Antioxidant, Anti-Inflammatory, and Postulated Cytotoxic Activity of Phenolic and Anthocyanin-Rich Fractions from Polana Raspberry (Rubus idaeus L.) Fruit and Juice-In Vitro Study.
ABSTRACT: In this study, the antioxidative and anti-inflammatory potential of crude extracts (CE), anthocyanin-rich fractions (ARF), and phenolic fractions (PF) from raspberry (R) and raspberry juice (J) were evaluated. The antioxidant properties were evaluated with three complementary assays: DPPH radical scavenging activity, chelating Fe(II) power, and ferric reducing power. The highest antioxidant activity was determined for the crude extract from raspberry pulp (RCE) in the case of all methods used. The anti-inflammatory activity was demonstrated by inhibitory effect on lipoxygenase (LOX) and cyclooxygenase-2 (COX-2) activity in vitro. The highest efficiency in inhibiting the activity of both enzymes was exhibited by RCE, 0.79 and 0.59 mg FW/mL, respectively. In turn, JARF had the lowest ability to inhibit LOX (EC50 = 4.5 mg FW/mL) and JPF caused the lowest COX-2 inhibition (1.75 mg FW/mL). Additionally, we have performed a pilot study of in vitro cytotoxic activity using two human leukemia cell lines: J45 and HL60. All examined extracts inhibited the viability of J45 cells more effectively than HL60. The highest cytotoxic effect was observed in the J45.01 cell line after exposure to RCE (EC50 = 0.0375 mg FW/mL).
Project description:Red raspberries (Rubus idaeus L.) are increasingly popular foods in contemporary diets due to their freshness, nutritional value and health claims. Among the existing cultivars, "Kweli" is one of the most productive and widely cultivated. In this study, the nutritional value and chemical composition of "Kweli" red raspberry were characterized by the official method of food analysis and chromatographic techniques, and its antioxidant and antimicrobial activities were tested against biological/biochemical oxidizable substrates and foodborne bacteria and fungi strains, respectively. Carbohydrates (including fructose and glucose, 14.3 and 12.6 g/100 g dw, respectively), proteins (6.8 g/100 g dw), and ashes (3.90 g/100 g dw) were major constituents. The fat content was quite low and constituted mainly by unsaturated fatty acids (58.3%), with a predominance of oleic acid. Fresh red raspberry also contained high levels of citric (2.7 g/100 g) and ascorbic (17 mg/100 g) acids. The anthocyanins (4.51 mg/g extract) cyanidin-O-hexoside and mostly cyanidin-O-sophoroside were identified in the red raspberry hydroethanolic extract, which was able to inhibit thiobarbituric acid reactive substances (TBARS) formation (EC50 of 122 µg/mL), oxidative hemolysis (IC50 of 298 µg/mL), and ?-carotene bleaching (EC50 of 18.7 µg/mL). In turn, the extract was more effective than the food additive E224 against Bacillus cereus. All these results highlighted the nutritional quality of "Kweli" red raspberry and showed some compositional differences in relation to other cultivars. Therefore, its inclusion in a daily diet can be helpful to obtain nutrients and antioxidants and bring health benefits.
Project description:This study evaluated the antioxidant capacity of Ricinus communis L. (RC) leaves and powder when used as a feed additive for laying hens. Results showed that the total phenolic content of the aqueous leaf extract of Ricinus communis L. (RCE) was 48.39 mg gallic acid equivalent (GAE) per gram dry weight (DW). The flavonoid content was 9.76 mg quercetin dihydrate equivalent (QE)/g DW. Ferrous chelating activity was approximately 56.2% with an RCE concentration of 1 mg/mL; the highest chelating activity was 91.2% with 4 mg/mL extract. The reducing power of 1 mg/mL RC was 1.17 times better than 1 mg/mL butylated hydroxytoluene (BHT). The Trolox equivalent antioxidant capacity (TEAC) value of 12.5 mg/mL RCE was equivalent to 3.09 mg/mL Trolox. RCE (10 mg/mL) had a lipid oxidative inhibition capacity of 35.3%. A total of 80 ISA brown laying hens at twenty-nine weeks of age were randomly allocated into the control or 1 of 3 treatment groups; the latter received 0.5%, 1% or 2% of RC, respectively, for 12 weeks. Results showed that the RC supplementation improved the feed conversion rate and 0.5% RC generated the best results. Additionally, the egg yolk score was significantly increased in all RC-supplemented groups. Moreover, there was no significant difference in serum characteristics between the treatment groups. Serum antioxidant enzyme activity showed that superoxide dismutase (SOD) activity increased in the RC-supplemented groups relative to the control but was not significantly different. mRNA expression levels of the antioxidant regulatory genes GCLC, GST, HO-1, SOD1, and SOD2 were significantly increased with 2% RC supplementation. In summary, RC is a suitable feed additive for laying hens and the addition of 0.5% RC leaf powder resulted in the greatest benefits.
Project description:Rubus coreanus Miquel (R. coreanus) is a unripen fruit of black raspberry native to eastern Asia. It is used as traditional oriental medicine and supplementary foods for centuries. Previous studies have shown that the R. coreanus extract (RCE) and its main constitute ellagic acid possess diverse biological activities. However, the effects of RCE on antitumor immunity and T cell function were not fully understood. The present study describes the anti-tumor effect of RCE in humanized PD-1 mice by blocking PD-1/PD-L1 interaction. Competitive enzyme-linked immunosorbent assay (ELISA) and pull down assay were performed to elucidate the binding properties of RCE in vitro. Cellular PD-1/PD-L1 blockade activities were measured by T cell receptor (TCR)-induced nuclear factor of activated T cells-luciferase activity in co-cultured cell models with PD-1/NFAT Jurkat and PD-L1/aAPC CHO-K1 cells. The in vivo efficacy of RCE was confirmed in humanized PD-1 mice bearing MC38 colorectal tumor. RCE and ellagic acid dose-dependently block the binding of PD-1 to PD-L1. Moreover, oral administration of RCE showed the potent anti-tumor activity similar to anti-PD-1 antibody. The present study suggests that RCE possesses potent anti-tumor effect via PD-1/PD-L1 blockade, and ellagic acid is the main compound in RCE. Thus, we provide new aspects of RCE as an immunotherapeutic agent.
Project description:Begonia nelumbiifolia is a traditional edible plant consumed and commercialized in the northern highlands of Puebla, México. The present study reports the seasonal variation in proximate analysis as well as organic acids, carotenoids and flavonoids content in both leaves and stalks of this plant. The stalks contained low concentrations of protein (~3%), fiber (~1.5%) and nitrogen free extract (~0.26%) during the time of study. Both organs showed contents of oxalic acid (91-103 mg 100 g-1 FW), ascorbic acid (50-65 mg 100 g-1 FW), lutein (1-2.5 mg 100 g-1 FW), ?-carotene (1-3 mg 100 g-1 FW), quercetin (1.3-2.7 mg 100 g-1 DW) and rutin (0.5-1.7 mg 100 g-1 DW). Antioxidant activity against DPPH was observed by the stalk extracts from 30% methanol (IC50, 0.21-0.37 mg L-1), pure methanol (IC50, 0.14-0.21 mg L-1) and hexane: acetone (IC50, 0.77-1.21 mg L-1). In vitro anti-HMG-CoA reductase (IC50, 0.07-0.36 mg L-1) and anti-alpha-glucosidase (IC50, 0.28-0.43 mg L-1) activities were observed in extracts from the edible stalks from pure methanol and 30% methanol. The leaf extracts from 30% methanol inhibited the growth of Pseudomonas syringae pv. tabaci TBR2004 (MIC, 254 µg mL-1), P. syringae pv. tomato DC3000 (MIC, 423 µg mL-1), P. syringae pv. glycinea (MIC, 605 µg mL-1) and Clavibacter michiganensis AB299158 (MIC, 162 µg mL-1). B. nelumbiifolia contained valuable phytochemicals associated to nutraceutical and biological properties. However, the consumption of the fresh stalks must be carefully considered because of the high oxalate content.
Project description:In vitro suspension culture techniques are cost effective for large-scale production of secondary metabolites. In the present study, firstly, suspension cultures of untransformed <i>Lactuca sativa</i> were prepared using different hormonal combinations and were subjected to different pH, temperature and salt concentrations. Maximum biomass was obtained for suspensions supplemented with 1.5 mg/L BAP and 0.1 mg/L NAA, at pH 5.8, temperature 28 °C and 0 mM NaCl concentration. Using these parameters, suspensions were produced for <i>rol ABC-</i> and <i>rol C</i>-transformed lines of <i>L. sativa</i>. All the transgenic lines showed prominent increase in fresh weight (FW) and dry weight (DW) with maximum values for <i>rol ABC</i>2 line producing 169.8 mg/mL FW and 25.3 mg/mL DW. The exudates of transformed and untransformed plants were tested for the antioxidant activity and in vivo assays on rats. Maximum phenolic content (261 ?g/mL) and flavonoid content (637.6 ?g/mL) were obtained for <i>rol C</i>1 transgenic line. Total antioxidant capacity was found maximum (1451.7 ?g/mL) for untransformed lettuce, whereas <i>rol C</i>1 showed maximum total reducing power activity (637.6 ?g/mL). In DPPH assay, maximum activity (104.7 ?g/mL) was shown by <i>rol ABC</i>3 line. In rats analgesic assay, maximum activity (74.9%) was shown by <i>rol C</i>2. Line <i>rol C</i>1 showed maximum anti-inflammatory activity (69.2%) and maximum antidepressant activity (minimum immobility time of 55 s). Maximum anticoagulant activity was observed for <i>rol ABC</i>2 with maximum clotting time of 130 s. The present study could help in using lettuce suspension culture as platform for the enhanced production of important metabolites.
Project description:Colorectal cancer is the fourth leading cause of cancer-related deaths worldwide. Here, we investigated the anticancer effect of Rhus coriaria extract (RCE) on HT-29 and Caco-2 human colorectal cancer cells. We found that RCE significantly inhibited the viability and colony growth of colon cancer cells. Moreover, RCE induced Beclin-1-independent autophagy and subsequent caspase-7-dependent apoptosis. Blocking of autophagy by chloroquine significantly reduced RCE-induced cell death, while blocking of apoptosis had no effect on RCE-induced cell death. Mechanistically, RCE inactivated the AKT/mTOR pathway by promoting the proteasome-dependent degradation of both proteins. Strikingly, we also found that RCE targeted Beclin-1, p53 and procaspase-3 to degradation. Proteasome inhibition by MG-132 not only restored these proteins to level comparable to control cells, but also reduced RCE-induced cell death and blocked the activation of autophagy and apoptosis. The proteasomal degradation of mTOR, which occurred only 3?hours post-RCE treatment was concomitant with an overall increase in the level of ubiquitinated proteins and translated stimulation of proteolysis by the proteasome. Our findings demonstrate that Rhus coriaria possesses strong anti-colon cancer activity through stimulation of proteolysis as well as induction of autophagic and apoptotic cell death, making it a potential and valuable source of novel therapeutic cancer drug.
Project description:This study focused on the effect of kaempferol, catechin, apigenin, sinapinic acid, and extracts from plants (i.e., parsley, cumin, mustard, green tea, and green coffee) on thyroid peroxidase (TPO) and lipoxygenase (LOX) activity, antiradical potential, as well as the result of interactions among them. Catechin, sinapinic acid, and kaempferol acted as a competitive TPO inhibitors, while apigenin demonstrated an uncompetitive mode of inhibitory action. Ethanol extracts from all plants acted as competitive TPO inhibitors, while, after in vitro digestion, TPO activation was found especially in the case of mustard (24%) and cumin (19.85%). Most importantly, TPO activators acted synergistically. The TPO effectors acted as LOX inhibitors. The most effective were potentially bioaccessible compounds from green tea and green coffee (IC50 = 29.73 mg DW/mL and 30.43 mg DW/mL, respectively). The highest free radical scavenging ability was determined for catechin and sinapinic acid (IC50 = 78.37 µg/mL and 84.33 µg/mL, respectively) and potentially bioaccessible compounds from mustard (0.42 mg DW/mL) and green coffee (0.87 mg DW/mL). Green coffee, green tea, cumin, and mustard contain potentially bioaccessible TPO activators that also act as effective LOX inhibitors, which indicate their potentially health-promoting effects for people suffering from Hashimoto's disease.
Project description:Russula senecis, a worldwide distributed mushroom, is exclusively popular among the tribal communities of West Bengal for food purposes. The present study focuses on its reliable taxonomic identification through macro- and micro-morphological features, DNA barcoding, confirmation of its systematic placement by phylogenetic analyses, myco-chemicals and functional activities. For the first time, the complete Internal Transcribed Spacer region of R. senecis has been sequenced and its taxonomic position within subsection Foetentinae under series Ingratae of the subgen. Ingratula is confirmed through phylogenetic analysis. For exploration of its medicinal properties, dried basidiocarps were subjected for preparation of a heat stable phenol rich extract (RusePre) using water and ethanol as solvent system. The antioxidant activity was evaluated through hydroxyl radical scavenging (EC50 5 µg/ml), chelating ability of ferrous ion (EC50 0.158 mg/ml), DPPH radical scavenging (EC50 1.34 mg/ml), reducing power (EC50 2.495 mg/ml) and total antioxidant activity methods (13.44 µg ascorbic acid equivalent/mg of extract). RusePre exhibited antimicrobial potentiality against Listeria monocytogenes, Bacillus subtilis, Pseudomonas aeruginosa and Staphylococcus aureus. Furthermore, different parameters were tested to investigate its chemical composition, which revealed the presence of appreciable quantity of phenolic compounds, along with carotenoids and ascorbic acid. HPLC-UV fingerprint indicated the probable existence of at least 13 phenolics, of which 10 were identified (pyrogallol > kaempferol > quercetin > chlorogenic acid > ferulic acid, cinnamic acid > vanillic acid > salicylic acid > p-coumaric acid > gallic acid). Result from the present work suggests that the fraction, RusePre, may open novel prospect as a functional ingredient in antioxidant supplements and in drugs to treat infectious disease.
Project description:Supercritical fluid extraction (SFE) was used to extract total alkaloids, peimisine, peimine and peiminine from the bulb of Fritillaria thunbergii Miq. The antioxidant capacity of the extracts was evaluated by DPPH radical scavenging activity (DPPH-RSA), ABTS radical scavenging activity (ABTS-RSA) and ferric reducing capacity (FRAP) assay. A central composite design (CCD) with four variables and five levels was employed for optimization of process parameters, and response surface plots were constructed in accordance with a second order polynomial model. Under optimal conditions of 3.0 h, 60.4 °C, 26.5 MPa and 89.3% ethanol, the highest yields were predicted to be 3.8 mg/g for total alkaloids, 0.5 mg/g for peimisine, 1.3 mg/g for peimine and 1.3 mg/g for peiminine, and the antioxidant capacity of extracts displayed EC50, DPPH value of 5.5 mg/mL, EC50, ABTS value of 0.3 mg/mL and FRAP value of 118.2 mg ascorbic acid equivalent (AAE)/100 g.
Project description:Black mulberry (<i>Morus nigra</i> L.) fruits are known due to their delicious, sweet and slightly acid flavor and high anthocyanin content. In the present study, the diversity of phytochemical, sensory, and antioxidant characteristics of the fruits of 20 black mulberry genotypes, from the Artvin region of Turkey, were evaluated. As important phytochemical assessments in fruits, we chromatographically (HPLC/DAD) determined glucose (7.22 to 11.10 g/100 g fresh weight (fw)) and fructose content (6.32 and 9.94 g/100 g fw), as well as predominant organic acid in black mulberry genotypes fruits-malic acid (6.02-11.44 g/100 g fw), followed by citric acid. Titrative determination was used for ascorbic acid, finding contents of 17.41-28.33 mg/100 g fw. There was found a great diversity of sensory (taste, juiciness, and aroma) characteristics, indicating a richness of the fruit germplasm. Antioxidant parameters, such as total phenolic (TP) and anthocyanin (TA) content, were assessed spectrometrically; antioxidant activity (AA) was assessed by DPPH and FRAP assays; individual flavonoids and phenolic acids were determined chromatographically (HPLC/DAD). Antioxidant characteristics of the fruits, defined by TP and TA content, ranged from 1951 to 2733 μg GAE/g fw and 508-712 μg C3GE/g fw, respectively. The most abundant compounds of flavonoids and phenolic acids groups were determined to be rutin (47.10-97.20 mg/100 g fw) and chlorogenic acid (51.3-90.8 mg/100 g fw). AA results, measured by the DPPH method as EC<sub>50</sub> value, ranged between 16.10 and 25.45 μg/mL; a FRAP assay revealed values of AA between 9.80 and 13.22 μmol TE/g fw. Significant differences in phytochemical and antioxidant qualities were observed among the analyzed <i>M. nigra</i> genotypes. Regarding the best values of phytochemical and antioxidant characteristics, three genotypes of <i>M. nigra</i> were selected to be recommended for fruit production. The results thus highlight the potential for the exploitation of local black mulberry genotypes through crop selection and breeding program.