JP2002138046A - Nk cell activator - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an NK(natural killer) cell activator containing a cabbage fermentation extract as a main component. SOLUTION: This cabbage fermentation extract activates the NK cells of patients developing cancers of stomach, large intestine, lung or uterus, leukemia or brain tumor, and of aged persons as well as able-bodies persons, and also enhances immunity. Accordingly, the extract can become a support food for activating the natural killer cells.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a food that activates natural killer cells (hereinafter referred to as NK cells).

[0002]

2. Description of the Related Art NK cells are defined as cells that have cytotoxicity to target cells in the unsensitized state. These NK cells are cells that specifically act on their own cancer or allogeneic cancer, and are attracting attention as cells that are responsible for carcinogenesis or resistance to the growth of cancer cells.

[0003] Such NK cells are thought to produce many cytokines by various stimuli, thereby stimulating and controlling cell proliferation, and regulating immune responses by T cells or B cells. It is considered that the activation can enhance the resistance to carcinogenesis or cancer cell growth. And substances that promote NK cell activation are being searched for, but most of them are used as drugs, and if they can be ingested as foods, they can be easily taken in daily life to increase immunity. Is expected.

[0004]

Therefore, there is a need for a food that activates NK cells and enhances immunity.

[0005]

Means for Solving the Problems The present inventors examined foods that activate NK cells, and surprisingly found that the cabbage fermented extract has an NK cell activating effect, and completed the present invention. I let it.

[0006] The present invention relates to an NK cell activator comprising a cabbage fermented extract as a main component.

[0007] In a preferred embodiment, the cabbage fermented extract is in the form of a liquid, paste or powder.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION The NK cell activator of the present invention comprises a cabbage fermented extract as a main component. Examples of the fermented cabbage extract include liquid extracts obtained by fermenting cabbage, pastes thereof, powders obtained from liquid extracts or pastes, and the like.

The cabbage used in the present invention is not limited as long as it is cabbage used for food. The cabbage variety does not matter. So-called red cabbage is also used in the present invention. Cabbage may be used alone or in combination of two or more different kinds.

The cabbage is first cut to an appropriate size with a cutter, slicer or the like, and then pulverized with a mixer, a juicer, a blender, a mass colloider or the like to form a slurry. Due to the slurrying, the cabbage becomes a thick porridge (a suspension of liquid and solid particles). The slurry is preferably ground as small as possible so that the nutrient components can be recovered in the extract.

When fermenting cabbage, salt may or may not be added. The salt is added when the cabbage is pulverized relatively large, in order to make the osmotic pressure adjusting action and the bacteriostatic action of bacteria other than lactic acid bacteria effectively function. In the slurried state, the need to add salt is low.

The cabbage slurried may be fermented as it is, or may be fermented once after removing insolubles by filtration, centrifugation, etc. (hereinafter sometimes referred to as squeezed juice). After the cabbage or squeezed juice obtained by slurrying is subjected to heat treatment (including sterilization), lactic acid bacteria and, if necessary, yeast may be added to carry out fermentation.

Prior to starting fermentation, it is also preferable to lower the pH of the liquid to be fermented. Lowering the pH creates an environment in which lactic acid bacteria can preferentially grow. The pH may be adjusted so that the desired bacteria can easily grow. For example, in Lactobacillus plantarum, if fermentation is started after adjusting the pH to about 4.0, the fermentation can be performed in a short period of time. Can end.

Further, before the start of fermentation, lactic acid bacteria metabolizable sugar can be added. This addition of sugar is useful when fermenting cabbage with a low sugar content. Therefore, when cabbage containing a sugar content of 3% by weight or more is fermented, it is not necessary to add sugar. However, the addition of sugar may have the effect of promoting fermentation and adding sweetness to the beverage, and may be added. The sugar to be added may be any sugar as long as it is used for the growth and fermentation of lactic acid bacteria, such as sucrose, glucose, fructose, and maltose, but is not limited thereto. These sugars are preferably added so that the sugar content is approximately 3% by weight in combination with the sugar content of the cabbage, but may be higher.

At the start of fermentation, it is not necessary to particularly add lactic acid bacteria, yeast and the like. This is because lactic acid bacteria and yeasts originally attached to cabbage can proliferate. Lactic acid bacteria may be added at the start of fermentation. This lactic acid bacterium can function as a starter, leading to a reduction in fermentation time. When heat sterilization is applied to slurried cabbage or squeezed juice before fermentation, the addition of lactic acid bacteria is essential. Yeast may be added as needed.

As the lactic acid bacteria to be added, Leuconostoc mesenteroide (Leuconostoc mesenteroide)
s), Lactobacillus plantarum (Lactobacillus p
lantarum) or Lactobacillus brevis (Lactob)
acillus brevis) and the like are used alone or in combination. When used alone, it is preferable to use Lactobacillus plantarum, since fermented cabbage extract which can be easily fermented and has high nutrient content can be obtained from the viewpoint of acid resistance, growth temperature and growth rate.

The yeast to be added as required is added in consideration of flavor, aroma, and the like. As the yeast, yeast used for fermentation or production of foods, such as brewer's yeast (beer yeast, sake yeast, etc.) and baker's yeast, are preferable. Preferably Saccharomyces cerevisiae (Saccharomyces
cerevisiae), Saccharomyces carlsbergensis, and the like.

The fermentation is carried out at 20 to 50 ° C., preferably at 25 ° C.
Perform in the range of ４５45 ° C. In order to promote lactic acid fermentation and to improve the flavor of the obtained cabbage fermented extract, it is preferable to perform the anaerobic condition. Anaerobic conditions include:
Deaeration of slurried cabbage or squeezed juice, sealing of the fermenter, filling of the fermenter with a gas such as nitrogen gas or carbon dioxide gas, reduced pressure, or a combination thereof.

The fermentation time is preferably 1 to 30 days. The fermentation time may be determined in combination with the fermentation temperature. When the acidity of the fermentation liquor is used as an index, it is preferable to terminate the fermentation when the acid in the fermentation liquor reaches about 0.5 to 2%, more preferably 1.0 to 1.5%. If it exceeds 2%, the acidity becomes too strong.

After the above fermentation, fermentation (low-temperature fermentation) may be further performed at 5 to 20 ° C., preferably 5 to 15 ° C. for 1 to 30 days.

The fermentation can be stopped by adding sugar. Examples of such sugars include sugar alcohols (eg, sorbitol) that cannot be metabolized by lactic acid bacteria, and oligosaccharides (eg, maltooligosaccharides, chitooligosaccharides, fructooligosaccharides), and the like. Such an oligosaccharide can contribute to improving the function of the fermented liquid as a beverage.

After completion of the fermentation, the cabbage fermented extract is recovered from the fermented liquid. A known method for recovering the extract, for example,
Centrifugation, filtration, etc. may be applied.

If necessary, the extract may be subjected to a heat treatment or a sterilization treatment to obtain a state suitable for storage. In the case of sterilization, it is preferable to sterilize at as low temperature as possible in order to maintain various nutrients.

Further, the obtained fermented extract can be concentrated to obtain a paste of cabbage fermented extract. Production of the paste from the fermented extract is performed by a concentration method usually used by those skilled in the art. For the concentration, various methods such as heat concentration, vacuum (reduced pressure) concentration, and freeze concentration without heating are used. Considering the decomposition of the active ingredient, it is preferable to avoid heating as much as possible, and a method such as vacuum concentration is preferably used.

Further, the cabbage fermented extract can be dried and powdered to obtain a cabbage fermented extract powder. Various methods commonly used by those skilled in the art are applied for drying. Of these, vacuum drying, freeze drying, and spray drying are preferably used. When performing spray drying, it is performed by adding excipients such as dextrin, cyclodextrin, starch, and maltose as necessary.

[0026] The cabbage fermented extract (liquid, pasty and powder) can be sterilized by a method used by those skilled in the art such as air sterilization, high pressure sterilization, and heat sterilization.

The obtained cabbage fermented extract (liquid, paste and powder) can activate NK cells, and is therefore used as an NK cell activator. This NK
When a cell activator is added to food, it is used as an NK cell activation supplement that activates NK cells containing cabbage fermented extract as a main component.

As the NK cell activation supplement, liquid cabbage fermented extract may be used as it is or various seasonings, for example, sweeteners such as granulated sugar, honey and sorbitol, alcohol, citric acid, malic acid, tartaric acid, and the like. Spices,
Beverages adjusted to a desired taste by adding a dye or the like are included.

If the obtained fermented cabbage extract of the present invention is mixed with other fermented juices or vegetable juices, such as ginseng juice, or mixed cabbage juices such as ginseng, celery, and parsley, the nutritional value is further improved. Can be juice. The mixing ratio is arbitrary, but NK
In consideration of the cell activating effect, it is preferable that the ratio of the cabbage fermented extract is high.

Also, for example, jelly can be mixed with agar or the like, and can also be made into sherbet, frozen yogurt or ice cream.

Further, the obtained paste of cabbage fermented extract or cabbage fermented extract powder is combined with other food materials such as royal jelly, vitamins, minerals, chitin / chitosan, lecithin, if necessary. Further, it is formed as capsules such as hard capsules and soft capsules, tablets or pills, or in the form of powder, granules, tea, tea bags or candy. These, depending on their shape or preference,
It can be taken as is, or dissolved in water, hot water or milk, or leached out of the ingredients.

By ingesting such an NK cell activation supplement (food containing a cabbage fermented extract) in such a form, NK cell activation can be achieved, and thus the food obtained by the present invention is effective in preventing cancer. It is likely to contribute to prevention of progress.

The fact that the cabbage fermented extract (liquid, pasty and powder) of the present invention has an NK cell activating effect can be measured, for example, by the following method. First, lymphocytes are separated from blood collected from a healthy person and used as effector cells. A cabbage fermented extract diluted to a predetermined concentration with physiological saline is added to the effector cells, cultured for several days, mixed with cancer cells in which ⁵¹ Cr has been previously incorporated as target cells, and the target cells (cancer cells) The degree of NK cell activation is determined by measuring the amount of ⁵¹ Cr released into the culture supernatant caused by the disruption of NK cells. The measurement can be performed with higher accuracy by using physiological saline as a negative control and IL-2 as a positive control.

[0034]

EXAMPLES The present invention will be described below, but it goes without saying that the present invention is not limited to these examples.

Example 1 After removing the outer leaves of the cabbage, the outer surface was washed, crushed by a cutter and a slicer, and then slurried using a mass colloider (manufactured by Masuko Sangyo). Cabbage 3K is added to the slurried cabbage.
1 g of water and 0.5 g of dry lactic acid bacterium (Lactobacillus plantarum) powder are added per gram, and stirred well to
Fermentation at 2 ° C. for 2 days. After completion of the fermentation, the fermented product was centrifuged to collect the supernatant, heated, concentrated, heat-sterilized, and 200 g of dextrin was added per 3 kg of cabbage, followed by spray-drying to obtain a cabbage fermented extract powder.

On the other hand, lymphocytes were separated from blood collected from healthy adult males by centrifugation to obtain effector cells. The above cabbage fermented extract powder was dissolved in physiological saline and sterilized through a filter. The cabbage fermented extract powder is added to the effector cells at a final concentration of 0.
It was added to be 25 mg / ml and 1.25 mg / ml. As a negative control, physiological saline was added to effector cells, and IL-
2 was added to a final concentration of 4 U / ml.

As a target cell, Da derived from human lymphoma was used.
Using udi cells, a sample in which ⁵¹ Cr had been incorporated in advance was prepared.

The E / T ratio between the cultured effector cells and target cells (number of effector cells / number of target cells)
= 50/1 and cultured for 4 hours. Thereafter, a culture supernatant was obtained and its radioactivity was measured. The radioactivity of all cells lysed with a surfactant was defined as 100, and the ratio to the radioactivity in each sample supernatant was determined and expressed as NK activity (%). Furthermore, the ratio (%) of the NK activity of each sample, when the NK activity of the negative control was set to 100, was calculated,
The NK activity enhancing effect was evaluated. Table 1 shows the results.

[0039]

[Table 1]

As a result of the evaluation of Daudi cells, an increase in NK activity was observed in all samples. Thereafter, 0.5 mg /
It was decided to use cabbage fermented extract powder having a concentration of ml.

(Example 2) As in Example 1, the outer leaves of the cabbage were removed, the outer surface was washed, crushed with a cutter and a slicer, and then slurried using a mass colloider (manufactured by Masuko Sangyo). . Using the cabbage slurried as a raw material, a cabbage fermented liquor obtained by treating under various conditions was centrifuged and filtered to remove solids to obtain a cabbage fermented extract (Production Examples 1 to 8).

In Production Example 1, fermentation was performed using an actual production apparatus, using Lactobacillus plantarum as a lactic acid bacterium, dextrin was added, spray-dried, and powdered. Fermentation using Lactobacillus plantarum as a lactic acid bacterium using an actual production apparatus, dextrin was added, and the mixture was concentrated under reduced pressure with an evaporator to obtain a powder. Fermented using Bacillus plantarum, dextrin was added, concentrated under reduced pressure with an evaporator and powdered, Production Example 4 was pretreated for 60 minutes on a boiling water bath, and Lactobacillus plantarum was used as a lactic acid bacterium. Fermented, dextrin added, concentrated under reduced pressure with an evaporator and powdered. Production Example 5 uses an experimental device to produce lactic acid bacteria. Fermentation using thermofils, dextrin added, concentrated under reduced pressure with an evaporator and powdered, Production Example 6 was pretreated on a boiling water bath for 60 minutes, fermented using thermofils as lactic acid bacteria, Dextrin was added, concentrated under reduced pressure with an evaporator, and powdered. In Production Example 7, fermentation was performed using red cabbage as a raw material, using lactobacillus plantarum as a lactic acid bacterium using an actual production apparatus, and adding dextrin. Spray dried and powdered, Production Example 8 fermented red cabbage using Lactobacillus plantarum as a lactic acid bacterium using an actual production apparatus, added dextrin, and concentrated under reduced pressure with an evaporator. It is powdered.

The NK activity of the obtained cabbage fermented extract was measured in the same manner as in Example 1 using lymphocytes obtained from three persons A, B and C as effector cells. Table 2 shows the results.

[0044]

[Table 2]

The results show that all the cabbage fermented extract powders showed an increase in NK activity relative to the negative control.

(Example 3) Next, using the cabbage fermented extract powder of Production Examples 2, 3, 5, and 7, which had relatively high activity in Example 2, the target cells were K5 derived from human chronic myeloid leukemia.
NK cell activity was measured in the same manner as in the case of 62 cells. Table 3 shows the results.

[0047]

[Table 3]

The results show that the cabbage fermented extract enhances NK activity and can also target K562 cells.

(Example 4) In the method of Production Example 3, two pastes, which were concentrated under reduced pressure by an evaporator and a powdered one, were used, and the final concentration was 0.5 mg / ml in terms of weight. Effector cells were treated as described. As effector cells, lymphocytes collected from the elderly (60 to 89 years old) and various cancer patients were used.

In the same manner as in Example 1, NK activity was measured using Daudi cells as target cells. 60 to 6 elderly
9-year-old, 70-79-year-old, 80-89-year-old divided into three groups, NK
Table 4 shows the results of examining the activity enhancing effect.

[0051]

[Table 4]

The results in Table 4 show that the paste and powder of the cabbage fermented extract enhanced NK activity relative to the negative control. In particular, the cabbage fermented extract paste appeared to have a higher NK activity effect than the powder.

Next, Table 5 shows the results of examining the NK activity enhancing effect of lymphocytes of patients with gastric cancer, colorectal cancer, and lung cancer, and Table 5 shows the results of examining the NK activity enhancing effects of lymphocytes of uterine cancer and brain tumor patients. 6 is shown.

[0054]

[Table 5]

[0055]

[Table 6]

The results show that the paste or powder of the cabbage fermented extract increased the NK activity enhancing effect on lymphocytes of various cancer patients, and that no difference in the effect was found between the types of cancer. Is shown. In particular, it was found that the paste of the cabbage fermented extract had a stronger NK activity enhancing effect than the powder. From these facts, it is expected that the elderly and various cancer patients will continuously ingest the cabbage fermented extract to enhance immunity and maintain high NK activity. Therefore, the cabbage fermented extract contains N
A food used as a K cell activator and containing a cabbage fermented extract can be an NK cell activation supplement.

[0057]

Industrial Applicability The fermented cabbage extract activates NK cells of not only healthy persons but also elderly persons and various cancer patients to increase immunity. Therefore, the cabbage fermented extract is used as an NK cell activator, and the food containing the cabbage fermented extract can be an NK cell activation supplement.

Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat II (reference) A61P 43/00 107 A61P 43/00 107 C12P 1/04 C12P 1/04 Z // (C12P 1/04 (C12P 1 / 04 Z C12R 1:25) C12R 1:25) (72) Inventor Shinji Tsuzaki 2-19-27 Hakata-ekimae, Hakata-ku, Fukuoka City, Fukuoka Toyo Shinyakunai Co., Ltd. (72) Kinya Takagaki, Hakata, Fukuoka City, Fukuoka Prefecture 2-19-19, Hakata-ku, Toku F-term (Reference) 4B018 LE03 LE04 LE05 MD53 MD81 MD86 ME08 MF01 MF04 MF06 MF07 MF13 4B064 AH19 CA02 CA06 CD22 CE16 CE20 DA01 DA10 4C088 AB15 AC05 AD22 CA11 CA12 CA17 MA25 MA28 MA44 MA52 NA14 ZB09 ZB22 ZB26

Claims (2)

[Claims] 1. An NK containing a cabbage fermented extract as a main component.
Cell activator. 2. The NK cell activator according to claim 1, wherein the cabbage fermented extract is in the form of a liquid, a paste, or a powder.