Astragalin Research Articles

Quantitative Changes of Flavonol Glycosides from Pine Needles by Cultivar, Harvest Season, and Thermal Process

Five flavonol glycosides including quercetin 3-O-β-D-glucoside (QG), kaempferol 3-O-β-D-glucoside (KG), quercetin 3-O-(6″-O-acetyl)-β-D-glucoside (QAG), kaempferol 3-O-(6″-O-acetyl)-β-D-glucoside (KAG), and quercetin 3-O-(3″-O-p-coumaroyl)-β-D-glucoside (QCG) were isolated and purified from red pine (Pinus densiflora Sieb. et Zucc.) nee-dles, and identified by nuclear magnetic resonance and mass spectrometer spectral analyses. In addition, the quantification of the five flavonol glycosides in pine needles was performed by high-performance liquid chromatography analysis according to cultivar, growing district, harvest season, and thermal processing. The red pine needles had higher amounts of the five flavonol glycosides than the black pine needles except for QCG. There were no large differences in flavonoid composition and content among pine needles grown in three different areas. Levels of the five flavonol glycosides in red pine needles harvested during Spring ranged from 6.13 to 27.03 mg/100 g dry weight. Levels of two flavonol glycosides, QG and KG, gradually decreased with increasing harvest time, whereas the acylated flavonol glycoside, QCG, a predominant flavo-noid in pine needles, increased gradually with increasing harvest time. Two acetyl flavonol glycosides, QAG and KAG, increased steadily through Spring to Autumn, and then decreased gradually by Winter. Meanwhile heat treatments, such as roasting and steaming, increased the five flavonol glycosides during heating for 3 min, but then slowly decreased these when heating for 10 min. Microwave processing increased to some extent the five flavonol glycosides when heating for 3 min, and remained unchanged during the 10 min heating. These results suggest that the pretreated red pine needles with enhanced flavonoid content may be useful as potential sources for nutraceuticals and cosmeceuticals.

Astragalin Alleviates Neuropathic Pain by Suppressing P2X4-Mediated Signaling in the Dorsal Root Ganglia of Rats.

Neurologic damage often leads to neuropathic pain, for which there are no effective treatments owing to its complex pathogenesis. The purinergic receptor P2X4 is closely associated with neuropathic pain. Astragalin (AST), a compound that is used in traditional Chinese medicine, has protective effects against allergic dermatitis and neuronal injury, but its mechanism of action is not well understood. The present study investigated whether AST can alleviate neuropathic pain in a rat model established by chronic constriction injury (CCI) to the sciatic nerve. The model rats exhibited pain behavior and showed increased expression of P2X4 and the activated satellite glial cell (SGC) marker glial fibrillary acidic protein in dorsal root ganglia (DRG). AST treatment partly abrogated the upregulation of P2X4, inhibited SGC activation, and alleviated pain behavior in CCI rats; it also suppressed ATP-activated currents in HEK293 cells overexpressing P2X4. These data demonstrate that AST relieves neuropathic pain by inhibiting P2X4 and SGC activation in DRG, highlighting its therapeutic potential for clinical pain management.

New Norneolignan and Bioactive Constituents of Clitoria ternatea

A new norneolignan derivative, clitorternalactone (1), has been isolated from the stems of Clitoria ternatea, together with six known compounds, clitorienolactone A (2), myricetin 3-glucoside (3), quercetin 3-glucoside (4), kaempferol 3-glucoside (5), taraxerol (6), and taraxerone (7). The structure of the new compound 1 was determined through spectroscopic and MS analyses. Among the isolates, compound 1 exhibited cytotoxicities with IC50 values of 2.54 ± 0.23, 3.68 ± 0.17, and 4.05 ± 0.43 μM, respectively, against DLD-1, CCRF-CEM, and IMR-32 cell lines. In addition, clitorternalactone (1), clitorienolactone A (2), quercetin 3-glucoside (4), and kaempferol 3-glucoside (5) showed potent inhibition with IC50 values of 3.05 ± 0.12, 5.44 ± 0.46, 10.20 ± 0.69, and 13.57 ± 0.36 μM, respectively, against LPS-induced NO generation.

Astragalin Attenuates Dextran Sulfate Sodium (DSS)-Induced Acute Experimental Colitis by Alleviating Gut Microbiota Dysbiosis and Inhibiting NF-κB Activation in Mice.

With the ulcerative colitis (UC) incidence increasing worldwide, it is of great importance to prevent and treat UC. However, efficient treatment options for UC are relatively limited. Due to the potentially serious adverse effects of existing drugs, there is an increasing demand for alternative candidate resources derived from natural and functional foods. Astragalin (AG) is a type of anti-inflammatory flavonoid, with Moringa oleifera and Cassia alata being its main sources. In this study, we investigated the therapeutic effects of AG on mice with dextran sulfate sodium (DSS)-induced colitis. Our results suggested that AG treatment reduced weight loss and the disease activity index (DAI), prevented colon shortening and alleviated colonic tissue damage. AG treatment reduced the expression of pro-inflammatory cytokines and related mRNAs (such as TNF-α, IL-6, and IL-1β), inhibited colonic infiltration by macrophages and neutrophils, ameliorated metabolic endotoxemia, and improved intestinal mucosal barrier function (increased expression levels of mRNAs such as ZO-1, occludin, and Muc2). Western blot analysis revealed that AG downregulated the NF-κB signaling pathway. Moreover, AG treatment partially reversed the alterations in the gut microbiota in colitis mice, mainly by increasing the abundance of potentially beneficial bacteria (such as Ruminococcaceae) and decreasing the abundance of potentially harmful bacteria (such as Escherichia-Shigella). Ruminococcaceae and Enterobacteriaceae (Escherichia-Shigella) were thought to be the key groups affected by AG to improve UC. Therefore, AG might exert a good anti-UC effect through microbiota/LPS/TLR4/NF-kB-related pathways in mice. The results of this study reveal the anti-inflammatory effect and mechanism of AG and provide an important reference for studying the mechanisms of natural flavonoids involved in preventing inflammation-driven diseases.

Inhibitors of α-Synuclein Fibrillation and Oligomer Toxicity in Rosa damascena: The All-Pervading Powers of Flavonoids and Phenolic Glycosides.

There is an intense search for natural compounds that can inhibit the oligomerization and fibrillation of α-synuclein (α-Syn), whose aggregation is key to the development of Parkinson's disease (PD). Rosa damascena is a medicinal herb widely used in Middle Eastern food, ceremonies, and perfumes. The herb is known to contain many different polyphenols. Here we investigated the existence of α-Syn fibrillation inhibitors in R. damascena extract. Different HPLC fractions of the extract were assessed in α-Syn fibrillation and toxicity assays. The most active fractions led to the formation of more α-Syn oligomers but with less toxicity to SH-SY5Y cells, according to MTT and LDH assays. LC-MS analysis identified gallic acid, kaempferol 3-glucoside, kaempferol-3-O-β-rutinoside, and quercetin which were subsequently shown to be strong α-Syn fibrillation inhibitors. Our results highlight the benefits of R. damascena extract to combat PD at the population level.

Astragalin alleviates ischemia/reperfusion‑induced brain injury via suppression of endoplasmic reticulum stress.

Excessive apoptosis and neuronal dysfunction are pathological features of ischemic stroke. Previous studies have demonstrated that astragalin(AST) exerted both anti‑​apoptotic and anti‑inflammatory effects in several types of disease, although its potential effect in ischemic stroke remains unclear. The purpose of the present study was to investigate the effects of AST on cerebral ischemia/reperfusion (I/R)‑induced brain injury and the underlying mechanisms. Brain injury was assessed in an experimental rat model using measurement of neurological scores and inflammatory factors. To assess the role of AST in I/R‑induced brain injury and the potential mechanism of action, SH5Y were treated with thapsigargin and AST. Apoptotic rate and ERstress levels were measured by western blotting, reverse transcription‑quantitative PCR and immunofluorescence staining. It was discovered that AST significantly improved long‑term neurological outcomes in rats following cerebral I/Rinjury, through the attenuation of the expression levels of apoptotic proteins (Bax and cleaved‑caspase‑3) and the release of inflammatory cytokines, as well as upregulating the expression levels of the anti‑apoptotic protein Bcl‑2. Furthermore, AST attenuated the expression levels of the endoplasmic reticulum(ER) stress‑related protein, glucose‑regulated protein, 78kDa, as well as its downstream apoptotic mediators (CHOP and caspase‑12). Thapsigargin‑induced ERstress activation and apoptosis were also attenuated by AST in an invitro neuronal cell culture model. In conclusion, these results suggested that AST may protect against I/R‑induced brain injury, thus, highlighting its therapeutic potential in patients with ischemic stroke.

Astragalin Exerted Antidepressant-like Action through SIRT1 Signaling Modulated NLRP3 Inflammasome Deactivation.

Inflammation plays a key role in the pathogenesis of depression and antidepressant therapies. Astragalin (AST) is a bioactive flavonoid that possesses an anti-inflammatory property. However, the antidepressant action of astragalin has not been addressed. In this study, we explored the antidepressant effects of astragalin and its underlying mechanism. Our results showed that AST significantly improved the behavioral defects in chronic unpredictable mild stress (CUMS) model, promoted SIRT1 expression, and decreased the protein levels of NF-κB p65, NLRP3, cleaved capase-1, cleaved IL-1β and cleaved gasdermin D in the hippocampus. Immunohistochemistry revealed AST mitigated CUMS-induced microglia overactivation. In vitro, AST profoundly increased the cell viability in lipopolysaccharides (LPS) and adenosine triphosphate (ATP) treated BV2 cells, with upregulated SIRT1 expression and downregulated protein levels of nuclear NF-κB p65, NLRP3, cleaved capase-1, and cleaved gasdermin D. Declined cleavage of gasdermin D was observed after AST administration in immunocytochemistry. Nevertheless, the in vivo and in vitro effects of AST were compromised by SIRT1 inhibitor EX-527. These results indicated that AST possessed an antidepressant property, which was dependent on SIRT1 signaling modulated NLRP3 inflammasome deactivation.

Spectroscopic methodologies and computational simulation studies on the characterization of the interaction between human serum albumin and astragalin

In the present work, the interactions of astragalin (AST) with human serum albumin (HSA) were studied systematically thought fluorescence spectra, ultraviolet-visible (UV-vis) absorption spectra, circular dichroism (CD), molecular docking and molecular dynamics (MD) simulations. Fluorescence spectra elaborated that AST reduced the intrinsic fluorescence of HSA through static quenching and non-radiative energy transfer with moderate binding constants in the order of 104 mol/L. Thermodynamic parameters and computational simulations elaborated that hydrogen bond, van der Walls force and hydrophobic interaction played a major role in the binding process of AST to HSA. UV-vis absorption, synchronous fluorescence, three-dimensional (3D) fluorescence and CD spectra illustrated that AST disturbed slightly the microenvironment of tryptophan (Trp) and tyrosine (Tyr) residues and decreased α-helical structure content. The effect of some biologically significant metal ions, such as Mg2+, Cu2+ and Fe3+, on the binding of AST to HSA was also investigated in detail. Binding displacement and docking studies revealed that AST was located in the binding site I of subdomain IIA in HSA. Finally, MD simulations evaluated the binding stability of the HSA-AST system in a simulated environment. Communicated by Ramaswamy H. Sarma

1H-NMR Metabolomics and LC-MS Analysis to Determine Seasonal Variation in a Cosmeceutical Plant Leucosidea sericea.

Leucosidea sericea is an evergreen shrub belonging to the Rosaceae family with previous studies that indicated that L. sericea extracts exhibited strong anti-bacterial properties against Propionibacterium acnes, showing potential as a cosmeceutical. The plant is traditionally used as a vermifuge, as an astringent and to treat conjunctivitis. Commercial production is, however, not possible as no information is available on cultivation and the effect of external environmental factors such as seasonal variation on the medicinal properties of the plant. Seasonal variation was investigated and it was found that significant differences were observed between the anti-acne (P. acnes) activity of plant material collected in different seasons. The best activity was found in winter with a mean minimum inhibitory concentration (MIC) of 5.20 μg mL–1 compared to spring at 26.04 μg mL–1. A 1H NMR-based untargeted metabolomic analysis was used to determine the differences in the chemical profiles of plant samples collected in different seasons. Principal component analysis (PCA) showed clear separation of the seasons and a supervised orthogonal partial least square discriminant analysis (OPLS-DA) was used to determine the compounds that differentiated the spring from the winter samples. The contribution plot indicated a strong positive association with the NMR regions from δ 1.2–1.6, 3.3–4.1, and 6.8–8.0 ppm indicative of a compound with an aromatic ring. Different LC-MS analyses were used in conjunction with a compound database, MAGMa and CSIFingerID, which led to the identification of the compound 2-(4-ethoxyphenyl)-5,6,7,8-tetramethoxy-4H-1-benzopyran-4-one and also confirmed the presence of tangeritin, rutin, quercetin glucoside, and kaempferol glucosides as well as several other compounds previously identified from the plant. This compound similar in structure to the anti-microbial flavonoid tangeritin, was only present in the winter samples. It is therefore recommended that seasonal variation be closely monitored during cultivation and commercial harvesting, and that winter is the preferred harvesting season to obtain the best anti-acne activity.

The regression of endometriosis with glycosylated flavonoids isolated from Melilotus officinalis (L.) Pall. in an endometriosis rat model

ObjectiveMelilotus officinalis (L.) Pall. is commonly used for treating bronchitis, painful menstruation, hemorrhoids, kidney stones, ulcers of the eyes, earache, and hardening and swelling of uterus. The European Medicines Agency reported the use of M. officinalis orally against stomach ache, gastric ulcer, and disorders of the liver and uterus in folk medicine. The present study aimed to appraise the activity of M. (L.) Pall. aerial parts in endometriosis rat model. Materials and methodsThe endometriosis rat model was used to evaluate the potential activity of M. officinalis aerial parts based on its folkloric usage. The aerial parts of M. officinalis were extracted with n-hexane, ethyl acetate (EtOAc), and methanol (MeOH), respectively. The adhesion scores, endometrial foci areas, and cytokine levels were measured in all treated groups. After the biological activity studies, phytochemical studies were performed on the active extract and the fractions obtained from the active extract. ResultsThe MeOH extract significantly decreased the endometrial foci areas and cytokine levels in rats with endometriosis. Fractionation was performed on the MeOH extract to achieve bioactive molecules. Following the fractionation, the fractions obtained from the MeOH extract were tested. Fraction C showed the highest activity in the rat endometriosis model. Phytochemical investigation of the active fraction (Fraction C) resulted in isolation and elucidation of some quercetin and kaempferol glucoside derivatives. ConclusionFraction C obtained from the MeOH extract of M. officinalis showed the highest activity, yielding four glycosylated flavonoids.

Polyphenolic Profiles of Yellow Bean Seed Coats and Their Relationship with Iron Bioavailability.

Previous work with Caco-2 cell cultures has shown that individual polyphenols can either promote or inhibit iron uptake. This investigation was designed to characterize the relationship between iron bioavailability and seed coat polyphenol composition in a panel of 14 yellow beans representing five market classes with the potential for fast cooking time and high iron content. The study included two white and two red mottled bean lines, which represent high and low iron bioavailability capacity in dry beans, respectively. Polyphenols were measured quantitatively by high-performance liquid chromatography-mass spectrometry (HPLC-MS)/UV and iron bioavailability of seed coat extracts was measured in Caco-2 assays. Thirteen of the yellow bean seed types contained high concentrations (up to 35.3 ± 2.7 μmol/g) of kaempferol 3-glucoside (k 3-g), a known promoter of iron uptake. A general association between the ratio of promoting to inhibiting polyphenols (P/I) and iron uptake was observed. The presence of iron uptake inhibiting condensed tannins proportionately countered the promotional effects of kaempferol compounds. Unidentified factors present in seed coats other than polyphenols also appeared to affect iron uptake.

Phytochemical and Antioxidant Studies on a Rare Rheum cordatum Losinsk. Species from Kazakhstan.

An optimisation of extraction towards an increased antioxidant capacity and the study on the extracts' composition by HPLC-ESI-Q-TOF-MS were performed on different organs of a rarely studied plant: Rheum cordatum Losinsk (Polygonaceae) growing in Kazakhstan. More than 20 compounds from anthraquinones and phenolics were identified in an optimised method. The plant was proven to contain a wide variety of phenolic compounds (catechins, flavonoids, and their glucosides and phenolic acids) in contrast to the anthraquinone composition, which was mainly represented by emodin and its analogues. The results of the studies could determine the plant as a rich source of pharmacologically precious polyphenols. It was evidenced that the extracting solvents, the time of collection, and the organs tested affected both the chemical content and the antioxidant potential of the extracts. Ethanol : water (50 : 50 v/v) was selected as the most beneficial extractant for all metabolites, and based on the principal component analysis of raw data, the radical scavenging potential of the plant was strictly related to the presence of epicatechin gallate (ECG), kaempferol glucoside, and epigallocatechin gallate (EGCG) occurring in this extract at the concentration of 1.69-5%, 0.16-0.47%, and 0.001-2.93%, respectively.

Identification and quantification of phenolic compounds of Western Siberia Astragalus danicus in different regions

The potential of phenolic compounds of medicinal plants including Astragalus danicus L is determined by but not limited to their antioxidant activity. Their anti-inflammatory, antitumor, and other useful properties are known, which allows using these phytochemicals within preventive activities to reduce the risk of many serious diseases. Chromatographic analysis of the Astragalus danicus L. biomaterial from the plant samples collected in three regions of the Kemerovo region (Western Siberia, Russia) established the presence of compounds of flavonols (isorhamnetin glucoside, kaempferol glucoside), flavones (apigenin 7-glucoside), phenylpropanoids (chlorogenic acid) in the aerial part of plants. The total content of phenolic compounds in plant samples ranged from 100.75 ± 3.87 mg/g (Yashkinsky district) to 190.95 ± 7.34 mg/g (Belovsky district). The content of chlorogenic acid in the studied samples was from 0.14 ± 0.01 mg/g to 1.16 ± 0.04 mg/g. Isorhamnetin glucoside was found only in samples of plants from two districts - Prokopievsky (41.39 ± 1.58 mg/g) and Belovsky (95.0 ± 3.66 mg/g). The content of glucosides of kaempferol ranged from 0.38 ± 0.01 mg/g to 0.55 ± 0.02 mg/g. Its content is almost twice as high as the content in the well-known analogues of Astragalus. Apigenin-7-glucoside was isolated in Astragalus samples for the first time, in a small amount (3.34 ± 0.13 mg/g) in a sample of plants of one growing zone. Studies have confirmed that the content of flavonoids in plants significantly depends not only on the genetic characteristics of plants, but also on the hydrothermal regime, the climatic conditions of different botanical and geographical areas of the habitat. This work shows that Astragalus danicus L. growing in Kemerovo region is a promising raw material for pharmacological preparations.

HPLC-DAD-MS/MS profiling of phenolics from different varieties of peach leaves and evaluation of their antioxidant activity: A comparative study

Peach (Prunus persica L.) leaves have been traditionally used in folk medicine for their several biological activities that are correlated to the presence of polyphenolic compounds. The aim of the present study is to characterize phenolic compounds present in foliar extracts of seven peach varieties cultivated in Algeria by HPLC–MS and the study of their antioxidant potential. Antioxidant capacity of the foliar extracts was assessed by several tests acting by different mechanisms: Oxygen Radical Absorbance Capacity (ORAC), 2,2-DiPhenyl-PicrylHydrazyl radical (DPPH), 2,-Azinobis(3-ethylBenzoThiazoline-6-Sulfonic acid) (ABTS), Potassium Ferricyanide Reducing Antioxidant Power (PFRAP) and Iron Chelating Activity (ICA). Fourteen phenolic compounds were identified in the peach leaf extracts including cinnamic acids and flavonols. Flavonols represent the main class of phenolic compounds accounting for an average percentage higher than 95% of the overall phenolics. Kaempferol 3-glucoside is the main phenolic compound in all peach leaf extracts with an average percentage higher than 32% followed by quercetin 3-glucoside (17.9%), quercetin 3-galactoside (17.1%) and kaempferol 3-galactoside (15.4%). Results showed that variety significantly affected the phenolic content of peach leaves. Romea and Red Top varieties present the higher concentration in phenolic compounds, Dixired, Flavorcrest and Tebana a moderate one and, Cardinal and Spring Belle the lowest content. The data obtained with DPPH, ABTS, ORAC and PFRAP assays showed that polyphenols present in the all foliar peach extracts were potent antioxidative agents. Except for ICA assay, good positive correlations were found between phenolic concentration and the different measured antioxidant capacities. That means that phenolic compounds present in peach leaf cultivars were major contributors of reducing power and scavenging radicals capacities (DPPH, ABTS and ORAC). All these results allowed us to conclude that peach leaves are a good source of phenolics with active properties, as antioxidant ones.

Four New Flavonoids Isolated from the Aerial Parts of Cadaba rotundifolia Forssk. (Qadab).

Cadaba rotundifolia (Forssk.) (family: Capparaceae; common name: Qadab) is one of four species that grow in the Red Sea costal region in the Kingdom of Saudi Arabia. The roots and leaves of C. rotundifolia is traditionally used to treat tumors and abscesses in Sudan. A previous phytochemical study of the roots yielded a quaternary alkaloid, but no report on chemical constituents of the aerial parts of the C. rotundifolia growing in Saudi Arabia has been issued so far. Oxidative stress and advanced glycation end products (AGEs) are thought as causal factors in many degenerative diseases, such as Alzheimer’s disease, diabetes, atherosclerosis and aging. In this study, a total of twenty compounds, including four previously undescribed acylated kaempferol glucosides, were isolated from the aerial parts of C. rotundifolia collected in Saudi Arabia. These new compounds were identified as kaempferol 3-O-[2-O-(trans-feruloyl)-3-O-β-d-glucopyranosyl]-β-d-glucopyranoside (1), kaempferol 3-O-β-neohesperidoside-7-O-[2-O-(cis-p-coumaroyl)-3-O-β-d-glucopyranosyl]-β-d-glucopyranoside (2), kaempferol 3-O-[2,6-di-O-α-l-rhamnopyranosyl]-β-d-glucopyranoside-7-O-[6-O-(trans-feruloyl)]-β-d-glucopyranoside (3) and kaempferol 3-O-[2,6-di-O-α-l-rhamnopyranosyl]-β-d-glucopyranoside-7-O-[6-O-(trans-p-coumaroyl)]-β-d-glucopyranoside (4). Their structures were established based on UV-visible, 1D, 2D NMR, and HR-ESI-MS analyses. Of the assayed compounds, 17 and 18 showed potent radical scavenging activity with IC50 values of 14.5 and 11.7 µM, respectively, and inhibitory activity toward AGEs together with compound 7 with IC50 values 96.5, 34.9 and 85.5 µM, respectively.

A Combined In Vitro (Caco-2 Cell) and In Vivo (Gallus gallus) Assessment Reveals the Iron Benefits of Consuming the Fast Cooking Manteca Yellow Bean (Phaseolus vulgaris L.) (P10-114-19)

ObjectivesThe common dry bean (Phaseolus vulgaris L.) is a globally produced pulse crop and an important source of protein and micronutrients for millions of people across Latin America and Africa. In these regions, energy for cooking is expensive or scarce and long cooking times deter consumers from purchasing beans. In addition, many of the preferred black and red seed types have phytate and polyphenols that limit the absorption of trace minerals. Yellow beans are unique because their seed coats are rich in kaempferol 3-glucoside, a recently discovered promoter of iron absorption. Several market classes of yellow beans are sold throughout Latin America and Africa, where they are marketed at premium prices for their fast cooking tendencies. Exploring the yellow bean’s unique heritage to develop new fast cooking varieties that deliver more absorbable iron would be useful for regions where inhabitants have limited access to fuelwood for cooking. This study compared the iron bioavailability of three fast cooking yellow beans from Africa with contrasting seed coat colors (Manteca, Amarillo, Njano) to slower cooking white and red kidney commercial varieties from North America (Table 1). MethodsCooked beans were formulated into diets with the complementary food crops of potato, rice and cabbage. Iron bioavailability was measured as ferritin formation in an in vitro digestion Caco-2 bioassay and the ability to maintain total body iron hemoglobin (Hb-Fe) during a 6 week in vivo (Gallus gallus) feeding trial. ResultsAnimals fed yellow bean diets had faster growth rates, accumulated more dietary iron and had higher Hb-Fe than animals fed either kidney bean diet (Figure 1). In contrast to yellow beans, the kidney beans had almost no kaempferol 3-glucoside (Table 2). When compared to the other four bean based diets, the fast cooking Manteca yellow bean diet had the highest Caco-2 ferritin formation in vitro (Table 3) and delivered the largest increase in Hb-Fe in vivo (Figure 1). ConclusionsThrough the added benefit of fast preparation times and improved iron quality after cooking, this study provides evidence that the Manteca market class is worthy of germplasm enhancement as a new convenience food to help alleviate trace mineral deficiencies in regions where beans are widely accepted as a dietary staple. Funding SourcesUSDA-NIFA. Supporting Tables, Images and/or Graphs▪▪▪▪

Extraction, purification by macrospores resin and in vitro antioxidant activity of flavonoids from Moringa oliefera leaves

To promote extraction of flavonoids compounds from Moringa oliefera leaves (MOL) by ultrasound-assisted extraction, Box behken desing as a kind of response surface methodology was utilized. Moreover, five macrospores resins were used to investigate the kinetics of purification of flavonoids from the MOL extract. As results, the optimal conditions for maximizing flavonoids extraction were determined as follows: ratio of aqueous ethanol solution to material 52 mL/g, extraction time 43 min and extraction temperature 76 °C. Nevertheless, AB-8 resin was the suitable resin for purification of flavonoids with the following conditions: initial flavonoids concentration 20 mg/mL, eluting solvent 70% aqueous ethanol solution and 1.5 mL/min eluting flow rate. Moreover, five flavonoids (rutin, kaempferol acetyl glycoside, quercetin-3-glucoside, quercetin-3-acetyl-glucoside and kaempferol 3-glucoside) were identified by HPLC-ESI-MS. Regarding to antioxidant activity purified flavonoids, the results showed that the scavenging activity against 2,2-diphenyl-1-picrylhydrazyl free radicals was 85.99% at a concentration of 100 μg/mL, while the scavenging activity against 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) radicals was 84.72% at 50 μg/mL. The results suggest that the flavonoids from MOL can be used in the development of functional foods.

Astragalin Promotes Osteoblastic Differentiation in MC3T3-E1 Cells and Bone Formation in vivo.

Astragalin (AG) is a biologically active flavonoid compound that can be extracted from a number of medicinal plants. However, the effects of AG on osteoblastic differentiation in mouse MC3T3-E1 cells and on bone formation in vivo have not been studied fully. In this study, we found that the activities of alkaline phosphatase (ALP) and mineralized nodules in MC3T3-E1 cells were both significantly increased after treatment with AG (5, 10, and 20 μM). Meanwhile, the mRNA and protein levels of osteoblastic marker genes in MC3T3-E1 cells after AG treatment were markedly increased compared with a control group. In addition, the levels of BMP-2, p-Smad1/5/9, and Runx2 were significantly elevated in AG-treated MC3T3-E1 cells. Moreover, we found that the protein levels of Erk1/2, p-Erk1/2, p38, p-p38, and p-JNK were also significantly increased in AG-treated MC3T3-E1 cells compared to those in the control group. Finally, in vivo experiments demonstrated that AG significantly promoted bone formation in an ovariectomized (OVX)-induced osteoporotic mouse model. This was evidenced by significant increases in the values of osteoblast-related parameters (BFR/BS, MAR, Ob.S/BS, and Ob.N/B.Pm) and bone histomorphometric parameters (BMD, BV/TV, Tb.Th, and Tb.N.) in OVX mice after AG treatment (5, 10, and 20 mg/kg). Collectively, these results demonstrated that AG may promote osteoblastic differentiation in MC3T3-E1 cells via the activation of the BMP and MAPK pathways and promote bone formation in vivo. These novel findings indicated that AG may be a useful bone anabolic agent for the prevention and treatment of osteoporosis.

Identification of bioactive compounds from Fraxinus angustifolia extracts with anti- NADH oxidase activity of bovine milk xanthine oxidoreductase.

Fraxinus angustifolia leaves and bark are used in traditional medicine against various inflammatory-related pathologies incumbent to reactive oxygen species (ROS) generation by the NADH oxidase activity of enzymes such as xanthine oxidoreductase (XOR). This study was designed to investigate the in vitro and in vivo inhibitory activities of this enzyme by Fraxinus angustifolia extracts. The leaf organic phase of ethyl acetate (LFA) and its bark aqueous counterpart (BFA) showed the strongest anti-NADH oxidase activity in vitro (IC50 = 38.51 and 42.04 µg mL-1, respectively). They consequently suppressed superoxide generation both enzymatically (53% and 19%, respectively) and nonenzymatically (34% and 19%, respectively). These results were corroborated in vivo, with high antiNADH oxidase potential of the leaves and bark extracts (75.32% and 51.32%, respectively) concomitant with moderate hypouricemic activities (36.84% and 38.59%, respectively). Bio-guided fractionation led to the identification, by LC-DAD-MS/MS, of esculin and calcelarioside in bark and kaempferol glucoside in leaves as the main compounds responsible for the anti-NADH oxidase activity of XOR. These results plead in favor of the use of F. angustifolia as a source of potentially interesting therapeutic substances.

Acylated Flavonoid Glycosides are the Main Pigments that Determine the Flower Colour of the Brazilian Native Tree Tibouchina pulchra (Cham.) Cogn.

Tibouchina pulchra (Cham.) Cogn. is a plant native to Brazil whose genus and family (Melastomataceae) are poorly studied with regards to its metabolite profile. Phenolic pigments of pink flowers were studied by ultra-performance liquid chromatography with a photodiode array detector and electrospray ionization quadrupole time-of-flight mass spectrometry. Therein, twenty-three flavonoids were identified with eight flavonols isolated by preparative high-performance liquid chromatography and analysed by one- and two-dimensional nuclear magnetic resonance. Kaempferol derivatives were the main flavonols, encompassing almost half of the detected compounds with different substitution patterns, such as glucoside, pentosides, galloyl-glucoside, p-coumaroyl-glucoside, and glucuronide. Concerning the anthocyanins, petunidin p-coumaroyl-hexoside acetylpentoside and malvidin p-coumaroyl-hexoside acetylpentoside were identified and agreed with previous reports on acylated anthocyanins from Melastomataceae. A new kaempferol glucoside was identified as kaempferol-(2′′-O-methyl)-4′-O-α-d-glucopyranoside. Moreover, twelve compounds were described for the first time in the genus with five being new to the family, contributing to the chemical characterisation of these taxa.