JP2006109754A - Food inducing expression of thioredoxin - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide food inducing expression of thioredoxin comprising safe drink and food inducing expression of thioredoxin, specifically, softening influence caused by oxidation stress. <P>SOLUTION: The drink and food contains sulforaphane comprising extract from crucifers. The extract contains 0.01-50 mg/g of the sulforaphane. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a food that induces the expression of thioredoxin including an extract or extract from a cruciferous plant.

  Thioredoxin is a molecule that performs a reversible oxidation-reduction reaction (redox control) using two cysteine residues in its active site as a catalyst for thioredoxin reductase, which is an NADPH-dependent enzyme (for example, Non-Patent Document 1). Thioredoxin is known to have a role of protecting the cell from damage caused by oxidative stress (free radical) and tumor necrosis factor α (TNF-α) while maintaining an intracellular reducing environment (for example, non-patented) References 2 and 3).

  Furthermore, thioredoxin transgenic mice show resistance to prolongation of life span, ischemic injury, acute lung failure, diabetes and the like (for example, Non-Patent Documents 4 to 8). Since these pathological conditions are closely related to oxidative stress, thioredoxin is considered to exhibit an important protective action against oxidative stress.

It is considered that the influence of oxidative stress can be mitigated by inducing the expression of thioredoxin using such characteristics of thioredoxin.
Holmgren A.M. et al. , Methods Enzymol 1995; 252: 199-208. Nakamura H. et al. et al. , Immunol Lett 1994; 42: 75-80. Matsuda M.M. et al. , J Immunol 1991; 147: 3837-3841. Mitsui A. et al. , Antioxidid Redox Signal 2002; 4: 693-696. Takagi Y. et al. et al. Proc Natl Acad Sci USA 1999; 96: 4131-4136. Hoshino T. et al. , Am J Respir Crit Care Med 2003; 168: 1075-1083. Hotta M.M. et al. J Exp Med 1998; 188: 1455-1451. Yoon BI. et al. , Arch Environ Contam Toxicol 2001; 41: 232-236.

  The main object of the present invention is to provide a safe food and drink that can induce the expression of thioredoxin and can alleviate the effects of, for example, oxidative stress (free radicals).

  The present inventors have found that sulforaphane induces thioredoxin expression, and have completed the present invention.

The present invention provides foods and drinks including the following sulforaphanes.
Item 1. A food or drink comprising a sulforaphane-containing extract.
Item 2. The food or drink according to Item 1, wherein the extract contains 0.01 to 50 mg / g of sulforaphane.
Item 3. The food or drink according to Item 1, wherein sulforaphane is added in an amount of 10 to 2000 μg / 100 g of the content composition.
Item 4. The food or drink according to Item 1, wherein the sulforaphane is derived from a cruciferous plant.
Item 5. The food or drink according to Item 1, wherein the cruciferous plant is at least one selected from the group consisting of broccoli sprout, silkworm radish, and purple brussels sprouts.
Item 6. The food or drink according to Items 1 to 5, which is for induction of thioredoxin expression.
Item 7. Claim | item 1-5 food / beverage products which are an object for oxidative stress relaxation.
Item 8. The manufacturing method of the food / beverage products containing a sulforaphane containing extract characterized by adding 10-2000 micrograms / 100g of content compositions of sulforaphane in food / beverage products.

  Sulforaphane is included in plants of the Brassicaceae plant, such as Brassica, Eluca Iberis, Raphanus, etc., and has anticancer activity, antioxidant activity, etc. However, the thioredoxin expression-inducing action has not been known. Various useful actions such as improvement of liver dysfunction, improvement of oxidative stress disorder, and anti-inflammatory action can be expected based on the action of sulforaphane to induce thioredoxin expression.

  The plant species used for extracting sulforaphane in the present invention is not particularly limited, and examples thereof include cabbage, purple cabbage, broccoli, kale, rocket vegetables, cauliflower, Japanese radish, Chinese cabbage, turnip, komatsuna, and chingensai. And are preferably shoots of these plants. Among them, broccoli sprouts (broccoli sprout), radish sprouts (kaiwared radish) and purple cabbage sprouts are particularly preferable.

  The site for extracting sulforaphane in these plants is not particularly limited as long as the effect as sulforaphane is obtained, for example, the seeds of the plant, roots of radish, turnips, leaves of cabbage, kale, Chinese cabbage, broccoli, Examples include cauliflower and other florets.

  When sulforaphane is obtained from the above-mentioned plant shoots, the content of sulforaphane in the shoots varies greatly depending on the number of days after germination. For example, in the case of broccoli sprout, about 1 to 5 days after germination, preferably 1 to 1 after germination. It is desirable to extract from shoots of about 3 days.

  The sulforaphane used in the present invention can be obtained from the above-mentioned plants by a method usually used by those skilled in the art. The extraction method is not particularly limited, but for example, pulverizing a raw material containing sulforaphane such as broccoli, freeze-drying if necessary, impregnating or extracting with water or a water-containing solvent, filtering, For example, a method of concentrating is used.

Although it does not specifically limit as a solvent used for extraction, For example, primary alcohol, such as water, ethanol, methanol, n-propanol, iso-propanol, n-butanol, iso-butanol, tert-butanol, ethyl acetate, etc. Examples include lower alkyl esters, hydrocarbons such as benzene and hexane, conventionally known solvents such as acetone and methylene chloride, or mixed solutions thereof. Among them, preferred solvents are water, ethanol or a mixed solution of ethanol and water. Alternatively, extraction may be performed with a supercritical solvent such as supercritical CO ₂ .

When performing extraction using these methods, the number of extractions may be single, or multiple times in order to increase the yield.

  When extracting, in order to hydrolyze glucosinolate with the enzyme contained in a plant and to obtain sulforaphane efficiently, you may self-digest after grind | pulverizing a raw material. The self-digestion may be carried out under conditions usually used by those skilled in the art, but it is desirable to carry out, for example, at about 10-50 ° C. for about 15-60 minutes.

  The extract thus obtained can be used as it is as a sulforaphane extract, and purification operations such as deodorization and decolorization can be added within a range not losing the effects of the present invention.

  As another method, for example, a separation / purification operation using a synthetic adsorbent may be performed. Separation / purification operation method in the case of using a synthetic adsorbent is not particularly limited and may be a conventionally known method. For example, after removing a residue by pulverizing a raw material containing sulforaphane, a synthetic adsorbent was filled Examples of the method include elution with water, ethanol (or a mixed solution thereof) and the like through a column or the like. Examples of the synthetic adsorbent include aromatic (cross-linked styrene) synthetic adsorbents, substituted aromatic synthetic adsorbents, and acrylic synthetic adsorbents.

  Moreover, after extracting the pulverized plant with suitable solvents, such as ethanol, you may use the method of filtering and concentrating. As a simpler method, there is a method in which the raw material plant is made into juice, and then the plant residue is removed and concentrated.

  In place of the concentration step, if necessary, water-soluble dietary fibers such as pectin and dextrin may be added to the filtrate as an excipient, dried by a known method such as a spray drying method, and used as a powder. . Further, the powdered extract can be used by re-dissolving in a solvent such as water, ethanol, propylene glycol, 1,3-butylene glycol, glycerin and the like.

  In the present invention, it is expected to improve the stability of sulforaphane by using the extract or the extract after drying.

  The sulforaphane-containing extract obtained by the above method desirably contains sulforaphane in an amount of about 0.001 to 200 mg / g, preferably about 0.005 to 80 mg / g, more preferably about 0.01 to 50 mg / g.

  If the amount of thioredoxin whose expression is induced by sulforaphane is, for example, 10 to 500 μg / 100 g of sulforaphane, the amount of thioredoxin contained in the cell is about 1.3 to 2 times the constant amount, preferably 1.5 times. About 3 times the amount, more preferably about 2 to 6 times the amount.

  Examples of the food and drink of the present invention include health foods, nutritional supplements (balance nutritional foods, supplements, etc.), functional nutritional foods, foods for specified health use, and the like. The method for producing these foods is not particularly limited as long as the desired effect of sulforaphane can be obtained, and may be a method used by those skilled in the art for each application.

Examples of foods and beverages include gums, candy, gummi, tablet confectionery, cookies, cakes, chocolate, ice cream, jelly, mousse, pudding, biscuits, corn flakes, chewable tablets, wafers, rice crackers, etc .; carbonated drinks, refreshing Beverages such as beverages, milk beverages, coffee beverages, tea beverages, fruit juice beverages, nutritional beverages, alcoholic beverages and mineral water; powdered beverages such as powdered juice and powdered soup; balanced nutritional foods; powders, capsules, tablets, etc. Examples of supplements include: seasonings such as dressings and sauces; breads; noodles; kneaded products such as kamaboko; and sprinkles.

  Although the amount of sulforaphane contained in the food or drink of the present invention is not particularly limited as long as it is an amount necessary for inducing thioredoxin expression, it is usually about 1 to 5000 μg / 100 g, preferably about 5 to 3000 μg / 100 g. More preferably, it is about 10 to 2000 μg / 100 g, and the extent that does not impair the flavor of the food or drink to be blended is desirable.

  When the food of the present invention is a cookie-like food such as cookies or biscuits, it is preferable to contain about 10 to 2000 μg / 100 g of sulforaphane.

  For example, in the case of beverages, it is preferable to contain about 10 to 2000 μg / 100 g of sulforaphane.

  In the case of a tablet tableted after granulating an extract extracted from a plant containing sulforaphane, it is preferable to contain about 2-140 μg / g of sulforaphane.

  In the case of mousse-like foods such as jelly, mousse and pudding, it is preferable to contain about 10 to 500 μg / 100 g of sulforaphane.

  In the case of dressing, it is preferable to contain about 10 to 2000 μg / 100 g of sulforaphane.

  In the process of manufacturing these foods, in addition to the sulforaphane-containing extract, raw materials containing sulforaphane such as broccoli (broccoli sprout), radish (kaiwaredaikon), murasaki cabbage (sprouts of murasaki cabbage) are added to make food You can also.

  The food / beverage products in this invention can be prepared according to the conventional method in the said food / beverage products form by mixing the active ingredient of this invention with the support | carrier containing a raw material or an additive. Examples of additives include sweeteners, colorants, antioxidants, vitamins, and fragrances.

  The intake of the above food and drink is appropriately selected depending on its usage, the age, sex, body weight, health condition, other conditions, degree of symptoms, etc. of the person who takes it, but usually the amount of sulforaphane as an active ingredient is an adult It is preferable that the amount per person per day is about 0.001 to 1000 μg / kg, preferably about 0.005 to 200 μg / kg, more preferably about 0.01 to 60 μg / kg. Other components can be appropriately selected based on the amount of the sulforaphane. Further, a raw material paste containing sulforaphane may be used as long as the desired effect can be obtained.

  The food / beverage products of this invention can also be manufactured as animal feed. That is, it can also be used as pet food or livestock feed.

  These may be in any form as long as they are used in pet food or feed, as in the case of the above-mentioned food, and are usually combined with additives or food materials usually contained in pet food or feed. It can be prepared in various forms according to the method. The dose can be appropriately selected with reference to the food.

  Since sulforaphane contained in the food and drink of the present invention induces the expression of thioredoxin, it can be used for the purpose of mitigating oxidative stress (erasing free radicals).

  Through the expression of thioredoxin, for example, improvement of liver disorders including hangover, improvement of menopause, reduction of blood cholesterol, reduction of blood neutral lipid, reduction of blood glucose level, reduction of blood pressure, prevention of arteriosclerosis, aging Prevention of such is expected.

  For example, arteriosclerosis is caused by accumulation of lipid peroxide, which is a kind of free radical. By ingesting the food or drink of the present invention, expression of TRX is induced, and it is expected that arteriosclerosis is improved by the free radical scavenging action of TRX itself or TRX-dependent peroxidase.

  In addition, it is considered that the excessive increase in blood glucose level is caused by the involvement of injury due to oxidative stress in insulin-producing cells of the pancreatic islet. By ingesting the food / beverage product of the present invention, expression of TRX is induced, and the blood glucose level is expected to decrease by inhibiting the damage of insulin-producing cells by the free radical scavenging action of TRX itself or TRX-dependent peroxidase. Is done.

  Moreover, the food / beverage products of this invention can also be used for the purpose of nutrition tonic, fatigue recovery, etc.

  According to the present invention, by inducing the expression of thioredoxin, for example, it is possible to provide a food and drink containing sulforaphane that is effective in improving liver disorders including hangover, improving menopause, and nutritional tonic. It is. Moreover, since sulforaphane used for this invention is derived from an edible plant, it can provide highly safe food and drink.

  EXAMPLES Hereinafter, although an Example and a test example are given and this invention is demonstrated in more detail, it cannot be overemphasized that this invention is not limited by these Examples and a test example.

Extract (1) Washed broccoli sprouts were pulverized with COMITOLOL (manufactured by Archer Japan), added with an equal amount of water, self-digested at 20 ° C for 1 hour, filtered through diatomaceous earth to remove the residue, and reduced pressure And concentrated by evaporation. The amount of sulforaphane contained in the extract thus obtained was 0.1 to 1 mg.
(2) Washed broccoli shoots with comitrol, filtered through diatomaceous earth to remove residues, and a mixture of ethanol and water using a crosslinked styrene synthetic adsorbent (DIAION HP-20: manufactured by Mitsubishi Chemical Corporation) ( Ethanol: water = 70: 30). The amount of sulforaphane contained in the extract thus obtained was 1.0 to 12 mg / g.
(3) Washed broccoli shoots with comitrol, filtered through diatomaceous earth to remove residues, and a mixture of ethanol and water using a cross-linked styrene synthetic adsorbent (DIAION HP-20: manufactured by Mitsubishi Chemical Corporation) ( (Ethanol: water = 50: 50), followed by vacuum drying. The amount of sulforaphane contained in the extract thus obtained was 0.5 to 10 mg / g.

Soft drink (1) (i) Broccoli and carrot were crushed with commitolol.

      (ii) The apple was crushed with a hammer crusher and then squeezed with a squeezer.

      (iii) (i) (ii) was passed through a strainer to remove residues such as leather.

(iv) Broccoli juice 100kg, carrot juice 50kg, apple 100kg, water 300k
g, after mixing 10 kg of lemon juice and 20 kg of fructose glucose solution, (2
10 kg of the extract prepared in (1) was added and mixed uniformly.

(v) Sterilized by plate type sterilization at 95 ° C. for 60 seconds, cooled, and then filled with 180 ml.
(2) 100 kg of the extract prepared in (1) of Example 1, 500 kg of water, 5 kg of citric acid, 50 kg of oligosaccharide, vitamin C, B1, B2, and perfume are mixed in appropriate amounts, passed through a strainer, and then plate-type sterilized. After sterilization at 98 ° C. for 30 seconds and cooling, 100 ml was filled into a container.

Dressing (i) After crushing 20 kg of celery, 50 kg of onion and 5 kg of garlic, 20 kg of the extract prepared in (1) of Example 1 was added to 100 kg of vinegar, 10 kg of lemon juice, 10 kg of salt, herbs and spices and mixed. .
(Ii) 55 ml of (i) was filled into a container together with 50 ml of vegetable oil.

The extract prepared in (1) of tablet Example 1 was granulated with a spray granulation dryer, and then 15 kg of maltodextrin was added to 35 kg of the granular extract and molded with a rotary tablet press.

mousse
(i) Milk (50 kg) and fresh cream (25 kg) were bubbled with a stirrer.
(ii) 25 kg of water was added to 10 kg of premixed sugar and 2.5 kg of agar and dissolved by heating. Then, 25 kg of broccoli paste and an appropriate amount of flavor were added and mixed.
(iii) After mixing (i) and (ii), 80 g was filled in the container.

Test example 1

  In order to examine whether the activation of thioredoxin gene expression in K562 cells, a human erythroblast-like leukemia cell line, is altered by extracts from various vegetables, a vector containing the entire expression regulatory region of the thioredoxin gene is used. The luciferase reporter assay was performed.

[Cultivated cells and culture systems]
K562 cells in RPMI 1640 (Life Technologies, Grand Island, NY) containing 5% CO ₂ in 10% FCS and supplemented with antibiotics (100 IU / ml penicillin, 100 μg / ml streptomycin) Incubated at 37 ° C. HepG2 cells, which are human cultured hepatoma cell lines, were cultured in DMEM, and RPE cells derived from human retinal pigment epithelial cells were cultured in F-2 (Sigma).

  Cells used throughout this test example were maintained under the culture conditions described above.

[Preparation of vegetable extract]
The vegetable extract was prepared with reference to the literature (Zhang Y. et al., Proc Natl Acad Sci USA 1992; 89: 2399-2403).

  Two times the amount of water was added to fresh vegetables, pulverized (after 7000 rpm for 5 minutes, then 9000 rpm for 3 minutes), self-digested at 37 ° C. for 3 hours, and lyophilized. 4 g of the lyophilized product was dissolved in 140 ml of acetonitrile, extracted by shaking at 4 ° C. for 6 hours. The extract was filtered through a membrane filter and dried at 30 ° C. using an evaporator. The dried product was dissolved in 1 ml of ethanol and used for later analysis.

[Plasmid]
The pTrxCAT plasmid was transformed into Taniguchi Y. et al. , Nucleic Acids Res 1996; 24: 2746-2752. It was produced as described in 1. That is, the HindIII-BamHI insert of the pTrxCAT vector was subcloned into pBluescriptIIKS (+) (pTRXblue vectors).

  The pTRX-Luc vector is Kim YC. et al. J Biol Chem 2001; 276, 18399-18406. And Kim YC. et al. , Oncogene 2003; 22, 1860-1865. As described. That is, a pTRX (-1148) -Luc vector was prepared by binding the KpnI / BamHI fragment of the pTRXblue vector to the KpnI / BglII site of the pGL3 basic vector (Promega, WI).

Thermo Sequenase II dye terminator cycle sequencing kit (Amersham)
Pharmacia) was used to confirm the nucleotide sequence by direct nucleotide sequencing.

[Transfection and luciferase assay]
A luciferase reporter expression vector (pTRX (-1148) -Luc) or a pGL3 basic vector was introduced into K562 cells using DMRIE-C (GIBCO) reagent according to the product description. For standardization of gene transfer efficiency, pRL-TK (Promega) was cotransfected to measure Renilla luciferase gene expression.

  After culturing the cells for 4 hours, vegetable extract (final concentration 1%) or ethanol (final concentration 1%) was added and allowed to react for 16 hours. 24 hours after gene transfer, luciferase gene expression was analyzed using an assay kit (Promega). The analysis was performed by performing independent gene transfer for each inducer using two separate wells, collecting cells separately from each well, and separately measuring luciferase activity.

  As shown in FIG. 1A, broccoli sprout extracts of two different varieties (Broccoli Sprout A or Broccoli Sprout B) activated the thioredoxin reporter gene 13.5 times and 4.1 times, respectively. On the other hand, the same test was performed on the pGL3-basic vector, but there was no change in activation. In contrast, Kale, Cabbage and Broccoli flower bud extracts activated the thioredoxin reporter gene compared to the control (Ethanol) but were not significantly different (FIG. 1B).

Test example 2

Broccoli is known to contain sulforaphane as an important component (Fahey JW. Et al., Proc Natl Acad S
ci USA 1997; 94: 10367-10372. ), Whether the activation of thioredoxin gene expression was altered by sulforaphane.

[Transfection and luciferase assay]
After pTRX (-1148) -Luc or pGL3 basic vector was transfected into K562 cells and cultured for 4 hours, DMSO (final concentration 1%), or sulforaphane 1 μM (SF1) or 10 μM (SF10) was applied to each cell. In addition, the reaction was allowed to proceed for 16 hours. 24 hours after transfection, luciferase assay was performed under the same conditions as in Test Example 1. For normalization of transfection efficiency, pRL-TK vector (Promega) was co-transfected and Renilla luciferase gene expression was measured.

  HepG2 cells were also transfected with pTRX (-1148) -Luc or pGL3 basic vector and cultured for 4 hours. Thereafter, DMSO final concentration of 1% and sulforaphane 10 μM (SF) were added and reacted for 16 hours. 24 hours after transfection, luciferase assay was performed in the same manner as in Test Example 1.

  For introduction of the luciferase reporter expression vector (pTRX (-1148) -Luc) into HepG2 cells, FuGENE6 transfection reagent (Roche Applied Science) was used.

  As shown in FIGS. 2A and 2B, sulforaphane induced activation of a reporter gene pTRX (-1148) -Luc containing the entire thioredoxin promoter region in K562 cells (FIG. 2A) and HepG2 cells (FIG. 2B), but pGL3-basic. There was no effect on the activation of the vector. In K562 cells, sulforaphane 10 μM (SF10) showed a significant enhancement in thioredoxin expression.

[Quantitative RT-PCR]
To analyze the effect of thioredoxin on mRNA expression, quantitative Real Time RT-PCR was performed.

  K562 cells were treated with 0.1% DMSO, sulforaphane (SF) 1 μM or 10 μM. Thereafter, after 24 or 48 hours, the cells were collected and washed with PBS. Total RNA was isolated by RNeasy Mini Kit (QIAGEN) according to the product description. Human thioredoxin forward and reverse primers and Taqman probe were designed according to the product description.

  Quantitative RT-PCR was performed on an ABI PRISM 7000 using TaqMan One-Step RT-PCR Master Mix Reagent (Applied Biosystems) in 96-well plates. The tested reaction solution contains 50 ng of total RNA in a final volume of 50 μl.

  The conditions for RT-PCR are as follows. The RT step was performed at 48 ° C. for 30 minutes, the PCR step at 95 ° C. for 15 seconds and 60 ° C. for 1 minute for 45 cycles. As an internal standard, 18S rRNA was amplified using TaqMan Ribosomal Control Reagent (Applied Biosystems) to standardize the quantification of the thioredoxin gene.

  In K562 cells, treatment with sulforaphane (SF) 10 μM resulted in significant induction of thioredoxin mRNA levels at 24 and 48 hours (FIG. 3).

[Western blot analysis]
The effect of sulforaphane on the expression level of thioredoxin protein was examined.

K562 cells were treated with 1% DMSO, Sulforaphan 5 μM
Alternatively, cells were collected by treatment with 10 μM sulforaphane for 48 hours. The collected cells were washed twice with chilled PBS and then lysed on ice for 30 minutes (10 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1% NP-40, 1 mM EDTA, 0.1 mM PMSF, 8 μg / ml aprotinin, 2 μg / ml leupeptin). The extract was clarified by centrifugation.

The K562 cell lysate (13.5 μg) was heat-treated at 95 ° C. for 5 minutes, and then separated by 15% SDS polyacrylamide gel electrophoresis. The separated protein was transferred to a PVDF membrane (polyvinylden difluoride) (Millipore Co., Bedford, MA). The PVDF membrane was treated overnight in 10% skim milk dissolved in T-PBS (PBS containing 0.05% Tween 20), incubated with anti-human thioredoxin monoclonal antibody (Redox Bioscience, Inc.) for 1 hour, Thereafter, the reaction was carried out for 1 hour with anti-mouse IgG conjugated with peroxidase (1: 5000 dilution) (Amershan Pharmacia Biotech). Color development is ECL Western Blot detection kit (Amershan Pharmacia)
Biotech) was used (FIG. 4A).

  The expression level of thioredoxin protein in RPE cells was also observed using the same method. The RPE cells were treated with 1% DMSO, 0.1 μM sulforaphane, 1 μM sulforaphane or 10 μM sulforaphane for 48 hours, the cells were collected, and an RPE cell lysate was prepared in the same manner as described above. Next, Western blot analysis was performed in the same manner as described above using 5 μg of the obtained RPE cell lysate (FIG. 4B).

  In K562 cells and RPE cells treated with 10 μM sulforaphane for 48 hours, it was confirmed that thioredoxin expression was significantly enhanced (FIGS. 4A and 4B).

Test example 3

Varieties (Brocoli sprout) in which the thioredoxin gene was strongly induced in Example 1
Using A), the effect of the concentration of broccoli extract on the activation of thioredoxin gene induction was examined.
[Preparation of broccoli sprout extract]
Broccoli sprout extract was prepared in the same manner as in Test Example 1.
[Transfection and luciferase assay]
After pTRX (-1148) -Luc or pGL3 basic vector was transfected into K562 cells and cultured for 4 hours, the final concentrations were 1%, 0.75%, 0.5% and 0.25%. Ethanol (control), broccoli sprout extract (Brocoli sprout), or sulforaphane (Sulfor
aphan) 10 μM was added to each cell and allowed to react for 16 hours. In this test example, when broccoli sprout extract was used, the amount of sulforaphane contained was 2.8 μM, 2.1 μM, 0.14 μM and 0.71 μM, respectively.

  24 hours after the transfection, a luciferase assay was performed under the same conditions as in Test Example 1. For normalization of transfection efficiency, pRL-TK vector (Promega) was co-transfected and Renilla luciferase gene expression was measured.

  As shown in FIG. 5, in K562 cells transfected with pTRX (-1148) -Luc, activation of the thioredoxin gene was observed in a concentration-dependent manner within a broccoli sprout extract concentration range of 1 to 0.25%. It was. On the other hand, there was no change in activation in the pGL3-basic vector.

The activation of the thioredoxin gene by two types of broccoli sprout extracts (Broccoli Sprout A or Broccoli Sprout B) of different varieties is shown. Activation of the thioredoxin gene by Kale, Broccoli and Cabbage extracts is shown. Figure 2 shows activation of the thioredoxin gene by sulforaphane in K562 cells. Figure 2 shows activation of the thioredoxin gene by sulforaphane in HepG2. The induction of thioredoxin mRNA expression by sulforaphane is shown. FIG. 6 shows induction of thioredoxin protein expression by sulforaphane. lane 1: shows the expression of thioredoxin protein in K562 cells treated with DMSO. lane 2: shows expression of thioredoxin protein in K562 cells treated with 5 μM sulforaphane for 48 hours. lane 3: shows expression of thioredoxin protein in K562 cells treated with 10 μM sulforaphane for 48 hours. FIG. 6 shows induction of thioredoxin protein expression by sulforaphane. lane 1: shows the expression of thioredoxin protein in RPE cells treated with DMSO. lane 2: shows thioredoxin protein expression in RPE cells treated with 0.1 μM sulforaphane for 48 hours. The expression of thioredoxin protein in RPE cells treated with lane 3: 1 μM sulforaphane for 48 hours is shown. lane 4: Expression of thioredoxin protein in RPE cells treated with 10 μM sulforaphane for 48 hours. Induction of thioredoxin expression in K562 cells incubated with 10 μM sulforaphane or 1%, 0.75%, 0.5% or 0.25% sulforaphane. The white bar in the figure indicates the control (ethanol).

Claims (8)

A food or drink comprising a sulforaphane-containing extract. The food or drink according to claim 1, wherein the extract contains sulforaphane of 0.01 to 50 mg / g. The food or drink according to claim 1, wherein 10 to 2000 µg / 100 g of the content composition is added to sulforaphane. The food or drink according to claim 1, wherein the sulforaphane is derived from a cruciferous plant. The food / beverage product according to claim 1, wherein the cruciferous plant is at least one selected from the group consisting of broccoli sprout, silkworm radish, and purple brussels sprouts. The food or drink according to claim 1, which is used for inducing thioredoxin expression. The food or drink according to claim 1, which is for reducing oxidative stress. The manufacturing method of the food / beverage products containing a sulforaphane containing extract characterized by adding 10-2000 micrograms / 100g of content compositions of sulforaphane in food / beverage products.

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