. By contrast, the bioactivity of its glucosylated derivative tricetin 4′-O-glucoside has not been extensively explored. We conducted metabolite profiling analysis of pomegranate (a Myrtales plant) floral tissues and revealed that tricetin and tricetin 4′-O-glucoside accumulate in anthers, but not petals. In addition, the comparative analysis of anther and petal transcriptomes identified 10 UGTs that are more highly expressed in anthers than petals. Of the 10 UGTs, PgUGT76Z1 and PgUGT73AL1 glucosylated specifically at the 4′-O position of tricetin to form tricetin 4′-O-glucoside. The phylogenetic analysis indicated that PgUGT76Z1 and PgUGT73AL1 belong to different plant UGT groups, suggesting a convergent evolution of these tricetin UGTs. Overall, identification and characterization of PgUGT76Z1 and PgUGT73AL1 not only provides evolutionary insights into tricetin glucosylation, but also offers an opportunity to produce tricetin 4′-O-glucoside in large quantities through microbial biotransformation or plant metabolic engineering, thus facilitating the investigation of tricetin 4′-O-glucoside bioactivities. Full article

Tyrosinase is generally known as a melanin-forming enzyme, facilitating monooxygenation of phenols, oxidation of catechols into quinones, and finally generating biological melanin. As a homologous form of tyrosinase in plants, plant polyphenol oxidases perform the same oxidation reactions specifically toward plant polyphenols. Recent studies reported synthetic strategies for large scale preparation of hydroxylated plant polyphenols, using bacterial tyrosinases rather than plant polyphenol oxidase or other monooxygenases, by leveraging its robust monophenolase activity and broad substrate specificity. Herein, we report a novel synthesis of functional plant polyphenols, especially quercetin and myricetin from kaempferol, using screened bacterial tyrosinases. The critical bottleneck of the biocatalysis was identified as instability of the catechol and gallol under neutral and basic conditions. To overcome such instability of the products, the tyrosinase reaction proceeded under acidic conditions. Under mild acidic conditions supplemented with reducing agents, a bacterial tyrosinase from Bacillus megaterium (BmTy) displayed efficient consecutive two-step monophenolase activities producing quercetin and myricetin from kaempferol. Furthermore, the broad substrate specificity of BmTy toward diverse polyphenols enabled us to achieve the first biosynthesis of tricetin and 3′-hydroxyeriodictyol from apigenin and naringenin, respectively. These results suggest that microbial tyrosinase is a useful biocatalyst to prepare plant polyphenolic catechols and gallols with high productivity, which were hardly achieved by using other monooxygenases such as cytochrome P450s. Full article

Bee pollen is commonly reputed as a rich source of nutrients, both for bees and humans. Its composition is well balanced and can be taken as a stand-alone food or as supplement, including for the elderly owing its low caloric value. However, storage conditions frequently lead to product degradation, namely due to the high moisture content that enable the proliferation of molds and bacteria. Herein, an infrared (IR)-based technology is proposed as a mean to determine moisture content, setting also a new scalable approach for the development of a drying technology to be used for bee pollen processing, which can be carried out in a short time, without impacting the phenolic and flavonoid content and associated bioactive effects. Proof-of-concept was attained with an IR moisture analyzer, bee pollen samples from Eucalyptus globulus Labill and Salix atrocinerea Brot. being selected as models. Impact of the IR radiation towards the phenolic and flavonoid profiles was screened by HPLC/DAD profiling and radical scavenging ability by the DPPH assay. The IR-based approach shows good reproducibility while simultaneously reducing drying time and energy consumption, thus implying a low environmental impact and being suitable for industrial scale-up once no degradation has been found to occur during the radiation process. Full article

Flavonoids are rather common plant phenolic constituents that are known for potent antioxidant effects and can be beneficial for human health. Flavonoids with a pyrogallol moiety are highly efficient reducing agents with possible pro- and antioxidant effects, depending on the reaction milieu. Therefore, the redox properties of myricetin and tricetin were investigated by differential pulse voltammetry and deoxyribose degradation assay. Tricetin proved to be a good antioxidant but only showed negligible pro-oxidant activity in one of the deoxyribose degradation assay variants. Compared to tricetin, myricetin showed pro- and antioxidant effects. The more efficient reducing properties of myricetin are probably caused by the positive mesomeric effect of the enolic 3-hydroxy group on ring C. It is evident that the antioxidant properties of structurally similar flavonoids can be converted to apparent pro-oxidant effects by relatively small structural changes, such as hydroxylation. Since reactive oxygen species (ROS) often serve as secondary messengers in pathological and physiological processes in animal and plant cells, the pro- and antioxidant properties of flavonoids are an important part of controlling mechanisms of tissue signal cascades. Full article

Noni (Morinda citrifolia L.) fruit juice has been used in Polynesia as a traditional folk medicine and is very popular worldwide as a functional food supplement. In this study, compounds present in Hawaiian Noni fruit juice, with anti-inflammatory activity in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells were identified. Five compounds were isolated using a bioassay-driven technique and phytochemical analysis of noni fruit juice: asperulosidic acid (1), rutin (2), nonioside A (3), (2E,4E,7Z)-deca-2,4,7-trienoate-2-O-β-d-glucopyranosyl-β-d-glucopyranoside (4), and tricetin (5). The structures of these five compounds were determined via NMR spectroscopy and LC/MS. In an anti-inflammatory assay, compounds 1–5 inhibited the production of nitric oxide (NO), which is a proinflammatory mediator, in LPS-stimulated macrophages. Moreover, the mechanisms underlying the anti-inflammatory effects of compounds 1–5 were investigated. Parallel to the inhibition of NO production, treatment with compounds 1–5 downregulated the expression of IKKα/β, I-κBα, and NF-κB p65 in LPS-stimulated macrophages. Furthermore, treatment with compounds 1–5 downregulated the expression of nitric oxide synthase and cyclooxygenase-2. Thus, these data demonstrated that compounds 1–5 present in noni fruit juice, exhibited potential anti-inflammatory activity; these active compounds may contribute preventively and therapeutically against inflammatory diseases. Full article