Combinatorial preparation and structural characterization of anthocyanins and aglycones from Purple-heart Radish for evaluation of physicochemical stability and pancreatic lipase inhibitory activity.

In this study, an efficient approach to preparation of different anthocyanins from Purple-heart Radish was developed by combining microwave-assisted extraction (MAE), macroporous resin purification (MRP) and ultrasound-assisted acid hydrolysis (UAAH) for evaluation of physicochemical stability and pancreatic lipase (PL) inhibitory activity. By optimization of MAE, MRP and UAAH processes, the anthocyanins reached the yield of 6.081 ± 0.106 mg/g, the purity of 78.54 ± 0.62 % (w/w) and the content of 76.29 ± 1.31 % (w/w), respectively. With high-resolution UHPLC-Q-Orbitrap/MS, 15 anthocyanins were identified as pelargonins with diverse glucosides and confirmed by pelargonidin standard. By glycosylation, pelargonins exhibited higher stability in different pH, temperature, light, metal ions environments than that of pelargonidin. However, PL inhibitory assay, kinetic analysis and molecular docking demonstrated that pelargonidin had higher PL inhibitory activity than pelargonins even though with similar binding sites and a dose-effect relationship. The above results revealed that the effect of glycosylation and deglycosylation on PL inhibitory activity and physicochemical stability.

Phytochemical Profile, Bioactive Properties, and Se Speciation of Se-Biofortified Red Radish (Raphanus sativus), Green Pea (Pisum sativum), and Alfalfa (Medicago sativa) Microgreens.

The impact of selenium (Se) enrichment on bioactive compounds and sugars and Se speciation was assessed on different microgreens (green pea, red radish, and alfalfa). Sodium selenite and sodium selenate at a total concentration of 20 µM (1:1) lead to a noticeable Se biofortification (40-90 mg Se kg-1 DW). In green pea and alfalfa, Se did not negatively impact phenolics and antioxidant capacity, while in red radish, a significant decrease was found. Regarding photosynthetic parameters, Se notably increased the level of chlorophylls and carotenoids in green pea, decreased chlorophyll levels in alfalfa, and had no effect on red radish. Se treatment significantly increased sugar levels in green pea and alfalfa but not in red radish. Red radish had the highest Se amino acid content (59%), followed by alfalfa (34%) and green pea (28%). These findings suggest that Se-biofortified microgreens have the potential as functional foods to improve Se intake in humans.

Novel insights into the anti-asthmatic effect of Raphanus sativus L. (Raphani Semen): Targeting immune cells, inflammatory pathways and oxidative stress markers.

ETHNOPHARMACOLOGICAL RELEVANCE: Raphanus sativus L. is a well-known medicinal plant with traditional therapeutic applications in various common ailments including inflammation and asthma. AIMS OF THE STUDY: This study aimed to evaluate the chemical composition and anti-asthmatic potential of the hydro-methanolic extract of the leaves of R. sativus L. (Rs.Cr) using various in vitro and in vivo investigations. MATERIALS AND METHODS: The Rs.Cr was subjected to preliminary phytochemical analysis and HPLC profiling. The safety was assessed through oral acute toxicity tests in mice. The antiasthmatic effect of the extract was studied using milk-induced leukocytosis and ovalbumin (OVA)-induced allergic asthma models established in mice. While mast cell degranulation and passive paw anaphylaxis models were established in rats. Moreover, effect of the extract was studied on various oxidative and inflammatory makers. The antioxidant effect of the extract was also studied by in vitro DPPH method. RESULTS: The HPLC profiling of Rs.Cr showed the presence of important polyphenols in a considerable quantity. In toxicity evaluation, Rs.Cr showed no sign of morbidity or mortality with LD50 < 2000 mg/kg. The extract revealed significant mast cell disruption in a dose-dependent manner compared to the intoxicated group. Similarly, treatment with Rs.Cr and dexamethasone significantly (p < 0.001) reduced paw edema volume. Subcutaneous injection of milk at a dose of 4 mL/kg, after 24 h of its administration, showed an increase in the leukocyte count in the intoxicated group. Similarly, mice treated with dexamethasone and Rs.Cr respectively showed a significant decrease in leukocytes and eosinophils count in the ovalbumin-induced allergic asthma model. The extract presented a significant (p˂0.001) alleviative effect on the levels of SOD and GSH, MDA, IL-4, IL-5, and IL-13 in a dose-dependent manner as compared to the intoxicated group. Furthermore, the histological evaluation also revealed a notable decrease in inflammatory and goblet cell count with reduced mucus production. CONCLUSION: The current study highlights mechanism-based novel insights into the anti-asthmatic potential of R. sativus that also strongly supports its traditional use in asthma.

Understanding the nutraceutical diversity through a comparative analysis of the taproot metabolomes of different edible radish types via UHPLC-Q-TOF-MS.

Radishes are root vegetables that are rich in bioactive compounds and provide numerous health benefits, but the overall metabolic profiles of radish taproots and the metabolic differences among different edible types are not fully understood. In this research, we used UHPLC-Q-TOF-MS to identify the metabolites in cooked, processed and fruit radishes of ten varieties. In total, 264 metabolites belonging to 18 categories were detected. A multivariate analysis revealed that the metabolite composition differed among the three radish groups, and a comparative analysis showed that the significantly differentially accumulated metabolites were mainly amino acids and derivatives, lipids, flavonoids, hydroxycinnamate derivatives and carbohydrates. The accumulation of metabolites, particularly flavonoids, was greater in fruit radishes than in cooked and processed radishes. This work provides novel insights into the radish metabolomic profiles for assessment of the nutritional value of different edible radish types for humans.

Raphani Semen (Raphanus sativus L.) Ameliorates Alcoholic Fatty Liver Disease by Regulating De Novo Lipogenesis.

In this study, we investigated the pharmacological effect of a water extract of Raphani Semen (RSWE) on alcoholic fatty liver disease (AFLD) using ethanol-induced AFLD mice (the NIAAA model) and palmitic acid (PA)-induced steatosis HepG2 cells. An RSWE supplement improved serum and hepatic triglyceride (TG) levels of AFLD mice, as well as their liver histological structure. To explore the molecular action of RSWE in the improvement of AFLD, we investigated the effect of RSWE on four major pathways for lipid homeostasis in the liver: free fatty acid transport, lipogenesis, lipolysis, and ß-oxidation. Importantly, RSWE decreased the mRNA expression of de novo lipogenesis-related genes, such as Srebf1, Cebpa, Pparg, and Lpin1, as well as the protein levels of these factors, in the liver of AFLD mice. That these actions of RSWE affect lipogenesis was confirmed using PA-induced steatosis HepG2 cells. Overall, our findings suggest that RSWE has the potential for improvement of AFLD by inhibiting de novo lipogenesis.

[Effects of alcohol extract of radish seed on blood lipid, glucose and liver steatosis in ApoE-/- mice].

Objective: To investigate the effects and mechanisms of alcohol extract of Raphani seed (AERS) on blood lipid, blood glucose and hepatic steatosis in ApoE-/- mice. Methods: Sixty ApoE-/- mice were randomly divided into control group (CG), normal saline group (NG), rosiglitazone group (RG) and AERS treatment groups (AERS-L / M / H). Except CG, other groups were fed with high-fat, high-sugar and high-salt diet for 9 weeks. The mice in RG were treated with rosiglitazone (1.33 mg·kg-1, 0.2 ml ·10 g-1) by gavage daily. The mice in CG and NG were treated with equal volume of NS by intragastric administration daily. The mice in AERS groups were treated with AERS at the doses of 100, 300 and 900 mg·kg-1 for 9 weeks, respectively. FPG and Fins were detected. Insulin resistance index (IRI) and liver coefficient (LC) were calculated. The levels of ALT, AST, Leptin (LEP), TNF - α and blood lipid (TC, FFA, etc.) were tested. The expressions of proteins related to liver lipid metabolism (HMG-R、LDL-R、LEP-R) were detected by Western blot. Results: Compared with NG or RG, CG showed significant changes in liver appearance (color, swelling, etc.) and pathology (steatosis, hepatocyte necrosis, etc.), while AERS-M/H groups were similar to CG. Compared with CG, the serum levels of FPG, Fins, IRI, ALT, AST, TNF-α, LC, TG, LDL-C, FFA and LEP were increased significantly (P＜0.05) . Compared with NG, AERS could decrease the above mentioned indicators in a dose-dependent manner (P＜0.05). Compared with RG, the levels of FPG and Fins of AERS-H were increased significantly, while the level of IRI was decreased (P＜0.05). Compared with NG and RG, the protein expressions of HMG-R and LEP-R in AERS groups were decreased, while the protein expression of LDL-R was increased in a dose-dependent manner (P＜0.05). Conclusion: AERS can prevent the increase of blood lipid, glucose and hepatic steatosis induced by high-fat and high-sugar diet in ApoE-/- mice. The mechanism is related to the decrease of FFA and LEP, the inhibition of TNF-α, HMG-R, LEP-R protein expressions and the promotion of LDL-R protein expression.

Radish sprout alleviates DSS-induced colitis via regulation of NF-kB signaling pathway and modifying gut microbiota.

In this study, we investigated the effects of radish sprout ethanol extract (RSE) on inflammatory responses in the macrophages and a mouse model of colitis. RSE administration was found to effectively inhibit the phosphorylation of IκB and, in turn, the production of pro-inflammatory enzymes and cytokines in lipopolysaccharide-stimulated macrophages. In dextran sulfate sodium (DSS)-colitis mice, RSE administration prevented body weight and colon length reduction, while decreasing inflammation and mucosal necrosis. The diversity of the fecal microbiota was significantly increased in the group treated with RSE. In addition, RSE administration decreased the relative abundance of the phylum Proteobacteria, which includes many pathogens, and increased the abundance of the genus Akkermansia. Beta diversity analyses showed that RSE administration restored the gut microbiota composition close to that of healthy mice. For the first time, we identified glycosides of sinapic acid as part of hydroxycinnamic acids in RSE with colitis-alleviating effects. Notably, 1,2-O-disinapoyl glucoside substantially decreased nitric oxide generation in LPS-stimulated macrophages.

Monitoring the infection process of Rhizopus stolonifer on strawberry fruit during storage using films based on chitosan/polyvinyl alcohol/polyvinylpyrrolidone and plant extracts.

Different formulations based on nanoparticles of chitosan-plant extracts were evaluated to detect the infection process from the earliest stage of the fungus Rhizopus stolonifer on strawberry fruit during storage. Chitosan/polyvinyl alcohol (Ch/PVA) and chitosan/polyvinylpyrrolidone (Ch/PVP) films enriched with nanoparticles (NPs) of chitosan blended with plant extracts were prepared. They were placed inside a plastic package containing inoculated fruits and stored at 25 °C for 72 h. The thickness values of the films were in the range of 0.10 to 0.25 mm. All samples showed a maximum absorbance peak of about 300-320 nm; however, the Ch/PVP films enriched with NPs of chitosan and 10% of radish extract had an evident decrease in the optical absorbance as the fungal infection progressed. Additionally, as observed by scanning electron microscopy, the cross-section and surface morphology of films were not modified during storage, and the growth of R. stolonifer was evident after 48 h. Therefore, the Ch/PVP films enriched with chitosan NPs blended with 10% radish extract could be a reliable indicator of this fungus's growth.

Structural changes in cell wall of Japanese radish accompanied by release of rhamnogalacturonan during pressure cooker heating.

Changes in the cell wall of Japanese radish due to heating at 100 °C or 117 °C for 3 h were examined. Signals in 13C cross polarization magic angle spinning solid-state NMR (which detects rigid components) showed no differences between heating temperatures. 13C pulse saturation transfer magic angle spinning NMR (which detects flexible components) showed clear temperature-dependent changes in the rhamnose side chains of rhamnogalacturonan. Alcohol-insoluble solids isolated from raw samples were heated in water at 100 °C or 117 °C for 3 h. The concentrations of dissolved sugars and metal ions measured after heating in water at 117 °C were greater than in samples heated at 100 °C, indicating that loosening of cell wall structures increased with temperature, likely via degradation and elution of rhamnogalacturonan followed by ß-elimination of homogalacturonan, and fewer interactions between cell wall components, including divalent metal ions. Vegetable shape was retained despite fewer interactions.

Application of Natural Deep Eutectic Solvents for the metal nanoparticles extraction from plant tissue.

The study aimed to use Natural Deep Eutectic Solvents (NADES) as an extractant of metal oxide NPs from plant material. The plant chosen for the study was radish after exposure, growing on media containing: copper(II) oxide, cerium(IV) oxide, and titanium(IV) oxide. The first step of the study was to investigate the influence of NADES on NPs. In the second step, selected NADES solvents were used as extractants of NPs present in radish after exposure. Single-particle Inductively Coupled Plasma Mass Spectrometry technique (SP-ICP-MS) was used to determine the number and size of NPs. As a result of the research, it was found that copper(II) oxide NPs, regardless of the solvent used, is not present in the extract. Only the ionic form of the element was present in the solution. Higher sized cerium(IV) oxide NPs were accumulated in the root, while smaller sized ones were found in radish leaves. The titanium(IV) oxide NPs were agglomerated and were present in a small amount in radish leaves, accumulating mainly in the root. Finally, it can be concluded that NADES allows the extraction of metal oxide NPs from the plant material.

Beetroot and radish powders as natural nitrite source for fermented dry sausages.

The aim of this study was to investigate the use of radish and beetroot powders as potential substitutes of nitrite in fermented dry sausages due to their high nitrate content (around 16,000 and 14,000 mg/kg, respectively). Six treatments were prepared and evaluated during the ripening process and storage time: C1 (control with 150 mg/kg sodium nitrite and 150 mg/kg sodium nitrate), C2 (control without sodium nitrite/nitrate), R05 (0.5% radish powder), R1 (1% radish powder), B05 (0.5% beetroot powder) and B1 (1% beetroot powder). The addition of vegetable powders influenced moisture content, weight loss and water activity of sausages. Nitrite was formed from radish and beetroot powders during the ripening process, especially in R1 and B1 treatments. Beetroot powder affected colour, pigments and lactic acid bacteria counts. The results of pH, colour, lipid oxidation, nitrite and nitrate analysis suggest R1 treatment as a potential nitrite replacer obtained from a simple and feasible drying process.

Effect of cold plasma and elicitors on bioactive contents, antioxidant activity and cytotoxicity of Thai rat-tailed radish microgreens.

BACKGROUND: Raphanus sativus var. caudatus or Thai rat-tailed radish (RTR) contains glucosinolates and isothiocyanates with chemopreventive effects; however, only mature plants have been investigated to date. Thus, the present study aimed to determine isothiocyanates, phenolic compounds and flavonoid compounds, antioxidant activity, cytotoxicity, and antiproliferative activity of RTR microgreens grown from seeds treated with cold plasma (21 kV for 5 min), organic elicitor (160 mmol L-1 NaCl, 10 mmol L-1 CaCl2 or 176 mmol L-1 sucrose) or both in combination. Seeds were germinated on vermiculite and sprayed with deionized water or elicitor for 7 days before harvest. RESULTS: Cold plasma had insignificant effect on growth, whereas NaCl and CaCl2 increased fresh weight. Plasma with CaCl2 led to the highest total isothiocyanate (ITC) content [1.99 g kg-1 dry weight (DW)] in RTR microgreens containing raphasatin as the only ITC detected. Plasma treatment gave the highest total phenolic content (7.56 mg gallic acid equivalents g-1 DW), antioxidant activity from a 2,2-diphenyl-1-picrylhydrazyl assay (7.70 mg trolox equivalents g-1 DW) and ferric reducing antioxidant power assay (21.72 mg Fe2+ g-1 DW). Microgreen extracts from plasma showed an IC50 value of 29.28 and 13.83 µg mL-1 towards MCF-7 and HepG2, respectively, with inhibitory properties on matrix metalloproteinase (MMP)-2 and MMP-9 proteins. Plasma enhanced Bax and Caspase-3 gene expression but reduced Bcl-2 and MMP-9 expression, indicating activation of apoptosis. CONCLUSION: Cold plasma shows promise as an innovative tool to enhance bioactive compounds with chemopreventive benefits in microgreens. © 2020 Society of Chemical Industry.

Characteristic Analysis of Trigonelline Contained in Raphanus sativus Cv. Sakurajima Daikon and Results from the First Trial Examining Its Vasodilator Properties in Humans.

Vascular disease poses a major public health problem worldwide. Trigonelline isolated from Raphanus sativus cv. Sakurajima Daikon (Sakurajima radish) induces nitric oxide production from vascular endothelial cells and enhances vascular function. Here, we investigated the characteristics of trigonelline and its effects on endothelial function after consumption of Sakurajima radish by humans. Our results show that Sakurajima radish contains approximately 60 times more trigonelline than other radishes and squashes. Additionally, no significant differences were observed between varieties of Sakurajima radish, suggesting that any type of Sakurajima radish can be ingested for trigonelline supplementation. The effects of cooking and processing Sakurajima radish were also evaluated, as were the effects of freezing, and changes in osmotic pressure and pH. A first-in-human trial using Sakurajima radish showed that ingestion of 170 g/day of Sakurajima radish for ten days increased blood trigonelline concentrations and significantly improved flow-mediated dilation, which is a measure of vascular endothelial function. Overall, our findings suggest that the trigonelline contained in Sakurajima radish may contribute to improved human vascular endothelial function. Hence, Sakurajima radish may enhance vascular endothelial function as a functional food.

Application of Natural Pigments in Ordinary Cooked Ham.

The possibility of obtaining a carmine or pink color on ordinary cooked ham by applying natural dyes from three plant species, namely red radish (Raphanus sativus L.), hibiscus (Roselle sabdariffa L.) and red beetroot (Beta vulgaris L.), was investigated. The extracts were evaluated for the stability at physical-chemical parameters and subjected to cytotoxicity assays in the gastric cell line AGS Encapsulation of the extracts in soybean lecithin liposomes and maltodextrin microcapsules was performed. Lyophilized extracts before and after encapsulation in maltodextrin were applied in the formulation of ordinary cooked ham and used in a pilot scale of production. The color of cooked ham samples from different assays was evaluated visually and by colorimetry. The results suggest that the coloration of ordinary cooked ham obtained with extracts of red beetroot is very promising for future applications in this type of meat product.

Protective Effects of Radish Extract Against Neurotoxicity in Mice and PC12 Cells.

A variety of natural compounds have been used to reduce the oxidative stress associated with Alzheimer's disease, and many of these defend cells from oxidative stress-induced neuronal toxicity. In this study, the protective effect of radish (Raphanus raphanistrum) extract was investigated in mice and PC12 cells. In vivo behavioral tests were completed to examine the protective effects of the extract on amyloid beta (Aß)-peptide1-42-induced learning and memory impairment in a mouse model. The extract increased spontaneous alternation behaviors and step-through latency in mice. We discovered that administration of the extract reduced lipid peroxidation and Aß aggregation in a biochemical study of mice brain tissues. Treatment with the extract also increased acetylcholine and catalase activity in the brain. Furthermore, the radish extract attenuated H2O2-induced oxidative stress in cells. Through sequential fractionation of the radish extract, the active compound was identified as oleamide. These results suggest that the radish extract could have a protective role against oxidative stress-induced neuronal toxicity, possibly owing to the antioxidative activity of oleamide.

Sulforaphane: Expected to Become a Novel Antitumor Compound.

Natural products are becoming increasingly popular in a variety of traditional, complementary, and alternative systems due to their potency and slight side effects. Natural compounds have been shown to be effective against many human diseases, especially cancers. Sulforaphane (SFE) is a traditional Chinese herbal medicine. In recent years, an increasing number of studies have been conducted to evaluate the antitumor effect of SFE. The roles of SFE in cancers are mainly through the regulation of potential biomarkers to activate or inhibit related signaling pathways. SFE has exhibited promising inhibitory effects on breast cancer, lung cancer, liver cancer, and other malignant tumors. In this review, we summarized the reports on the activity and functional mechanisms of SFE in cancer treatment and explored the efficacy and toxicity of SFE.

Raphanus sativus L. seed extracts induce apoptosis and reduce migration of oral squamous cell carcinoma KB and KBCD133+cells by downregulation of ß-catenin.

Although, oral cancer therapies have been developed for decades, patient survival rates have not changed. Side effects of chemotherapy and radiotherapy reduce quality of life of patients and it remains difficult to treat oral cancers due to the presence of cancer stem cells (CSCs) that cause recurrence and metastasis. Therefore, we search for natural products that affect oral cancer cells including oral cancer stem cells. In the present study, we investigated the anticancer effects of Raphanus sativus L. seed (RSLS) extracts on oral squamous cell carcinoma KB cells and CSC-like KBCD133+ cells. CD133 plays an important role in CSCs and physically binds to ß-catenin to activate the ß-catenin signaling targets. Therefore, a natural extract that can inhibit ß-catenin act in may be effective anticancer drug acquiring CSC. Of the natural product extract candidates, RSLS extracts induced apoptosis in KB and KBCD133+ cells and inhibited nuclear translocation of ß-catenin cell migration and invasion rates. Treatment of RSLS extracts resulted in increases of Axin and it leds to reductions of ß-catenin in KB and KBCD133+ cells. Hence, the result suggests that RSLS are potential candidate for anticancer drug against oral cancer cells and CSCs.AbbreviationsCSCcancer stem cellsOSCCsquamous cell carcinoma cellsRSLSRaphanus sativus L. seed.

Biocompatible film sensors containing red radish extract for meat spoilage observation.

pH-sensitive films were developed based on biocompatible materials and natural pH sensitive dye. The films were successfully fabricated using starch/gelatin and red radish anthocyanin. The colors of films could be differentiated by naked eye within 5 min changing from orange to grey-purple at pH 2-12 and captured by a smartphone. The color parameters were evaluated by the Image J software. In addition, the color change of films was observed in ammonia gas atmosphere. The color stability of sensing films was evaluated and the results indicated that the films had great stability and were able to store more than two weeks. The results from intra-day and inter-day color response study showed good precision. Finally, the pH-sensitive films could be applied to real samples for real-time meat spoilage observation.

Valorisation of the green waste parts from turnip, radish and wild cardoon: Nutritional value, phenolic profile and bioactivity evaluation.

The recovery of bio-wastes to obtain high added value compounds is of great interest for the pharmaceutical, medicinal and food industries. Therefore, the aerial parts of turnip (Brassica rapa L.), radish (Raphanus sativus L.) and leaf blade of wild cardoon (Cynara cardunculus L. var. sylvestris (Lamk) Fiori) were characterized regarding their nutritional composition, as also their content in soluble sugars, organic acids, fatty acids, and tocopherols. Furthermore, their hydroethanolic extracts and infusion preparations, were profiled regarding individual phenolic compounds by HPLC-DAD/ESI-MS and their antioxidant, antibacterial and hepatotoxic activities were evaluated. Regarding the nutritional content, wild cardoon revealed the best results, however it was radish and turnip that showed higher values for organic acids and phenolic compounds. The hydroethanolic extract and infusion preparation of wild cardoon stood out for its antioxidant and antibacterial activity. Overall, the hydroethanolic extracts seemed more effective (regarding antioxidant and antibacterial activity) than the infusions. Total phenolic acids proved to be strongly correlated with the antioxidant and antibacterial (against Morganella morganii) activities. This study showed that the discarded parts of these plants can be used as an important natural source of valuable nutrient content and new and safe bioactive compounds, beneficial for human health. Moreover, the extraction of those compounds from underused parts of turnip, radish and cardoon could be used to preserve foods, avoiding artificial additives and thus, contributing to the development of new natural ingredients.

Protective Effect of Selenium-Enriched Red Radish Sprouts on Carbon Tetrachloride-Induced Liver Injury in Mice.

This study aimed to investigate the effect of Se (Selenium) treatment on nutritional quality in radish sprouts. The results showed that 15 µM sodium selenite significantly increased phenolics compounds, flavonoids compounds, anthocyanins, and some essential amino acid content, while improving the total antioxidant capacity of radish sprouts. Besides, the Se-enriched radish sprouts significantly alleviated the liver damage caused by carbon tetrachloride (CCl4 ) in mice and improved the antioxidant capacity of the liver in mice, whereas the Se-enriched radish sprouts alleviated the inflammatory reaction and apoptosis caused by CCl4 . These results imply that Se-enriched radish sprouts have a positive impact on mice with CCl4 -induced liver injury, and that in future Se-enriched radish sprouts could be developed into an effective food and health care product for the liver injury prevention. PRACTICAL APPLICATION: Because selenium is an essential trace element in the human body, selenium-enriched sprouts can help eliminate free radicals in the body, relieve aging, and selenium-deficient diseases. They are easy to grow and have low costs. Hence, selenium-enriched sprouts have a great potential of being widely consumed.