JP6209153B2 - Powder formulation - Google Patents

Description

  The present invention relates to a powder blend in which a powder using shiitake mushroom is blended.

  Shiitake mushrooms (fruit mushroom fruit bodies) are widely used as edible and dried shiitake mushrooms and dried shiitake mushrooms. In addition, lentinan contained in shiitake mushrooms (fruits of shiitake mushrooms) has been shown to have an immune enhancing effect. In addition, an anticancer agent (Patent Document 1) consisting of an extract obtained from a medium before the occurrence of shiitake fungus fruit bodies in which mycelia of shiitake mushrooms are cultured, and an immunosuppressive function and antiallergy using shiitake mushrooms (shiitake fungus fruit bodies) as a raw material An extract having an effect (Patent Document 2) has been proposed.

JP 2008-255057 A International Publication WO2005 / 107496

  By the way, when a fruit body is formed from a medium of shiitake mushroom, first, a bud of shiitake mushroom fruit body is formed on the medium, and this bud grows into a shiitake mushroom fruit body (so-called shiitake mushroom). The buds of shiitake mushroom fruit bodies contain a high proportion of cells that are growing vigorously, such as actively dividing cells, and other parts are considered to contain various active ingredients that are absent or relatively small. . On the contrary, as described in Patent Document 1, when a medium before the generation of shiitake fungus fruit bodies in which mycelia of shiitake mushrooms are cultured is used as a raw material, the proportion of cells with remarkable growth is low. In addition, as described in Patent Document 2, even when shiitake mushroom fruiting bodies that have finished growing are used as a raw material, the proportion of cells with remarkable growth is low.

  The present invention has been made in view of such points, and an object of the present invention is to provide a powder formulation capable of efficiently ingesting bud parts of shiitake mushroom fruiting bodies containing a high proportion of cells with remarkable growth. To do.

The solution is (1) a powder blend in which buds of shiitake mushroom fruit bodies are crushed and plant lactic acid bacteria are blended.

This powder blend is formed by blending bud powder obtained by pulverizing the shoots of shiitake mushroom fruit bodies (hereinafter also simply referred to as bud powder). Can be ingested efficiently. By ingesting this bud powder, various physiological activities such as an immune activation effect, a tumor growth inhibitory effect (antitumor effect), and a hair loss inhibitory effect are obtained. It is also possible to increase cytokines (IL-2, IL-4, IL-10, IL-12, IFN-γ).
IL-2, IL-4 and IFN-γ are involved in the activation of lymphocytes and NK cells. IL-10 is involved in maintaining the quality of life (QOL) of the sick and the elderly. Furthermore, IL-12 is involved in the suppression of malignant tumor growth.
In addition, plant lactic acid bacteria have an action of enduring gastric acid and reaching the intestinal tract to adjust the intestinal environment. In addition to various physiological activities such as immunity activation effect, tumor suppression effect (antitumor effect), hair loss suppression effect, etc. possessed by bud powder, by taking it together with bud powder, combined with the action of plant lactic acid bacteria Can be obtained.
Examples of plant lactic acid bacteria include Lactobacillus brevis subspecies coagulans, trail S1 bacteria (Lactobacillus plantarum s1), F-1 bacteria (Lactobacillus piantarum Formula-1), and the like.

This powder formulation is a powder, granule, tablet, capsule, paste, syrup combined with powder of bud powder and plant lactic acid bacteria , or other ingredients (dry powder of lactic acid bacteria other than plant lactic acid bacteria, various vitamin powders, etc.) Can be commercialized as. These can be taken as supplements such as nutritional supplements and health supplements, or taken as pharmaceuticals and quasi drugs.
Also, liquid or jelly-like drinks with added shiitake mushroom fruit bud powder and plant lactic acid bacteria , or other ingredients (dried lactic acid bacteria other than plant lactic acid bacteria, various vitamin powders, etc.) It can also be converted. Examples include Japanese tea, black tea, coffee, cocoa, alkaline ionized water, natural water, soft drinks, sake, dobowl, makgeolli, shochu, fruit liquor and other drinks, and jelly-type drinks (fluid foods).
Furthermore, it can also be commercialized as various foods to which bud powder and plant lactic acid bacteria have been added, or to which other components (dried powder of lactic acid bacteria other than plant lactic acid bacteria, various vitamin powders, etc.) have been added. Examples of such foods include, but are not limited to, fresh food such as meat, seafood, vegetables and fruits, processed livestock products such as ham and sausage, processed fishery products such as hampen, kamaboko and fish sausage, jam , Processed fruits such as dried fruits, processed vegetables such as pickles, dairy products such as milk, butter, cream, cheese, yogurt, oil and fats such as rapeseed oil, palm oil, sunflower oil, shortening, tofu, fried chicken, natto, etc. Processed soy foods, soy sauce, miso, sauce, ketchup and other seasonings, breads, cakes, sweets such as Japanese confectionery, Western confectionery, noodles such as udon, soba, somen and spaghetti.

In addition, shiitake mushroom fruit bodies are shiitake mushrooms. The shoots of shiitake mushroom fruit bodies are produced from a medium in which shiitake mushrooms are infested (fungi beds made of raw wood such as hodwood, sawdust, etc.), and shiitake mushroom fruit bodies (shaft part (stem part) and umbrella part). It refers to the part that grows. The shoots of shiitake mushroom fruit bodies can be easily separated from the medium by harvesting them from the culture medium such as the fungus bed on which the buds are grown or by hooking fingers or instruments. Also, in order to greatly grow shiitake mushroom fruit bodies, the number of shiitake mushroom fruit body buds that occur in the fungus bed is reduced, so-called “bud-out” (thinning out), so that shiitake mushroom fruit body buds are mushroomed. It can also be obtained (harvested) as a by-product.
Examples of the powder obtained by pulverizing the buds of shiitake mushroom fruit bodies include dry powders of shiitake mushroom fruit bodies, undried and semi-dried powders.
As a method for cultivating shiitake mushroom fruit bodies, the same technique as that for shiitake mushroom fruit bodies (shiitake cultivation) can be employed. Specifically, inoculation of soda-boiled seeds (seed piece) in soda-tree cultivation method of soda-shii mushroom cultivation and so-called shiitake-fungus spread on a fungus bed mixed with sawdust, etc. Cultivation techniques such as mycelia shiitake cultivation can be used. In particular, the fungal bed cultivation is preferable because the cultivation period can be shortened and the handling of the fungus bed is easier than that of the wood, so that the harvest of shiitake mushroom fruit bodies can be facilitated. As a method for cultivating the fungus bed, a known method such as a shelf cultivation method or a fungus bed hanging method can be employed.

  (2) The powder blend according to (1) above, wherein the bud powder is a powder blend that is a dried powder of the shoots of the shiitake mushroom fruiting bodies.

Since the bud powder in this powder blend is a dried powder of shiitake mushroom fruit body buds, storage and handling of the bud powder and powder blend is less than when blending undried or semi-dried bud powder. Easy.
As dried powder, buds of shiitake mushroom fruit bodies or bud fragments cut from buds of shiitake mushroom fruit bodies are warm air dried, cold air dried, infrared dried, far infrared dried, hot air / far infrared combined dried, frozen Examples thereof include drying (vacuum freeze drying), drying by a technique such as drying with a desiccant such as silica gel, and pulverization after drying.

( 3 ) Furthermore, it is good to use the powder formulation as described in (1) or (2) , wherein the plant lactic acid bacteria is Lactobacillus piantarum Formula-1.

The plant lactic acid bacterium F-1 (Lactobacillus piantarum Formula-1) has the action of enduring gastric acid and reaching the intestinal tract to adjust the intestinal environment.
Moreover, a synergistic effect with shiitake mushroom fruit body bud powder can be obtained with respect to various physiological activities such as an immune activation effect, a tumor suppression effect (antitumor effect), and a hair loss suppression effect. Cytokines (IL-2, IL-4, IL-10, IL-12, IFN-γ) can also be increased synergistically.

( 4 ) Furthermore, the powder composition according to any one of (1) to ( 3 ), wherein the buds of the shiitake mushroom fruit body are collected from a fungus bed cultivated by fungus bed shiitake cultivation. And good.

In addition, shiitake mushroom cultivation makes it easier to collect buds of shiitake mushroom fruit bodies and does not mix with sardine parts (such as the bark of the tree). It can be made into a powder formulation using the shoots of the entity.
As a method for cultivating the fungus bed, as described above, a known method such as a shelf cultivation method or a fungus bed hanging method can be employed.

( 5 ) The powder blend described in any one of (1) to ( 4 ) above is a powder blend that is a food, drink, pharmaceutical product, quasi-drug, mammalian pharmaceutical product, or mammalian food. good.

Since the powder formulation of the present invention is a food, drink, pharmaceutical product, quasi-drug, mammalian pharmaceutical product, or mammalian food blended with bud powder, it can be taken by humans or non-human mammals. Various physiological activities such as an immunity activation effect, a tumor suppression effect (antitumor effect), and a hair loss suppression effect of the powder can be obtained.
That is, since the food and drink that is the powder composition of the present invention is blended with bud powder, by taking this food and drink, the immune ability activation effect, tumor suppression effect (antitumor effect) of the bud powder has ), Various physiological activities such as hair loss inhibiting effect can be obtained.
Examples of such foods and drinks include nutritional supplements in the form of powders, granules, tablets, capsules, pastes, syrups and the like. In addition, as described above, for example, a liquid or jelly-like drink to which bud powder is added may be mentioned. In addition, various foods with bud powder added, for example, fresh foods such as meat, seafood, vegetables, fruits, processed livestock products, processed marine products, processed fruits, processed vegetables, dairy products, fats and oils, processed soybeans Examples include foods, seasonings, breads, cakes, confectionery, and noodles.

In addition, since the medicinal product that is the powder blend of the present invention is blended with bud powder, by taking this medicinal product, from the physiological activity of the bud powder, the effect of immunity activation, the effect of tumor suppression, And the effect of hair loss suppression can be acquired.
Examples of pharmaceutical dosage forms include powders, granules, tablets, capsules, pastes, syrups and the like.

In addition, since the quasi-drug that is the powder formulation of the present invention is blended with bud powder, by taking this quasi-drug, the physiological activity of the bud powder, the effect of immunity activation Moreover, the effect of tumor suppression and the effect of hair loss suppression can be obtained.
In addition, examples of dosage forms for quasi-drugs include powders, granules, tablets, capsules, pastes, syrups, and the like.

Furthermore, since the medicinal product for mammals which is the powder blend of the present invention is blended with bud powder, the effect of immunity activation can be obtained from the physiological activity of the bud powder by ingesting the mammal medicinal product into the mammal. Moreover, the effect of tumor suppression and the effect of hair loss suppression can be obtained.
In addition to the powder, the dosage form of a pharmaceutical product for mammals includes forms such as granules, tablets, capsules, pastes, and syrups.

Furthermore, since the feed for mammals which is the powder blend of the present invention is blended with bud powder, the immunity activation effect, tumor suppression, which bud powder has by giving this mammal feed to mammals Various physiological activities such as effects (antitumor effects) and hair loss suppressing effects can be obtained.
Examples of such food for mammals include pet food given to pets (companion animals) and feed given to mammals such as pigs and cows to be raised. For example, as a form of food for mammals, there is a liquid or jelly animal drink to which bud powder is added. In addition, various pet foods and feeds to which bud powder is added may be mentioned.

It is a flowchart which shows the manufacture procedure of the powder formulation which mix | blended the bud powder. It is a graph which shows the body weight change after an experiment start about the cancer-free mouse | mouth of each group in NT1 group. It is a graph which shows the body weight change after an experiment start about the tumor bearing mouse | mouth of each group in T5 group. It is a graph which shows the change of the tumor volume after an experiment start about the tumor bearing mouse | mouth of each group in T5 group. It is a graph which shows the density | concentration of blood cytokine IL-2 measured about 10 days after the experiment start measured about the cancer-free mouse | mouth of each group in NT1 group. It is a graph which shows the density | concentration of the blood cytokine IL-4 on the 10th after the experiment start measured about the cancer-free mouse | mouth of each group in NT1 group. It is a graph which shows the density | concentration of blood cytokine IL-10 measured about 10 days after the experiment start measured about the cancer-free mouse | mouth of each group in NT1 group. It is a graph which shows the density | concentration of the blood cytokine IL-12 measured about 10 days after the experiment start measured about the cancer-free mouse | mouth of each group in NT1 group. It is a graph which shows the density | concentration of the blood cytokine IFN-gamma measured 10 days after the experiment start measured about the cancer-free mouse | mouth of each group in NT1 group. It is a graph which shows the hair loss area measured about the mouse | mouth of the NT1-C group and the NT1-F + M group among the cancer-free mice 90 days after the experiment start. It is a photograph which shows the fur line (hair loss state) of the NT1-F + M group mouse | mouth (4 animals) 45 days after the experiment start. It is a photograph which shows the fur line (hair loss state) of the NT1-C group mouse | mouth (4 animals) 45 days after the experiment start. It is a graph which shows the change of the tumor volume after an experiment start about the tumor bearing mouse | mouth of each group in NT2 group, T3 group, and T1.5 group.

  First, shiitake mushroom fruit body bud powder (hereinafter also referred to as “bud powder”) and plant lactic acid bacteria (specifically, Lactobacillus piantarum Formula-1: Louis Pasteur Medical Research Center, Molecular Immunology Institute). The following experiment was conducted in order to confirm the physiological action and synergistic action of “F1 lactic acid bacteria”.

(Tumor transplant)
Mouse lung metastases tumor cells (LLC cells) stored frozen at -80 ° C were washed with PBS solution, cultured in RPMI-1640 (10% FBS) culture solution for 2 days, and pipetted with 0.025% trypsin solution. Then, the proliferated cells were peeled off, centrifuged at 1000 rpm for 5 minutes, and the supernatant was aspirated and discarded. A PBS solution cooled on ice at 4 ° C. was added to the precipitated cells, centrifuged again at 1000 rpm for 5 minutes, and the supernatant was aspirated and discarded. The precipitated cells were diluted with physiological saline to prepare a tumor cell solution having a concentration of 5.0 × 10 ⁵ cells / 0.05 mL.

Next, C57BL / 6 mice (male, 7 weeks old) were implanted subcutaneously into the thigh by 0.05 mL (5.0 × 10 ⁵ equivalents). The first experiment described later was started one week after transplantation when the tumor grew to 2 to 3 mmφ.
Separately, a tumor cell solution having a concentration of 3.0 × 10 ⁵ cells / 0.05 mL and 1.5 × 10 ⁵ cells / 0.05 mL was prepared by the same method, and the thigh of C57BL / 6 mice was prepared. 0.05 mL (corresponding to 3.0 × 10 ⁵ or 1.5 × 10 ⁵ ) was injected and transplanted subcutaneously into the part. A second experiment described later was started one week after transplantation when the tumor grew to 2 to 3 mmφ.

In the first experiment, a group of mice transplanted with 5.0 × 10 ⁵ tumor cells is designated as T5 group, and a mouse that has not undergone tumor transplantation is designated as NT1 group.
In the second experiment, a group of mice transplanted with 3.0 × 10 ⁵ mice is designated as T3 group, and a group of mice transplanted with 1.5 × 10 ⁵ mice is designated as T1.5 group. Moreover, let the mouse | mouth which has not performed tumor transplantation be NT2 group.

(Preparation of bud powder feed, lactic acid bacteria water)
Harvest shoots of shiitake mushroom fruit bodies grown on the fungus bed cultivated by the fungus bed hanging method at the agricultural association INS, dried by hot air and far-infrared combined drying below 50 ° C, pulverized, shiitake mushrooms A powder (bud powder) obtained by grinding the shoots of the substance was obtained.
50 wt% of this bud powder, 20 wt% of pulverized powder of mouse breeding solid feed (MF), 20 wt% of kinako flour, 5 wt% of cheese and 5 wt% of rice flour for hardening are mixed with tap water, allowed to air dry, A solid feed containing powder (hereinafter also referred to as “bud powder feed”) was prepared.
Also, F1 lactic acid bacteria (Lactobacillus piantarum Formula-1) cultured at the Institute for Molecular Immunology of Louis Pasteur Medical Research Center were diluted with physiological saline to a concentration of 1.0 × 10 ⁷ cells / 0.3 mL. "Lactic acid bacteria water" was prepared separately.

(First experiment)
The T5 group mice (LLC tumor-bearing mice) that had undergone the above-described tumor transplantation were divided into 4 groups, and were reared in the same manner as the C57BL / 6 mice (non-cancer-bearing mice) that had not undergone the 4 group tumor transplantation, and the body weight change Tumor volume change and blood cytokine concentration were measured. In each group, 15 C57BL / 6 mice were bred per group.
<Non-cancer-bearing mice: NT1 group>
NT1-C group: cancer-free mouse / control group (general feed / water administration group)
・ NT1-F group: Cancer-free mouse / Lactic acid bacteria single administration group (General feed / Lactic acid bacteria water administration group)
NT1-M group: cancer-free mouse / bud powder alone administration group (bud powder feed / water administration group)
NT1-F + M group: cancer-free mouse / bud powder + lactic acid bacteria combination group (bud powder feed / lactic acid bacteria water administration group)
<Tumor bearing mouse: T5 group (5.0 × 10 ⁵ transplanted)>
-T5-C group: Cancer-bearing mice / control group (general feed / water administration group)
・ T5-F group: Tumor-bearing mouse / lactic acid bacteria alone administration group (general feed / lactic acid bacteria water administration group)
T5-M group: Tumor-bearing mouse / bud powder alone administration group (bud powder feed / water administration group)
・ T5-F + M group: Tumor-bearing mouse / bud powder + lactic acid bacteria combination group (bud powder feed / lactic acid bacteria water administration group)

In addition, the mouse | mouth breeding solid feed (MF) and water (tap water) which are general feeds were administered to the mouse | mouth of the control group (NT1-C group, T5-C group), and was bred.
The mice of the lactic acid bacteria water administration group (NT1-F group, NT1-F + M group, T5-F group, T5-F + M group) were forcibly orally administered daily with 0.3 mL of the lactic acid bacteria water using a metal sonde. 0.0 × 10 ⁷ cells / day / animal of F1 lactic acid bacteria were ingested.
In addition, for the mice in the bud powder feed administration group (NT1-M group, NT1-F + M group, T5-M group, T5-F + M group), the above-mentioned bud powder feed was freely taken every day (0.8 to 1). 2 g / day / animal).

(Weight measurement)
About NT1 group (uncancer-bearing mouse) and T5 group (cancer-bearing mouse), the body weight was measured every day from the start of the experiment. The change is shown in FIGS. As can be seen from FIG. 2, the body weight change of each non-cancer-bearing mouse in the NT1 group is significantly different in the whole experimental period between administration / non-administration of bud powder feed (bud powder) and lactic acid bacteria water (F1 lactic acid bacteria). There wasn't. Therefore, no toxicity is observed for bud powder feed (bud powder) or lactic acid bacteria water (F1 lactic acid bacteria) from the change in body weight. The same can be said from the change in body weight of each tumor-bearing mouse in the T5 group shown in FIG. However, in the T5-C group (tumor bearing mouse / control group), the body weight decreased after peaking on the 10th day after the start of the experiment. This is because the amount of food (feed intake) decreased with the progression of cancer.

(Tumor volume measurement)
Tumor growth status was quantified by tumor volume. Assuming that the tumor has a rugby ball shape (spheroid: major axis D1, minor axis D2), the volume V of the tumor is V = 3π / 4 · (D1 / 2) · (D2 / 2) ² = π / 6 · D1 · D2 ² ≈0.523D1 · D2 ² Therefore, the major axis D1 and the minor axis D2 when the tumor of each mouse of the T5 group was viewed as a spheroid were measured, and the volume V of the tumor was calculated using the above equation.

(Evaluation of antitumor effect)
For the 4 groups of mice in the T5 group, the tumor volume V was measured every day, and the antitumor effect in the case of lactic acid bacteria single administration, bud powder single administration, and bud powder + lactic acid bacteria combination was evaluated. The results are shown in FIG. The data for each group is an average of 15 animals.

In terms of the number of days required for the tumor volume V to reach 200 mm ^3, it was 6.3 days for the mice in the T5-C group (tumor bearing mouse / control group). In the T5-F group (cancer-bearing mouse / lactic acid bacterium alone administration group), it was 9.5 days, 1.5 times. In contrast, in the T5-M group (tumor bearing mouse / bud powder single administration group), it was 15.6 days, 2.5 times. Furthermore, in the T5-F + M group (cancer-bearing mouse / bud powder + lactic acid bacteria combination group), it was 3.1 times 19.4 days.

  Thus, although anti-tumor effects are observed even when lactic acid bacteria are administered alone, T5-M group (tumor bearing mouse / bud powder alone administration group) and T5-F + M group (tumor bearing mouse / In the bud powder + lactic acid bacteria combination group), a high antitumor effect is observed. Moreover, the T5-F + M group (cancer-bearing mouse / bud powder + lactic acid bacteria combination group) using bud powder and lactic acid bacteria in combination has a particularly high antitumor effect, and a synergistic effect between bud powder administration and lactic acid bacteria administration is observed. This antitumor effect is presumed to occur not only in mice but also in mammals including humans.

(Measurement of cytokines)
14 days after the start of the experiment, blood (about 1.5 to 2.5 mL) collected from the heart of the mouse was centrifuged at 3500 rpm for 5 minutes in a cryogenic centrifuge (treatment temperature: 4 ° C.), and the supernatant serum was collected. 0.7-1.0 mL was collected. The collected serum was immediately stored at −80 ° C., and after treatment of all mice, each serum was thawed, and the cytokine concentration was measured twice for each mouse using Bio-Plex (NAVIOS: manufactured by BECKMAN COULTER). . The measured cytokines are IL-2, IL-4, IL-10, IL-12, and IFN-γ, and each group is an average value of 3 animals (6 measured values).

  The cytokine IL-2 is produced from T cells (lymphocytes) that have differentiated in the thymus and have acquired specificity, and have the effect of activating lymphocyte chemotaxis and macrophages. Cytokine IL-4 is produced by activated Th2T cells and has the effect of suppressing Th1T cells and inducing B cell activation and proliferation differentiation. Cytokine IL-10 is an excess of inflammatory cytokines (IL-1, IL-8, TNF-α) produced by CD4 T cells and macrophages and cytokines (IL-2, IFN-γ) involved in cytotoxic reactions. It suppresses the activity and is involved in the QOL of the sick and elderly. Furthermore, the cytokine IL-12 is produced from dendritic cells and macrophages, has the function of maintaining the proliferation of activated T cells, directing the expression of killer T cells, and the activity of NK cells to proliferate, and is an indicator for knowing the progress of malignant tumors Become. IFN-γ has an antitumor action and an immune enhancing action, and activates NK cells.

FIG. 5 is a graph showing the blood concentration of cytokine IL-2 measured for four NT1 group mice (unbearing mice) 14 days after the start of the experiment.
Compared with the blood concentration (0.85 pg / ml) of the NT1-C group (unbearing mouse / control group), the NT1-F group (unbearing mouse / lactic acid bacterium alone administration group) is 1.1 times Concentration.
In contrast, in the NT1-M group (non-cancerous mouse / bud powder single administration group), IL-2 was increased 2.3 times compared to the NT1-C group. From this, the bud powder feed (bud powder) has an increased effect of cytokine IL-2, which is better than lactic acid bacteria water (F1 lactic acid bacteria).
In particular, in the NT1-F + M group (non-cancerous mouse / bud powder + lactic acid bacteria combination group), IL-2 is 2.53 times (= 1.1 × 2.3) compared to the NT1-C group. Also increased by a factor of 4.2. From this, the bud powder feed (bud powder) has a particularly good increase effect due to the synergistic effect on the cytokine IL-2 when used in combination with lactic acid bacteria water (F1 lactic acid bacteria).

FIG. 6 is a graph showing the blood concentration of cytokine IL-4 measured for four NT1 group mice (unbearing mice) 14 days after the start of the experiment.
Compared with the blood concentration (0.22 pg / ml) of the NT1-C group (unbearing mouse / control group), the NT1-F group (unbearing mouse / lactic acid bacterium alone administration group) is 1.3 times Concentration.
On the other hand, IL-2 increased 1.6 times in the NT1-M group (unbearing cancer mouse / bud powder single administration group) compared to the NT1-C group. From this, the bud powder feed (bud powder) has an increased effect of cytokine IL-4, which is better than lactic acid bacteria water (F1 lactic acid bacteria).
In particular, in the NT1-F + M group (non-cancerous mouse / bud powder + lactic acid bacteria combination group), IL-4 is 2.08 times (= 1.3 × 1.6) compared to the NT1-C group. Also increased by a large 4.6 times. From this, the bud powder feed (bud powder) has a particularly good increase effect due to synergistic action on the cytokine IL-4 when combined with lactic acid bacteria water (F1 lactic acid bacteria).

FIG. 7 is a graph showing blood concentrations of cytokine IL-10 measured for four NT1 group mice (uncarried cancer mice) 14 days after the start of the experiment.
Compared with the blood concentration (5.8 pg / ml) of the NT1-C group (unbearing mouse / control group), the NT1-F group (unbearing mouse / lactobacillus alone administration group) is 1.5 times. Concentration.
On the other hand, IL-2 increased 1.6 times in the NT1-M group (unbearing cancer mouse / bud powder single administration group) compared to the NT1-C group. From this, bud powder feed (bud powder) has a slightly better cytokine IL-10 increasing effect than lactic acid bacteria water (F1 lactic acid bacteria).
In particular, in the NT1-F + M group (uncancerous mouse / bud powder + lactic acid bacteria combination group), IL-10 is 2.4 times (= 1.5 × 1.6) compared to the NT1-C group. Also increased by a large 3.8 times. From this, the bud powder feed (bud powder) shows a particularly good increase effect due to the synergistic action of cytokine IL-10 when used in combination with lactic acid bacteria water (F1 lactic acid bacteria).

FIG. 8 is a graph showing the blood concentration of cytokine IL-12 measured for four NT1 group mice (unbearing mice) 14 days after the start of the experiment.
Compared with the blood concentration (9.7 pg / ml) of the NT1-C group (unbearing mouse / control group), the NT1-F group (unbearing mouse / lactobacillus alone administration group) is 1.8 times. Concentration.
In contrast, in the NT1-M group (unbearing mouse / bud powder single administration group), IL-12 was increased 2.7 times compared to the NT1-C group. From this, the bud powder feed (bud powder) has a favorable effect of increasing cytokine IL-12 as compared with lactic acid bacteria water (F1 lactic acid bacteria).
In particular, in the NT1-F + M group (non-cancer-bearing mouse / bud powder + lactic acid bacteria combination group), IL-12 is 4.86 times (= 1.8 × 2.7) compared to the NT1-C group. Was also increased by a factor of 5.1. From this, the bud powder feed (bud powder) shows a particularly good increase effect due to the synergistic effect on cytokine IL-12 when used in combination with lactic acid bacteria water (F1 lactic acid bacteria).

FIG. 9 is a graph showing the blood concentration of cytokine (interferon) IFN-γ measured for four NT1 group mice (unbearing mice) 14 days after the start of the experiment.
Compared with the blood concentration (1.6 pg / ml) in the NT1-C group (unbearing mouse / control group), the NT1-F group (unbearing mouse / lactic acid bacterium alone administration group) is 1.3 times higher. Concentration.
In contrast, in the NT1-M group (non-cancer-bearing mouse / bud powder single administration group), IFN-γ increased 1.5 times as compared with the NT1-C group. From this, the bud powder feed (bud powder) has a better increasing effect of interferon IFN-γ than lactic acid bacteria water (F1 lactic acid bacteria).
In particular, in the NT1-F + M group (uncarried cancer mouse / bud powder + lactic acid bacteria combination group), IFN-γ is 1.95 times (= 1.3 × 1.5) compared to the NT1-C group. Also increased by a large 2.7 times. From this, the bud powder feed (bud powder) shows a particularly good increase effect due to the synergistic action of interferon IFN-γ when used in combination with lactic acid bacteria water (F1 lactic acid bacteria).

  From these, the intake of bud powder feed (bud powder) shows a good increasing action for any of cytokines IL-2, IL-4, IL-10, IL-12, and IFN-γ. Immune ability activation effect by intake of feed (bud powder) is recognized. In particular, when used in combination with lactic acid bacteria water (F1 lactic acid bacteria), a synergistic immune ability activation effect is observed. In addition, it is guessed that this immune ability activation effect similarly arises not only for mice but also for mammals including humans.

(Confirmation of hair loss suppression effect)
In general, when C57BL / 6 mice are bred at about 14 weeks of age, mice with hair loss or white hair appearing on the back or flank appear.
Of the mice in the NT1-F + M group (non-cancer-bearing mouse / bud powder + lactic acid bacteria combination group), mice were photographed on the 45th day (14 weeks old) from the start of the experiment, and the shape was corrected on the enlarged photo. The hair loss area (mm ² ), which is the area of the part that became hair loss or white hair, was measured. Of the mice in the NT1-C group (cancer-free mouse / control group), the hair loss area was similarly measured for mice on the 45th day (14 weeks old) from the start of the experiment. The number of samples is 7 respectively.

The result is shown in FIG. According to the graph of FIG. 10, in the NT1-C group (uncancerous mouse / control group) mice, the average hair loss area was 27 ± 6 mm ² , whereas in the NT1-F + M group (uncancerous mice). In the case of the mouse / bud powder + lactic acid bacteria combination group), it was 2.2 ± 0.4 mm ² of 1/12 or less.
FIG. 11 shows a photograph of mice in the NT1-F + M group, and FIG. 12 shows a photograph of mice in the NT1-C group. As can be understood by comparing these photographs, the NT1-C mice in FIG. 12 clearly have white hair on the back and hair loss on the flank. It can be seen that the fur is also worse than the NT1-F + M group of mice (see FIG. 11). On the other hand, it can be seen that the mice in the NT1-F + M group in FIG. 11 have smooth fur and little hair loss or white hair. Although not shown in the photograph or the hair loss area, the NT1-M group (uncarried cancer mouse / bud powder single administration group) mice are not as large as the NT1-F + M group mice, but the NT1-C group mice. Compared to hair loss, hair loss and white hair are suppressed, and the appearance of the hair is improved.

  As is clear from these results, in the NT1-M group and NT1-F + M group mice in the bud powder feed administration group, hair loss and the generation of white hair can be suppressed, and the fur level can be improved. Furthermore, it can be seen that hair loss and white hair can be remarkably suppressed in the NT1-F + M group of mice in the bud powder + lactic acid bacteria combination group, and that the fur can be kept very well. In addition, it is guessed that this hair loss inhibitory effect similarly arises not only for mice but also for mammals including humans.

(Second experiment)
In the first experiment described above, the transition of the tumor volume V was observed for mice (LLC tumor-bearing mice) in which a relatively large amount (5.0 × 10 ⁵ ) of tumors had been transplanted.
In contrast, in the second experiment, a relatively small amount (3.0 × 10 ⁵ , 1.5 × 10 ⁵ ) of mice transplanted with tumors (LLC tumor-bearing mice) was used. The suppression state of the tumor volume V by administration and administration of lactic acid bacteria water was observed.

Specifically, the T3 group (LLC tumor-bearing mice) subjected to the above-described tumor transplantation is divided into two groups, the T1.5 group mice (LLC tumor-bearing mice) are divided into two groups, and the two groups of tumor transplantation C57BL / 6 mice that had not been treated (uncancerous mice) were bred in the same manner, and tumor volumes were measured. Each group is 15 C57BL / 6 mice / group.
<Non-cancer-bearing mice: NT2 group>
NT2-C group: cancer-free mouse / control group (general feed / water administration group)
NT2-F + M group: cancer-free mouse / bud powder + lactic acid bacteria combination group (bud powder feed / lactic acid bacteria water administration group)
<Tumor bearing mice: T3 group (3 × 10 ⁵ transplants)>
-T3-C group: Cancer-bearing mice / control group (general feed / water administration group)
・ T3-F + M group: Tumor-bearing mouse / bud powder + lactic acid bacteria combination group (bud powder feed / lactic acid bacteria water administration group)
<Tumor bearing mice: T1.5 group (1.5 × 10 ⁵ transplants)>
-T1.5-C group: cancer-bearing mouse / control group (general feed / water administration group)
・ T1.5-F + M group: Tumor-bearing mouse / bud powder + lactic acid bacteria combination group (bud powder feed / lactic acid bacteria water administration group)

As in the first experiment, mice in the control group (NT2-C group, T3-C group, T1.5-C group) include mouse-feeding solid feed (MF), which is a general feed, and water (tap water). Water) was administered and reared. In addition, for the mice in the bud powder feed / lactic acid bacteria water administration group (NT2-F + M group, T3-F + M group, T1.5-F + M group), 0.3 mL of the aforementioned lactic acid bacteria water was forcibly orally administered daily using a metal sonde. and, 1.0 × 10 ⁷ cells / day / animal of F1 lactobacilli 10 ⁷ equivalent) addition was ingested, the aforementioned bud powder feed, daily, ad libitum (1.3～1.5G / day / animal )

(Tumor volume measurement)
Tumor growth status was quantified by tumor volume. As in the first experiment, the tumor volume V was calculated by measuring the major axis D1 and the minor axis D2 when the tumor of each mouse was viewed as a spheroid.

(Evaluation of antitumor effect)
For each mouse of the T3 group and the T1.5 group, the tumor volume V was measured every day, and the temporal change of the volume V, and thus the antitumor effect in the case of bud powder + lactic acid bacteria combination was evaluated. The results are shown in FIG. In addition, the data of each group are average values of 15 animals.

In terms of the number of days required for the tumor volume V to reach 36 mm ^3, it was 9.3 days for the mice in the T3-C group (tumor bearing mouse / control group). On the other hand, in the T3-F + M group (cancer-bearing mouse / bud powder + lactic acid bacteria combination group), it was 17.0 days, which is 1.8 times that of the T3-C group.
On the other hand, in the T1.5-C group (cancer-bearing mouse / control group) with a small number of transplanted tumor cells, it was 13.5 days. On the other hand, in the T1.5-F + M group (cancer-bearing mouse / bud powder + lactic acid bacteria combination group), it was 1.8 times 24.0 days compared to the T1.5-C group.
From these, in the T3-F + M group and T1.5-F + M group (tumor bearing mouse / bud powder + lactic acid bacteria combination group) that use bud powder and lactic acid bacteria in combination, high antitumor effects are the same as in T5 group (first experiment). Is recognized.

In particular, in the T1.5-F + M group (tumor bearing mouse / bud powder + lactic acid bacteria combination group) mice transplanted with a relatively small amount of 1.5 × 10 ⁵ tumor cells, the tumor grew until 24 days after the start of the experiment. Since then, the tumor has shrunk. After 37 days, mice with disappeared tumors appeared, and in other mice in the T1.5-F + M group, a marked antitumor effect, ie, tumor shrinkage was observed.

  From these, it can be seen that when bud powder feed (bud powder) and lactic acid bacteria water (F1 lactic acid bacteria) are used in combination, a very good antitumor effect can be obtained. In particular, it can be seen that a remarkable antitumor effect (tumor shrinkage, tumor extinction) is obtained at an early stage where the number of tumor cells is small. Such an antitumor effect is presumed to occur not only in mice but also in mammals including humans.

(Production method of powder compound)
Next, the production of the bud powder of the present case and the powder blend containing the bud powder will be described with reference to FIG. First, as shown in step S1, shoots of shiitake mushroom fruit bodies grown on a culture medium (in this example, a fungus bed cultivated by a fungus bed suspension method) are harvested. Next, in step S2, the shoots of shiitake mushroom fruit bodies are cut into small pieces, dried by combined drying with hot air and far-infrared rays at 50 ° C. or less, and pulverized to obtain a powder (bud powder) obtained by crushing the shoots of shiitake mushroom fruit bodies. Got .

  Furthermore, in step S3, a food and drink, specifically, a tablet as a dietary supplement is formed. Specifically, in addition to bud powder and dry powder of F1 lactic acid bacteria, binders (such as starch paste and gum arabic paste), disintegrants (such as starch and celluloses), and lubricants (such as magnesium stearate) Wax, talc, etc.) are mixed and compressed into tablets. A powder formulation in the form of a tablet is thus obtained.

In the above, the present invention has been described with reference to the embodiments. However, the present invention is not limited to the above embodiments, and it is needless to say that the present invention can be appropriately modified and applied without departing from the gist thereof.
For example, when forming a tablet, the tablet may be formed using an excipient (such as lactose or starch) together. Alternatively, an enteric tablet can be prepared by coating with a gastric acid insoluble coating agent. Moreover, in said embodiment, the bud powder and the dry powder of F1 lactic acid bacteria were mixed and shape | molded into the tablet. However, it may be formed into a tablet using only bud powder as an active ingredient. Conversely, vitamins, minerals and other active ingredients may be included in addition to the bud powder and the dry powder of F1 lactic acid bacteria.

  Further, the present invention is not limited to tablet dosage forms, but may be used in the form of granules, or in the form of granules, capsules, pastes, or syrups. Moreover, it can also be set as food and drink of various forms, such as a fish sausage to which bud powder was added, a soft drink to which bud powder and F1 lactic acid bacteria were added, for example. However, in view of the synergistic effect, in addition to bud powder, it is preferable to use a food and beverage containing F1 lactic acid bacteria, a pharmaceutical product, a quasi-drug, a pharmaceutical product for mammals, or a powder blend as a food for mammals.

Claims (5)

A powder blend containing bud powder obtained by pulverizing buds of shiitake mushroom fruit bodies and plant lactic acid bacteria . A powder formulation according to claim 1,
The bud powder is
A powder formulation that is a dry powder of the shoots of the fruit mushroom fruit bodies. A powder formulation according to claim 1 or claim 2 ,
A powder formulation in which the plant lactic acid bacterium is Lactobacillus piantarum Formula-1. A powder formulation according to any one of claims 1 to 3 ,
The buds of shiitake mushroom fruit bodies are a powder formulation obtained from a fungus bed cultivated by fungus shiitake cultivation. The powder blend according to any one of claims 1 to 4 ,
A powder formulation that is a food or drink, a pharmaceutical product, a quasi-drug, a pharmaceutical product for mammals, or a food for mammals.