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Kaiware Daikon (Raphanus sativus L.) Extract: A Naturally Multipotent Chemopreventive Agent #
Abstract
Brassica vegetables are attracting major attention as healthy foods because of their content of glucosinolates (GLs) that release the corresponding isothiocyanates (ITCs) upon myrosinase hydrolysis. A number of studies have so far documented the chemopreventive properties of some ITCs. On the other hand, single nutrients detached from the food itself risk being somewhat "reductive", since plants contain several classes of compounds endowed with a polyhedral mechanism of action. Our recent finding that 4-methylthio-3-butenyl isothiocyanate (GRH-ITC) and 4-methylsulfinyl-3-butenyl isothiocyanate (GRE-ITC), released by the GLs purified from Japanese (Kaiware) Daikon (Raphanus sativus L.) seeds and sprouts, had selective cytotoxic/apoptotic activity on three human colon carcinoma cell lines prompted further research on the potential chemopreventive role of a standardized Kaiware Daikon extract (KDE), containing 10.5% w/w GRH and 3.8% w/w GRE, compared to its isolated components. KDE administered in combination with myrosinase at doses corresponding to 50 microM GRH-ITC plus 15 microM GRE-ITC (50 microM KDE-ITC) to three human cancer cell lines (LoVo, HCT-116 and HT-29) significantly reduced cell growth by 94-96% of control in six days (p < 0.05), outperforming pure GRH-ITC or GRE-ITC at the same dose. On the other hand, the same treatment had no significant toxicity on normal human T-lymphocytes. A 50 microM concentration of KDE-ITC had relevant apoptosis induction in all tested cancer cell lines, as confirmed by annexin V assay (e.g., 33% induction in LoVo compared to control, p < 0.05), Bax protein induction (e.g., +20% in HT-29, p < 0.05), and Bcl2 downregulation (e.g.-20% in HT-29, p < 0.05), and induced caspase-1 and PARP-1 activation in all cancer cells as shown by Western blot analysis. Unlike pure GRH or GRH-ITC, KDE also had significant chain-breaking antioxidant activity, retarding the AAPH-initiated autoxidation of methyl linoleate in SDS micelles at concentrations as low as 4.4 ppm (-50% in oxygen consumption rate), as monitored by Clark-type microelectrode oxygen-uptake kinetics, and induced very fast quenching of DPPH. radical in methanol with t(1/2) (s) = (1.47 +/- 0.25) x 10(-2)/[KDE; (g/L)], measured by stopped-flow UV-vis kinetics at 298 K. The potential chemopreventive role of KDE is discussed.
... 4,5 However, the interest is further boosted by the involvement of free radical and red-ox processes both in normal biological functions and in the pathophysiology of several diseases. 6,7 The role of so-called oxidative stress in human health as well as in the aging process has suggested that natural or dietary antioxidants may have beneficial properties and potential applications as drugs or health-oriented products. 3,7 Clearly, the necessity to test many structurally diverse compounds or even to screen crude plant extracts has given impetus to the development of an impressing variety of testing methods. ...
... 6,7 The role of so-called oxidative stress in human health as well as in the aging process has suggested that natural or dietary antioxidants may have beneficial properties and potential applications as drugs or health-oriented products. 3,7 Clearly, the necessity to test many structurally diverse compounds or even to screen crude plant extracts has given impetus to the development of an impressing variety of testing methods. This, however, creates a rather confusing scenario, as many such methods owe their popularity mainly to their simplicity and lack the necessary soundness, often leading to inconsistent or misleading results because they are applied without consideration of the chemistry behind the antioxidant activity and of what they are actually measuring. ...
... The bimolecular rate constant for the reaction between DPPH· and various antioxidants allows us to clarify the structure−activity relationship of families of antioxidants such as flavonoids 26 and their reaction mechanisms. 7 Additionally, a single-point measurement of DPPH· discoloration provides the stoichiometry of radical trapping 27 or can be used to titrate the effective content of antioxidants when testing raw extracts of ...
Measurement of antioxidant properties in plant-derived compounds require appropriate methods that address the mechanism of antioxidant activity and focus on the kinetics of the reactions involving the antioxidants. Methods based on inhibited autoxidations are the most suited for chain-breaking antioxidants and for termination-enhancing antioxidants, while different specific studies are needed for preventive antioxidants. A selection of chemical testing methods is critically reviewed highlighting their advantages and limitations, and discussing their usefulness to investigate both pure molecules and raw extracts. The influence of the reaction medium on antioxidants' performance is also addressed.
... This study indicated that the antioxidant and antityrosinase capacity of these vegetables in Thailand, including white R. sativus, garlic and ginger due to the presence of radical scavenging, phenolic flavonoids and antityrosinase compounds [15]. Antioxidant activity is generally based on the number and location of hydroxyl groups present as well as the presence of a 2-3 double bond and 4-oxofunction [16]. Flavonoids inhibit enzymes such as prostaglandin synthase, lipoxygenase, and cycloxygenase, closely related to tumorigenesis, and may induce detoxifying enzymes such as glutathione S-transferase [17]. ...
... Saponin-also found in cruciferous plants-was reported to cause some necrotic cell death [62]. Isothiocyanates are reported to inhibit carcinogenesis by various mechanisms [16]. Sulforaphane increases many phase II metabolic enzymes responsible for the reduction of oxidative stress molecules including carcinogens [63]. ...
Raphanus sativus (Radish) is a widely used vegetable belonging to the family Brassicaceae, generally grown as annual or biennial plants, with a taproot which is much enlarged when it is cultivated. In Yemenite folk medicines, R. sativus juice is used in eliminating kidney stones. Few people, specifically in the Middle East, prefer to drink its juice in pursuit of certain health benefits. In Unani, Greeko-Arab, and Indian folk medicine, It is used as a home remedy for the treatment of many diseases such as jaundice, gallstone, liver diseases, rectal disorder, indigestion, and other gastric pains. This article has reviewed the information available on R. sativus ethnopharmacology, geographical distribution, chemical composition, and pharmacological uses. The information on botanical description, distribution, traditional uses, chemical composition, bioactive components, and therapeutic investigations was gathered from a comprehensive literature search of electronic databases such as Science Direct, PubMed, Web of Science, Wiley, ACS, Springer, Google Scholar, and SCOPUS until 2020 for publications. An elaborative study has been done on botanical characterization, traditional uses, chemical composition, and various pharmacological or therapeutic uses. Mainly this plant is fully loaded with polyphenolic compounds which exert a promising antioxidant property. This plant possesses various therapeutic benefits such as anti-inflammatory, anti-microbial, anti-tumorogenesis, anti-cancer, anti-diabetic, and anti-nephrotoxicity activity. The comprehensive literature analysis shows that a wide range of populations has utilized various parts of R. sativus around the globe. The above information shows that the plant holds a variety of hidden and unknown potentials which can be studied extensively for its phytoconstituents and therapeutic outcomes. However, while searching through the literature available, it was observed that there is a lack of information on its phytochemical profiling and its corresponding pharmacological benefits. It is believed that this review will help lay the groundwork for encouraging pharmacological and pharmaceutical studies.
... Bax can act on the mitochondria to induce mitochondrial permeability transition, resulting in the release of various components, including cytochrome c [26]. Meanwhile, Bcl-2 normally suppresses apoptosis by inhibiting the release of mitochondrial proteins and phosphatidyl serine (PS) exposure [27]. That is to say, the ratio of Bax/Bcl-2 is believed to play an important role in determining apoptosis [28]. ...
... In addition, the results demonstrated that Bax was translocated to mitochondria, leading to cytochrome c release from mitochondria into the cytosol in response to EEDH [26], subsequently, enhances activation of caspase-9, and then binds to Apaf-1, thereby leading to the activation of caspase-3 and PARP cleavage [29]. Although, the protein expression of caspase-9 and Apaf-1 were not investigated in the current study, EEDH induced a remarkable expression of caspase-3 activity and enhancement of PARP cleavage, suggesting that apoptosis might be initiated through an internal mitochondrial pathway [27]. In other words, the over-expressions of caspase-3, PARP cleavage, and pro-apoptotic Bax, accompanied by under-expressed anti-apoptotic Bcl-2, indicated that EEDH induced HepG2 cell apoptosis via a mitochondrial pathway [30]. ...
The antiproliferative effect and mediation of apoptosis in human hepatoma HepG2 cells induced by djulis husk and its bioactive compounds was investigated. The ethanolic extracts of djulis husk (EEDH) at 50, 250, and 500 µg/mL induced remarkable cytotoxicity on HepG2 cells. By flow cytometry analysis, EEDH slowed down the cell cycle at the Sub-G0 phase after 24 h of incubation. Moreover, all EEDH treatment induced an apoptotic response in HepG2 cells. EEDH-induced apoptosis was associated with the attenuation of mitochondrial transmembrane potentials (ΔΨm), an increase in Bax/Bcl-2 ratio, activation of caspase-3, and poly(ADP-ribose)polymerase (PARP) cleavage, as well as an increase in reactive oxygen species (ROS) generation. According to the HPLC-DAD and HPLC-MS/MS analysis, quercetin and kaempferol derivatives and another sixteen compounds were present in EEDH. Quercetin and kaempferol at 25–150 μM showed antiproliferative action and induced apoptosis on HepG2 cells, which may in part account for the anticancer activity of EEDH. Overall, EEDH may be a potent chemopreventive agent due to apoptosis in HepG2 cells.
... A further protective effect associated with oxidative stress was revealed in experimental models (Guerrero-Beltrána et al., 2012). Sulforaphene (SE) (Fig. 1)-with an additional double bond to SF-has been reported to reduce cancer cell proliferation in a dose-dependent manner and induce apoptosis in: (a) colon carcinoma cell lines (Barillari et al., 2008;Papi et al., 2008); (b) human and murine erythroleukemic cells; (c) human T-lymphoid cells; and, (d) human cervix carcinoma cells (Nastruzzi et al., 2000). SE (at 1 lM) was a potent inducer of hepatic enzymes involved in the detoxification of chemical carcinogens (Razis, Nicola, Pagnotta, Iori, & Ioannides, 2012). ...
... A further protective effect associated with oxidative stress was revealed in experimental models (Guerrero-Beltrána et al., 2012). Sulforaphene (SE) (Fig. 1)-with an additional double bond to SF-has been reported to reduce cancer cell proliferation in a dose-dependent manner and induce apoptosis in: (a) colon carcinoma cell lines (Barillari et al., 2008;Papi et al., 2008); (b) human and murine erythroleukemic cells; (c) human T-lymphoid cells; and, (d) human cervix carcinoma cells (Nastruzzi et al., 2000). SE (at 1 lM) was a potent inducer of hepatic enzymes involved in the detoxification of chemical carcinogens (Razis, Nicola, Pagnotta, Iori, & Ioannides, 2012). ...
A simple, rapid and precise HPLC assay was developed for the well-known anti-cancer isothiocyanates—sulforaphene (SE) and sulforaphane (SF).The analytical system comprised RP-C18 column with isocratic 5%, THF-95% and water. High resolution was obtained (and eluted) of two distinct HPLC peaks of similar structures SE and SF analytes (at 23.01±0.02 and 25.65±0.03 min, respectively). The respective LOD vs. LOQ for SE and SF was 0.34 and 0.36 μg/ml vs. 1.02 and 1.08 μg/ml. This assay had the best linearity and accuracy. The recoveries were in the range of 96.83 to 101.17%. SF and SE were quantified in the Raphanus sativus L. var. caudatus Alef extracts (2,253.05±246.18 and 111.94±16.49 μg/g in the crude extract, respectively), while only SE was detected in the stem (1,105.14±243.10 μg/g crude extract), as SF was lower than the detection limit. The validated method thus minimized and expedited simultaneous SE and SF analysis.
... Furthermore, decreased or increased cell viability could be unrelated to the actual antioxidant activity and depend on diff erent pharmacological activities [88]. This is particularly the case when vegetable extracts of an uncertain or complex composition are tested as potential antioxidants. ...
... Clearly, kinetic studies referring to an extract possibly containing many active molecules cannot provide real rate constants or the stoichiometry of their reaction, but they could provide apparent " specifi c " rate constants ( e.g. rate constants per mg/ mL of extract) [88]. ...
Abstract Owing to the importance of antioxidants in the protection of both natural and man-made materials, a large variety of testing methods has been proposed and applied. These include methods based on inhibited autoxidation studies, which are better followed by monitoring the kinetics of oxygen consumption or that of formation of hydroperoxides, the primary oxidation products. Analytical determination of secondary oxidation products (e.g. carbonyl compounds) has also been used. The majority of testing methods, however, does not involve substrate autoxidation. They are based on the competitive bleaching of a probe (e.g. ORAC assay, β-carotene and crocine bleaching assays, luminol assay), on reaction with a different probe (e.g. spin-trapping and TOSC assay), or they are indirect methods based on the reduction of persistent radicals (e.g. galvinoxyl, DPPH and TEAC assays), or of inorganic oxidizing species (e.g. FRAP, CUPRAC and Folin-Ciocalteu assays). Yet other methods are specific for preventive antioxidants. The relevance, advantages and limitations of these methods are critically discussed stepping from their chemistry and the mechanisms of antioxidant activity. A variety of cell-based assays has also been proposed to investigate the biological activity of antioxidants. Their importance and critical aspects are discussed, along with arguments for the selection of the appropriate testing methods, according to the different needs.
... White radish is mostly consumed as soup in Asia, it can be processed into pickling, drying and fermentation food products (Jung et al., 2022). Several research studies revealed that methylisogermabullone bioactive compounds isolated from radishes were found to stimulate small bowel mobility by activating the acetylcholinergic receptors, while 4-(methylthio)-3-butenyl isothiocyanate was found to induce apoptosis in human colon cancer cells, thereby reducing abnormal cell growth (Barillari et al., 2008). Furthermore, the spicy flavor of radish is due to some chemical substances like myrosinase, glucosinolate and isothiocyanate (Nakamura et al., 2008). ...
This study investigated the physicochemical quality, antioxidant activity, instrumental sensory characteristics of nurungi prepared from rice and dried-radish root bugak powder (DRBP). DRBP was made from dried-radish root bugak (DRB) coated with 80% superfine glutinous and 20% non-glutinous rice powders. The nurungji food products were prepared using 80% non-glutinous rice and 20% glutinous rice, and DRBP, which was added at different levels of 2, 4, 6, 8, and 10%, respectively (NR-2, NR-4, NR-6, NR-8, and NR-10). The total polyphenol and flavonoid contents were increased from 21.34 to 27.87 mL, and 7.46 to 20.05 mgNE/mL respectively, as the amounts of DRBP increased. Color L*-value, pH, and hydration properties were decreased with the increased amounts of DRBP. The electronic tongue results showed that NR-6 sample had high umami scores and sourness compared to the control sample (NR-0). Furthermore, SEM images indicated different shapes between the control (NR-0) and other treatments. However, no differences among nurungji treated with DRBP. Therefore, the use of DRB coated with superfine glutinous and non-glutinous rice powders is a viable ingredient for nurungji preparation. This study shows that adding 6% DRBP produced nurungji with good-quality physicochemical and sensory characteristics.
... White radish is mostly consumed as soup in Asia, it can be processed into pickling, drying and fermentation food products (Jung et al., 2022). Several research studies revealed that methylisogermabullone bioactive compounds isolated from radishes were found to stimulate small bowel mobility by activating the acetylcholinergic receptors, while 4-(methylthio)-3-butenyl isothiocyanate was found to induce apoptosis in human colon cancer cells, thereby reducing abnormal cell growth (Barillari et al., 2008). Furthermore, the spicy flavor of radish is due to some chemical substances like myrosinase, glucosinolate and isothiocyanate (Nakamura et al., 2008). ...
This study investigated the physicochemical quality, antioxidant activity, instrumental sensory characteristics of nurungi prepared from rice and dried-radish root bugak powder (DRBP). DRBP was made from dried-radish root bugak (DRB) coated with 80% superfine glutinous and 20% non-glutinous rice powders. The nurungji food products were prepared using 80% non-glutinous rice and 20% glutinous rice, and DRBP, which was added at different levels of 2, 4, 6, 8, and 10%, respectively (NR-2, NR-4, NR-6, NR-8, and NR-10). The total polyphenol and flavonoid contents were increased from 21.34 to 27.87 mL, and 7.46 to 20.05 mgNE/mL respectively, as the amounts of DRBP increased. Color L*-value, pH, and hydration properties were decreased with the increased amounts of DRBP. The electronic tongue results showed that NR-6 sample had high umami scores and sourness compared to the control sample (NR-0). Furthermore, SEM images indicated different shapes between the control (NR-0) and other treatments. However, no differences among nurungji treated with DRBP. Therefore, the use of DRB coated with superfine glutinous and non-glutinous rice powders is a viable ingredient for nurungji preparation. This study shows that adding 6% DRBP produced nurungji with good-quality physicochemical and sensory characteristics.
... For example, a biologically active compund named as methylisogermabullone (C23H31O5NS, MW 433) is extracted from radish help in activation of acetyl cholinergic receptors that ultimately leads to small bowel motility (Jeong et al., 2005). 4-(Methylthio)-3-butenyl isothiocyanate induce apoptosinin to reduce the risk of human colon cancer (Barillari et al., 2008) . ...
To check and analyze the effect of nickel sulphate toxicity on radish plant, a research was conducted in the old Botanical Garden at University of Agriculture, Faisalabad. In different concetrations of nickel sulphate solution (10, 20, 30 mM), Raphanussativus specie of radish (Mooli Day-40) was grown to study the morphological parameters like shoot and root length, shoot nad root weight, number of leaves, total carotenoids and chloropyll. Ni-stress remarkably reduced the growth attributes of radish plant. Results were reported after data collection and applying statistical anaylsis by using latest software packages. A noticeable drop in physiological and morphological attributes was recorded. However in biochemical attributes level of chlorophyll a and b was reduced while subsequent elevation was noticed in carotenoid concentration. Index Terms-Chlorophyll a and b, Carotenoid, Nickel sulphate, Toxicity
... These ingredients help lower high blood pressure and reduce the risk of heart disease. Additionally, R. sativus contains a large quantity of glucose-nolates-sulfur compounds that protect cells against mutations that cause cancer [13,14]. ...
The aim of the study was to determine the effect of ozone gas fumigation on the mechanical, chemical, and microbiological parameters of radish tubers. Radish plants were grown in the ground in accordance with the principles of good agricultural practice and condition suitable for the soil and climatic conditions of south-eastern Poland. At the end of the growing season, 24 h before harvest, radish plants were exposed to a variable factor, i.e., fumigation with ozone gas at various doses (1 ppm for 1 and 5 min; 5 ppm for 1 and 5 min) in order to modify selected metabolic pathways of bioactive compounds. Then, 24 h after ozonation, radish tubers were harvested and placed in a climatic chamber with controlled conditions, i.e., 2 °C and 90% humidity. Laboratory analyses were performed during storage on days 1, 5, and 10. The ozonation used did not significantly improve the mechanical properties and water content of radish tubers. There was a beneficial effect of selected gaseous ozone doses (1 ppm for 1 and 5 min; 5 ppm for 1 min on the 10th day of storage) on the biosynthesis of selected bioactive compounds, i.e., ascorbic acid content, total polyphenol content, and antioxidant potential during storage. The most beneficial effects of the use of gaseous ozone were observed in the storage process in reducing the microbiological load of radish tubers. Among the ozonation doses used, the dose of 5 ppm for 5 min had the most beneficial effect on reducing the microbiological load. It reduced the number of yeasts and molds by 14.2% and aerobic mesophilic bacteria by 20.9% compared to the control sample on the last day of storage. Additionally, between the 5th and 10th day of storage, a significant effect of each ozone dose applied on reducing the occurrence of yeasts, molds, and mesophilic aerobic bacteria during tuber storage was noted.
... The bioactivity of glucosinolates and their hydrolysis products have been reported in the Brassicaceae family [64][65][66][67]. Although the direct antioxidant action of glucosinolates is controversial, due to the fact that their antioxidant activity is mainly associated with transcription triggering of phase II metabolic enzymes [68], the direct antioxidant potential of glucoerucin in arugula extract (Eruca sativa) [69], glucobrassicin in cauliflower (Brassica oleracea L.) [70], and glucoraphasatin in radish (Raphanus sativus L.) [71] has been demonstrated. ...
Kale sprouts contain health-promoting compounds that could be increased by applying plant nutrients or exogenous phytohormones during pre-harvest. The effects of selenium (Se), sulfur (S), and methyl jasmonate (MeJA) on lutein, glucosinolate, and phenolic accumulation were assessed in kale sprouts. Red Russian and Dwarf Green kale were chamber-grown using different treatment concentrations of Se (10, 20, 40 mg/L), S (30, 60, 120 mg/L), and MeJA (25, 50, 100 μM). Sprouts were harvested every 24 h for 7 days to identify and quantify phytochemicals. The highest lutein accumulation occurred 7 days after S 120 mg/L (178%) and Se 40 mg/L (199%) treatments in Red Russian and Dwarf Green kale sprouts, respectively. MeJA treatment decreased the level of most phenolic levels, except for kaempferol and quercetin, where increases were higher than 70% for both varieties when treated with MeJA 25 μM. The most effective treatment for glucosinolate accumulation was S 120 mg/L in the Red Russian kale variety at 7 days of germination, increasing glucoraphanin (262.4%), glucoerucin (510.8%), 4-methoxy-glucobrassicin (430.7%), and glucoiberin (1150%). Results show that kales treated with Se, S, and MeJA could be used as a functional food for fresh consumption or as raw materials for different industrial applications.
... Raphani Semen contains an extensive variety of pharmaceutically active compounds [11]. In fact, advanced information about the pharmacological role of Raphani Semen in relation to several gastrointestinal diseases has been revealed in a variety of animal experiment models, e.g., ulcerative colitis, intestinal motility, and colon cancer [12,13]. In addition, Raphani Semen and its active compounds have been shown to have potential for antioxidant and anti-inflammatory activity [14,15]. ...
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 β-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.
... italica) and radish (Raphanus sativus L.) sprouts, contain substantial amounts of antioxidants, vitamin C and health-promoting compounds such as glucosinolates and phenolic compounds 2,3 . It was shown that radish sprouts have anticancer and antioxidant activities both in vivo and in vitro 4,5 . To consider the health benefits of radish sprouts, it is important to increase radish sprouts production 6 . ...
The use of low-temperature plasma for the pre-sowing seed treatment is still in the early stage of research; thus, numerous factors affecting germination percentage, seedling growth, and yield remains unknown. This study aimed to estimate how two critical factors, such as harvest year and seed coat color, affect the percentage of germination and seedling growth after plasma treatment. Radish seeds stored for 2 and 1 year after harvesting (harvested in 2017 and 2018) were sorted into two colors (brown and grey) to investigate the plasma effect on harvest year and seed coat color. We analyzed the amounts of seed phytohormones and antioxidant (γ-tocopherol) were analyzed using mass spectrometry, and physical changes were studied using SEM, EDX, and EPR to understand the mechanism of plasma-induced changes in radish seeds. The obtained results revealed that plasma treatment on seeds affects the germination kinetics, and the maximal germination percentage depends on seed color and the time of seed storage after harvest. Through this study, for the first time, we demonstrated that physical and chemical changes in radish seeds after plasma treatment depends upon the seed color and harvest year. Positive effects of plasma treatment on growth are stronger for sprouts from seeds harvested in 2017 than in 2018. The plasma treatment effect on the sprouts germinated from grey seeds effect was stronger than sprouts from brown radish seeds. The amounts of gibberellin A3 and abscisic acid in control seeds strongly depended on the seed color, and plasma induced changes were better in grey seeds harvested in 2017. Therefore, this study reveals that Air scalar-DBD plasma's reactive oxygen and nitrogen species (RONS) can efficiently accelerate germination and growth in older seeds.
... For example, a biologically active compund named as methylisogermabullone (C23H31O5NS, MW 433) is extracted from radish help in activation of acetyl cholinergic receptors that ultimately leads to small bowel motility (Jeong et al., 2005). 4-(Methylthio)-3-butenyl isothiocyanate induce apoptosinin to reduce the risk of human colon cancer (Barillari et al., 2008) . ...
To check and analyze the effect of nickel sulphate toxicity on radish plant, a research was conducted in the old Botanical Garden at University of Agriculture, Faisalabad. In different concetrations of nickel sulphate solution (10, 20, 30 mM), Raphanussativus specie of radish (Mooli Day-40) was grown to study the morphological parameters like shoot and root length, shoot nad root weight, number of leaves, total carotenoids and chloropyll. Ni-stress remarkably reduced the growth attributes of radish plant. Results were reported after data collection and applying statistical anaylsis by using latest software packages. A noticeable drop in physiological and morphological attributes was recorded. However in biochemical attributes level of chlorophyll a and b was reduced while subsequent elevation was noticed in carotenoid concentration. Index Terms-Chlorophyll a and b, Carotenoid, Nickel sulphate, Toxicity
... Some glucosinolates contribute to the antioxidant capability of the plant. Glucoraphasatin, which is the main glucosinolate in radish sprouts, displayed antioxidant activity and contributed to the total antioxidant capacity of radish sprout extract 30 . Glucoerucin and 4-methoxyglucobrassicin are good antioxidants because of their ability to decompose hydroperoxides and hydrogen peroxide 20,31 . ...
Chinese kale (Brassica oleracea var. alboglabra) has high nutritional value. This study investigated the contents of glucosinolates, antioxidants (chlorophylls, carotenoids, vitamin C, and total phenolics), and antioxidant capacity in five organs from six varieties of Chinese kale. The highest concentrations of individual and total glucosinolates were in the roots and inflorescences, respectively. The highest levels of antioxidants and antioxidant capacity were in inflorescences and leaves. Plant organs played a predominant role in glucosinolate and antioxidant accumulation. Glucoiberin, glucoraphanin, and glucobrassicin, the main anticarcinogenic glucosinolates, could be enhanced simultaneously because of their high positive correlations. The relationship between glucosinolates and antioxidant capacity indicated that glucobrassicin might contribute to the total antioxidant capacity. These results provide useful information related to consumption, breeding of functional varieties, and use of the non-edible organs of Chinese kale.
... For example, a biologically active compund named as methylisogermabullone (C23H31O5NS, MW 433) is extracted from radish help in activation of acetyl cholinergic receptors that ultimately leads to small bowel motility (Jeong et al., 2005). 4-(Methylthio)-3-butenyl isothiocyanate induce apoptosinin to reduce the risk of human colon cancer ( Barillari et al., 2008) . ...
... The study utilized two imbibing conditions. First, the seeds were imbibed in tap water and maintained at 25 ℃ for various durations (1,3,6,8,12, and 24 hr), leaving some seeds untreated as a control (0 hr). Secondly, the seeds were imbibed in water of various temperatures (4,16,20, 25, and 30 ℃) for 3 hr, leaving some seeds were not imbibed in water as a control and to measure the coefficients of uniformity, days of germination, and germination rate. ...
Sprouts are expected to be a promising crop for closed plant factories that use artificial light, because of the fact that sprouts can be grown using only freshwater and low-light conditions. Therefore, we focused on Tartary buckwheat (Fagopyrum tataricum Gaertn.), which has high nutritive value and is currently uncultivated as a sprout for industrial or commercial purposes. This study aimed to understand the effects of imbibition time and water temperature on germination and growth of Tartary buckwheat sprouts. The effects of imbibition time and water temperature on seed germination were evaluated by measuring variations in the coefficients of uniformity, days of germination, and germination rate. The study utilized two imbibing conditions. First, the seeds were imbibed in tap water and maintained at 25 ℃ for various durations (1, 3, 6, 8, 12, and 24 hr), leaving some seeds untreated as a control (0 hr). Secondly, the seeds were imbibed in water of various temperatures (4, 16, 20, 25, and 30 ℃) for 3 hr, leaving some seeds were not imbibed in water as a control and to measure the coefficients of uniformity, days of germination, and germination rate. As a result, the coefficients of uniformity and days of germination of the seeds were improved by the imbibing treatment consisting of a water temperature of 25 ℃ for 3 hr compared to the control. Furthermore, hypocotyl length and fresh weight of the sprouts tended to increase when an imbibing treatment was used in comparison to the control. On the other hand, the germination rate and dry weight of the imbibed seeds were not significantly different from those of the control seeds. Thus, our results suggest that imbibing of seeds could be effective for promoting germination and growth in Tartary buckwheat sprouts.
... The control efficacy of Carwoodi Radish is at least 90% on M. chitwoodi and G. rostochiensis (Young-Mathews, 2016) and reduced M. incognita egg mass production by 93-99% (Ros et al., 2016). It is reported that Japanese (Kaiware) daikon Raphanus sativus seeds and sprouts released 4-methylthio-3-butenyl isothiocyanate and 4-methylsulfinyl-3-butenyl isothiocyanate, which had selective cytotoxic/apoptotic activity (Barillari et al., 2008). ...
The nematotoxic potential of water extracts and green manures prepared from three plant species, daikon Raphanus sativus var. longipinnatus L., purslane Portulaca oleracea L. and chinaberry Melia azedarach L., on Globodera rostochiensis was examined in vitro and in microplots. Significant alteration in J2 (second stage juvenile) activity and their hatching from eggs was observed with different exposure times by all plant species; more than 99% of the J2s were inactivated after 72h and the same degree of inhibition in hatching of the eggs occurred after exposure to the plant extracts. In microplots, the numbers of newly formed cysts and final nematode multiplication rates were reduced in unsterilized soil at 1, 3 and 5% (w:w) rates of amendment with fresh plant materials, and the infestation rates of potato plants did not differ significantly from those in soil treated with metham sodium 37%. The rate of emergence of J2 from cyst inocula declined by 36% in soil treated with chinaberry and purslane and by 71% in soil treated with daikon. The reduced availability of J2 in soil must be one of the reasons for decrease in nematode multiplication rates of 65% and 86% where soil was amended with chinaberry/purslane and daikon, respectively. In terms of plant growth improvement and nematode control, daikon amendment outperformed other treatments, including metham sodium.
... The presence of antioxidant is needed to prevent oxidation process and maintain the quality of food products from rancidity, discoloration, and other physical spoilage (Tamat et al., 2007). Several researches done by Barillari et al. (2008), Hanindita (2011), and Ghasemzadeh et al. (2012), showed that radish extract had 10.5% (w/v) glucosinolate glucoraphasatin (GRH) activity and 154.5% anti radical power (ARP) by the presence of 0.098 flavonoid and 24.32 µg/ml phenolic compound. ...
... Table 4 Some widely studied plant extracts where anti-cancer effects are mainly due to non-polyphenol phytochemicals. Chang et al., 2018Chang et al., , 2003Lee et al., 2008;Manosroi et al., 2016;Son et al., 2017) Peganum harmala L. (Barillari et al., 2008;Beevi et al., 2010) Allium sativum L. (Auyeung et al., 2012;Tseng et al., 2016;Wu et al., 2017;You et al., 2012) Crocus sativus L. (Bhandari, 2015;D'Alessandro et al., 2013;Khorasanchi et al., 2018;Patel et al., 2017) Ginkgo biloba L. Ahmed et al., 2017;Cao et al., 2017;Dias et al., 2008;Han et al., 2016;Park et al., 2013;Ye et al., 2007) Panax ginseng C.A.Mey. Ahuja et al., 2018;Pan et al., 2013;Qi et al., 2010;Sharma and Goyal, 2015;Wang et al., 2016) A. B. Oyenihi, C. Smith Journal of Ethnopharmacology 229 (2019) 54-72 Tables 2 and 3. ...
Ethnopharmacological relevance:
Given the severe side effects associated with most of the conventional cancer medications, as well as the expanding body of evidence indicating secondary toxicity of these drugs, individuals with cancer are increasingly turning to natural alternatives. Similarly, the pharmaceutical industry is in search of natural products to treat cancer. An understanding of the specific active components in plant products with which anti-cancer efficacy is achieved is required for this research to move forward.
Aim of the study:
To integrate data from cancer-related studies on plant-derived products or extracts, to elucidate whether these products may have similar active ingredients and/or mechanisms of action, that can explain their efficacy. This review also includes a discussion of the methodological complexities and important considerations involved in accurate isolation and characterisation of active substances from plant material.
Conclusions:
From the literature reviewed, most plant products with consistently reported anti-cancer efficacy contains high levels of polyphenols or other potent antioxidants and their mechanisms of action correlate to that reported for isolated antioxidants in the context of cancer. This suggests that natural products may indeed become the panacea against this chronic disease - either as therapeutic medicine strategy or to serve as templates for the design of novel synthetic drugs. The recommendation is made that antioxidant activity of plant actives and especially polyphenols, should be the focus of anti-cancer drug discovery initiatives. Lastly, researchers are advised to exploit current techniques of chemical compound characterization when investigating polyphenol-rich plants to enable the easy consolidation of research findings from different laboratories.
... The composition of the primary glucosinolates, glucoraphenin and glucoraphasatin, is an important feature to distinguish radish from other cruciferous vegetables. Sulforaphene, a degradation product of glucoraphenin, inhibits the proliferative growth of various carcinoma cells (Barillari et al. 2008;Kuang et al. 2013;Papi et al. 2008). Because of its high medicinal properties, improving the yield and quality of radish crops is an important area for investigation. ...
High-throughput small RNA sequencing and degradome analysis were used in this study to thoroughly investigate the role of miRNA-mediated regulatory network in tuberous root development of radish. Samples from the early seedling stage (RE) and the cortex splitting stage (RL) were used for the construction of six small RNA libraries and one degradome library. A total of 518 known and 976 novel miRNAs were identified, of which, 338 known and 18 novel miRNAs were expressed in all six libraries, respectively. A total of 52 known and 57 novel miRNAs were identified to be significantly differentially expressed between RE and RL, and 195 mRNAs were verified to be the targets of 194 miRNAs by degradome sequencing. According to the degradome analysis, 11 differentially expressed miRNAs had miRNA-mRNA targets, and 13 targets were identified for these 11 miRNAs. Of the 13 miRNA-mRNA targets, 4 genes (RSG11079.t1, RSG11844.t1, RSG16775.t1, and RSG42419.t1) were involved in hormone-mediated signaling pathway, 2 gens (RSG11079.t1 and RSG16775.t1) were related to post-embryonic root development, and 1 gene (RSG23799.t1) was involved in anatomical structure morphogenesis, according to the GO function analysis for biological process. Target Genes participated in these processes are important candidates for further studies. This study provides valuable information for a better understanding of the molecular mechanisms involved in radish tuberous root formation and development.
... We also found that SFE on A549 cells induced the cleavage of caspase-3, -8, and -9 involved in apoptosis mechanism ( [30,31]. However, given its rapid degradation, RH has only a weak effect in fighting cancer cells. ...
Sulforaphene (SFE), a major isothiocyanate in radish seeds, is a close chemical relative of sulforaphane (SFA) isolated from broccoli seeds and florets. The anti-proliferative mechanisms of SFA against cancer cells have been well investigated, but little is known about the potential anti-proliferative effects of SFE. In this study, we showed that SFE purified from radish seeds inhibited the growth of six cancer cell lines (A549, CHO, HeLa, Hepa1c1c7, HT-29, and LnCaP), with relative half maximal inhibitory concentration (IC50) values ranging from 1.37 to 3.31 g/mL. Among the six cancer cell lines evaluated, SFE showed the greatest growth inhibition against A549 lung cancer cells. In A549 cells, SFE induced apoptosis via changes in the levels of poly (adenosine diphosphate ribose) polymerase and caspase-3, -8, and -9. Our results indicate that SFE from radish seeds may have significant anti-proliferative potency against a broad range of human cancer cells via induction of apoptosis.
... Its extracts are known to show an inhibitory effect on tumor growth. For example, the root of RS extracts showed a pro-apoptotic activity in human colon carcinoma cells and breast cancer cells (10,11). In addition, they inhibited metastasis in melanoma cells (12). ...
Raphanus sativus L. (RS) is a cruciferous vegetable that is widely consumed in Korea. The anticancer activity of leaves of RS (RSL) extract has been investigated; however, no studies focused on its anti-inflammatory effects. Therefore, the aim of the current study was to evaluate the anti-inflammatory effects of RSL extract. In brief, RSL powder was fractionated into n-hexane, chloroform, ethyl acetate, n-butanol, and water-soluble fractions. Lipopolysaccharide (LPS)-stimulated RAW264.7 cells were treated with each fraction for initial screening. It was found that the chloroform fraction significantly inhibited nitric oxide release in LPS-stimulated RAW264.7 cells with a half maximal inhibitory concentration value of 196 μg/mL. In addition, the mRNA and protein expression levels of inducible nitric oxide synthase, measured using reverse transcriptase-polymerase chain reaction and western blotting, respectively, were reduced in a concentration-dependent manner. Moreover, the inflammatory cyclooxygenase-2 enzyme expression decreased. Furthermore, the expression of nuclear factor-kappa B (NF-κB), the key regulator of the transcriptional activation of the inflammatory cytokine genes, was reduced by the RSL chloroform fraction. Therefore, the results of our study suggest that RSL exhibits anti-inflammatory effects in LPS-stimulated macrophages via NF-κB inactivation.
... In the present study, cherry radish sprouts were chosen as the materials, which contain substantial amounts of antioxidants, vitamin C and health-promoting compounds such as glucosinolates and phenolic compounds [42][43][44] . Consumption of radish sprouts has been confirmed to reduce the risk of cancer and oxidative stress both in vivo and in vitro [45][46][47] . Furthermore, with red hypocotyls resulting from anthocyanin accumulation, cherry radish sprouts could provide visual evidence for the biosynthesis of anthocyanins, and consequently, the high concentration of anthocyanin accumulated in cherry radish sprouts could underpin their high antioxidant activity. ...
The cross talk among hydrogen peroxide (H2O2), nitric oxide (NO) and UV RESISTANCE LOCUS8 (UVR8) in UV-B-induced anthocyanin accumulation in the hypocotyls of radish sprouts was investigated. The results showed that UV-B irradiation significantly increased the anthocyanin accumulation and the expression of UVR8, and a similar trend appeared in radish sprouts subjected to cadmium, chilling and salt stresses regardless of light source. However, these responses disappeared under dark exposure. These results suggest that abiotic stress-induced anthocyanin accumulation and UVR8 expression were light-dependent. Moreover, abiotic stresses all enhanced the production of H2O2 and exogenous H2O2 addition significantly increased the anthocyanin concentration and UVR8 transcription, while these increases were severely inhibited by addition of dimethylthiourea (DMTU, a chemical trap for H2O2). It seems to suggest that H2O2 played an important role in the anthocyanin biosynthesis. Furthermore, addition of 0.5 mM sodium nitroprusside (SNP, a NO-releasing compound) substantially induced the anthocyanin accumulation, and H2O2-induced anthocyanin accumulation and UVR8 expression were significantly suppressed by co-treatment with 2-phenyl-4,4,5,5-tetramethylimidazoline-3-oxide-1-oxyl (PTIO, a NO scavenger), which was parallel with the expression of anthocyanin biosynthesis-related transcription factors and structural genes. All these results demonstrate that both H2O2 and NO are involved in UV-B-induced anthocyanin accumulation, and there is a crosstalk between them as well as a classical UVR8 pathway.
... Dried RSL seeds are also ethnogynecologically used in Udhampur district for the treatment of menstrual disorders, in which one teaspoon of the seeds is taken with water several times for 2-3 days depending on the case of disorders (Bhatia et al., 2015). A preliminary study showed that the seeds of RSL exerted antioxidant and chemopreventive potentials (Barillari et al., 2003). A recent report revealed the anti-inflammatory and anti-tumor activities of the seeds (Kim et al., 2014). ...
... Recently studies have demonstrated that the extract of radish's taproot have potential in inhibiting the abnormal growth of vascular smooth muscle cells [13], protecting the cell membranes against lipid peroxidation and intervening in dysfunction of gastrointestinal motility [14]. 4-(Methylthio)-3-butenyl isothiocyanate (MTBITC), the pungent principle and bioactive chemical of radish which is produced via myrosinase-catalyzed hydrolysis of 4-(Methylthio)-3-butenyl glucosinolates, has been studied as a potential phytochemical that has the effect of inducing the apoptosis and reducing the growth of tumour cells by affecting it's signaling pathways [15,16]. Also, MTBITC alone was proved to be safe and have the chemoprotective effects against the toxicity of Zearalenone [17,18]. ...
The aim of this study is to develop a high fat diet and over nutrition induced nonalcoholic fatty liver disease(NAFLD) in rat, and to investigate the effect of 4-(Methylthio)-3-butenyl isothiocyanate (MTBITC) on ameliorating the NAFLD. Twenty Sprague-Dawley (SD) rats were equally divided into 4 groups (C, M, E1 & E2). Control group (C) were treated with standard restricted diet; Model group (M) were given high fat liquid diet ad libitum; Experimental group (E1 & E2) were treated with high fat liquid diet ad libitum and MTBITC by gavage. The experiment last 9 weeks, and serum chemistry and liver histology were assessed. The rats of M group showed severe lipid deposition and peroxidation in liver. When compared with group C, group M also showed significantly higher serum concentration of low-density lipoprotein, tumor necrosis factor-α and glucose. Histopathologic sections demonstrated lipid accumulation and macrovascular steatosis with ballooning degeneration in the livers of M. Group E2 presented significantly better conditions when assessed based on the parameters of NAFLD. The data suggested that MTBITC might significantly attenuate fat liquid diet induced NAFLD.
... Raphasatin is an antioxidant (3,4), a potent inducer of phase II enzymes in precision-cut rat liver slices (5), cytotoxic to multiple cancer cell lines (6)(7)(8)(9), and has antimutagenic properties (10). Sulforaphene has antimicrobial, antiviral, and antioxidant properties (4,10,11). ...
The biologically active compounds raphasatin and sulforaphene are formed during the hydrolysis of radishes by an endogenous myrosinase. Raphasatin is very unstable, and it is generated and simultaneously degraded to less active compounds during hydrolysis in aqueous media. This study determined the hydrolysis conditions to maximize the formation of raphasatin and sulforaphene by an endogenous myrosinase and minimize their degradation during the hydrolysis of radish roots. The reaction parameters, such as the reaction medium, reaction time, type of mixing, and reaction temperature were optimized. A stability test for raphasatin and sulforaphene was also performed during storage of the hydrolyzed products at 25°C for 10 days. The formation and breakdown of raphasatin and sulforaphene in radish roots by endogenous enzymolysis was strongly influenced by the reaction medium, reaction time, and type of mixing. The production and stabilization of raphasatin in radishes was efficient in water and dichloromethane with shaking for 15 min at 25°C. For sulforaphene, the favorable condition was water as the reaction medium without shaking for 10 min at 25°C. The maximum yields of raphasatin and sulforaphene were achieved in a concurrent hydrolysis reaction without shaking in water for 10 min and then with shaking in dichloromethane for 15 min at 25°C. Under these conditions, the yields of raphasatin and sulforaphene were maximized at 12.89 and 1.93 μmol/g of dry radish, respectively. The stabilities of raphasatin and sulforaphene in the hydrolyzed products were 56.4% and 86.5% after 10 days of storage in water and dichloromethane at 25°C.
... The markers sulforaphane and sulforaphene in Thai rat-tailed radish extract might therefore directly contribute to the apoptosis induction effect. Our study agrees with a previous report regarding the cytotoxic and apoptotic activities of isothiocyanates extracted from R. sativus in cancer cell lines (Barillari et al., 2008; Papi et al., 2008). ...
The cancer preventive effect of Raphanus sativus L. var. caudatus Alef (Thai rat-tailed radish) extract was evaluated against the HCT116 colon cancer cell line. HPLC and GC-MS were used to identify the phytoconstituents in Thai rat-tailed radish extract. Thai rat-tailed radish extract showed high cytotoxicity against HCT116 with an IC50 at 9.42 +/- 0.46 mu g/ml. Apoptosis induced by Thai rat-tailed radish extract was confirmed (a) by DAPI staining which demonstrated nuclei morphological changes, chromatin condensation and nuclear fragmentation and (b) by Annexin V/PI flow cytometry. GC-MS analysis of Thai rat-tailed radish extract revealed two isothiocyanates; sulforaphane and sulforaphene, whose cytotoxicity and apoptosis induction effect were evident, giving Thai rat-tailed radish extract its anticancer attributes. This is the first report on the cancer preventive effect of Thai rat-tailed radish extract against HCT116 colon cancer cell line, through apoptosis induction.
... The bioactive potencies of RH and SFE, which are the major ITCs in radish roots, have not been widely reported.Scholl et al. (2011)showed that the RH from GRH in Spanish black radish roots induced apoptosis of Hep1c1c7 cancer cells. The SFE from GRE in radish seeds or roots had anticancer and antioxidant effects (Barillari et al., 2005Barillari et al., , 2008). The chemical stability of ITCs is critical for their development as anticancer agents or as dietary foods (Papi et al., 2008;Hanlon and Barnes, 2011). ...
... Other natural substances containing divalent sulfur atoms may similarly act as preventive antioxidants by decomposing hydroperoxides by a nonradical pathway. Glucosinolates from daikon (Raphanus sativus L.) 28, 29 and rocket (Eruca sativa Mill.) 30 are able to reduce H 2 O 2 and organic hydroperoxides to water and alcohols, respectively, at the expense of a S(II) atom, which is oxidized to sulfoxide SO (see Scheme 6). The same chemistry is responsible for the direct (preventive) antioxidant activity of volatile isothiocyanates (ITCs) released from Brassicaceae upon chopping or grinding the vegetables or seeds, by enzymatically (myrosinase) induced decomposition of glucosinolates. ...
Essential oils (EOs) are liquid mixtures of volatile compounds obtained from aromatic plants. Many EOs have antioxidant properties and the use of EOs as natural antioxidants is a field of growing interest since some synthetic antioxidants like BHA and BHT are now suspected to be potentially harmful to human health. Addition of EOs to edible products, either by direct mixing or in active packaging and edible coatings, may therefore represent a valid alternative to prevent the autoxidation and prolong their shelf-life. The evaluation of the antioxidant performance of EOs is however a crucial issue, since many commonly used "tests" are inappropriate and give contradictory results that may mislead future research. The chemistry explaining EOs antioxidant activity is discussed along with an analysis of the potential in food protection. Literature methods to assess EOs' antioxidant performance are critically reviewed.
Essential oils (EOs) are mixtures of volatile molecules endowed with health-promoting biological activities that go beyond their role as aromas and natural preservatives and can be exploited to develop functional foods and diet supplements. Their composition is briefly addressed along with regulatory aspects. The potential health benefit of human diet supplementation with EOs is outlined through a review of the recent literature on available clinical trials and preclinical research concerning EOs activity towards: (1) irritable bowel syndrome; (2) inflammatory bowel disease; (3) regulation of microbiota; (4) gastroprotection; (5) hepatoprotection; (6) protection of the urinary tract and diuresis; (7) management of metabolic disorders including hyperglycemia and hyperlipidemia; (8) anti-inflammatory and pain control; (9) immunomodulation and protection from influenza; and (10) neuroprotection and modulation of mood and cognitive performance. The emerging potential in such activities of selected EOs is given focus, particularly green and black cumin, bergamot, orange, myrtle, peppermint, sage, eucalyptus, lavender, thyme, lemon balm, ginger, and garlic.
Plant volatiles as bio-fumigants are effective tools for food protection and infestation control. Effective applications of plant volatiles depend on the stabilization and controlled release of these compounds. Tremendous efforts have been devoted in this research area. This chapter reviews: (1) their functions and modes of actions, (2) the approaches to stabilize and control their release, and (3) characterization methods used in the literature in developing these systems. Examples of these systems in various forms are presented to illustrate how these volatile compounds can be stabilized and delivered.
Sulforaphene prepared from glucoraphenin by myrosinase is one of the main active ingredients of radish, which has various biological activities and brilliant potential for food and pharmaceutical applications. In this paper, a recombinant food-grade yeast transformant 20-8 with high-level myrosinase activity was constructed by over-expressing a myrosinase gene from Arabidopsis thaliana in Yarrowia lipolytica. The highest myrosinase activity produced by the transformant 20-8 reached 44.84 U/g dry cell weight when it was cultivated in a 10 L fermentor within 108 h. Under the optimal reaction conditions, 6.1 mg of sulforaphene was yielded from 1 g of radish seeds under the catalysis of the crude myrosinase preparation (4.95 U) at room temperature within 1.5 h. What is more is that when the whole yeast cells harboring myrosinase activity were reused 10 times, the sulforaphene yield still reached 92.53% of the initial level. Therefore, this efficient approach has broad application prospects in recyclable and large-scale preparation of sulforaphene.
Radish (Raphanus raphanistrum) is an annual and a biennial cool-season crop of ancient origin believed to be originated from Mediterranean South Asian region and grown in Southeast Asian countries. Radish is a root crop particularly grown for its taproot that varies in colour from white, pink, red, purple, and black and has appetizing flavour. The taproot edible as raw as it is palatable, juicy and crunchy with a pungent peppery pleasing taste. All botanical parts of radish such as leaves, stem, seeds, and pods are utilized in food in one or another way. Besides its consumption as fresh juice and in salad form, it is processed to traditional pickles, and its leaves and pods are used in traditional foods. It is also well known to cure various ailments and used from centuries in Southeast Asian countries as traditional medicine. With increasing interest of researchers in this root over last few decades to confirm its health claims, the natural molecules such as glucosinolates, isothiocyanates, phenols, flavonoids, anthocyanins and other bioactive substances are believed to be responsible for its medicinal activities. Leaves, seeds, peel and sprouts of radish are rich sources of many functional molecules that may have possible medicinal values. This chapter aims to summarize the history, production, botanical classification, chemical components and health beneficial properties of radish.
The autoxidation kinetics of stripped sunflower oil (SSO), squalene (SQ) and p-cymene (p-C) initiated by 2,2′-azobis(isobutyronitrile) at 303 K were investigated under controlled conditions by differential oximetry so to build reference model systems that are representative of the natural variability of oxidizable materials, for quantitative antioxidant testing. Rate constants for oxidative chain-propagation kp and chain-termination 2kt and the oxidizability (kp/√2kt) were measured using 2,6-di-tert-butyl-4-methoxyphenol, 2,2,5,7,8-pentamethyl-6-chromanol, BHT, and 4-methoxyphenol as reference antioxidants. Measured values of kp (M-1s-1) / 2kt (M-1s-1) / oxidizability (M-1/2s-1/2) at 303K in chlorobenzene were 66.9 / 3.45106 / 3.6×10-2, 68.0 / 7.40106 / 2.5×10-2, and 0.83 / 2.87106 / 4.9×10-4, respectively for SSO, SQ and p-C. Quercetin, magnolol, caffeic acid phenethyl ester and 2,4,6-trimethylphenol, were investigated to validate calibrations. The distinctive usefulness of the three substrates in testing antioxidants is discussed.
Isothiocyanates (ITCs) have gained increasing attention since they have been attributed the merits for the potential beneficial effects of cruciferous vegetable dietary consumption on cancer. The aim of the present study was to determine the cytotoxic effects of 3-butenyl ITC (3-BI) on prostate cancer (PC) cells under in vitro conditions. Two androgen-insensitive human PC cell lines, PC-3 and DU145, were assayed. Cells were cultured in the presence of increasing concentrations of 3-BI (5, 10, 30 and 50 µM) in the absence or presence of the chemotherapeutic drug docetaxel (DOCE) (1 and 2 nM). The cytotoxic effects of these compounds were analyzed using the trypan blue exclusion assay at 24, 48 and 72 h. Apoptosis and migration assays were also performed. The results showed that 3-BI induced a dose-dependent cytotoxic effect on PC-3 cells at 24, 48 and 72 h. These effects were significantly higher than those found with DOCE at 72 h of culture. Moreover, 3-BI also potentiated the effects of DOCE in a dose-dependent manner. Additionally, 3-BI showed inhibition of the migration of PC-3 cells. Nevertheless, 3-BI was not effective in the DU145 PC cell line. These results show a promising role for the 3-BI compound as a co-adjuvant agent in DOCE-based therapy in certain types of PC.
For more than three decades, various in vitro and in vivo studies have linked radishes with diabetes, though this link has not been discussed. This review systematically addresses and summarizes the effect of radishes on diabetes. We searched the Web of Science, PubMed, and EMBASE databases for English language articles from June 1987 through May 2017 using the key words " radish " and " diabetes, " and the references from particular reports were also considered if relevant. In summary, radish has been identified as having antidiabetic effects, making it favorable for those with diabetic conditions. This may be due to its ability to enhance the antioxidant defense mechanism and reduce the accumulation of free radicals, affect hormonal-induced glucose hemostasis, promote glucose uptake and energy metabolism, and reduce glucose absorption in the intestine. However, this summary requires further confirmation in research in vivo studies and clinical trials.
The antioxidant and anti-inflammatory effects of roasted dried radish (RDR) against renal oxidative stress were examined in high-fat diet (HFD)-fed mice. The HFD was prepared by adding lard to chow diet to provide 50% of the calories from fat. Hot water extracts of dried radish (DR) or RDR were administered orally to mice at 237 mg/kg bw/day, whereas distilled water was administered as a vehicle for 12 weeks. Compared to the control group, renal reactive oxygen species, peroxynitrite, and thiobarbituric acid reactive substance level in the DR or RDR group were significantly decreased, whereas the glutathione level was increased (p<0.05). Protein expressions of antioxidant factors such as nuclear factor erythroid 2-related factor-2, heme oxygenase-1, glutathione S-transferase, superoxide dismutase, catalase, and glutathione peroxidase were significantly increased in the DR and RDR groups; however, nuclear factor-kappa B expression was suppressed (p<0.05). These antioxidant and anti-inflammatory effects of RDR were found to be significantly greater than those of DR.
Gastric cancer migration and invasion considered as main causes of this cancer-related death around the world. Sulforaphene (4-isothiocyanato-4R-(methylsulfinyl)-1-butene), a structural analog of sulforaphane, has been found to exhibit anticancer potential against different cancers. Our aim was to investigate whether dietary isothiocyanate sulforaphene (SFE) can promote human gastric cancer (AGS) cells apoptosis and inhibit migration. Cells were treated with various concentrations of SFE and cell viability, morphology, intracellular ROS, migration and different signaling protein expressions were investigated. The results indicate that SFE decreases AGS cell viability and induces apoptosis in a dose-dependent manner. Intracellular ROS generation, dose- and time-dependent Bax/Bcl2 alteration and signaling proteins like cytochrome c, Casp-3, Casp-8 and PARP-1 higher expression demonstrated the SFE-induced apoptotic pathway in AGS cells. Again, SFE induced apoptosis also accompanied by the phosphorylation of mitogen-activated protein kinases (MAPKs) like JNK and P-38. Moreover, dose-dependent EGFR, p-ERK1/2 down-regulation and cell migration inhibition at non-toxic concentration confirms SFE activity in AGS cell migration inhibition. Thus, this study demonstrated effective chemotherapeutic potential of SFE by inducing apoptisis as well as inhibiting migration and their preliminary mechanism for human gastric cancer management.
A novel electrochemical method has been developed for the determination of the oxygen solubility in cetyltrimethylammonium bromide (CTAB) micelles. The electroreduction of oxygen is studied in aqueous phosphate buffer solutions, using a microelectrode. The addition of micelle forming surfactants to solutions pre-saturated with oxygen leads to a reduction of the oxygen signal allowing the oxygen uptake by the micelles to be measured. For CTAB micelles, a concentration of oxygen of 6.7 ± 0.72 mM was observed, and shown to remain constant with increasing CTAB concentration in the bulk solution. The method has general applicability.
Chemical investigation of the dichloromethane extract of freeze-dried Raphanus sativus roots led to the isolation of 4-methylthio-3-butenyl isothiocyanate or raphasatin (1a), 4-(methylthio)butyl isothiocyanate or erucin (1b), β- sitosterol (2) and unsaturated triglycerides (3). The structures of 1a and 1b were elucidated by extensive 1D and 2D NMR spectroscopy, while those of 2 and 3 were identified by comparison of their NMR data with those reported in the literature.
To explore if a natural isothiocyanate, sulforaphene (SFE), sensitizes ovarian cancer cells to the chemotherapy drug cisplatin (CDDP).
We studied reactive oxygen species (ROS), mitochondrial membrane depolarization and cell-cycle distribution in two ovarian cancer cell lines SKOV3 and SNU 8 treated with SFE and cisplatin. We further analyzed the expression of caspases 3, 8, and 9, Phosphoinositide 3-kinase (PI3K) and Phosphatase and tensin homolog (PTEN) by western blotting.
SFE sensitized cells to cisplatin by enhancing ROS and mitochondrial membrane depolarization that released cytochrome c and activated caspase 9 and caspase 3 in the mitochondrial pathway. It also inhibited extrinsic pathway protein caspase 8, growth-related protein PI3K and further activated PTEN in combination with cisplatin.
SFE synergistically inhibited proliferation and induced apoptosis of SKOV3 and SNU8 cells in combination with cisplatin by activating multiple apoptotic pathways. Therefore, we suggest sulforaphene as a chemo-enhancing adjuvant to improve the efficacy of cisplatin in ovarian cancer treatment.
Copyright© 2015 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.
The effects of ultrasonic pressure on the growth and antioxidant activity of Raphanus sativus L. sprouts (Japanese radish sprouts) were studied. Sprouts were grown under four different ultrasonic pressures (40 kHz; 0, 33.4, 78.9 and 136.4 kPa). The fresh weight per plant decreased with the increase in ultrasonic pressure, resulting from the decrease in water content. However, the total polyphenol content and 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging activity are independent of ultrasonic pressure. Therefore, both values increase with the increase in ultrasonic pressure for per fresh weight of sprout. In conclusion, the application of ultrasound to the roots of Japanese radish sprouts is an effective method to increase the total polyphenol content.
The seeds of Raphanus sativus L. (RSL) have long been used as anti-inflammatory traditional medicine. However, scientific bases for the purported potential of the medicine and the associated mechanisms were barely defined. This study investigated the effects of RSL seeds on lipopolysaccharide (LPS)-stimulated inflammatory responses in vitro and in vivo. Treatment with 100 μg/ml ethyl acetate fraction (REF), which was isolated from water extract of the seeds, significantly inhibited LPS-stimulated production of nitric oxide (P < 0.05), interleukin-6 (P < 0.001), and tumor necrosis factor (TNF)-α (P < 0.001) in RAW264.7 cells. Oral supplementation with 30 mg/kg REF protected mice by 90% against LPS-induced septic death and prevented the increases of serum TNF-α and interferon-γ levels in LPS-injected mice. When REF was divided into four sub-fractions (REF-F1–F4), REF-F3 showed the greatest activity to suppress LPS-stimulated production of inflammatory mediators. We subsequently isolated an active fraction from the REF-F3 and identified sinapic acid as the main constituent. The addition of 50 μg/ml active fraction markedly inhibited LPS-stimulated production of inflammatory mediators by suppressing p38 MAPK and nuclear factor-κB activation. Furthermore, supplementation with the active fraction (10 mg/kg) improved the survival rate of LPS-injected mice by 80% of the untreated control. Additional experiments revealed that sinapic acid was the active component responsible for the anti-inflammatory potential of RSL seeds. Collectively, our current results suggest that both RSL seeds and sinapic acid may be attractive materials for treating inflammatory disorders caused by endotoxins.
Glucosinolates, which are abundant in Brassicaceae crops as secondary metabolites, play an important role in both plant growth and self-defense. The present study was carried out to find out the temporal relationship between aliphatic glucosinolates content in tissue and level of expression of the genes involved in the biosynthesis during early seedling growth and herbivory of S. exigua in radish. The major glucosinolates found in radish were glucoraphenin in seed and glucoraphasatin in tissues other than seed. Glucoraphenin content was high in radish seeds, showing 7-fold compared to the level of glucoraphasatin, and decreased fast during germination. Glucoraphasatin in radish seedlings increased abruptly up to 7 days during germination, ranging 8.5 to 10.8 mg/g dry weight, thereafter, decreased consistently. RT-PCR study of the genes involved in the biosynthesis of glucosinolates in radish revealed that the expression of CYP79F1, CYP83A1 are gradually increased after germination, however, the glucoraphenin sharply decreased. Herbivory with S. exigua increased the amount of glucoraphasatin by 1.3 fold in all three cultivars tested.
The radish is a root vegetable of the Brassicaceae family that is mainly consumed in Far Eastern Asian countries. Because chemical indexes indicating the quality of a radish have not been established, we investigated the variations in the contents of the soluble sugars and organic acids that influence taste among seven radish cultivars. In terms of the total soluble sugar content, which is a strong index of sweetness, Koshin, Kouto, and Shogoin were the three highest among the seven cultivars. Sobutori, which is the most common radish cultivar in Japan, was the lowest. The total organic acid contents varied among the seven cultivars, although they were five to thirteen times lower than the total soluble sugar contents. These results indicate that there are variations in the soluble sugar and organic acid contents among radish cultivars; therefore, these contents may be used as chemical indexes for the quality of radishes.
The purpose of this study was optimization of the conditions for mixing different amounts of red radish (Raphanus sativus L.) sprout powder, butter, and sugar when baking domestic wheat cookies prepared using red radish sprout powder. Response surface methodology, with a central composite design comprising 5 levels and 3 variables, was used to identify the best possible combination of amounts of red radish sprout powder (), butter (), and sugar (). The physical and mechanical properties of each of the 20 samples analyzed, including color L (p
The vegetative, or above ground, portion of radishes are not typically consumed and have not been characterized in terms of their phytochemical content or biological activity. The antioxidant capacity of the freeze-dried vegetative portion of mature Spanish black radishes (harvested 2months after planting) was 3.7-, 3.6- and 2.9-fold greater than freeze-dried roots as assayed by ferric ion reducing antioxidant power (FRAP), total phenolics and oxygen radical absorbance capacity (ORAC), respectively. No carotenoids were detected in the roots, however the vegetative portion contained 905.0 and 421.2μg/g of lutein and β-carotene, respectively. Despite differences in glucosinolate composition, a class of compounds related to the induction of detoxification enzymes, the aqueous extracts of roots and the vegetative portion similarly induced quinone reductase activity and the expression of multiple phase I and II detoxification enzymes in the HepG2 human hepatoma cell line. When juiced, the resulting freeze-dried juice and pulp of the vegetative portion maintained the ability to induce detoxification enzymes, although there were some differences in the phytochemical contents of these two dried materials. These data demonstrate that the ability of the vegetative portion of Spanish black radish, which has significantly greater antioxidant potential than roots, to induce detoxification enzyme expression was similar to that of the roots.
Raphanus sativus, a common cruciferous vegetable has been attributed to possess a number of pharmacological and therapeutic properties. This
present study evaluated the protective effect of different parts of R. sativus such as root, stem and leaf obtained with a variety of extraction solvents against cell death and oxidative DNA damage induced
by hydrogen peroxide (H2O2) in normal human lymphocytes. R. sativus extracts as such showed no cytotoxicity and genotoxicity to the lymphocytes at the tested concentrations. Of the different
extracts, hexane extract of root and methanolic extract of stem and leaf showed significant protective effect against oxidative
damage induced by 200 μM H2O2 in a dose dependent manner, as compared to cells exposed only to H2O2. Our results suggest that the protective effect afforded by R. sativus extract could be related to the presence of isothiocyanates and polyphenolics, as they possess significant capacity to remove
reactive species by virtue of their ability to scavenge free radicals and induce antioxidant enzyme system in the cells.
The restriction of phosphatidylserine (PtdSer) to the inner surface of the plasma membrane bilayer is lost early during apoptosis. Since PtdSer is a potent surface procoagulant, and since there is an increased incidence of coagulation events in patients with systemic lupus erythematosus (SLE) who have anti-phospholipid antibodies, we addressed whether apoptotic cells are procoagulant and whether anti-phospholipid antibodies influence this. Apoptotic HeLa cells, human endothelial cells, and a murine pre-B-cell line were markedly procoagulant in a modified Russell viper venom assay. This procoagulant effect was entirely abolished by addition of the PtdSer-binding protein, annexin V, confirming that it was PtdSer-dependent. The procoagulant effect was also abolished by addition of IgG purified from the plasma of three patients with anti-phospholipid antibody syndrome, but not IgG from normal controls. Confocal microscopy of apoptotic cells stained with fluorescein-isothiocyanate-conjugated-annexin V demonstrated (Ca2+)-dependent binding to the surface of membrane blebs o apoptotic cells, but not to intracellular membranes. Recent data indicate that the surface blebs of apoptotic cells constitute an important immunogenic particle in SLE. We propose that the PtdSer exposed on the outside of these blebs can induce the production of anti-phospholipid antibodies, which might also enhance the immunogenicity of the bleb contents. When apoptosis occurs in a microenvironment in direct contact with circulating plasma, the unique procoagulant consequences of the apoptotic surface may additionally be expressed. This might explain the increased incidence of pathological intravascular coagulation events that occur in some lupus patients who have anti-phospholipid antibodies.
Unlabelled:
Cytotoxic CD8+ cells play an important role in determining host response to tumor, thus chemotherapy is potentially dangerous as it may lead to T cells depletion. The purpose of this study was to elucidate the propensity of quiescent and proliferating human CD8+ cells to undergo cell death upon treatment with curcumin, a natural dye in Phase I of clinical trials as a prospective chemopreventive agent.
Methods:
We treated human quiescent or proliferating CD8+ cells with 50 microM curcumin or irradiated them with UVC. Cell death symptoms such as decreased cell viability, chromatin condensation, activation of caspase-3 and specific DFF40/CAD endonuclease and oligonucleosomal DNA fragmentation were analyzed using MTT test, microscopic observation, Western blotting and flow cytometry.
Results:
Curcumin decreased cell viability, activated caspase-3 and decreased the level of DFF45/ICAD, the inhibitor of the DFF40/CAD endonuclease. However, this did not lead to oligonucleosomal DNA degradation. In contrast, UVC-irradiated proliferating, but not quiescent CD8+ cells revealed molecular and morphological changes characteristic for apoptosis, including oligonucleosomal DNA fragmentation. Curcumin can induce cell death in normal human lymphocytes both quiescent and proliferating, without oligonucleosomal DNA degradation which is considered as a main hallmark of apoptotic cell death. Taking into account the role of CD8+ cells in tumor response, their depletion during chemotherapy could be particularly undesirable.
Industrial grade cardanol is a yellow oil containing mostly cardanol with smaller percentages of cardol and methylcardol, obtained by thermal treatment and subsequent distillation of cashew nut shell liquid available in large amounts in some developing countries. In order to find useful applications of this widely available and renewable raw material in fine chemical processes we undertook a kinetic investigation of the antioxidant activity of some hydrogenated cardanol and cardol derivatives, compared with that of analogous commercial products.
Rate constants for hydrogen atom abstraction from two water-soluble α-tocopherol (α-TOH, vitamin E) analogues by tert-butoxyl radicals have been measured at room temperature in four solvents, including water. The results imply that H-atom abstraction from α-TOH is 3.9 times slower in water than in benzene and yield a “reliable” Kamlet−Taft β solvent parameter for water of 0.31. Literature rate constants for H-atom abstraction by peroxyl radicals from α-TOH in SDS micelles and in phospholipid micelles are, respectively, about 100 and 1000 times lower than in styrene. Since only a small fraction of the observed rate diminution can be attributed to hydrogen bonding of α-TOH to water it is concluded that in these heterogeneous systems much of the α-TOH is physically inaccessible to the attacking radicals. Whether this is also true for α-TOH in the biological membranes in living animals remains to be determined.
An EPR investigation of the kinetics of the exit, k
-, and entrance, k
+, processes in micelles of sodium dodecyl sulfate, hexadecyltrimethylammonium bromide and polyoxyethylene(6)decanol of a family of para-substituted benzyl tert -butyl nitroxides and para-substituted benzyl hydroxyalkyl nitroxides is reported. The inclusion of nitroxide probes in the hydrophobic environment of the micelle gives rise to a reduction of the value of both nitrogen and β-proton splittings, with the result that the resonance fields for the MI(2Hβ
) = ±1lines of the free and included species are significantly different. The rate constants were obtained by analyzing the EPR line shape variations as function of surfactant concentration and temperature. The experimental value of k
+ obtained from the study of benzyl tert-butyl nitroxide indicates that the association reaction is very close to being controlled by diffusion. The value of the exit rate, k
-, instead, depends on the probe hydrocarbon chain length. A comparison of our results with those obtained by luminescence quenching techniques is also reported.
Epidemiological and animal studies on vitamin A and its analogues support the hypothesis that β-carotene can prevent cancer in humans¹. However, chemoprevention trials have unexpectedly shown that β-carotene, either alone or in combination with vitamin A or vitamin E, actually increases lung-cancer incidence and mortality in heavy smokers and asbestos workers2, 3, ⁴. We find that β-carotene in rat lung produces a powerful booster effect on phase I carcinogen-bioactivating enzymes, including activators of polycyclic aromatic hydrocarbons (PAHs), and that this induction is associated with the generation of oxidative stress. Our findings might explain why β-carotene supplementation increases the risk of lung cancer in smokers.
The effects of glucosinolates (GLs) (sinigrin, gluconapin, progoitrin, epi-progoitrin, sinalbin, glucotropaeolin, glucoerucin, glucocheirolin, and glucoraphenin) and their enzymatic hydrolysis-derived products (EHDPs) have been studied in controlling the proliferation of cancer cell lines. The results of this study indicate the following: (i) neither myrosinase nor intact GLs have any effect on tumor cell growth when used up to 36 U/mL and 500 μM, respectively; (ii) all EHDPs show a clear inhibition of human erythroleukemic K562 cell proliferative growth, which is particularly evident for EHDPs from sinigrin, glucotropaeolin, glucoerucin, and glucocheirolin (IC50 < 20 μM); (iii) the EHDP production by in situ or pre-mix procedures gives rather similar antiproliferative effects; and finally, (iv) the EHDPs from glucoraphenin are active toward several other tumor cells, viz. FL (murine erythroleukemic cells), Jurkat (human T-lymphoid cells), HeLa (human cervix carcinoma cells), H9 (human T-lymphoid cells), and H3-T1-1 cells (obtained by transfection of HeLa with a LTR-HIV-1-CAT plasmid). Keywords: Crucifers; glucosinolates; isothiocyanates; myrosinase; antitumor activity
Rate constants have been measured at 25 degrees C in 13 solvents (S) for abstraction of the phenolic hydrogen atom from alpha-tocopherol (TOH) by tert-butoxyl (BO.), k(TOH/BO)(S), and by 2,2-diphenyl-1-picrylhydrazyl (DPPH.), k(TOH/DO)(S), and in eight solvents for abstraction of the phenolic hydrogen atom from phenol by cumyloxyl, k(PhOH/CumO)(S), and DPPH., k(PhOH/DPPH)(S). Over the range of solvents examined k(TOH/BO)(S) and k(TOH/DPPH)(S) vary by a factor of Ca. 65, and k(PhOH/CumO)(S) and k(PhOH/DPPH)(S) vary by a factor of ca. 120. in accordance with a prediction(5) the kinetic solvent effect is essentially identical for the same substrate and is independent of the attacking radical. That is, for almost any pair of solvents, A and B (k(TOH/BO)(A)/k(TOH/BO)(B))/(k(TOH/DPPH)(A)/k(TOH/DPPH)(B)) approximate to 1.0. The same applies with phenol as the substrate. Exceptions to this 1:1 relationship occur when one of the reactions becomes partly diffusion-controlled and in the solvent tert-butyl alcohol in which DPPH. shows a larger reactivity than would be expected. The absolute magnitudes of the alkoxyl and DPPH rate constants in the same solvent differ by a factor of over 1 000 000 (10(6)) for alpha-tocopherol and by 10 000 000 000 (10(10)) for phenol! We have therefore confirmed, under extreme conditions, a new, unifying principle for free radical chemistry in solution.
An improved high yielding procedure for the purification of myrosinase from Sinapis alba L. consisting of concanavalin A affinity chromatography followed by a chromatofocusing step is presented. The purified enzyme was homogeneous as judged by sodium dodecyl sulfate—gel electrophoresis and by analytical ultracentrifugation although the presence of at least three isoenzymes, with pI values from 5.05 to 5.15, was detected by isoelectric focusing. It was found that the enzyme has a molecular weight of 135.1 kg mol−1 and consists of two, possibly identical, subunits of molecular weight 71.7 kg mol−1. The structure of myrosinase was studied by circular dichroism. Contin analysis of the CD data indicates a mixed α-helix and β-sheet conformation for the native protein a with ~19% α-helix and ~35% β-sheet content. Denaturation with guanidinium chloride was found to be irreversible although the enzyme has excellent storage characteristics in aqueous solution.
Consumption of vegetables, especially crucifers, reduces the risk of developing cancer. Although the mechanisms of this protection are unclear, feeding of vegetables induces enzymes of xenobiotic metabolism and thereby accelerates the metabolic disposal of xenobiotics. Induction of phase II detoxication enzymes, such as quinone reductase [NAD(P)H:(quinone-acceptor) oxidoreductase, EC 1.6.99.2] and glutathione S-transferases (EC 2.5.1.18) in rodent tissues affords protection against carcinogens and other toxic electrophiles. To determine whether enzyme induction is responsible for the protective properties of vegetables in humans requires isolation of enzyme inducers from these sources. By monitoring quinone reductase induction in cultured murine hepatoma cells as the biological assay, we have isolated and identified (-)-1-isothiocyanato-(4R)-(methylsulfinyl)butane [CH3-SO-(CH2)4-NCS, sulforaphane] as a major and very potent phase II enzyme inducer in SAGA broccoli (Brassica oleracea italica). Sulforaphane is a monofunctional inducer, like other anticarcinogenic isothiocyanates, and induces phase II enzymes selectively without the induction of aryl hydrocarbon receptor-dependent cytochromes P-450 (phase I enzymes). To elucidate the structural features responsible for the high inducer potency of sulforaphane, we synthesized racemic sulforaphane and analogues differing in the oxidation state of sulfur and the number of methylene groups: CH3-SOm-(CH2)n-NCS, where m = 0, 1, or 2 and n = 3, 4, or 5, and measured their inducer potencies in murine hepatoma cells. Sulforaphane is the most potent inducer, and the presence of oxygen on sulfur enhances potency. Sulforaphane and its sulfide and sulfone analogues induced both quinone reductase and glutathione transferase activities in several mouse tissues. The induction of detoxication enzymes by sulforaphane may be a significant component of the anticarcinogenic action of broccoli.
A rapid, sensitive and automated assay procedure was developed for the in vitro evaluation of anti-HIV agents. An HTLV-I transformed T4-cell line, MT-4, which was previously shown by Koyanagi et al. (1985) to be highly susceptible to, and permissive for, HIV infection, served as the target cell line. Inhibition of the HIV-induced cytopathic effect was used as the end point. The viability of both HIV- and mock-infected cells was assessed spectrophotometrically via the in situ reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The procedure was optimized as to make optimal use of multichannel pipettes, microprocessor-controlled dispensing and optical density reading. The absorbance ratio of the mock-infected control to the HIV-infected samples was about 20. This allowed an accurate determination of the 50% effective doses, as demonstrated for 3'-azido-2',3'-dideoxythymidine (AZT), 2',3'-dideoxycytidine (ddCyd), dextran sulfate and heparin. The technique significantly reduced labor time as compared to the trypan blue exclusion method, and permits the evaluation of large numbers of compounds for their anti-HIV activity.
A colorimetric chemosensitivity test was investigated using sulforhodamine B (SRB), which stains protein synthesized by cells, as an end-point marker. Four cultured cell lines, 9 human tumor xenografts serially transplanted into nude mice, and 14 fresh surgical specimens were subjected to this assay. The optimal conditions for the assay were 3-5 x 10(4) cells per well in a 96-microplate, an SRB concentration of 4%, and an incubation time of more than 10 minutes. When mitomycin C, doxorubicin, cisplatin, and 5-fluorouracil were assessed by the SRB assay, the concentration-effect curves revealed a sharp slope between plateaux at low and high concentrations, suggesting that this assay has an excellent sensitivity which can assess the effect of drugs as "all or none." Although this high sensitivity resulted in good reproducibility of the assay for cultured cell lines, the predictive rate of the SRB assay for the chemosensitivity of human tumor xenografts in vivo was limited to 63.9%. As a result, this SRB assay is thought to be useful for evaluating the chemosensitivity of cultured cells as all or none, since it can assess directly cellular protein synthesis, which is one of the most important parameters of cell renewal, with excellent sensitivity.
REVIEW
Bcl-2 and related cytoplasmic proteins are key regulators of apoptosis, the cell suicide program critical for development,
tissue homeostasis, and protection against pathogens. Those most similar to Bcl-2 promote cell survival by inhibiting adapters
needed for activation of the proteases (caspases) that dismantle the cell. More distant relatives instead promote apoptosis,
apparently through mechanisms that include displacing the adapters from the pro-survival proteins. Thus, for many but not
all apoptotic signals, the balance between these competing activities determines cell fate. Bcl-2 family members are essential
for maintenance of major organ systems, and mutations affecting them are implicated in cancer.
Gastric infection with Helicobacter pylori is a cosmopolitan problem, and is especially common in developing regions where there is also a high prevalence of gastric cancer. These infections are known to cause gastritis and peptic ulcers, and dramatically enhance the risk of gastric cancer. Eradication of this organism is an important medical goal that is complicated by the development of resistance to conventional antimicrobial agents and by the persistence of a low level reservoir of H. pylori within gastric epithelial cells. Moreover, economic and practical problems preclude widespread and intensive use of antibiotics in most developing regions. We have found that sulforaphane [(-)-1-isothiocyanato-(4R)-(methylsulfinyl)butane], an isothiocyanate abundant as its glucosinolate precursor in certain varieties of broccoli and broccoli sprouts, is a potent bacteriostatic agent against 3 reference strains and 45 clinical isolates of H. pylori [minimal inhibitory concentration (MIC) for 90% of the strains is <or=4 microg/ml], irrespective of their resistance to conventional antibiotics. Further, brief exposure to sulforaphane was bactericidal, and eliminated intracellular H. pylori from a human epithelial cell line (HEp-2). In complementary experiments, sulforaphane blocked benzo[a]pyrene-evoked forestomach tumors in ICR mice. This protection resulted from induction of phase 2 detoxication and antioxidant enzymes, and was abrogated in mice lacking the nrf2 gene, which regulates phase 2 enzymes. Thus, the dual actions of sulforaphane in inhibiting Helicobacter infections and blocking gastric tumor formation offer hope that these mechanisms might function synergistically to provide diet-based protection against gastric cancer in humans.
A study on the regeneration of alpha-tocopherol (vitamin E) by phenolic co-antioxidants in homogeneous hydrocarbon solution is reported. The behavior of some relevant phenols such as BHA, BHT, and trans-resveratrol appears to be nicely predicted by a model based on the knowledge of kinetic and thermochemical data concerning the various reactants. Despite its good reputation as an antioxidant, trans-resveratrol was found only moderately effective (k(inh) = 2.0 x 10(5) M(-1) s(-1) in chlorobenzene at 303 K) and unable to recycle vitamin E.
Methanol extracts of 11 kinds of commonly available vegetables were examined for hydroxyl radical scavenging potency using the bleomycin-Fe method. In this method, the iron ion and bleomycin in water form hydroxyl radicals, and the scavenging activity is monitored by the modified thiobarbituric acid method. All extracts showed scavenging capacity, even though the activity of some of them was lower than that of l-ascorbic acid. Those vegetables were classified into three groups according to their activity, groups showing strong activity, moderate activity, and weak activity, as compared to the activity of l-ascorbic acid at the same concentration. Among them, the methanol extract of radish sprout (Japanese name "kaiware-daikon") exhibited the highest potency (1.8 times as l-ascorbic acid). Then, we investigated the constituents of the methanol extract of radish sprout and the contribution to the overall activity of each compound by examining their activity. As the result, several kinds of sinapinic acid esters and flavonoids were isolated with high radical scavenging potency, which must contribute substantially to the activity.
We investigated proliferation and apoptosis induction in Jurkat T-leukemia cells by the new isothiocyanate 4-(methylthio)butylisothiocyanate (MTBITC). To help elucidate whether the effects of MTBITC are specific for cancer cells, we tested MTBITC on freshly isolated, non-transformed human peripheral T lymphocytes. The effects of MTBITC are leukemic-cell-specific and consist of derangements in a critical point of cell-cycle control (G2/M transition). In fact, an increase in the proportion of G2 cells (from about 18% to 50%) was apparent following 24 h of treatment, associated with a decrease in the protein expression of cyclin B1. The expression of cyclin-dependent kinase (CDK) 1 was more mildly attenuated by MTBITC. Our results demonstrated that high concentrations of MTBITC can also induce apoptosis, through an increase of p53 and bax, but not bcl-2, protein expression. No effects of MTBITC were demonstrated on non-transformed T lymphocytes. Taking into account its in vitro antineoplastic activity and selectivity toward leukemia cells, MTBITC can be viewed as a conceptually promising agent in cancer therapy.
Chemoprevention is a cancer preventive strategy to inhibit, delay or reverse carcinogenesis using naturally occurring or synthetic chemical agents. Numerous epidemiological studies as well as experimental animal studies clearly demonstrate that high intake of cruciferous vegetables protects against tumorigenesis. Thus, cruciferous vegetables have been of great interest for potential use in the chemoprevention of cancer. Cruciferous vegetables are rich source of glucosinolates, which are degraded into isothiocyanates by enzymatic action of plant-specific myrosinase or intestinal flora in the body. It appears that significant portion of the chemopreventive effects of isothiocyanates may be associated with the inhibition of the metabolic activation of carcinogens by cytochrome P450s (Phase I), coupled with strong induction of Phase II detoxifying and cellular defensive enzymes. Inductions of Phase II cellular enzymes are largely mediated by the antioxidant responsive element (ARE), which is regulated by the transcriptional factor, Nrf2. Additional potent regulatory mechanisms of Nrf2 include the different signaling kinase pathways (MAPK, PI3K, PKC and PERK) as well as other non-kinase dependent mechanisms. Moreover, apoptosis and cell cycle perturbations appear to be yet another potential chemopreventive mechanisms elicited by isothiocyanates, especially with respect to the effects on pre-initiated or initiated tumor cells. Finally, modulation of other critical signaling mediators, including the NF-kappaB and AP-1 by a wide array of chemopreventive agents including isothiocyanates may also contribute to the overall chemopreventive mechanisms.
This review summarizes the multifaceted aspects of antioxidants and the basic kinetic models of inhibited autoxidation and analyzes the chemical principles of antioxidant capacity assays. Depending upon the reactions involved, these assays can roughly be classified into two types: assays based on hydrogen atom transfer (HAT) reactions and assays based on electron transfer (ET). The majority of HAT-based assays apply a competitive reaction scheme, in which antioxidant and substrate compete for thermally generated peroxyl radicals through the decomposition of azo compounds. These assays include inhibition of induced low-density lipoprotein autoxidation, oxygen radical absorbance capacity (ORAC), total radical trapping antioxidant parameter (TRAP), and crocin bleaching assays. ET-based assays measure the capacity of an antioxidant in the reduction of an oxidant, which changes color when reduced. The degree of color change is correlated with the sample's antioxidant concentrations. ET-based assays include the total phenols assay by Folin-Ciocalteu reagent (FCR), Trolox equivalence antioxidant capacity (TEAC), ferric ion reducing antioxidant power (FRAP), "total antioxidant potential" assay using a Cu(II) complex as an oxidant, and DPPH. In addition, other assays intended to measure a sample's scavenging capacity of biologically relevant oxidants such as singlet oxygen, superoxide anion, peroxynitrite, and hydroxyl radical are also summarized. On the basis of this analysis, it is suggested that the total phenols assay by FCR be used to quantify an antioxidant's reducing capacity and the ORAC assay to quantify peroxyl radical scavenging capacity. To comprehensively study different aspects of antioxidants, validated and specific assays are needed in addition to these two commonly accepted assays.
Rocket (Eruca sativa Mill. or Eruca vesicaria L.) is widely distributed all over the world and is usually consumed fresh (leafs or sprouts) for its typical spicy taste. Nevertheless, it is mentioned in traditional pharmacopoeia and ancient literature for several therapeutic properties, and it does contain a number of health promoting agents including carotenoids, vitamin C, fibers, flavonoids, and glucosinolates (GLs). The latter phytochemicals have recently gained attention as being the precursors of isothiocyanates (ITCs), which are released by myrosinase hydrolysis during cutting, chewing, or processing of the vegetable. ITCs are recognized as potent inducers of phase II enzymes (e.g., glutathione transferases, NAD(P)H:quinone reductase, epoxide hydrolase, etc.), which are important in the detoxification of electrophiles and protection against oxidative stress. The major GL found in rocket seeds is glucoerucin, GER (108 +/- 5 micromol g(-)(1) d.w.) that represents 95% of total GLs. The content is largely conserved in sprouts (79% of total GLs), and GER is still present to some extent in adult leaves. Unlike other GLs (e.g., glucoraphanin, the bio-precursor of sulforaphane), GER possesses good direct as well as indirect antioxidant activity. GER (and its metabolite erucin, ERN) effectively decomposes hydrogen peroxide and alkyl hydroperoxides with second-order rate constants of k(2) = 6.9 +/- 0.1 x 10(-)(2) M(-)(1) s(-)(1) and 4.5 +/- 0.2 x 10(-)(3) M(-)(1) s(-) , respectively, in water at 37 degrees C, thereby acting as a peroxide-scavenging preventive antioxidant. Interestingly, upon removal of H(2)O(2) or hydroperoxides, ERN is converted into sulforaphane, the most effective inducer of phase II enzymes among ITCs. On the other hand, ERN (and conceivably GER), like other ITCs, does not possess any chain-breaking antioxidant activity, being unable to protect styrene from its thermally (37 degrees C) initiated autoxidation in the presence of AMVN. The mechanism and relevance of the antioxidant activity of GER and ERN are discussed.
Forget drugs carefully designed to hit one particular molecule - a better way of treating complex diseases such as cancer may be to aim for several targets at once, says Simon Frantz.
The most promising among glucosinolates (GLs) are those bearing in their aglycon an extra sulfur function, such as glucoraphasatin (4-methylthio-3-butenyl GL; GRH) and glucoraphenin (4-methylsulfinyl-3-butenyl GL; GRE). The GRE/GRH redox couple is typically met among secondary metabolites of Raphanus sativus L. and, whereas GRE prevails in seeds, GRH is the major GL in full-grown roots. During the 10 days of sprouting of R. sativus seeds, the GRE and GRH contents were determined according to the Eurpean Union official method (ISO 9167-1). In comparison to the seeds, the GRE content in sprouts decreased from about 90 to about 12 micromol g(-1) of dry weight (dw), whereas a 25-fold increase--from about 3 to 76 micromol g(-1) of dw--of the GRH content was measured. An efficient pure GRH gram-scale production process from R. sativus (kaiware daikon) sprouts resulted in significant yield improvement of up to 2.2% (dw basis). The reaction of GRH with both H2O2 and ABTS*+ radical cation was investigated. Whereas H2O2 oxidation of GRH readily resulted in complete transformation into GRE, ABTS*+ caused complete decay of the GL. Even though not directly related to its radical scavenging activity, the assessed reducing capacity of GRH suggests that R. sativus sprouts might possess potential for health benefits.
Epidemiological and animal studies linking high fruit and vegetable consumption to lower cancer risk have strengthened the belief that long-term administration of isolated naturally occurring dietary constituents could reduce the risk of cancer. In recent years, metabolites derived from phytoalexins, such as glucoraphanin found in broccoli and other cruciferous vegetables (Brassicaceae), have gained much attention as potential cancer chemopreventive agents. The protective effect of these micronutrients is assumed to be due to the inhibition of Phase-I carcinogen-bioactivating enzymes and/or induction of Phase-II detoxifying enzymes, an assumption that still remains uncertain. The protective effect of glucoraphanin is thought to be due to sulforaphane, an isothiocyanate metabolite produced from glucoraphanin by myrosinase. Here we show, in rat liver, that while glucoraphanin slightly induces Phase-II enzymes, it powerfully boosts Phase-I enzymes, including activators of polycyclic aromatic hydrocarbons (PAHs), nitrosamines and olefins. Induction of the cytochrome P450 (CYP) isoforms CYP1A1/2, CYP3A1/2 and CYP2E1 was confirmed by Western immunoblotting. CYP induction was paralleled by an increase in the corresponding mRNA levels. Concomitant with this Phase-I induction, we also found that glucoraphanin generated large amount of various reactive radical species, as determined by electron paramagnetic resonance (EPR) spectrometry coupled to a radical-probe technique. This suggests that long-term uncontrolled administration of glucoraphanin could actually pose a potential health hazard.
The antioxidant activity of several phenolic acids and esters has been investigated both in organic solutions and in large unilamellar phosphatidylcholine vesicles. In solution these compounds behaved as good antioxidants, with the exception of protocatechuic acid, due to the presence of the catechol moiety. Because their antioxidant activity followed an inverse dependence on the magnitude of their O-H bond dissociation enthalpies (BDE), the key mechanism of the chain-breaking action was attributed to hydrogen atom transfer (HAT) from the phenolic OH to peroxyl radicals. In unilamellar vesicles the antioxidant activity was strongly dependent on the pH of the buffer solution. In acid media (pH 4) all of the examined phenolic acids or esters behaved as weak inhibitors of peroxidation, whereas, with increasing pH, their antioxidant activity increased substantially, becoming comparable to or even better than that of Trolox. At pH 8 they also gave rise to lag phases 2-3 times longer than that of Trolox. The increased activity being observed in proximity of the pK(a) value corresponding to the ionization of one of the catecholic hydroxyl groups, this effect has been attributed to the high antioxidant activity of the phenolate anion.
Brassica vegetables and glucosinolates contained therein are supposed to reduce the risk of cancer and to possess health-promoting properties. The benefits of a Brassica-based diet may be particularly expressed by eating sprouts, in which the glucosinolate content is higher than in mature vegetables. With this in mind, a first objective of this study was to evaluate the antioxidant properties of radish (Raphanus sativus L.) sprouts (Kaiware Daikon) extract (KDE), in which the glucosinolate glucoraphasatin (GRH), showing some antioxidant activity, is present at 10.5% w/w. The contribution of GRH to KDE's antioxidant activity was considered in two chemical assays (Trolox equivalent antioxidant capacity and Briggs-Rauscher methods). The total phenol assay by Folin-Ciocalteu reagent was performed to quantify the reducing capacity of KDE. Finally, on the basis of the putative choleretic properties of antioxidant plant extracts, the effect on the bile flow of KDE administration was investigated in an animal experimental model. The findings showed that KDE has antioxidant properties and significantly induced bile flow in rats administered 1.5 g/kg of body weight for 4 consecutive days.
Brassica vegetables are an important dietary source of glucosinolates (GLs), whose breakdown products exhibit anticancer activity. The protective properties of Brassicaceae are believed to be due to the inhibition of Phase-I or induction of Phase-II xenobiotic metabolizing enzymes (XMEs), thus enhancing carcinogen clearance. To study whether GLs affect XMEs and the role of their chemical structure, we focused on two alkylthio GLs differing in the oxidation degree of the side chain sulfur. Male Sprague-Dawley rats were supplemented (per oral somministration by gavage) with either glucoraphasatin (4-methylthio-3-butenyl GL; GRH) or glucoraphenin (4-methylsulfinyl-3-butenyl GL; GRE), at 24 or 120 mg/kg body weight in a single or repeated fashion (daily for four consecutive days), and hepatic microsomes were prepared for XME analyses. Both GLs were able to induce XMEs, showing different induction profiles. While the inductive effect was stronger after multiple administration of the higher GRH dosage, the single lower GRE dose was the most effective in boosting cytochrome P-450 (CYP)-associated monooxygenases and the postoxidative metabolism. CYP3A1/2 were the most affected isoforms by GRH treatment, whereas GRE induced mainly CYP1A2 supported oxidase. Glutathione S-transferase increased up to approximately 3.2-fold after a single (lower) GRE dose and UDP-glucuronosyl transferase up to approximately 2-fold after four consecutive (higher) GRH doses. In conclusion, the induction profile of these GLs we found is not in line with the chemopreventive hypothesis. Furthermore, the oxidation degree of the side chain sulfur of GLs seems to exert a crucial role on XME modulation.
There is high current interest in the chemopreventive potential of Brassica vegetables (cruciferae), particularly due to their content in glucosinolates (GL), which upon myrosinase hydrolysis release the corresponding isythiocyanates (ITC). Some ITCs, such as sulforaphane (SFN) from broccoli ( Brassica oleacea italica), have been found to possess anticancer activity through induction of apoptosis in selected cell lines, as well as indirect antioxidant activity through induction of phase II detoxifying enzymes. Japanese daikon ( Raphanus sativus L.) is possibly the vegetable with the highest per capita consumption within the Brassicaceae family. Thanks to a recently improved gram scale production process, it was possible to prepare sufficient amounts of the GL glucoraphasatin (GRH) as well as the corresponding ITC 4-methylthio-3-butenyl isothiocyanate (GRH-ITC) from its sprouts. This paper reports a study on the cytotoxic and apoptotic activities of GRH-ITC compared with the oxidized counterpart 4-methylsulfinyl-3-butenyl isothiocyanate (GRE-ITC) on three human colon carcinoma cell lines (LoVo, HCT-116, and HT-29) together with a detailed kinetic investigation of the direct antioxidant/radical scavenging ability of GRH and GRH-ITC. Both GRH-ITC and GRE-ITC reduced cell proliferation in a dose-dependent manner and induced apoptosis in the three cancer cell lines. The compounds significantly ( p < 0.05) increased Bax and decreased Bcl2 protein expression, as well as producing caspase-9 and PARP-1 cleavage after 3 days of exposure in the three cancer cell lines. GRH-ITC treatment was shown to have no toxicity with regard to normal human lymphocytes (-15 +/- 5%) in comparison with SFN (complete growth inhibition). GRH and GRH-ITC were able to quench the 2,2-diphenyl-1-picrylhydrazyl radical, with second-order rate constants of 14.0 +/- 2.8 and 43.1 +/- 9.5 M(-1) s(-1), respectively (at 298 K in methanol), whereas the corresponding value measured here for the reference antioxidant alpha-tocopherol was 425 +/- 40 M (-1) s (-1). GRH reacted with H2O2 and tert-butyl hydroperoxide in water (pH 7.4) at 37 degrees C, with rate constants of 1.9 +/- 0.3 x 10(-2) and 9.5 +/- 0.3 x 10(-4) M(-1) s (-1) (paralleling recently developed synthetic antioxidants) being quantitatively (>97%) converted to GRE. It is demonstrated that GRH-ITC has interesting antioxidant/radical scavenging properties, associated with a selective cytotoxic/apoptotic activity toward three human colon carcinoma cell lines, and very limited toxicity on normal human T-lymphocytes.
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Revised manuscript received Financial support from the University of Bologna (Italy) and MIUR (Rome, Italy), as well as from the Research Centre for Industrial Crops (Bologna, Italy) and Agricultural Research Council (Rome, Italy), is gratefully acknowledged
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Received for review April 10, 2008. Revised manuscript received May 28, 2008. Accepted June 19, 2008. Financial support from the University of Bologna (Italy) and MIUR (Rome, Italy), as well as from the Research Centre for Industrial Crops (Bologna, Italy) and Agricultural Research Council (Rome, Italy), is gratefully acknowledged. JF8011213 7830 J. Agric. Food Chem., Vol. 56, No. 17, 2008 Barillari et al.
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Revised manuscript received Financial support from the University of
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of Bologna (Italy) and MIUR (Rome, Italy), as well as from the Research
Centre for Industrial Crops (Bologna, Italy) and Agricultural Research
Council (Rome, Italy), is gratefully acknowledged.
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