Species within the genus Centaurium readily hybridize and polyploid complexes are often seen in natural populations. We describe phytochemical profiles of newly discovered allohexaploid hybrid, here ...named Centaurium pannonicum, and its parental tetraploid species C. erythraea and rare C. littorale ssp. compressum. Our aim was to examine chemodiversity of these taxa in the area of Vojvodina (North Serbia) and to perform metabolomics search for chemical classifiers which would provide high resolution discrimination of parental and hybrid individuals. In sum, UHPLC-MS/MS Orbitrap metabolomics fingerprinting revealed seventy compounds in methanol extracts. Despite the lack of qualitative chemical novelty in hybrid plants, UHPLC-qqqMS targeted metabolomics approach, aimed at three secoiridoid compounds and seventeen phenolics, pointed to considerable differences in quantitative composition of these dominant compounds among the plant taxa studied. In addition to the difference in the ploidy levels, the hybrid taxon was well distinguished from both parental species based on metabolite profiles, and, for most individuals, positioned intermediately to the parental taxa in both PCA and hierarchical clustering. After optimizing and comparing several statistical learning methods, it was possible to narrow the number of taxonomic classifiers to five (three xanthones, one secoiridoid glycoside, and one phenolic acid), while increasing the differentiation resolution. The presented metabolomics approach will certainly, along with morphometrics and molecular genetics studies, have high impact on further elucidation of complex relationships among taxa within the genus Centaurium.
LDA based on the concentration of three secoiridoid and 17 phenolic compounds for Centaurium erythraea, C. littorale ssp. compressum and their interspecies hybrid C. pannonicum. Display omitted
•New interspecies hybridogenic taxon Centaurium pannonicum displayed hexaploid nature.•Metabolic fingerprints of hybrids and tetraploid parental taxa revealed 70 compounds.•Metabolic profiling was targeted towards 20 major secoiridoids and phenolics.•Targeted metabolomics approach fairly differentiated hybrids from parental taxa.•Chemometrics highlighted five compounds as high-resolution taxonomic classifiers.
Centaurium erythraea Rafn (CE) is used as a traditional medicinal plant in Serbia to treat different ailments due to its antidiabetic, antipyretic, antiflatulent and detoxification effects.
...Elucidation of the mechanisms that underlie the antioxidant and pro-survival effects of the CE extract (CEE) in beta-cells and pancreatic islets from streptozotocin (STZ)-treated diabetic rats.
Diabetes was induced in rats by multiple applications of low doses of STZ (40 mg/kg intraperitoneally (i.p.), for five consecutive days). CEE (100 mg/kg) was administered orally, in the pre-treated group for two weeks before diabetes induction, during the treatments with STZ and for four weeks after diabetes onset, and in the post-treatment group for four weeks after diabetes induction. The impact of CEE on diabetic islets was estimated by histological and immunohistochemical examination of the pancreas. Molecular mechanisms of the effects of CEE were also analyzed in insulinoma Rin-5F cells treated with STZ (12 mM) and CEE (0.25 mg/mL). Oxidative stress was evaluated by assessing the levels of DNA damage, lipid peroxidation, protein S-glutathionylation and enzymatic activities and expression of CAT, MnSOD, CuZnSOD, GPx and GR in beta-cells. The presence and activities of the redox-sensitive and islet-enriched regulatory proteins were also analyzed.
Treatment with CEE ameliorated the insulin level and glycemic control in STZ-induced diabetic rats by improving the structural and functional properties of pancreatic islets through multiple routes of action. The disturbance of islet morphology and islet cell contents in diabetes was reduced by the CEE treatment and was associated with a protective effect of CEE on the levels of insulin, GLUT-2 and p-Akt in diabetic islets. The antioxidant effect of CEE on STZ-treated beta-cells was displayed as reduced DNA damage, lipid peroxidation, protein S-glutathionylation and alleviation of STZ-induced disruption in MnSOD, CuZnSOD and CAT enzyme activities. The oxidative stress-induced disturbance of the transcriptional regulation of CAT, MnSOD, CuZnSOD, GPx and GR enzymes in beta-cells was improved after the CEE treatment, and was observed as readjustment of the presence and activities of redox-sensitive NFκB-p65, FOXO3A, Sp1 and Nrf-2 transcription factors. The observed CEE-mediated induction of proliferative and pro-survival pathways and insulin expression/secretion after STZ-induced oxidative stress in beta-cells could be partially attributed to a fine-tuned modulation of the activities of pro-survival Akt, ERK and p38 kinases and islet-enriched Pdx-1 and MafA regulatory factors.
The results of this study provide evidence that CEE improves the structural and functional properties of pancreatic beta-cells by correcting the endogenous antioxidant regulatory mechanisms and by promoting proliferative and pro-survival pathways in beta-cells.
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•Centauries are an appreciative source of biologically active compounds.•Phenolics, flavonoids, xanthones and secoiridoid glycosides were quantified.•Considerable antioxidant activities and ferric ...reducing capacity was recorded.•Appreciative antibacterial and strong antifungal activities were demonstrated.•Centauries could be recommended as food preservation additives.
Methanol extracts of aerial parts and roots of five centaury species (Centaurium erythraea, C. tenuiflorum, C. littorale ssp. uliginosum, C. pulchellum, and Schenkia spicata) were analysed for their main secondary metabolites: secoiridoid glycosides, a group of monoterpenoid compounds, and phenolics (xanthones and flavonoids), and further investigated for antioxidant capacity and antimicrobial activity. The results of ABTS, DPPH, and FRAP assays showed that above ground parts generally displayed up to 13times higher antioxidant activity compared to roots, which should be related to higher phenolics content, especially flavonoids, in green plant organs. Secoiridoid glycosides showed no antioxidant activity. All the tested extracts demonstrated appreciative antibacterial (0.05–0.5mgml−1) and strong antifungal activity (0.1–0.6mgml−1). Our results imply that above ground parts of all centaury species studied, could be recommended for human usage as a rich source of natural antioxidants and also in food industry as strong antimicrobial agents for food preservation.
The aim of this study was to determine intra- and interspecies variation in the qualitative and quantitative composition of methanol-soluble metabolites in the leaves of three
species (
,
, and
) ...from the central Balkans. Despite the steady use of foxglove constituents for human health as valuable medicinal products, populations of the genus
(
) have been poorly investigated to describe their genetic and phenetic variation. Following untargeted profiling using UHPLC-LTQ Orbitrap MS, by which we identified a total of 115 compounds, 16 compounds were quantified using the UHPLC(-)HESI-QqQ-MS/MS approach. In total, 55 steroid compounds, 15 phenylethanoid glycosides, 27 flavonoids, and 14 phenolic acid derivatives were identified across the samples with
and
showing a great similarity, while 15 compounds were characteristic only for
. The phytochemical composition of methanol extracts, considered here as complex phenotypes, are further examined along multiple levels of biological organization (intra- and interpopulation) and subsequently subjected to chemometric data analysis. The quantitative composition of the selected set of 16 chemomarkers belonging to the classes of cardenolides (3 compounds) and phenolics (13 compounds) pointed to considerable differences between the taxa studied.
and
were found to be richer in phenolics as compared to cardenolides, which otherwise predominate in
over other compounds. PCA revealed lanatoside C, deslanoside, hispidulin, and
-coumaric acid to be the main compounds contributing to the differences between
on one side and
and
on the other, while
-coumaric acid, hispidulin, and digoxin contribute to the diversification between
and
. However, quantitative variation in the metabolite content within species was faint with mild population diversification visible in
and particularly in
This pointed to the highly conserved content and ratio of targeted compounds within the analyzed species, which was not severely influenced by the geographic origin or environmental conditions. The presented metabolomics approach might have, along with morphometrics and molecular genetics studies, a high information value for further elucidation of the relationships among taxa within the genus
.
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•Persistent oxidative stress leads to the development of diabetic complications.•Centaurium erythraea extract (CEE) reduced protein glycosylation in diabetic liver and kidney.•CEE ...reduced lipid peroxidation, DNA damage, and protein glutathionylation in diabetic liver and kidney.•CEE improved liver and kidney function by boosting the endogenous antioxidant system.
The use of medicinal herbs can mitigate oxidative stress-induced diabetic complications and organ failure. This study investigated hepato- and reno-protective effects of methanol extract of Centaurium erythraea Rafn (CEE) in STZ-diabetic rats pre-treated (2 weeks) and/or post-treated (4 weeks) with CEE (orally, 100 mg/kg/day). Both CEE treatments markedly improved liver and kidney functionality in diabetic rats observed as reduced aspartate and alanine aminotransferase activities and decreased creatinine and blood urea nitrogen levels. CEE pre-treatment reduced the level of glycosylated proteins in diabetic liver more efficiently than post-treatment. Lowered levels of lipid peroxidation, DNA damage and protein glutathionylation, elevated ratio of reduced to oxidized glutathione, and mitigated disturbance of antioxidant enzyme activities reflected the antioxidant effect of CEE in diabetic liver and kidney. Although CEE pre-treatment was more effective, the obtained results indicate that both treatments protect the liver and kidney from oxidative damage by boosting the endogenous antioxidant defense system.
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•Four Iranian Nepeta species screened for phenolic compounds.•Five dominant phenolics and five flavonoids targeted for UHPLC/-HESI-MS/MS analysis.•Endemic Nepeta kotschyi highly ...productive for analyzed compounds.•Domestication and cultivation of N. kotschyi for pharmacology uses is recommended.
This study was aimed at exploring the possibilities for the exploitation of valuable germplasm of four Iranian Nepeta species, highlighting the opportunities for their economic development as crops. Selection of the most promising species for introducing into cultivation was based on the content of medicinally important phenolic compounds. Twenty nine accessions of four Iranian species, Nepeta kotschyi, N. menthoides, N. crassifolia, and N. cataria, cultivated under western Tehran (Iran) environmental conditions were assessed. Quantitative analysis of five targeted phenolic acids and five targeted flavonoids in methanol extracts was performed using UHPLC/-HESI-MS/MS method. Multivariate analyses of chemometric data (PCA and UPGMA) segregated N. kotschyi as a separate group with unique phenolics composition, characterized by high amounts of phenolic acids, especially chlorogenic acid. As expected, rosmarinic acid has been determined as the dominant phenolic compound in analyzed N. cataria, N. menthoides and N. crassifolia accessions. N. kotschyi is recognized as the most promising candidate species for an alternative crop, and “N16” and “N17” accessions as the most suitable for domestication and cultivation practices under specific environmental conditions provided here.
Centaurium erythraea, used as medical plant from the earliest times, is an immense depot of quite rare bioactive compounds. Secoiridoids (sweroside, swertiamarin, and gentiopicrin) and xanthones ...(methylbellidifolin and decussatin) are predominant bioactive compounds in C. erythraea. The present study aims at providing new insights into how the content of these bioactive principles can be related with plant ploidy level by characterizing possible differences in their biosynthesis and accumulation between diploid and tetraploid genotypes from both spatial and temporal aspects. In general, shoots are determined as the major site of secoiridoids’ and xanthones’ accumulation, whose ratio vary during the development. Genes involved in iridoid and xanthone metabolic pathways were found to be coordinately regulated at the transcriptional level both during the development and among organs. Biosynthetic gene expression levels were found highly correlated with the content of major compounds from these two classes. Diversification in chemical profiles between tetraploid and diploid genotypes may result from the expression difference between homologous loci correspondent to several key biosynthetic genes, which trigger changes in the two metabolic routes. Thus, enhanced expression of genes coding for geraniol synthase (GES), 8-hydroxygeraniol oxidoreductase (8HGO), and 7-deoxyloganic acid hydrolase (7DLH2) is strongly associated with intensive production of iridoids. Interestingly, transcript levels of beta-glucosidase (CebGLU), a candidate to catalyze the first step in the secoiridoid catabolism, is significantly positively correlated with the content of major secoiridoids. Elevated expression of genes coding for benzophenone synthase (BS) and 3-hydroxybenzoate:CoA ligase (3HBL) appear to account for enhanced production of hexa-substituted xanthones. Regarding content of iridoids and xanthones, a diploid genotype appeared to be more productive than a tetraploid genotype under controlled in vitro conditions, therewithal displaying significantly higher biomass.
•Iridoids and xanthones content and ratio varied during early development in vitro.•Two-month-old diploid shoots and roots are the most promising source of xanthones.•Three-month-old shoots are the most convenient for the secoiridoid production.•Enhanced expression of key genes associates with iridoid and xanthone production.•Iridoid and xanthone biosynthetic genes are regulated at the transcriptional level.
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•Enzymatic hydrolysis alternates phytochemical profile of Centaurium erythraea extract.•Aglycones erythrocentaurin and gentiopicral predominated in the hydrolyzed extract.•Extracts ...and pure secoiridoids displayed strong antimicrobial and antioxidant activity.•Hydrolysis boosted swertiamarin and sweroside antioxidant potential.•Secoiridoid glucosides were efficiently metabolized by Penicillium funiculosum.
The present study was principally aimed at ascertaining the differences in metabolomics profiles and biological activities between non-hydrolyzed (ME) and hydrolyzed methanol extract (HME) of Centaurium erythraea Rafn. UHPLC–MS/MS Orbitrap analysis showed that the enzymatic hydrolysis of the extract caused changes in β-d-glycoside/aglycone ratio of both flavonoid and secoiridoid compounds. UHPLC/DAD/+HESI−qqqMS characterization and/or quantification of secoiridoid glucosides (SGs) and their aglycones in both ME and HME revealed gentiopicral and erythrocentaurin as the major aglycones, the same metabolic products which appear after the hydrolysis of pure swertiamarin, the dominant secoiridoid glucoside of C. erythraea. SGs played an antioxidant role only in ABTS assay, whilst the remarkable antioxidant potential of C. erythraea methanol extract is ascribed chiefly to phenolics detected in it. Interestingly, antioxidant activities of swertiamarin and sweroside recorded in ABTS assay increased after the compounds have been hydrolyzed, which highlighted their possible antioxidant role during ingestion. Strong antimicrobial activities of ME and HME against a vast array of pathogens, which exceed the effects of the reference antibiotics and antimycotics, largely depended on the amount of secoiridoids in either of the glycosylation forms. Extracts and pure secoiridoids were especially effective against most of the tested Penicillium species. On the other hand, P. funiculosum has evolved an efficient mechanism of detoxification of sub-lethal concentrations of secoiridoid glucosides, involving their biotransformation and complete digestion. The presented findings will contribute to clarify the fate and role of the SGs after C. erythraea ingestion within in vivo systems, and to further promote this remarkable plant as a food preservation additive with significant health benefits.
Centaurium erythraea Rafn (CE) is a traditional medicinal herb in Serbia with antidiabetic, digestive, antipyretic and antiflatulent effects
To investigate the potential protective effects of the ...methanol extract of the aerial parts of CE against glyco-oxidative stress in red blood cells (RBCs) in rats with experimentally induced diabetes.
Diabetes was induced in Wistar rats by intraperitoneal (i.p.) injection of multiple low-dose streptozotocin (STZ) (40mg/kg, for five consecutive days), with the 1st day after the last STZ injection taken as the day of diabetes onset. The methanol extract of CE (100mg/kg) was administered orally and daily, two weeks before the first STZ injection, during the 5-day treatment with STZ, and for four weeks after the STZ injections (pre-treated group) or for four weeks after diabetes onset (post-treated group). The effect of CE extract administration on the redox status of RBCs was evaluated by assessing lipid peroxidation, the ratio of reduced/oxidized glutathione (GSH/GSSG), the level of S-glutathionylated proteins (GSSP) and the enzymatic activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) in RBCs four weeks after diabetes onset. The major biochemical parameters of diabetes, protein glycation/glycosylation of erythrocytes and parameters which correlate with their aggregation and deformability were also evaluated.
Daily application of CE extract to STZ-induced diabetic rats provided important antidiabetic effects, observed in both pre-treated and post-treated groups of diabetic rats as elevated serum insulin concentration, reduction of blood glucose and glycated hemoglobin concentrations and an improved lipid profile. Antioxidant effects of CE extract were detected in RBCs of diabetic rats and observed as decreased lipid peroxidation and ameliorated oxidative damage as a result of increased SOD, CAT and GR activities, an improved GSH/GSSG ratio and reduced GSSP levels. Moreover, the CE extract protected RBC proteins from hyperglycemia-induced damage by reducing non-enzymatic glycation and enzymatic glycosylation processes. CE extract was more effective when applied before diabetes induction (pre-treated group).
The results of this study show that the Centaurium erythraea methanol extract protects RBCs in diabetic animals from oxidative damage. They provide additional support for the application of this traditionally used plant in diabetes management.
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