In vitro inhibitory effects of cranberry bean (Phaseolus vulgaris L.) extracts on aldose reductase, α‐glucosidase and α‐amylase

The in vitro inhibitory activities of different seed extracts prepared from cranberry bean mutant SA‐05 and its wild‐type variety Hwachia against aldose reductase, α‐glucosidase and α‐amylase were examined. The results indicated that the polyphenolics‐rich extracts obtained using 800 g kg^?1 methanol and 500 g kg^?1 ethanol demonstrated inhibitory activities against aldose reductase (IC₅₀ of 0.36–0.46 mg mL^?1) and α‐glucosidase (IC₅₀ of 1.32–1.94 mg mL^?1). The 500 g kg^?1 ethanol extracts also showed α‐amylase inhibitory activities (IC₅₀ of 70.11–80.22 μg mL^?1). Subsequent extracts, prepared further with NaCl and H₂O from precipitates of 800 g kg^?1 methanol or 500 g kg^?1 ethanol extracts, exhibited potent α‐amylase inhibitory activities (IC₅₀ of 17.68–38.68 μg mL^?1). A combination of 500 g kg^?1 ethanol extraction plus a subsequent H₂O extraction produced highest polyphenolics and α‐amylase inhibitors. The SA‐05 α‐amylase inhibitor extracts showed greater inhibitory activities than that of Hwachia. Thus, cranberry bean mutant SA‐05 is an advantageous choice for producing anti‐hyperglycaemic compounds.     

Screening grape seeds recovered from winemaking by‐products as sources of reducing agents and mammalian α‐glucosidase and α‐amylase inhibitors

Grape seeds collected from vinification of various grape varieties were extracted by supercritical CO₂ for oil recovery. The defatted residues thus obtained were considered as a re‐utilisable co‐product and assessed for phenolic content, reducing capacity and inhibitory activities against mammalian α‐amylase and α‐glucosidase enzymes. Supercritical CO₂ treatment led to higher recovery of anthocyanins. Reducing capacity of phenolic extracts reached up to ~2200 mmolFe(II) kg^?1, much higher than that of various natural phenolic sources. The anthocyanin‐rich extracts showed the highest inhibitory effectiveness towards α‐glucosidase (I₅₀ value equal to ~40 μg gallic acid equivalents (GAE)/mL ~ half than acarbose). Inhibitory effectiveness towards α‐amylase activity was similar among grape varieties, with I₅₀ values comparable to that of acarbose and correlated with proanthocyanidin contents. These results could pave the way for an efficient processing of grapes, including cascade processes, namely: winemaking, oil extraction from recovered grape seeds and phenolic extraction from defatted grape seeds as potential cost‐effective nutraceuticals.     

Inhibitory potential of phenylpropanoid glycosides from Ligustrum purpurascens Kudingcha against α‐glucosidase and α‐amylase in vitro

The leaves of Ligustrum purpurascens are used in a Chinese traditional tea called small‐leaved kudingcha, which is rich in phenylpropanoid glycosides (PPGs) and has many beneficial properties. Two critical exoacting glycoside hydrolase enzymes (glucosidases) involved in carbohydrate digestion are α‐glucosidase and α‐amylase. We investigated the properties of PPGs from L. purpurascens for inhibiting α‐amylase and α‐glucosidase activity in vitro and found IC₅₀ values of 1.02 and 0.73 mg mL^?1, respectively. The patterns of inhibiting both α‐amylase and α‐glucosidase were mixed‐inhibition type. Multispectroscopy and molecular docking studies indicated that the interaction between PPGs and α‐amylase and α‐glucosidase altered the conformation of enzymes, with binding at the site close to the active site of enzymes resulting in changed enzyme activity. Our studies may help in the further health use of small‐leaved kudingcha.     

Phytosterols in banana (Musa spp.) flower inhibit α‐glucosidase and α‐amylase hydrolysations and glycation reaction

Three phytosterols were isolated from Musa spp. flowers for evaluating their capabilities in inhibiting glucosidase and amylase activities and glycation of protein and sugar. The three phytosterols were identified as β‐sitosterol (PS1), 31‐norcyclolaudenone (PS2) and (24R)‐4α, 14α, 4‐trimethyl‐5α‐cholesta‐8, 25(27)‐dien‐3β‐ol (PS3). IC₅₀ values (the concentration of inhibiting 50% of enzyme activity) of PS1, PS2 and PS3 against α‐glucosidase were 283.67, 11.33 and 43.10 μg mL^?1, respectively. For inhibition of α‐amylase, the IC₅₀ values of PS1, PS2 and PS3 were 52.55, 76.25 and 532.02 μg mL^?1, respectively. PS1 was an uncompetitive inhibitor against α‐amylase with K_m at 5.51 μg mL^?1, while PS2 and PS3 exhibited a mixed‐type inhibition with K_m at 52.36 and 2.49 μg mL^?1, respectively. PS1 and PS2 also significantly inhibited the formation of advanced glycation end products (AGEs) in a BSA–fructose model. The results suggest that banana flower could possess the capability in prevention of the diseases associated with abnormal blood sugar and AGEs levels, such as diabetes.     

α-Glucosidase- and α-amylase-inhibitory activities of phlorotannins from Eisenia bicyclis

BACKGROUND: In an effort to develop alternative therapeutic agents, strong inhibitory activity against α‐glucosidase and α‐amylase was detected in Eisenia bicyclis methanolic extract. RESULTS: In this study, two phlorotannins were isolated from E. bicyclis and characterised by chromatography and nuclear magnetic resonance. The active substances were identified as fucofuroeckol A (FF) and dioxinodehydroeckol (DD). To the authors' knowledge, this is the first report of the identification of these substances in E. bicyclis. However, to date, no antidiabetic activity of FF and DD has been reported. Both phlorotannins demonstrated significant inhibitory activity against α‐glucosidase and α‐amylase. FF showed potent antidiabetic activity, with IC₅₀ values of 131.34 nmol L^?1 against α‐glucosidase and 42.91 µmol L^?1 against α‐amylase. The corresponding IC₅₀ values of DD were 93.33 nmol L^?1 and 472.7 µmol L^?1. Furthermore, kinetic analysis revealed that FF and DD exhibited non‐competitive inhibitory activity against α‐glucosidase. CONCLUSION: These results suggest that FF and DD may be candidates for the development of an antidiabetic pharmaceutical agent or food additive. Copyright © 2012 Society of Chemical Industry     

α‐Glucosidase Inhibitory Activities of Fatty Acids Purified from the Internal Organ of Sea Cucumber Stichopus japonicas

α‐Glucosidase inhibitory activities of the various solvent fractions (n‐hexane, CHCl₃, EtOAc, BuOH, and water) of sea cucumber internal organ were investigated. 1,3‐Dipalmitolein (1) and cis‐9‐octadecenoic acid (2) with potent α‐glucosidase inhibitory activity were purified from the n‐hexane fraction of sea cucumber internal organ. IC₅₀ values of compounds 1 and 2 were 4.45 and 14.87 μM against Saccharomyces cerevisiae α‐glucosidase. These compounds mildly inhibited rat‐intestinal α‐glucosidase. In addition, both compounds showed a mixed competitive inhibition against S. cerevisiae α‐glucosidase and were very stable at pH 2 up to 60 min. The K_I values of compounds 1 and 2 were 0.48 and 1.24 μM, respectively. Therefore, the internal organ of sea cucumber might be a potential new source of α‐glucosidase inhibitors suitably used for prevention of obesity and diabetes mellitus.     

Identification of α‐glucosidase inhibitors from a new fermented tea obtained by tea‐rolling processing of loquat (Eriobotrya japonica) and green tea leaves

BACKGROUND: A new fermented tea produced by tea‐rolling processing of loquat (Eriobotrya japonica) leaf with green tea leaf (denoted as LG tea) showed a potent antihyperglycaemic effect in maltose‐loaded rats. The aim of this study, therefore, was to identify α‐glucosidase inhibitors in the antihyperglycaemic tea product. RESULTS: LG tea had a threefold higher maltase‐inhibitory activity (IC₅₀ 0.065 mg dried extract mL^?1) than either the constituent loquat leaf or green tea alone. In addition, LG tea favourably inhibited maltase action rather than sucrase action. As a result of bio‐guided high‐performance liquid chromatography separations of LG tea, theasinensin A, theasinensin B, strictinin and 1,6‐digalloylglucose were newly identified as maltase inhibitors with IC₅₀ values of 142, 225, 398 and 337 µmol L^?1 respectively, along with previously identified catechins and theaflavins. CONCLUSION: Judging from the magnitude of the α‐glucosidase‐inhibitory contribution of each isolated compound to the overall inhibition of LG tea, catechins were the main candidates responsible for α‐glucosidase or maltase inhibition in LG tea, followed by theaflavins, theasinensins, strictinin and 1,6‐digalloylglucose. Copyright © 2010 Society of Chemical Industry     

Interaction of aqueous extracts of two varieties of Yam tubers (Dioscorea spp) on some key enzymes linked to type 2 Diabetes in vitro

This study sought to assess the effect of processing of yam flour [from white yam (Dioscorea rotundata) and water yam (Dioscorea alata)] to paste, on the antioxidant properties and inhibition of key enzymes linked to type‐2 diabetes (α‐amylase and α‐glucosidase). The phenolic content (phenol and flavonoid), antioxidant properties, α‐amylase and α‐glucosidase inhibitory properties of the aqueous extracts of the yam products were determined. Processing into paste caused significant (P < 0.05) percentage decrease in their total phenol and flavonoid content of the yam varieties. However, significant increase (P < 0.05) in the antioxidant properties of the yam varieties was observed after processing. Furthermore, the yam extracts inhibited α‐amylase and α‐glucosidase activities in vitro in a dose‐dependent parttern (1–4 mg mL^?1), however, the pasting process caused significant (P < 0.05) decrease in the α‐amylase inhibitory and α‐glucosidase inhibitory activities. Therefore, processing the yam varieties into paste (browned) could potentially increase their antioxidant capacity as well as decreasing their α‐amylase and α‐glucosidase inhibitory properties.     

Inhibitory activities of kaempferol,galangin, carnosic acid and polydatin against glycation and α‐amylase and α‐glucosidase enzymes

The activities of four natural phenolics, kaempferol, galangin, carnosic acid and polydatin in scavenging free radicals, inhibiting advanced glycation end‐product (AGE) formation, α‐amylase and α‐glucosidase and trapping methylglyoxal (MGO), were evaluated in this study. Carnosic acid and galangin had the highest activity in scavenging free radicals. Kaempferol and galangin had the greatest activity in preventing bovine serum albumin (BSA) against glycation and reducing glycated proteins. Polydatin had the greatest performance in trapping MGO to reduce glycation reaction. However, there was no significant difference for kaempferol, galangin and carnosic acid in inhibiting AGE formation by BSA‐MGO reaction. Kaempferol, galangin and carnosic acid were the competitive inhibitors for α‐amylase, while kaempferol and carnosic acid were noncompetitive inhibitors for α‐glucosidase. However, polydatin showed as a mixed noncompetitive inhibitor for both α‐amylase and α‐glucosidase. The results indicated that the four natural phenolics have potential in inhibiting AGE production and the digestive enzymatic activity with different mechanisms.     

α‐Amylase inhibitors from an Indonesian medicinal herb,Phyllanthus urinaria

BACKGROUND: Diabetes mellitus and associated diseases are an increasing problem around the world. One of the hyperglycemic remedies is glucose absorption reduction by suppressing carbohydrate digestion due to utilization of α‐amylase inhibitors. RESULTS: Prospective herbs were analyzed by in vitro enzyme assay to evaluate their inhibitory activity against porcine pancreatic amylase (PPA), and it was found that Phyllanthus urinaria and three other herbs to showed a potent inhibitory activity. A 50% aqueous methanol‐soluble extract of the leaves of P. urinaria was chromatographed using a silica gel column. The active fractions were further purified by preparative high‐performance liquid chromatography to isolate active principles against PPA. Structural determination revealed that these isolated compounds were gallic acid, corilagin, and macatannin B, and showed mild activity against PPA (activity in 1 mmol L^?1 concentration: 23%, 21%, 33%, respectively). CONCLUSION: P. urinaria extracts show inhibitory activity against PPA. This activity, together with the information on isolated compounds, may benefit further exploration of P. urinaria utilization in the management of borderline diabetes patients. Copyright © 2011 Society of Chemical Industry     

Beneficial effect of the unsaponifiable matter from rice bran on oxidative stress in vitro compared with α‐tocopherol

Unsaponifiable matter (UM) was prepared from rice bran using n‐hexane extraction followed by removal of its fatty acid methyl ester with supercritical CO₂ under heat‐stable conditions. The UM was made up of 1% of vitamin E isomers, 28% of γ‐oryzanol and 71% of uncharacterized compounds. The aim of this study was to determine the antioxidant activities of the UM, using α‐tocopherol (α‐T) as a positive control, by measuring the Fe³⁺‐reducing antioxidant power (FRAP), 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH^?) free‐radical‐scavenging property and lipid peroxidation in rat liver microsomes. In addition, the effects of the UM on the tert‐butyl hydroperoxide (t‐BOOH)‐induced cytotoxicity in cultured rat hepatocytes were also investigated. In FRAP assay and DPPH^? free‐radical‐scavenging assay, the results were expressed g^?1α‐T or g^?1 UM. The amount of UM used in lipid peroxidation assay and cytotoxicity assay was the amount required to have equal amounts of total vitamin E isomers in the sample and the control α‐T. The UM, as well as α‐T, exhibited significant antioxidant activities in FRAP, radical‐scavenging and membrane‐lipid oxidation. The FRAP value for total vitamin E isomers in the UM (TVEIUM) was 9.1 times higher than that for α‐T. In terms of their capacities to perform radical‐scavenging and lipid peroxidation, both TVEIUM and α‐T showed similar antioxidant activities. In experiments using cultured rat hepatocytes, the t‐BOOH‐induced lactate dehydrogenase release was significantly inhibited by the addition of 63.5 and 160 µg ml^?1 of TVEIUM treatments (84 and 89%, respectively), and that of 63.5 and 160 µg ml^?1 of α‐T treatment (88 and 93%, respectively). The antioxidant function against oxidative stress of the UM prepared from rice bran may extend its use to being a potential dietary supplement. Copyright © 2004 Society of Chemical Industry     

A preliminary study of monosaccharide composition and α‐glucosidase inhibitory effect of polysaccharides from pumpkin (Cucurbita moschata) fruit

In this work, crude polysaccharide extracts were extracted from pumpkin (Cucurbita moschata) fruit by hot water. After removal of proteins and purification, polysaccharides of pumpkin fruit (PP1‐1) were subjected to structural identification. Gas chromatography analysis indicated that PP1‐1 comprised of galactose (86.4%), and glucose (13.6%). The molecular weight of PP1‐1 was measured to be 0.87 × 10⁴ Da by gel permeation chromatography. The inhibitory kinetic evaluation showed that it was non‐competitive inhibition of PP1‐1 on the α‐glucosidase‐catalysed hydrolysis of PNPG. The Michaelis–Menten constant (K_m) was 0.106 m , and the inhibitory constants (K_i), 0.435 mg.     

Glycoalkaloids in potato tubers: the effect of variety and drought stress on the α‐solanine and α‐chaconine contents of potatoes

Six varieties of Solanum tuberosum L potato grown in the Bolivian highlands under drought stress, with and without irrigation, were analysed for their content of glycoalkaloids (GAs). The plant material consisted of three drought‐tolerant varieties from a local breeding programme (PROINPA), Potosina, Chapaquita and Pampeña, and three control cultivated varieties, Malcacho, Sani Imilla and Desiree, either susceptible or relatively tolerant to drought. α‐Solanine and α‐chaconine were quantified in both the peel and flesh of the tubers. A significant increase in GA concentration (α‐solanine + α‐chaconine) was observed under drought stress conditions in most varieties; average concentration increases of 43 and 50% were registered in the improved and control cultivars respectively. In all tested cultivars, however, the GA concentration remained lower than the recommended food safety level (200 mg kg⁻¹ fresh tubers). It ranged from 52.4 to 100 mg kg⁻¹ fresh tubers in the improved cultivars and from 55.6 to 122.3 mg kg⁻¹ fresh tubers in the controls. In the improved and control varieties the α‐solanine content averaged 42.6 and 35.4% of the total potato GAs respectively and was not significantly affected by drought stress, except in Desiree. In all conditions the peel contained the greatest proportion of total GAs. The hybrid variety Pampeña (new drought‐tolerant variety) contained the lowest amounts of GAs, which were lower than those of the control varieties, with and without irrigation. © 2000 Society of Chemical Industry     

Change in α‐tocopherol contents,lipid oxidation and fatty acid profile in eggs enriched with linolenic acid or very long‐chain ω3 polyunsaturated fatty acids after different processing methods

The influences of dietary supplementation with α‐tocopheryl acetate (α‐TA) and of processing (by hard‐boiling and scrambling) of eggs enriched with ω3 fatty acids, either very long‐chain ω3 polyunsaturated fatty acids (VLC ω3 PUFAs) or linolenic acid (LNA), on fatty acid composition, α‐tocopherol content and lipid oxidation (thiobarbituric acid (TBA) values) were studied. Four dietary treatments were formulated from a basal diet containing 40 g kg^?1 linseed oil (LO) or fish oil (FO) combined with either 0 or 100 mg α‐TA kg^?1 of feed. Eggs from LO treatments were enriched with LNA and those from FO treatments were rich in VLC ω3 PUFAs. Neither processing nor dietary supplementation with α‐TA modified greatly the fatty acid profile of eggs. Dietary supplementation with α‐TA increased the α‐tocopherol content of eggs (187.2 versus 407.9 µg g^?1 dry matter). Eggs from FO treatments showed lower α‐tocopherol content than those from LO treatments (273.5 versus 321.6 µg g^?1 dry matter), and processing of eggs enriched with VLC ω3 PUFA reduced the α‐tocopherol content by a significant 16%. Moreover, processing of eggs increased lipid oxidation two‐ to nine‐fold. Oxidation levels of hard‐boiled eggs were 30.4% higher than those of scrambled eggs. TBA values in hard‐boiled and scrambled eggs were significantly reduced when 100 mg α‐TA kg^?1 of feed supplemented the diet only in those eggs enriched with VLC ω3 PUFA (from FO treatments). Copyright © 2003 Society of Chemical Industry     

Extraction,optimisation and characterisation of phenolics from Thymus vulgaris L.: phenolic content and profiles in relation to antioxidant,antidiabetic and antihypertensive properties

The objectives of this study were to examine varying extraction conditions of Thymus vulgaris L. as related to phenolic content and profiles of the extracts and their antioxidant, antihypertensive and antidiabetic properties. Phenolics were extracted under various conditions pertaining to free and bound phenolics, solvent type and combination of extraction time and temperature, and these extracts were evaluated in terms of their antioxidant activities and inhibitory activities of angiotensin‐converting enzyme (ACE), α‐glucosidase and α‐amylase. The acetone–water solvent mixture (1:1; v/v) produced the extract with the greatest phenolic content, antioxidant activity and inhibitory activities of ACE and α‐glucosidase. The optimal extraction temperature for maximum phenolic content and antioxidant activity associated with methanol extraction was 60 °C, whereas a lower temperature at 40 °C was required to maximise inhibitory activities for ACE, α‐glucosidase and α‐amylase. An inverse relationship was seen between antioxidant and glucosidase inhibitory activities vs. the ACE and α‐amylase inhibitory activities, which suggests the need for extractions to be directed to specific bioactivities of thyme extracts. Generally, the results indicate major differences in phenolic profiles among the tested extraction conditions with thymol as the predominant phenolic seen in most extractions, while gallic acid, rosmarinic acid or diosmin also predominated in other extracts. Extracts with the same predominant phenolic compound and similar phenolic content showed major disparities in their ACE, glucosidase and α‐amylase inhibitory activities, indicating that the major phenolic profiles of thyme extracts may not be necessarily related to the degree of inhibition of ACE, glucosidase and α‐amylase enzymes.     

Anti‐acetylcholinesterase,antidiabetic, anti‐inflammatory,antityrosinase and antixanthine oxidase activities of Moroccan propolis

Biological properties of Moroccan propolis have been scarcely studied. In the present work, the total phenols and flavonoids from 21 samples of propolis collected in different places of Morocco or 3 supplied in the market were determined, as well as the in vitro capacity for inhibiting the activities of acetylcholinesterase, α‐glucosidase, α‐amylase, lipoxygenase, tyrosinase, xanthine oxidase and hyaluronidase. The results showed that samples 1 (region Fez‐Boulemane, Sefrou city) (IC₅₀ = 0.065, 0.006, 0.020, 0.050, 0.014 mg mL^?1) and 23 (marketed) (IC₅₀ = 0.018, 0.002, 0.046, 0.037, 0.008 mg mL^?1) had the best in vitro capacity for inhibiting the α‐amylase, α‐glucosidase, lipoxygenase, tyrosinase and xanthine oxidase activities, respectively. A negative correlation between IC₅₀ values and concentration of phenols, flavones and flavanones was found. These activities corresponded to the generally higher amounts of phenols and flavonoids. In the same region, propolis samples have dissimilar phenol content and enzyme inhibitory activities.     

α‐Eleostearic acid is more effectively metabolized into conjugated linoleic acid than punicic acid in mice

BACKGROUND: The objective of this study was to investigate the metabolism of α‐eleostearic acid (α‐ESA, cis9,trans11,trans13‐18:3) and punicic acid (PA, cis9,trans11,cis13‐18:3) and to compare the relative conversion efficiency of these fatty acids into conjugated linoleic acids (CLAs) in mice. RESULTS: Twenty‐four male ICR mice were fed either a control diet or one of two experimental diets supplemented with 10 g kg^?1α‐ESA or PA in the form of triacylglycerols for 6 weeks. The accumulation of PA in all tissues examined was significantly higher than α‐ESA; in both groups it was found that cis9,trans11‐conjugated linoleic acid (cis9,trans11‐18:2) was detected in all tissues examined. The relative conversion rate of α‐ESA into cis9,trans11‐18:2 was significantly higher than that of PA. The highest conversion rate of α‐ESA was found in adipose tissue (91.8%), spleen (91.4%) and kidney (90.7%), and the lowest in the heart (84.6%). The highest conversion rate of PA occurred in the liver (76.2%) and the lowest occurred in the heart (54.5%). CONCLUSION: The present results indicate that both α‐ESA and PA are effectively metabolized into cis9,trans11‐18:2 in mice. These conjugated dietary fatty acids might be useful sources of CLAs in tissues owing to the natural abundance of α‐ESA and PA in some seeds. Copyright © 2009 Society of Chemical Industry     

In vitro inhibition of α‐chymotryptic activity by phenolic compounds

α‐Chymotrypsin was modified by covalent attachment of selected phenolic and related compounds (caffeic acid, chlorogenic acid, ferulic acid, gallic acid, quinic acid, m‐/o‐/p‐dihydroxybenzene and p‐benzoquinone) at pH 9. The derivatives formed were characterised in terms of their activity and selected physicochemical properties. In vitro experiments showed that the proteolytic digestion of food proteins with α‐chymotrypsin derivatives was adversely affected. This decrease depended on the reactivity of the phenolic and related substances tested as well as on the kind of substrate applied. The derivatisation was accompanied by a reduction in the amount of free lysine and tryptophan residues. Moreover, the solubility of the derivatives decreased over a broad pH range, with a parallel increase in the hydrophobicity. The isoelectric point was shifted to a lower pH value, and formation of high‐molecular‐weight fractions was documented by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS‐PAGE). © 2001 Society of Chemical Industry     

Changes of porcine pancreas α‐amylase in activity and secondary conformations under inhibition of tea polyphenols

To investigate two‐sided functions of tea polyphenols (TP) in antinutrition and energy balance modulation, TP were extracted from Chinese green tea and used to complex porcine pancreas α‐amylase (PPA). Changes of PPA in activity and secondary conformations were analysed. Porcine pancreas α‐amylase was found sensitive to TP treatment. Tea polyphenols exhibited IC₅₀ at 0.41 mg mL^?1 against PPA and maximum inhibitory rate (98.17%) at 3.0 mg mL^?1. Tea polyphenols inhibition was concluded as noncompetitive pattern based on its unchanged Km value (0.98 mg mL^?1) for soluble starch substrate. Tea polyphenols inhibition arose from pH 1.5 to 10.14, covering gastric and intestinal environments inside body. Circular dichroism spectra analysis revealed regular changes of PPA in secondary conformations (increased proportions of α‐helix and β‐sheet) prior to its inactivation at low TP concentrations. Tea polyphenols‐inhibited PPA had distinct double‐negative peaks at 204 nm and 208 nm. Porcine pancreas α‐amylase was inactivated by TP in ways of complexation and modification of secondary conformations.