Phenolic compounds and antioxidant activity in sweet potato after heat treatment.

BACKGROUND: The ability of heat treatment with a soaking solvent to increase soluble phenolic compounds due to the liberation or breakdown of the cell matrix has been investigated in various plants. This study investigated the changes in phenolic compounds and antioxidant activities of 12 sweet potato cultivars after heat treatment with distilled water or prethanol A. RESULTS: The highest total polyphenol content (134.67 mg gallic acid equivalents/g extract residue) and flavonoid content (65.43 mg catechin equivalents/g extract residue) was observed in the 'Jami' (JM) cultivar after heat treatment with prethanol A. Higher polyphenol and flavonoid content was generally observed in the purple sweet potato cultivars. Salicylic acid was the major phenolic acid, followed by protocatechuic acid or chlorogenic acid in almost all untreated sweet potato cultivars. The salicylic acid, vanillic acid, gallic acid, and caffeic acid content of the sweet potatoes increased after the heat treatment, whereas the protocatechuic acid and chlorogenic acid content decreased. The highest 1,1-Diphenyl-2-picrylhydrazyl (DPPH) and 2,2-azinobis(3-ethyl benzothiazoline)-6-sulfonic acid (ABTS) radical scavenging activity levels were observed in the JM cultivar subjected to heat treatment with prethanol A (48.15 and 80.00 mg TE/g extract residue, respectively). CONCLUSION: These results suggest that heat treatment with a soaking solvent is an efficient method to enhance the antioxidant characteristics of Korean sweet potato cultivars. © 2019 Society of Chemical Industry.

Oat germination and ultrafiltration process improves the polyphenol and avenanthramide contents with protective effect in oxidative-damaged HepG2 cells.

Oat is the nutritious crop containing various compounds with antioxidant properties, such as polyphenols. In this study, we investigated the effect of germination and ultrafiltration process on polyphenol and avenanthramide contents in oat as well as their cytoprotective effect. Germination of oat for 48 hr significantly increased avenanthramide (5.5 to 11.3 mg/g) and polyphenol (115 to 155 mg GAE/g) contents. The compounds were more concentrated after ultrafiltration using 10 kDa membranes (polyphenol, 206 GAE/g; avenanthramide, 18 mg/g). In addition, oat extracts significantly reduced the cellular ROS level against tert-butyl hydroperoxide (t-BHP) stimulation in HepG2 cells. In the mechanistic study, oat extracts induced Nrf2 translocation to the nucleus by inhibition of Keap1 expression, resulting into upregulation of γ-GCS and NQO1. In conclusion, oat germination and ultrafiltration processes increased the polyphenol content, including that of avenanthramide. These extracts protected cells from t-BHP by radical scavenging activities and induced Nrf2 pathway activation. PRACTICAL APPLICATIONS: This study presents the method for avenanthramide-concentrated extract which is unique bioactive compounds in oat. In addition, antioxidant activity and their mechanisms of the avenanthramide-enriched extracts were evaluated. The polyphenol compounds including avenanthramide were found to increase after germination and ultrafiltration, thereby improving the radical scavenging ability. These results can be utilized as data for the development of health-promoting materials using oats.

Antidiabetic Effects of Vigna nakashimae Extract in Humans: A Preliminary Study.

BACKGROUND: Vigna nakashimae (VN) extract has been shown to have antidiabetic and antiobesity effects in various animal studies; however, to our knowledge, no data on such effects exist in humans. METHODS: We performed a randomized placebo-controlled study to investigate the antidiabetic effects of VN extract treatment for 12 weeks in humans. A total of 18 Korean patients with type 2 diabetes were enrolled in this study and were allocated randomly to either the VN extract group (1 g thrice daily) or control group (placebo tablets) for 12 weeks. We investigated blood glucose levels, body weight, lipid profiles, and adverse events after 12 weeks of treatment. Fifteen subjects were included in the final analysis. RESULTS: There was no difference in age, sex, fasting glucose levels, or lipid profiles between the VN extract and control groups at baseline. However, the baseline glycosylated hemoglobin (HbA1c) levels of the control group were lower than those of the VN extract group. After treatment with VN extract for 12 weeks, the body weight and lipid profile of the VN extract group remained unchanged; however, the HbA1C levels decreased by 0.36% ± 0.33% (p = 0.027). In contrast, the HbA1C levels of the control group did not change after 12 weeks (p = 0.228). During the 12-week treatment with VN extract, no serious adverse events were reported. CONCLUSION: Our data indicate that VN extract has implications for glucose lowering in type 2 diabetic patients.

Comparative Biochemical and Proteomic Analyses of Soybean Seed Cultivars Differing in Protein and Oil Content.

This study develops differential protein profiles of soybean (Glycine max) seeds (cv. Saedanbaek and Daewon) varying in protein (47.9 and 39.2%) and oil (16.3 and 19.7%) content using protamine sulfate (PS) precipitation method coupled with a 2D gel electrophoresis (2DGE) approach. Of 71 detected differential spots between Daewon and Saedanbaek, 48 were successfully identified by MALDI-TOF/TOF. Gene ontology analysis revealed that up-regulated proteins in Saedanbaek were largely associated with nutrient reservoir activity (42.6%), which included mainly seed-storage proteins (SSPs; subunits of glycinin and ß-conglycinin). Similar results were also obtained in two cultivars of wild soybean (G. soja cv. WS22 and WS15) differing in protein content. Western blots confirmed higher accumulation of SSPs in protein-rich Saedanbaek. Findings presented and discussed in this study highlight a possible involvement of the urea cycle for increased accumulation of SSPs and hence the higher protein content in soybean seeds.

Effects of Mung Bean (Vigna radiata L.) Ethanol Extracts Decrease Proinflammatory Cytokine-Induced Lipogenesis in the KK-Ay Diabese Mouse Model.

Rapid increase in the prevalence of obesity-related metabolic inflammatory diseases has led to research focused on nutraceuticals for their treatment. This study investigated the effects of the ethanol extracts of mung bean testa (MBT) on the metabolic inflammation-induced lipogenesis in gastrocnemius muscle of KK-Ay diabese mice. Ethanol extracts of MBT were orally administered to diabese KK-Ay mice for 4 weeks after diet-induced obesity model was generated by feeding a 60% high-fat diet for 3 weeks. Although there were no changes in body weight gain, MBT treatments decreased total weight of white adipose tissue. MBT also decreased triacylglycerol and total cholesterol levels in the muscle by 30%, which was correlated with suppression of lipogenic genes such as ACC, C/EBP alpha, PGC-1 alpha, and PPAR gamma. In particular, decreased levels of p-ERK1/2, PPAR gamma, and C/EBP alpha in the MBT-treated groups suggest that MBT might inhibit adipogenesis and decrease differentiation via the MEK/ERK pathway. Furthermore, significantly lower amounts of plasma interleukin (IL)-6 and intramuscular tumor necrosis factor (TNF)-alpha and monocyte chemoattractant protein-1 (MCP-1) were detected in MBT groups, confirming the anti-inflammatory effect of mung bean. In addition, our in vitro pilot study with 3T3-L1 cells showed that vitexin, the functional chemical in MBT, inhibited inflammation-induced lipogenesis with significantly lower amounts of IL-6 and MCP-1 after 14 days of vitexin treatment. Thus, the functional compounds in the mung bean ethanol extracts such as vitexin and isovitexin may regulate intracellular lipogenesis and adipogenesis via anti-inflammatory mechanisms and MEK/ERK pathway in the KK-Ay mouse model.

Attenuation by a Vigna nakashimae extract of nonalcoholic fatty liver disease in high-fat diet-fed mice.

A Vigna nakashimae (VN) extract has been shown to have antidiabetic and anti-obesity effects. However, the mechanism underlying the effect of a VN extract on hepatic inflammation and endoplasmic reticulum (ER) stress remains unclear. In the present study, we investigated how a VN extract protects against the development of non-alcoholic fatty liver disease (NAFLD). A VN extract for 12 weeks reduced the body weight, serum metabolic parameters, cytokines, and hepatic steatosis in high-fat diet (HFD)-fed mice. A VN extract decreased HFD-induced hepatic acetyl CoA carboxylase and glucose transporter 4 expressions. In addition to the levels of high-mobility group box 1 and receptor for advanced glycation, the hepatic expression of ATF4 and caspase-3 was also reduced by a VN extract. Thus, these data indicate that a chronic VN extract prevented NAFLD through multiple mechanisms, including inflammation, ER stress, and apoptosis in the liver.

Inhibition of tyrosinase activity by polyphenol compounds from Flemingia philippinensis roots.

Flemingia philippinensis is used as a foodstuff or medicinal plant in the tropical regions of China. The methanol (95%) extract of the roots of this plant showed potent tyrosinase inhibition (80% inhibition at 30µg/ml). Activity-guided isolation yielded six polyphenols that inhibited both the monophenolase (IC50=1.01-18.4µM) and diphenolase (IC50=5.22-84.1µM) actions of tyrosinase. Compounds 1-6 emerged to be three new polyphenols and three known flavanones, flemichin D, lupinifolin and khonklonginol H. The new compounds (1-3) were identified as dihydrochalcones which we named fleminchalcones (A-C), respectively. The most potent inhibitor, dihydrochalcone (3) showed significant inhibitions against both the monophenolase (IC50=1.28µM) and diphenolase (IC50=5.22µM) activities of tyrosinase. Flavanone (4) possessing a resorcinol group also inhibited monophenolase (IC50=1.79µM) and diphenolase (IC50=7.48µM) significantly. In kinetic studies, all isolated compounds behaved as competitive inhibitors. Fleminchalcone A was found to have simple reversible slow-binding inhibition against monophenolase.

α-Glucosidase inhibition and antihyperglycemic activity of prenylated xanthones from Garcinia mangostana.

An ethanol extract of the fruit case of Garcinia mangostan, whose most abundant chemical species are xanthones, showed potent α-glucosidase inhibitory activity (IC(50)=3.2 µg/ml). A series of isolated xanthones (1-16) demonstrated modest to high inhibition of α-glucosidase with IC(50) values of 1.5-63.5 µM. In particular, one hitherto unknown xanthone 16 has a very rare 2-oxoethyl group on C-8. Kinetic enzymatic assays with a p-nitrophenyl glucopyranoside indicated that one of them, compound (9) exhibited the highest activity (K(i)=1.4 µM) and mixed inhibition. Using, a physiologically relevant substrate, maltose, as substrate, many compounds (6, 9, 14, and 15) also showed potent inhibition which ranged between 17.5 and 53.5 µM and thus compared favorably with deoxynojirimycin (IC(50)=68.8 µM). Finally, the actual pharmacological potential of the ethanol extract was demonstrated by showing that it could elicit reduction of postprandial blood glucose levels. Furthermore, the most active α-glucosidase inhibitors (6, 9, and 14) were proven to be present in high quantities in the native seedcase by a HPLC chromatogram.

Isolation of cholinesterase-inhibiting flavonoids from Morus lhou.

Cholinesterases are key enzymes that play important roles in cholinergic transmission. Nine flavonoids displaying cholinesterase inhibitory activity were isolated from the root bark of Morus lhou L., a cultivated edible plant. The isolated compounds were identified as a new flavone (1), 5'-geranyl-5,7,2',4'-tetrahydroxyflavone (2), kuwanon U (3), kuwanon E (4), morusin (5), morusinol (6), cyclomorusin (7), neocyclomorusin (8), and kuwanon C (9). All compounds apart from compound 6 inhibited cholinesterase enzyme in a dose-dependent manner with K(i) values ranging between 3.1 and 37.5 µM and between 1.7 and 19.1 µM against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzymes, respectively. The new compound was charactierized as 5'-geranyl-4'-methoxy-5,7,2'-trihydroxyflavone (1). It showed the most potent inhibitory activity (K(i) = 3.1 µM for AChE, K(i) = 1.74 µM for BChE). Lineweaver-Burk and Dixon plots and their secondary replots indicated that flavones (5-9) with prenyl substitution on C-3 were noncompetitive inhibitors, whereas those unsubstituted (1-4) at C-3 were mixed inhibitors of both AChE and BChE. In conclusion, this is the first study to demonstrate that alkylated flavonoids of M. lhou have potent inhibitory activities against AChE and BChE.

Polyphenols from Broussonetia papyrifera displaying potent alpha-glucosidase inhibition.

The organic extract of the roots of Broussonetia papyrifera showed extremely high alpha-glucosidase inhibitory activity with an IC50 of around 10 microg/mL. Due to its potency, subsequent bioactivity-guided fractionation of the chloroform extract led to 12 polyphenols, 1-12, 4 of which were identified as chalcones (1-4), another 4 as flavans (5-8), 2 as flavonols (9 and 10), and 2 others as the novel species benzofluorenones (11 and 12). Broussofluorenone A (11) and broussofluorenone B (12) emerged as new compounds possessing the very rare 5,11-dioxabenzo[b]fluoren-10-one skeleton. These compounds (1-12) were evaluated for alpha-glucosidase inhibitory activity to identify their inhibitory potencies and kinetic behavior. The most potent inhibitor, 10 (IC50=2.1 microM, Ki=2.3 microM), has an inhibitory activity slightly higher than that of the potent alpha-glucosidase inhibitor deoxynojirimycin (IC50=3.5 microM). The novel alpha-glucosidase inhibitors 11 (IC50=27.6 microM) and 12 (IC50=33.3 microM) are similar in activity to sugar-derived alpha-glucosidase inhibitors such as voglibose (IC50=23.4 microM). Interestingly, major constituents (1, 2, 6, 7, 9, and 10) of B. papyrifera displayed significant inhibitory activity with IC50 values of 5.3, 11.1, 12.0, 26.3, 3.6, and 2.1 microM, respectively. In kinetic studies, chalcones (1-4) exhibited noncompetitive inhibition characteristics, whereas the others (5-12) showed mixed behavior.

Xanthones from Cudrania tricuspidata displaying potent alpha-glucosidase inhibition.

We have proven that xanthones 1-8 isolated from the root of C. tricuspidata possess highly potent alphaalpha-glucosidase inhibition properties. Compound 1 was identified as a new isoprenylated tetrahydroxy xanthone, 1,3,6,7-tetrahydroxy-2-(3-methylbut-2-enyl)-8-(2-methylbut-3-en-2-yl)-9H-xanthen-9-one (1). These are the first natural xanthones documented to exhibit such inhibition. The IC(50) values of compounds 1-8 inhibiting alpha-glucosidase activity were determined to be up to 16.2microM. Mechanistic analysis showed the xanthones 1-8 exhibited full mixed inhibition.

Cudraflavanone A purified from Cudrania tricuspidata induces apoptotic cell death of human leukemia U937 cells, at least in part, through the inhibition of DNA topoisomerase I and protein kinase C activity.

A chloroform extract of the root bark of Cudrania tricuspidata showed an inhibitory effect on mammalian DNA topoisomerase I. The topoisomerase I inhibitory compound was purified and identified as 2S-2',5,7-trihydroxy-4',5'-(2,2-dimethylchromeno)-6-prenyl flavanone (cudraflavanone A). Cudraflavanone A was shown to inhibit the activity of topoisomerase I with approximately 0.4 mmol/l 50% inhibitory concentration. A concentration of 6 micromol/l cudraflavanone A caused a 50% growth inhibition of human cancer cell U937. Cudraflavanone A-induced cell death was characterized by the cleavage of poly(ADP-ribose) polymerase and pro-caspase-3. Furthermore, cudraflavanone A induced the fragmentation of DNA into multiples of 180 bp (an apoptotic DNA ladder), indicating that the inhibitor triggered apoptosis. This induction of apoptosis by cudraflavanone A was also confirmed using flow-cytometry analysis. In addition, this compound inhibited protein kinase C activity with approximately 150 micromol/l 50% inhibitory concentration. Taken together, these results suggest that cudraflavanone A may function by inhibiting oncogenic disease, at least in part, through the inhibition of protein kinase C and topoisomerase I activity.

2',5,7-Trihydroxy-4',5'-(2,2-dimethylchromeno)-8-(3-hydroxy-3-methylbutyl) flavanone purified from Cudrania tricuspidata induces apoptotic cell death of human leukemia U937 cells.

Cellular DNA topoisomerase I is an important target in cancer chemotherapy. A chloroform extract of the root barks of Cudrania tricuspidata showed an inhibitory effect on mammalian DNA topoisomerase I. The topoisomerase I inhibitory compound was purified and identified as 2',5,7-trihydroxy-4',5'-(2,2-dimethylchromeno)-8-(3-hydroxy-3-methylbutyl) flavanone. The compound, temporarily designated as PKH-3, was shown to inhibit the activity of topoisomerase I with IC50 about 1.0 mM. Concentration of 10 microM PKH-3 caused 50% growth inhibition of human cancer cell U937. PKH-3-induced cell death was characterized with the cleavage of poly(ADP-ribose) polymerase (PARP) and pro-caspase 3. Furthermore, PKH-3 induced the fragmentation of DNA into multiples of 180 b.p. (an apoptotic DNA ladder), indicating that the inhibitor triggered apoptosis. This induction of apoptosis by PKH-3 was also confirmed using flow cytometry analysis. Taken together, these results suggest that PKH-3 may function by inhibiting oncogenic disease, at least in part, through the inhibition of topoisomerase I activity.

LDL-antioxidant pterocarpans from roots of Glycine max (L.) Merr.

The methanolic root extract of Glycine max (L.) Merr. was chromatographed, which yielded 10 flavonoids, including three isoflavones 1-3, five pterocarpans 4-8, one flavonol 9, and one anthocyanidin 10. All isolated compounds were examined for LDL-antioxidant activities using four different assay systems on the basis of Cu2+-mediated oxidation. Among them, seven compounds showed potent LDL-antioxidant activities in the thiobarbituric acid reactive substances (TBARS) assay, the lag time of conjugated diene formation, relative electrophoretic mobility (REM), and fragmentation of apoB-100 on copper-mediated LDL oxidation. Three pterocarpans 4, 6, and 7, never reported as LDL-antioxidant, showed potent activities with IC50 values of 19.8, 0.9, 45.0 microM, respectively, in comparison with probucol (IC50 = 5.6 microM) as positive control. Interestingly, coumestrol 6 (IC50 = 0.9 microM) showed 20 times more activity in the TBARS assay than genistein (IC50 = 30.1 microM) and daidzein (IC50 = 21.6 microM), representative antioxidants in soybean. Moreover, coumestrol 6 had an extended lag time of 190 min at 3.0 microM in measuring conjugated diene formation, while both genistein (120 min) and daidzein (93 min) lag times were extended to less than 120 min at the same concentration.

Cytotoxic xanthones from Cudrania tricuspidata.

The new isoprenylated tetrahydroxyxanthone, 2,3,6,8-tetrahydroxy-1-(3-methylbut-2-enyl)-5-(2-methylbut-3-en-2-yl)-9H-xanthen-9-one (1), was isolated from the root bark of Cudrania tricuspidata together with macluraxanthone B (2) and cudraxanthone L (3), which were fully characterized by NMR spectroscopic and X-ray crystallographic analyses.