Bioassay-guided fractionation and identification of α-amylase inhibitors from Syzygium cumini leaves

Context: Pancreatic α-amylase and α-glucosidase inhibitors serve as important strategies in the management of blood glucose. Even though Syzygium cumini (L.) Skeels (Myrtaceae) (SC) is used extensively to treat diabetes; scientific evidence on antidiabetic effects of SC leaves is scarce.

Objective: SC leaf extract was investigated for α-amylase inhibitory effect and continued with isolation and identification of α-amylase inhibitors.

Materials and methods: Bioassay-guided fractionation was conducted using in vitro α-amylase inhibitory assay (with 20–1000?μg/mL test material) to isolate the inhibitory compounds from ethyl acetate extract of SC leaves. Structures of the isolated inhibitory compounds were elucidated using ¹H NMR and ¹³C NMR spectroscopic analysis and direct TLC and HPLC comparison with authentic samples. Study period was from October 2013 to October 2015.

Results: An active fraction obtained with chromatographic separation of the extract inhibited porcine pancreatic α-amylase with an IC₅₀ of 39.9?μg/mL. Furthermore, it showed a strong inhibition on α-glucosidase with an IC₅₀ of 28.2?μg/mL. The active fraction was determined to be a 3:1 mixture of ursolic acid and oleanolic acid. Pure ursolic acid and oleanolic acid showed IC₅₀ values of 6.7 and 57.4?μg/mL, respectively, against α-amylase and 3.1 and 44.1?μg/mL respectively, against α-glucosidase.

Discussion and conclusions: The present study revealed strong α-amylase and α-glucosidase inhibitory effects of ursolic acid and oleanolic acid isolated from SC leaves for the first time validating the use of SC leaves in antidiabetic therapy.     

Inhibition of key enzymes linked to type 2 diabetes by compounds isolated from Aframomum melegueta fruit

Context: The use of Aframomum melegueta K. Schum. (Zingiberaceae) fruit for treatment of diabetes has recently been established in Nigeria. However, compounds responsible for the antidiabetic action have not been identified.

Objective: The present study carried out the bioassay-guided isolation of possible bioactive compounds responsible for the antidiabetic action of A. melegueta fruit.

Materials and methods: The A. melegueta fruit was sequentially extracted using ethyl acetate (EtOAc), ethanol and water, and the most active extract (EtOAc) was subjected to column chromatography on a silica gel column using solvent gradient systems of hexane (HEX):EtOAc and EtOAc:MeOH and the isolation of compounds was guided by α-glycosidase and α-amylase inhibitory activities at various concentrations (30–240?μg/mL).

Results: According to the results, 3 arylalkanes, 6-paradol (1), 6-shogaol (2) and 6-gingerol (3) and a pentacyclic triterpene, oleanolic acid (4) were isolated from A. melegueta fruit. All the compounds exhibited inhibitory effects against α-amylase and α-glucosidase. 6-Gingerol (3) and oleanolic acid (4) showed higher inhibitory activity against α-amylase (IC₅₀: 6-gingerol: 81.78?±?7.79?μM; oleanolic acid: 91.72?±?1.63?μM) and α-glucosidase (IC₅₀: 6-gingerol: 21.55?±?0.45?μM; oleanolic acid: 17.35?±?0.88?μM) compared to the standard drug, acarbose and other isolated compounds. The kinetics of the enzyme action of the compounds showed a noncompetitive mode of inhibition.

Conclusion: The data of this study suggest that the 6-gingerol (3) and oleanolic acid (4) showed higher α-amylase and α-glucosidase inhibitory action and therefore could be responsible for the antidiabetic activity of A. melegueta fruit.     

Kinetics of α-glucosidase inhibition by different fractions of three species of Labiatae extracts: a new diabetes treatment model

Context: Glucosidases are a group of enzymes playing crucial roles in digestion of carbohydrates. Glucosidase inhibitors can reduce carbohydrate digestion rate and have the potential to prevent development of type 2 diabetes. The Labiatae is one of the largest plant families grown globally and many studies that have isolated new pharmaceutical compounds. In folk medicine, some of Labiatae plants such as Zataria multiflora Boiss, Salvia mirzayanii Rech. F. &; Esfand, and Otostegia persica Boiss are consumed for the treatment of diabetes.

Objectives: This study investigates the inhibitory effects of different fractions of three mentioned species extracts on α-glucosidase.

Materials and methods: Ethanol extracts of these plants leaves were fractionated using petroleum ether, chloroform, ethyl acetate, and n-butanol solutions. The duration of this study was 12?months. To measure enzyme inhibition, 5?μL of the enzyme, 20?μL of substrate and samples were used and for evaluation mode of inhibition, constant amounts of α-glucosidase were incubated with rising concentrations of substrate (PNPG).

Results: The results revealed that the ethyl acetate fraction of Zataria multiflora (IC₅₀ =?0.35?±?0.01?mg/mL) and petroleum ether fraction of Salvia mirzayanii (IC₅₀ =?0.4?±?0.11?mg/mL) were the most potent inhibitors of α-glucosidase in comparison with the other samples and acarbose as the standard (IC₅₀ =?7?±?0.19?mg/mL). All of the samples exhibited noncompetitive-uncompetitive inhibition.

Discussion and conclusion: It can be inferred from this study that α-glucosidase inhibitory potential of the studied extracts may be a marker of antidiabetic potential of these extracts.     

Phytochemical analysis,antiproliferative and antioxidant activities of Chrozophora tinctoria: a natural dye plant

Context: Chrozophora tinctoria (L.) A. Juss. (Euphorbiaceae) is known as ‘dyer’s-croton’ and used to obtain dye substances. Recently, natural antioxidants and colorants have been of interest because of their safety and therapeutic effects.

Objective: This study investigates the antiproliferative and antioxidant activities of the various extracts and fractions from C. tinctoria and analyzes their phytochemical contents.

Materials and methods: The aerial parts of C. tinctoria were extracted with water, ethyl acetate, n-butanol, and methanol/chloroform. Phenolic compounds and other constituents of the extracts were analyzed by HPLC/TOF-MS. The ethyl acetate extract (EA) was fractionated by flash chromatography. The extracts, fractions, and major phenolic compounds were investigated for their antiproliferative activities on human cervical adenocarcinoma (HeLa) cell line at the concentrations of 5–100?μg/mL by using BrdU ELISA assay during 24?h of incubation. DPPH radical scavenging activities (5–150?μg/mL) and total phenolic contents of the samples were also evaluated.

Results: 4-Hydroxybenzoic acid (268.20?mg/kg), apigenin-7-glucoside (133.34?mg/kg), and gallic acid (68.92?mg/kg) were the major components of EA. CT/E-F6 (IC₅₀?=?64.59?±?0.01?μg/mL) exhibited the highest antiproliferative activity. CT/E-F2 (IC₅₀=?14.0?±?0.0?μg/mL) and some fractions displayed higher radical scavenging activity compared to synthetic antioxidant BHT (IC_50?=_?23.1?±?0.0?μg/mL). Among the main phenolics, gallic acid exhibited the highest antiproliferative and radical scavenging abilities (IC_50?<_?5?μg/mL).

Conclusion: In this study, we have determined the biologically active fractions and their high effects may be attributed to the presence of gallic acid.     

Unlocking the in vitro anti-inflammatory and antidiabetic potential of Polygonum maritimum

Context: Several Polygonum species (Polygonaceae) are used in traditional medicine in Asia, Europe and Africa to treat inflammation and diabetes.

Objective: Evaluate the in vitro antioxidant, anti-inflammatory and antidiabetic potential of methanol and dichloromethane extracts of leaves and roots of the halophyte Polygonum maritimum L.

Material and methods: Antioxidant activity was determined (up to 1?mg/mL) as radical-scavenging activity (RSA) of 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), copper (CCA) and iron (ICA) chelating activities and iron reducing power (FRAP). NO production was measured in lipopolysaccharide (LPS)-stimulated macrophages for 24?h at concentrations up to 100?μg/mL and antidiabetic potential was assessed by α-amylase and α-glucosidase inhibition (up to 10?mg/mL) assays. The phytochemical composition of the extracts was determined by gas chromatography-mass spectrometry (GC-MS).

Results: The methanol leaf extract had the highest activity against DPPH? (IC₅₀ =?26?μg/mL) and ABTS⁺? (IC₅₀ =?140?μg/mL), FRAP (IC₅₀ =?48?μg/mL) and CCA (IC₅₀ =?770?μg/mL). Only the dichloromethane leaf extract (LDCM) showed anti-inflammatory activity (IC₅₀ =?48?μg/mL). The methanol root (IC₅₀ =?19?μg/mL) and leaf (IC₅₀ =?29?μg/mL) extracts strongly inhibited baker’s yeast α-glucosidase, but LDCM had higher rat’s α-glucosidase inhibition (IC₅₀ =?2527?μg/mL) than acarbose (IC₅₀ =?4638?μg/mL). GC-MS analysis identified β-sitosterol, stigmasterol, 1-octacosanol and linolenic acid as possible molecules responsible for the observed bioactivities.

Conclusions: Our findings suggest P. maritimum as a source of high-value health promoting commodities for alleviating symptoms associated with oxidative and inflammatory diseases, including diabetes.     

Prunus mume leaf extract lowers blood glucose level in diabetic mice

Context Diabetes is a common metabolic disease with long-term complications. Prunus mume Sieb. et Zucc. (Rosaceae) fruits have shown to ameliorate glucose intolerance. However, the antidiabetic effects of P. mume leaves have not been investigated.

Objective This study evaluated the effects of P. mume leaf 70% ethanol extract (PMLE) on alleviating diabetes in vivo and in vitro.

Materials and methods PMLE was fractionated into n-hexane, dichloromethane (CH₂Cl₂), ethyl acetate (EtOAc), n-butanol (BuOH) and water. Polyphenol and flavonoid contents in PMLE fractions were determined using Folin–Ciocalteu reagent and the aluminium chloride colorimetric method, respectively. We evaluated α-glucosidase inhibition using a microplate reader at 400?nm. Adipocyte differentiation by lipid accumulation was measured using Nile Red staining. Male imprinting control region (ICR) mice were injected with streptozotocin (STZ, 100?mg/kg, i.p.). High-fat diets were provided for three weeks prior to PMLE treatments to induce type 2 diabetes. PMLE (0, 5, 25 or 50?mg/kg) was administrated for four weeks with high-fat diets.

Results The EtOAc fraction of PMLE inhibited α-glucosidase activity (IC₅₀?=?68.2?μg/mL) and contained 883.5?±?14.9?mg/g of polyphenols and 820.1?±?7.7?mg/g of flavonoids. The 50?mg/kg PMLE supplement reduced 40% of blood glucose level compared to obese/diabetes mice. Obese/diabetic mice treated with 50?mg/kg PMLE showed a lower level of triacylglycerol (320.7?±?20.73?mg/dL) compared to obese/diabetes mice (494.9?±?14.80?mg/dL).

Conclusion The data demonstrate that P. mume leaves exert antidiabetic effects that may be attributable to high concentrations of polyphenols and flavonoids.     

LC-MS–MS and GC-MS analyses of biologically active extracts and fractions from Tunisian Juniperus phoenice leaves

Context: Despite some studies related to Juniperus phoenicea L. (Cupressaceae), phytochemical and biological investigations of this plant remain unexplored.

Objective: This work is the first report dealing with the identification and characterization of volatile components and flavonoids in hexane and methanol extracts from J. phoenicea leaves

Materials and methods: Antioxidant activity of hexane, and methanol extracts from J. phoenicea leaves were determined by DPPH-radical scavenging assay. α-Amylase inhibitory activity was evaluated by enzyme inhibition using in vitro assay (each extract was dissolved in DMSO to give concentrations of 50, 100 and 200?mg/mL). The chemical composition of fractions (Fr1-Fr3) from methanol extract was determined by high-performance liquid chromatography coupled with mass spectroscopy (HPLC-MS) analysis.

Results and discussion: The hexane extract was analyzed by GC-MS technique which allowed the identification of 32 compounds. The main constituents were α-humulene (16.9%), pentadecane (10.2%) and α-cubebene (9.7%). Fraction Fr 2 exhibited a strong DPPH radical-scavenging activity (IC_50?=_?20.1?μg/mL) compared to that of BHT as well as the highest α-amylase inhibitory activity (IC_50?=_?28.4?μg/mL). Three flavonoids were identified in these fractions using HPLC-MS analysis: Quercetin 3-O-glucoside, isoscutellarein 7-O-pentoside and quercetin 3-O-pentoside. In addition, the more active fraction (Fr 2) was purified with semi-preparative HPLC affording one pure compound (amentoflavone) using ¹H NMR analysis. This compound exhibited powerful DPPH radical-scavenging (IC_50?=_?14.1?μg/mL) and α-amylase inhibition (IC_50?=_?20.4?μg/mL) effects.

Conclusion: This study provides scientific support to some medicinal uses of J. phoenicea found in North Africa.     

Matrix metalloproteinase,hyaluronidase and elastase inhibitory potential of standardized extract of Centella asiatica

Abstract

Context: Centella asiatica (L.) Urban (Apiaceae), a valuable herb described in Ayurveda, is used in the indigenous system of medicine as a tonic to treat skin diseases.

Objective: Centella asiatica methanol extract and its ethyl acetate, n-butanol and aqueous fraction, were subjected for the evaluation of skin care potential through the in vitro hyaluronidase, elastase and matrix metalloproteinase-1 (MMP-1) inhibitory assay.

Materials and methods: The C. asiatica plant was extracted with methanol and fractionated with ethyl acetate, n-butanol and water. The enzymatic activities were evaluated using ursolic acid and oleanolic acid as standards. Isolate molecule asiaticoside was quantified in the crude extract and fractions through high-performance liquid chromatography (HPLC) and structural was characterized by liquid chromatography–mass spectroscopy (LC–MS) and ¹H nuclear magnetic resonance (NMR). Isolated compound was also evaluated for in vitro enzyme assays.

Results: Extract exhibited anti-hyaluronidase and anti-elastase activity with IC₅₀ of 19.27?±?0.37 and 14.54?±?0.39?µg/mL, respectively, as compared to ursolic acid. Centella asiatica n-butanol fraction (CAnB) and isolated compound showed significant hyaluronidase (IC₅₀?=?27.00?±?0.43 and 18.63?±?0.33?µg/mL) and elastase (IC₅₀?=?29.15?±?0.31 and 19.45?±?0.25?µg/mL) inhibitory activities, respectively, and also showed significant MMP-1 inhibition (p?<?0.05 and p?<?0.01).

Discussion and conclusion: n-Butanol fraction was found to be most effective among the all fractions from which asiaticoside was isolated and further quantified by HPLC. This work concludes that the asiaticoside from C. asiatica may be a prospective agent for skin care.     

A phlorotannin constituent of Ecklonia cava alleviates postprandial hyperglycemia in diabetic mice

Context: 2,7″-Phloroglucinol-6,6′-bieckol is a type of phlorotannin isolated from brown algae, Ecklonia cava Kjellman (Phaeophyceae; Laminareaceae). 2,7″-Phloroglucinol-6,6′-bieckol mediates antioxidant activities. However, there has been no research on improving postprandial hyperglycaemia using 2,7″-phloroglucinol-6,6′-bieckol.

Objective: This study investigated the inhibitory effects of 2,7″-phloroglucinol-6,6′-bieckol on activities of α-glucosidase and α-amylase as well as its alleviating effect on postprandial hyperglycaemia in streptozotocin-induced diabetic mice.

Materials and methods: α-Glucosidase and α-amylase inhibitory assays were carried out. The effect of 2,7″-phloroglucinol-6,6′-bieckol on hyperglycaemia after a meal was measured by postprandial blood glucose in streptozotocin-induced diabetic and normal mice. The mice were treated orally with soluble starch (2?g/kg BW) alone (control) or with 2,7″-phloroglucinol-6,6′-bieckol (10?mg/kg bw) or acarbose (10?mg/kg BW) dissolved in 0.2?mL water. Blood samples were taken from tail veins at 0, 30, 60, and 120?min and blood glucose was measured by a glucometer.

Results: 2,7″-Phloroglucinol-6,6′-bieckol showed higher inhibitory activities than acarbose, a positive control against α-glucosidase and α-amylase. The IC₅₀ values of 2,7″-phloroglucinol-6,6′-bieckol against α-glucosidase and α-amylase were 23.35 and 6.94?μM, respectively, which was found more effective than observed with acarbose (α-glucosidase IC₅₀ of 130.04?μM; α-amylase IC₅₀ of 165.12?μM). In normal mice, 2,7″-phloroglucinol-6,6′-bieckol significantly suppressed the postprandial hyperglycaemia caused by starch. The 2,7″-phloroglucinol-6,6′-bieckol administration group (2349.3?mmol·min/L) had a lower area under the curve (AUC) glucose response than the control group (2690.83?mmol·min/L) in diabetic mice.

Discussion and conclusion: 2,7″-Phloroglucinol-6,6′-bieckol might be used as an inhibitor of α-glucosidase and α-amylase as well as to delay absorption of dietary carbohydrates.     

Acetylcholinesterase inhibition effects of marine fungi

Context: To this day, there are no reports that marine compounds isolated from microorganisms of the Lianyungang area of China have been used for the treatment of Alzheimer’s disease.

Objective: The present study was to isolate fungi from the sea sediment of the Lianyungang area and screen for acetylcholineseterase inhibition activities of ethyl acetate extracts.

Materials and methods: Fungi were isolated from the sea sediment and fermented. After centrifugation, the supernate was extracted with ethyl acetate. The ethyl acetate extract was then fractionated into five fractions. Acetylcholinesterase inhibition activities of the ethyl acetate extracts and five sub-fractions were tested at a concentration of 500?μg/mL with the Ellman’s method.

Results: Forty-three marine fungi were isolated; 15 extracts inhibited acetylcholinestrease >50% and 3 extracts inhibited the acetylcholinesterase >80% at the concentration of 500?μg/mL. The 3 extracts (L1705, S1101, SH0701) inhibited AChE dose-dependently with IC₅₀ values of 11.3?±?1.2, 72.1?±?2.3, and 7.8?±?2.8?μg/mL, respectively. After the extract of SH0701 was fractionated into five fractions, the ethyl acetate fraction possessed the highest acetylcholinesterase inhibitory activity with an inhibition rate of 71.55% at the concentration of 10?μg/mL. The fungus SH0701 was identified as Aspergillus ochraceus SH0701 according to morphology and molecular identification.

Discussion and conclusion: The present results indicates that some ethyl acetate extracts of marine fungi isolated from Lianyungang area of China could inhibit AChE potently. Therefore, some novel AChE inhibitors might exist in those extracts.     

Indirect modulation of the endocannabinoid system by specific fractions of nutmeg total extract

Context: Nutmeg [Myristica fragrans Houtt. (Myristicaceae)] has a long-standing reputation of psychoactivity. Anecdotal reports of nutmeg use as a cheap marijuana substitute, coupled to previous studies reporting a cannabimimetic-like action, suggest that nutmeg may interact with the endocannabinoid system.

Objective: The study evaluates nutmeg fractions for binding capacity with various CNS receptors and their potential interaction with the endocannabinoid system.

Materials and methods: Dichloromethane (DF) and ethyl acetate (EF) fractions were prepared from the methanol extract of powdered whole nutmeg. The HPLC-profiled fractions were assayed by the NIMH Psychoactive Drug Screening Program (PDSP) in a panel of CNS targets at a 10?μg/mL concentration. The fractions were also screened for fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL) inhibition, initially at a concentration of 500?μg/mL, then by concentration-dependent inhibition studies.

Results: None of the tested fractions showed significant binding to CNS receptors included in the PDSP panel. However, both fractions exerted significant inhibition of the FAAH and MAGL enzymes. The DF fraction inhibited FAAH and MAGL enzymes at IC₅₀ values of 21.06?±?3.16 and 15.34?±?1.61?μg/mL, respectively. Similarly, the EF fraction demonstrated FAAH and MAGL inhibition with IC₅₀ values of 15.42?±?3.09 and 11.37?±?6.15?μg/mL, respectively.

Discussion and conclusion: The study provides the first piece of evidence that nutmeg interacts with the endocannabinoid system via inhibition of the endocannabinoid catabolizing enzymes. This mechanism provides insight into reported cannabis-like action as well as expands the potential therapeutic utility of nutmeg.     

Antimalarial and free radical scavenging activities of rhizomes of Polygonatum verticillatum supported by isolated metabolites

Antimalarial activity of the crude extract of Polygonatum verticillatum rhizomes and its sequentially partitioned fractions were investigated against Plasmodium falciparum. The crude extract possessed notable activity (IC₅₀: 21.67?μg/mL) that enhanced reasonably upon fractionation. The antiparasitic potency of the n-hexane fraction was maximum (IC₅₀: 2.33?μg/mL) followed by chloroform (IC₅₀: 4.62?μg/mL). However, the remaining fractions showed insignificant activity in the assay. The extracts of the plant showed marked scavenging activity on stable free radical, DDPH. The most potent antioxidant was the chloroform fraction (IC₅₀: 90?μg/mL) followed by ethyl acetate (IC₅₀: 93?μg/mL) and n-butanol (IC₅₀: 95?μg/mL) fractions. In the brine shrimps lethality test, the extracts were found nontoxic with the exception of ethyl acetate fraction (LD₅₀: 492.846?μg/mL). The bioactivity-guided isolation resulted into 5-hydroxymethyl-2-furaldehyde (HMF) and diosgenin which strongly supports the present experimental findings.     

Flavonoids,cytotoxic, antioxidant and antibacterial activities of Evax pygmaea

Context: Phytochemical study and biological potential of Evax pygmaea (L.) Brot. (Asteraceae) are reported for the first time.

Objective: To identify the secondary metabolites of Evax pygmaea and to determine its antioxidant, antibacterial and cytotoxic activities.

Materials and methods: Dried aerial parts (1?kg) were macerated in 70% MeOH (5?L) during 72?h. The concentrated hydromethanolic extract was subjected to extractions with chloroform (3?×?300?mL), ethyl acetate (3?×?300?mL) and n-butanol (3?×?300?mL), successively. VLC of combined ethyl acetate (EAEP) and n-butanol (BEP) fractions was followed by column purifications. Antioxidant activity was investigated using DPPH, CUPRAC, and metal chelating, β-carotene/linoleic acid and ABTS assays. Agar method was used in the antibacterial study. Cytotoxic activity was determined by Brine shrimp lethality test in DMSO and ethanol, at varying concentrations (2, 1 and 0.2%) and (1, 0.2 and 0.1%) successively.

Results: Quercetin (1), isorhamnetin 3-O-β-d-xyloside (2), isorhamnetin 3-O-β-d-glucoside (3), quercetin 3-O-β-d-glucoside (4), quercetin 7-O-β-D-glucoside (5), patuletin 3-O-β-d-glucoside (6) were isolated from for the first time from Evax genus. The EAEP was the most active in ABTS (IC₅₀: <3.125?μg/mL) assay whereas the BEEP exhibited the highest activity in the β-carotene/linoleic acid assay (IC₅₀: <3.125?μg/mL). The EAEP and BEP exhibited good antibacterial activity (MIC: 40–80 µg/mL). The plant did not show any toxicity (LD₅₀>80 µg/mL).

Discussion and conclusions: Six flavonoids were isolated for the first time from Evax pygmaea which exhibited good antioxidant and antibacterial activities.     

Aldose reductase and protein tyrosine phosphatase 1B inhibitory active compounds from Syzygium cumini seeds

Abstract

Context: Syzygium cumini (L.) Skeels (Myrtaceae), commonly known as jamun, is an Indian plant, traditionally well known for its medicinal properties including antidiabetic activity.

Objective: To isolate the antidiabetic compounds from Syzygium cumini seeds and evaluate their activity using aldose reductase (AR) and protein-tyrosine phosphatase 1B (PTP1B) inhibition assays.

Materials and methods: The dried seeds were extracted with methanol and partitioned with ethyl acetate, butanol, and water. The extracts were screened for antidiabetic activity at a concentration of 100?µg/mL using in vitro AR and PTP 1B inhibition assays.

Results and discussion: The highly enriched fractions obtained from broad ethyl acetate fraction yielded maslinic acid (1), 5-(hydroxymethyl) furfural (2), gallic acid (3), valoneic acid dilactone (4), rubuphenol (5), and ellagic acid (6). Structures were elucidated by ¹H-NMR and ¹³C-NMR. The initial ethyl acetate fraction showed AR inhibitory activity with the IC₅₀ value of 2.50?μg/mL and PTP1B enzyme inhibition with the IC₅₀ value of 26.36?μg/mL. Compounds 3, 4, 5, and 6 were found to inhibit AR with IC₅₀ values of 0.77, 0.075, 0.165, and 0.12?μg/mL while the compounds 4, 5, and 6 inhibited PTP1B with IC₅₀ values of 9.37, 28.14, and 25.96?μg/mL, respectively.

Conclusion: The results of this study demonstrate that the isolated constituents show promising in vitro antidiabetic activity and, therefore, can be candidates for in vivo biological screening using relevant models to ascertain their antidiabetic activity.     

Compounds from the pods of Astragalus armatus with antioxidant,anticholinesterase, antibacterial and phagocytic activities

Context: The phytochemical study and biological activities of Astragalus armatus Willd. subsp. numidicus (Fabaceae) pods, an endemic shrub of Maghreb, are reported.

Objective: This study isolates the secondary metabolites and determines the bioactivities of Astragalus armatus pods.

Materials and methods: The chloroform, ethyl acetate and n-butanol extracts of hydro-ethanolic extracts were studied. Antioxidant activity was investigated using DPPH and ABTS radical scavenging, CUPRAC and ferrous chelating assays at concentrations ranging from 3 to 200?μg/mL. Anticholinesterase activity was determined against acetylcholinesterase and butyrylcholinesterase enzymes at 50, 100 and 200?μg/mL. Antibacterial activity was performed according to minimum inhibitory concentration (MIC) method. Carbon clearance method in albino mice was used for the phagocytic activity at concentrations 50, 70 and 100?mg/kg body weight. Spectroscopic techniques were used to elucidate the compounds.

Results: Ethyl acetate extract afforded a flavonoid (1) while the n-butanol extract gave four flavonoids (2–5), a cyclitol (6) and a cycloartane-type saponin (7). The ethyl acetate extract exhibited highest antioxidant activity in DPPH (IC₅₀: 67.90?±?0.57?μg/mL), ABTS (IC₅₀: 11.30?±?0.09?μg/mL) and CUPRAC (A_0.50: 50.60?±?0.9?μg/mL) assays. The chloroform extract exhibited the best antibacterial activity against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa, each with 80?μg/mL MIC values. The n-butanol extract enhanced phagocytic activity.

Discussion and conclusion: Isorhamnetin (1), isorhamnetin-3-O-α-l-rhamnopyranosyl-(1 → 6)-β-d-galactopyranoside (2), isorhamnetin-3-O-β-d-apiofuranosyl-(1 → 2)-[α-l-rhamnopyranosyl-(1 → 6)]-β-d-galactopyranoside (3), kaempferol-3-O-(2,6-di-O-α-l-rhamnopyranosyl)-β-d-galactopyranoside (4), kaempferol-3-O-(2,6-di-O-α-l-rhamnopyranosyl)-β-d-glucopyranoside (5), pinitol (6) and cyclomacroside D (7) were isolated whereas 1, 2, 6 and 7 are reported for the first time from A. armatus.     

In vitro antioxidant and cytotoxic properties of ethanol extract of Alpinia oxyphylla fruits

Abstract

Context. Alpinia oxyphylla Miquel (Zingiberaceae) is a traditional Chinese herbal medicine widely used for the treatment of intestinal disorders, urosis and diuresis. However, information about antioxidant and cytotoxic properties of its fruits remains to be elucidated.

Objective: The ethanol crude extract (CE) and its fractions [petroleum ether fraction (PF), ethyl acetate fraction (EF), n-butanol fraction (BF) and water fraction (WF) extracted by petroleum ether, ethyl acetate, n-butanol and water, respectively] of A. oxyphylla fruits were investigated for their antioxidant activity and cytotoxicity.

Materials and methods: The total phenolic content (TPC) and antioxidant activity of the extracts were determined by Folin–Ciocalteu reagent, 1,1-diphenyl-2-picrylhydrazyl (DPPH^?), Trolox equivalent antioxidant capacity and reducing power assay. Cytotoxicity of the extracts (0–200?μg/mL) was tested on six human cancer cell lines (breast cancer cell line, cervix carcinoma cell line, lung adenocarcinoma cell line, liver carcinoma cell line, gastric cancer cell line and colon cancer cell line) using the sulforhodamine B assay.

Results: The TPC of extracts varied from 8.2 to 20.3?mg gallic acid equivalents/g dry weight. DPPH radical scavenging effect of extracts decreased in the order of EF?>?BF?>?CE?>?PF?>?WF, with IC₅₀ values ranging from 74.7 to 680.8?μg/mL. 2,2-azo-bis(3-Ethylbenzothiazoline-6-sulfoic acid) diammonium salt scavenging activity ranged from 0.118 to 0.236?mmol Trolox equivalence/mg extract. The extracts exhibited concentration-dependent reducing power, and EF showed the highest reducing ability. A satisfactory correlation (R²?>?0.826) between TPC and antioxidant activity was observed. In addition, EF, PF and CE exhibited potent anticancer effects on six cancer cell lines with IC₅₀ values ranging from 40.1 to 166.3?μg/mL.

Discussion and conclusion: The ethanol extract of A. oxyphylla fruit, especially the EF, was found to possess potent antioxidant and anticancer activities, and thus a great potential for the application in food and drug products.     

Antimicrobial,anticancer, and antioxidant compounds from Premna resinosa growing in Saudi Arabia

Context: Premna resinosa (Hochst.) Schauer (Lamiaceae) is used in many places to treat bronchitis, respiratory illness and convulsions of the rib cage.

Objective: This study evaluates the anticancer, antimicrobial and antioxidant activities of P. resinosa, and isolates some responsible constituents.

Materials and methods: The methanol extract of P. resinosa aerial parts and its fractions (n-hexane, dichloromethane, ethyl acetate and n-butanol) were tested. Antimicrobial activity was tested using microdilution method against three Gram-positive and four Gram-negative bacteria. The tested concentrations ranged from 4000 to 7.8?μg/mL and MIC values were determined after 24?h incubation. Anticancer activity was evaluated against three human cancer cell lines (Daoy, HepG2 and SK-MEL28) using MTT assay. Antioxidant activity was investigated by DPPH scavenging method and β-carotene-linoleic acid assay.

Results: The greatest antimicrobial activity was exhibited by n-hexane fraction (MIC 10?μg/mL) against Staphylococcus aureus, Enterococcus faecalis, and Shigella flexneri. The n-hexane fraction induced the greatest cytotoxic activity against Daoy, HepG2, and SK-MEL28 cell lines with IC₅₀ values of 9.0, 8.5 and 13.2, respectively. Moreover, the dichloromethane and ethyl acetate fractions showed the highest antioxidant potential. A bioassay-guided fractionation led to the isolation and characterization of seven compounds for the first time, namely, quercetin (1), 3-methoxy quercetin (2), kaempferol (3), 3-methoxy kaempferol (4), myricetin 3,7,3′-trimethyl ether (5), lupeol (6), and stigmasterol (7).

Conclusion: Our results indicate that P. resinosa is a source for antimicrobial and cytotoxic compounds. However, further work is required to isolate other active principles and to determine the mechanism of action.     

Photoprotective effects of cranberry juice and its various fractions against blue light-induced impairment in human retinal pigment epithelial cells

Context: Cranberry has numerous biological activities, including antioxidation, anticancer, cardioprotection, as well as treatment of urinary tract infection (UTI), attributed to abundant phenolic contents.

Objective: The current study focused on the effect of cranberry juice (CJ) on blue light exposed human retinal pigment epithelial (ARPE-19) cells which mimic age-related macular degeneration (AMD).

Materials and methods: Preliminary phytochemical and HPLC analysis, as well as total antioxidant capacity and scavenging activity of cranberry ethyl acetate extract and different CJ fractions (condensed tannins containing fraction), were evaluated. In cell line model, ARPE-19 were irradiated with blue light at 450?nm wavelength for 10?h (mimic AMD) and treated with different fractions of CJ extract at different doses (5–50?μg/mL) by assessing the cell viability or proliferation rate using MTT assay (repairing efficacy).

Results: Phytochemical and HPLC analysis reveals the presence of several phenolic compounds (flavonoids, proanthocyanidin, quercetin) in ethyl acetate extract and different fractions of CJ. However, the condensed tannin containing fraction of ethyl acetate extract of CJ displayed the greater (p?p?p?Discussion and conclusion: In conclusion, this study distinctly proved that condensed tannin containing fraction of CJ probably exhibits better free radicals scavenging activity and thereby effectively protected the ARPE-19 cells and thus, hampers the progress of AMD.     

Antidiabetic components of Cassia alata leaves: Identification through α-glucosidase inhibition studies

Context: Cassia alata Linn. [syn. Senna alata (L.) Roxb.] (Caesalpiniaceae) is used for treating various disease conditions including diabetes but its mechanism(s) of action and active principles remain to be elucidated.

Objective: The antidiabetic principles were identified using an in vitro α-glucosidase inhibition study.

Materials and methods: The methanol extract of leaves of C. alata, which showed potent α-glucosidase inhibitory activity (IC₅₀, 63.75?±?12.81 µg/ml), was fractionated. Active fractions were taken for further analysis by a variety of techniques including HPLC and Combiflash chromatography. The identity of the isolated compounds was established by spectroscopic analysis while their potential antidiabetic activity was assessed by in vitro enzyme inhibition studies.

Results: The α-glucosidase inhibitory effect of the crude extract was far better than the standard clinically used drug, acarbose (IC₅₀, 107.31?±?12.31 µg/ml). A subsequent fractionation of the crude extract was made using solvents of ascending polarity (petroleum ether, chloroform, ethyl acetate, n-butanol and water). The ethyl acetate (IC₅₀, 2.95?±?0.47 µg/ml) and n-butanol (IC₅₀, 25.80?±?2.01 µg/ml) fractions which contained predominantly kaempferol (56.7?±?7.7 µM) and kaempferol 3-O-gentiobioside (50.0?±?8.5 µM), respectively, displayed the highest carbohydrate enzyme inhibitory effect.

Discussion: One of the possible antidiabetic mechanisms of action of C. alata is by inhibiting carbohydrate digestion. This is the first report on α-glucosidase activity of kaempferol 3-O-gentiobioside.

Conclusion: Considering the activity profile of the crude extract and isolated bioactive compounds, further in vivo and clinical studies on C. alata extracts and compounds are well merited.