WO2002049661A1 - Preventievs/remedies for diabetes and functional foods containing the same - Google Patents

Abstract

Preventives/remedies for diabetes originating in a fungus which is a natural material, having a high safety, a low cost and stable qualities and exhibiting effects of inhibiting an increase in blood glucose level and improving glucose tolerance in type 1 diabetes and type 2 diabetes; and functional foods containing these preventives/remedies for diabetes which have effects of preventing and ameliorating diabetes. The preventives/remedies for diabetes having the effects of inhibiting an increase in blood glucose level and improving glucose tolerance are prepared by using, as the active ingredient, processed fruit body of a fungus belonging to the family Auriscalpiaceae, for example, fruit body of Mycoleptodonoides aitchisonii having been cultured in a mushroom bed. Further, functional foods having an antitumor effect are prepared by adding these preventives/remedies for diabetes to various foods. The effect of inhibiting an increase in blood glucose level can be confirmed by measuring the glucose level in blood after fasting for 16 to 20 hours, while the effect of improving glucose tolerance can be confirmed by carrying out a glucose-load test.

Description

 Description Diabetes prevention / improvement agent and functional food containing it

 The present invention relates to a diabetes preventive and ameliorating agent and a functional food having a diabetes preventive and ameliorating effect, and more particularly to a functional food obtained from a fruit body of a mushroom belonging to the family Climacodontaceae, such as Bunakaritake (Mycoleptodonoides aitchisonii). The present invention relates to a diabetes preventive or ameliorating agent having an effect of suppressing an increase in blood glucose level and an effect of improving glucose tolerance against type 1 and type 2 diabetes, and a functional food having a diabetes preventing or ameliorating effect including such a diabetes preventing or ameliorating agent. . Background art

 Diabetes is a group of diseases mainly associated with chronic hyperglycemia due to lack of insulin action and associated with various characteristic metabolic abnormalities. Its onset involves both genetic and environmental factors. Diabetes is broadly divided into insulin-dependent diabetes (type 1 diabetes) and non-insulin-dependent diabetes (type 2 diabetes), and is known to be a risk factor for stroke and heart disease. Deterioration of quality of life due to complications such as retinopathy and kidney damage is also a serious problem. In addition, the number of people with diabetes has increased sharply in recent years, with 690,000 people who are strongly suspected of diabetes and 137,000 people who cannot deny the possibility of diabetes. It is said. This is one of the most important health problems of the people as a lifestyle-related disease.

In the deep beech forest of the Tohoku region, beech agaric grows on fallen beech trees and dead trees in late autumn, and umbrellas are 3 to 10 cm in diameter, semicircular white, and later. Tint yellow, back white Needle-like, later yellowish on the surface as well as the surface, the meat is white, soft and water-absorbing, and is known to have a unique aroma.As a mushroom prized as mountain meat by mountain villagers Is also known. As methods for artificially cultivating beech hachiritake, Nagano Prefectural Forest Research Report No. 3, pages 32 to 37 (1989) and Fukushima Prefectural Forest Research Report No. 23, 81 On page 101 (1990), a report on research on cultivation using raw wood such as beech and cherry blossoms was published in “Food Flow Technology” Vol. 18, No. 1, 17- On page 1 (19989), there is a report on log cultivation and research on cultivation methods using sawdust. According to these reports, although log cultivation can grow into fruiting bodies, many log Must be supplied, and it is difficult to cultivate seed pieces with wood chips because of the strong decay of mycelia.On the other hand, container cultivation using ogakusu has many malformations, and It is difficult to grow fruiting bodies by bag cultivation using cultivation. It has been considered is the impossible.

 It is an object of the present invention to provide a diabetes preventive / ameliorating agent having a blood glucose level-inhibiting effect and an glucose tolerance improving effect on type 1 diabetes and type 2 diabetes, and a diabetes preventing / ameliorating effect including such a diabetes preventing / improving agent. To provide a functional food having

The present inventors have previously conducted an inoculation step of inoculating a seed medium of B. agaricus inoculated on a medium supplemented with a specific nutrient source, and culturing the medium inoculated with a seed of B. agaricus, and forming a fungal bed in which the mycelia grew on the medium. A pre-culturing step for obtaining the fungal bed; a middle culturing step for culturing the bacterial bed to obtain a P. aeruginosa primordium that has not been exposed to a physical space capable of growing fruit bodies; and exposing the medium to a physical space capable of growing the fruit bodies. A post-culturing process in which a primordium selected from the original primordial bamboo shoots is cultivated under conditions that expose the physical space in which the primordial cultivars of Buna-haritake can grow into fruiting bodies, to obtain fruiting bodies of Bunakaritake mushrooms Grows naturally after growing beech It has been confirmed that a B. agaricus fruit body that is larger than this can be obtained stably throughout the year, industrially, and inexpensively (see JP-A-2000-3000). The present inventors have been conducting various studies on the pharmacological action of the fruit body of B. aeruginosa, which has been available stably for the whole year, and found that the body of B. aeruginosa has a blood sugar level-inhibiting action and a glucose tolerance improving action. The inventors have found that the components shown are present, and have completed the present invention. Disclosure of the invention

That is, the present invention relates to a preventive and ameliorating agent for diabetes comprising a fruit body of a mushroom belonging to the family Crimsonaceae (Climacodontaceae) or a processed product thereof (claim 1), a mushroom belonging to the family Crimsonaceae (Climacodontaceae) belonging to the genus Bunahachitake ( The diabetes preventive and ameliorating agent according to claim 1, which is a mushroom of Mycoleptodonoides) or the mushroom of Mycoleptodonoides, which is a mushroom of Mycoleptodonoides, wherein the mushroom is Mycoleptodonoides aitchisonii. The agent (Claim 3) or the treated product is a dry powder. The preventive and ameliorating agent for diabetes (Claim 4) according to any one of Claims 1 to 3, and the treated product is a solvent extract. The diabetes preventive and ameliorating agent according to any one of claims 3 to 5, and the fruit body of the fungus is a fruit body of a fungus obtained by artificial cultivation. The diabetes preventive and ameliorating agent (Claim 6) and the diabetes preventive and ameliorating agent (Claim 7) according to claim 6, characterized in that the artificial cultivation is bacteria bed cultivation. A sterilization step of sterilizing a medium in which water is mixed with a cultivation nutrient including at least one of dried okara and beer lees; an inoculation step of inoculating the sterilized medium with a B. agaricus inoculum; A pre-culture step of culturing the inoculated medium to obtain a bacterial bed on which the mycelia have grown, and a bunahari that has not been exposed to a physical space capable of culturing the bacterial bed and growing into fruiting bodies. A medium culture step for obtaining bamboo primordia, and a primordium selected from among the Buna hachitake primordium which is not exposed to the physical space where it can grow into the fruiting body. 8. A preventive and ameliorating agent for diabetes mellitus according to claim 7, which comprises a post-culturing step of culturing under conditions exposed to a physical space capable of obtaining a fruit body of Buna-chitake. 9. The diabetes preventive and ameliorating agent (claim 9) according to any one of claims 3 to 8, wherein the fruiting body is a fruit body of Beechachi Ritake (FERMBP-6669). 10. A functional food having a diabetes prevention / amelioration effect, comprising the diabetes prevention / amelioration agent according to any one of 1 to 9 (claim 10). BRIEF DESCRIPTION OF THE FIGURES

 FIG. 1 is a diagram for explaining a method of cultivating a fungus bed of Bunahari yuga, which forms fruiting bodies using a translucent cultivation container. BEST MODE FOR CARRYING OUT THE INVENTION

 The mushroom fruit body used in the present invention is

(Mycoleptodonoides), Mushrooms belonging to the genus Climacodontaceae (Climacodon), Nebarino, and Rinkita (Donkia) are not particularly limited as long as they are fruit bodies of mushrooms belonging to the family Climacodontaceae. Ritake (Mycoleptodonoides aitchisonii)

, Climacodon septentrionalis), Aka small noritake, Climacodon roseo-maculatum) and the like. Preferably, it is used.

The fruit body of the Beech Eighth Ritake may be either a natural fruit body that grows naturally in the natural world or a fruit body that is artificially cultivated. Stable artificially cultivated beech agaric fruiting bodies are preferred. The natural beech Hachitake mushroom fruit that grows in the natural world varies in component content depending on the habitat and climate, and cannot be harvested stably throughout the year. Is not affected by the habitat or climate, and can be harvested stably throughout the year. The method of artificial cultivation is not particularly limited, but fungal bed cultivation, which is capable of inexpensively and stably harvesting beech agaricus having stable quality such as component content, is preferable to log cultivation. Here, the fungal bed cultivation refers to a method of inoculating a seed material into a material composed of a water retaining body and a nutrient source without using a log, and cultivating in a controlled environment of temperature, humidity, illuminance, and the like.

 As preferred embodiments of such fungal bed cultivation, a water retaining body, a culture source for cultivation containing a nutrient source including at least one of dried okara and beer lees, and a sterilization step of sterilizing a medium mixed with water, An inoculation step in which a sterilized medium is inoculated with a fungus inoculated with a fungus, a preculture step in which the medium inoculated with a fungus inoculated with a fungus is obtained, and a culture bed in which hypha grows in the medium is obtained. A medium cultivation step for obtaining a P. agaricus primordium that is not exposed to a physical space capable of growing the fruiting body; and a cultivation primordial primrose that is not exposed to the physical space capable of growing the fruiting body. An artificial cultivation comprising a post-culturing step of cultivating a primordium that is exposed to a physical space in which the fungus can grow into a fruiting body and obtaining a fruiting body of the fungus can be specifically exemplified. . Here, the physical space that can grow into a fruiting body refers to a space in which the fruiting body grows outward from the culture medium.For example, when the culture medium is sealed with a plastic bag or the like, the physical space outside the sealed container can be obtained. Means space.

The B. aeruginosa strain used in the present invention may be a commercially available strain obtained from a natural strain as long as it is a strain belonging to the B. aeruginosa containing a component exhibiting an excellent effect of suppressing the increase in blood glucose level and improving glucose tolerance in the fruit body of the B. aeruginosa. Strains may be any strains including the to be, but it is particularly preferable to use Bunahari Yuke B NH- 3 strain mycelium viability and primordia formation ability are good, the Bunaharitake B NH- 3 strains, Minami Akita In the mountains of Akita-gun, the present inventors isolated purely from the fruiting body that had naturally grown on the dead tree, and the culture on the potato dextrose agar slant medium can be stored at a temperature of about 4 ° C. Yes, it is desirable to transfer this stock strain in about 3 to 6 months. The morphological characteristics of the fruiting bodies, hyphae and spores of this strain are as follows. Fruiting bodies are clusters, umbrellas are fan-shaped to spatula-shaped, 3-8 X 3-10 cm. The surface is hairless and smooth, white to slightly yellowish. The meat is white, 2-5 mm thick.緣 is thin and slightly tooth-like. The hypha consists of two hyphae, a thick hypha with a width of 4 to 10 m and a thin hypha with a swelling and twist with a width of 3.5 to 5 m. Spores are intestinal, colorless, 2-2.5 x 5-6.5 m.

 Based on the morphological characteristics described above, Ryoya Imaseki and Tsuguo Hongo identified the new fungus encyclopedia of Japanese primary colors II (published first edition on May 31, 1989), and found that this fungus was found in Bunakaritake. Clearly belong. This bacterium was established on April 7, 2001 under the Budapest Treaty on International Recognition of the Deposit of Microorganisms for Patent Procedures on April 7, 2011. Deposited as FE RM BP-66697 at the National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary.

 In the fungus bed cultivation of Beech agaric, the water-retaining bodies used in the culture medium include coniferous oakakus derived from conifers such as cedar, hinoki and pine, and oakakus derived from broadleaf trees such as beech, oak and kunugi. In addition to corn cob used as a product (corn corn crushed product), commercially available bacterial bed materials and the like can be exemplified. These materials may be used alone or as a mixture of two or more. It can also be used.

In addition, in the fungus bed cultivation of the fungus beech, As a source, an essential nutrient of either beer lees or dried okara is used in combination with other nutrients other than the selected essential nutrient. Examples of other nutrient sources include rice bran, general bran, dedicated bran, corn bran and the like which are usually used for cultivation of mushrooms. When a nutrient for cultivation that does not contain both beer lees and dried okara is used in the culture medium, not only the growth of mycelia is slow, but also it becomes difficult to obtain fruit bodies that are excellent in size and shape.

 The mixing ratio of the water retaining body and the nutrient for cultivation is preferably in the range of 10: 0.7 to 10: 4, and more preferably in the range of 10: 2 to 3, in terms of fresh weight ratio. Further, the water content may be adjusted to 60 to 70% per final medium, but is more preferably about 65%. Further, as a medium component, soybean hulls, dried yeast, a pH adjuster, and the like, which are usually used for mushroom cultivation, can be added as a medium component.

 As described above, the fungal bed cultivation of B. agarita consists of a pre-culture step, a middle culture step, and a post-culture step, in which the mycelia of B. agaricus grows sufficiently in the medium under specific culture conditions, and the fruiting body is formed. A pre-culture step of obtaining a bacterial bed for formation, a medium culture step of culturing the bacterial bed under a specific culture condition to form a B. agaricus primordium, and culturing the formed B. agaricus primordium under a specific culture condition However, three culturing steps of a post-culturing step of growing to fruiting bodies are employed, and thereby it is possible to obtain fruiting bodies of B. agaricus having a stable content component and the like.

In the pre-culture step, a medium containing a water retention body, a nutrient for cultivation, and water is sterilized under pressure, and then an inoculum of B. agaricus is inoculated at a temperature of 15 to 35, preferably 21 to 27 ° C. Cultivation under dark conditions at a humidity of 40 to 80%, preferably 60 to 70%, spreads mycelia in the medium, and accumulates nutrients for the generation of fruiting bodies. It is a process. Culture ripening days, ie whole medium The number of days required for mycelia to spread and the number of days required for nutrients to accumulate in the mycelium is preferably 25 to 90 days when a 1.2 kg bag is used, and usually less than 25 days. No fruiting body is generated, or it requires a significant number in the later medium culture process. At this time, it goes without saying that the number of days required for the pre-culture step varies depending on the size of the culture vessel used and the amount of the inoculum inoculated.

 The medium culturing step is, as described above, a step performed to form a beech bamboo shoot primordium on the bacterial bed after the completion of the pre-culturing step, and the bacterial bed obtained in the pre-culturing step is heated to a temperature of 8 to 2 hours. 2 ° C, preferably 12 to 16 ° (Humidity 80 to: L 00%, preferably 85 to 95%, illuminance 50 lux or more, preferably 50 to 500 lux When cultivation is continued for 5 to 60 days, for example, Buna hachitake primordium which is not exposed to a physical space capable of growing fruit bodies, such as between the bacterial bed and the inner surface of the container, is formed.

 As described above, the post-culture step is a step performed to grow a P. aeruginosa primordium, which has not been exposed to the physical space that can grow into the fruit body after the completion of the middle culture step, into a fruit body. A beech bamboo shoot primordium is formed which is not exposed to the physical space that can grow on the fruiting body obtained in step (1). For example, remove the surrounding area of the container, temperature 8 ~ 22 ° C, preferably 12 ~ 16 ° C, humidity 80 ~ 100%, preferably 85 ~ 95%, illuminance 50 lux or more Culturing for 5 to 20 days, preferably at 50 to 500 lux, and under conditions where the primordium beech is exposed to a physical space where it can grow into fruiting bodies, Ke primordia grow into fruiting bodies.

For cultivation of Buna mushrooms, cultivation vessels are usually used because of the simplicity of sterilization of the culture medium, and cultivation vessels include cultivation bags. As shown in FIG. 1, the cultivation container 1 is preferably a container made of a transparent or translucent material so that the formed beech agaric primordium 2 can be visually observed from the outside of the container. In addition, grain formed between the fungal bed 3 and the inner surface of the cultivation vessel 1 Cultivating one Bunakari Yuga primordium 2a with a diameter of about 3 cm selected from among the Bunakaritake primordia that are not exposed to the physical space that can grow in the body. It is preferable to obtain <4.Furthermore, the cultivation container 1 having a rectangular cross section or the like is turned sideways so that the Bunakaritake primordium 2a is at the top, and the peripheral portion 5 of the cultivation container in which the It is preferable to harvest the large fruit body 4 of uniform shape by cultivating the fungus body 2a under conditions that expose the physical space where the fungus can grow into fruiting bodies.

 The agent for preventing and / or improving diabetes mellitus of the present invention includes a fruit body of a mushroom belonging to the family Azotaceae or a processed product thereof, preferably a fruit body of a mushroom belonging to the genus Funeraria or a treated product thereof, and more preferably a fruit body of a fungus or a treated product thereof. Any functional food may be used as long as it is an active ingredient. The functional food of the present invention is not particularly limited as long as it contains such a diabetes preventive / ameliorating agent and has a diabetes preventive / ameliorating action. Not done. Here, diabetes prevention and amelioration effect (effect) refers to the effect of suppressing the increase in blood glucose level by suppressing the increase in fasting blood glucose level (effect) and the effect of improving glucose tolerance by suppressing the rapid increase in blood glucose level immediately after eating (effect) The effect of suppressing an increase in blood glucose level can be confirmed, for example, by measuring the concentration of glucose contained in blood after a fast of 16 hours to 20 hours. The improvement effect can be confirmed by performing a glucose tolerance test.

The processed substance of the above-mentioned fruiting bodies is not particularly limited. For example, ground fruiting bodies, room-temperature water, hot water, hexane, getyl ether, acetone, black mouth form, methanol, ethanol Extracts of fruiting bodies using, etc., various enzyme-treated products of fruiting bodies, dried fruiting bodies by air drying, hot air drying, heat drying, freeze drying, microwave drying, etc., or dried powder crushed after drying Granules, capsules or tablets of the body or the dried powder by a conventional method Drying of the fruiting body in terms of suppressing the increase in blood glucose level in the fruiting body and improving the yield of components having an effect of improving glucose tolerance, preventing inactivation, and the use form as a food material, etc. Powders, solvent extracts of fruiting bodies and the like are preferred. The drying method for preparing the dry powder is not particularly limited, but a drying method using a hot air dryer is economically superior, and the drying temperature in such a hot air drying method is 40%. It is preferable to heat at 90 ° C to 90 ° C, especially 60 ° C to 70 ° C for several hours.Drying in this temperature range produces a burning odor and blood glucose level due to the action of enzymes contained in fresh mushrooms. Inhibition of increase · A decrease in glucose tolerance improving activity can be suppressed. If the harvested fruiting body is not processed immediately, it is preferable to store it at a low temperature of 10 ° C or less, for example, at 4 to 5 ° C.

 The diabetes preventive and ameliorating agent of the present invention can reduce fasting blood glucose and suppress a rapid increase in blood glucose immediately after ingestion, thereby preventing and improving diabetes such as suppression of blood sugar elevation and suppression of glucose tolerance. Since it has an action, it is advantageously used as a preventive or symptom-improving agent for diabetes, and as a pharmacological composition material for adding the compound to food to make it a functional food having a preventive / improving action for diabetes. be able to. The diabetes preventive ameliorating agent of the present invention usually has a diabetes preventive and ameliorating effect by taking 100 mg to 20 g / kg body weight in terms of dry fruit body per day. The intake can be adjusted appropriately according to the age, etc. When used as a preventive and / or symptom-ameliorating agent for diabetes, for example, it can be mixed with excipients generally used in pharmaceutical applications and used as an orally administered drug.

The functional food having the diabetes preventive and ameliorating action of the present invention, characterized by containing the diabetes preventive and ameliorating agent of the present invention, uses such a diabetes preventive and ameliorating agent as a part of a raw material for food and drink, Or the manufacturing process or after manufacturing It can be obtained by adding and blending in. Such functional foods are not particularly limited, and include baked goods such as cookies, breads, cakes, rice crackers, etc., tablet confections such as ramune confectionery, Japanese sweets such as yokan, pudding, jelly, ice cream, etc. Confectionery such as frozen desserts, chewing gum, candy, etc., snacks such as crackers, chips, etc., 麵 such as udon, buckwheat, etc., fish paste products such as kamaboko, ham, fish meat sausage, cheese, butter, etc. Dairy products, miso, soy sauce, dressing, mayonnaise, sweeteners, and other seasonings, tofu, konjac, other tsukudani, gyoza, croquettes, salads, soups, stews, etc. , Milk, soy milk, alcoholic beverages, coffee, black tea, green tea, green tea, sports drinks, etc. It can be specifically exemplified. For example, it is possible to produce tablet confections by pulverizing the dried fruit body of the fungus body, and tableting the fine powder according to a conventional method. it can. It is also possible to pulverize the dried fruit body of the fungus body, and mix it with lactose, dextrin, dried yeast and the like.

 Hereinafter, the present invention will be described more specifically with reference to examples, but the scope of the present invention is not limited to these examples.

Example 1 [Bacterial bed cultivation of Bunakaritake]

A total of 100 g, which is a mixture of 380 g of Bunagaguzu, 37.5 g of dried beer lees, 37.5 g of dried okara, and 545 g of tap water, is added to a 1.2 kg PP (poly) Propylene) The medium was prepared by filling the bag. The bag is covered with a polypropylene cap sandwiching a filter, sterilized under pressure at 121 ° C for 50 minutes, and the sterilized medium is cooled. The seeds of Bunakaritake (FERMBP—66997) , And pre-cultured in a dark place at a temperature of 23 to 25 ° C and a humidity of 60 to 80% for 40 days, and then shifted to a medium culture step. Medium culture In this process, cultivation of B. agarita primordium was carried out for 30 days at a temperature of 13 to 15 ° C, a humidity of 80 to 95%, and an illuminance of 200 lux. One portion of the bag where the primordium with a diameter of about 3 cm was observed was cut off, and the process was shifted to the post-culture step. In the post-culture step, the strain grew to a single fruit body of B. agaricus after culturing for 17 days under the conditions of a temperature of 13 to 15 ° C, a humidity of 80 to 95%, and an illuminance of 200 lux. The obtained fruit body of B. agaricus was 232.5 g, and the total number of days required for cultivation was 87 days.

Example 2 [Preparation of dried powder of fruit body of B. agaricus]

 Immediately after harvesting, 3 kg of the fruit body of Beech Hachiritake obtained from the above-described fungal bed cultivation is stored refrigerated at 4 ° C for up to 1 week, and then heated at 65 ° C for 6 hours using a hot air dryer. Drying was carried out to obtain 0.3 kg of a dried product of the fruit body of the fungus. The dried beechari yuga fruit body was pulverized using a crusher to obtain a dried powder of the beech body.

Example 3 [Effect on Type 1 Diabetes]

Using a model rat of streptozotocin (STZ), a type 1 diabetes mellitus model animal, diabetes mellitus model rat, the preventive effect of the fruit body of B. aeruginosa on diabetes mellitus was examined as follows. Approximately 4 weeks old male SD rats were intraperitoneally administered 65 mg / kg streptozotocin to induce diabetes. One week later, rats that showed hyperglycemia, urinary glucose, heavy eating, and heavy drinking were subjected to the following experiment. The STZ-diabetic model rats thus prepared were grouped into 8 rats, and a basic diet group to which CRF-1 (Charles River Japan) was administered as a basic diet, and a beech eight described in Example 2 described above so that the basic diet was 5%. They were grouped into a mixed food group of beech agaric mushrooms mixed with the dried powder of mushroom body. They were reared for 4 weeks in an environment in which food and water were freely available at a room temperature of 23 ° C and a 12-hour light / dark cycle. <Every 1 week from the start of rearing, they were fed from the rat tail vein after a 20-hour fast. Blood was collected and blood glucose was measured. In addition, a glucose tolerance test was performed at weeks 0, 2, and 4. I got it. That is, after a 20-hour fast, 2 g / kg glucose was orally administered using a sonde. Blood was collected from the tail vein before glucose administration (0 hour) and 0.5 hours, 1 hour, 1.5 hours, and 2 hours after administration, and blood glucose levels were measured. The blood glucose level was measured using Glucose CII One Test Co., Ltd. (Wako Pure Chemical Industries, Ltd.).

 The changes in fasting blood glucose during the test period are shown in Table 1, and the results of the glucose tolerance test are shown in Table 2. As can be seen from Table 1, the increase in fasting blood glucose was suppressed in the Beech Hachiriyuga mixed diet group compared to the basic diet group, and the blood glucose level was significantly decreased at the end of the test. Also, from Table 2, there is no difference between the two groups at the start of the test, but in the second and fourth weeks, the rise in blood glucose immediately after glucose administration was suppressed, and the recovery of blood glucose was quick, and therefore, the basic diet It can be seen that the glucose tolerance is improved in the mixed group of beech and garland mushrooms compared to the group. From the above, it was shown that B. agarita has preventive and ameliorating effects on type 1 diabetes.

table 1

表 2 血糖値 (ms/dl)

Table 2 Blood glucose (ms / dl)

 Duration Treatment group Blood sampling time (hours)

 0.0 0.5 1.0 1.5 2.0

Week 0 Basic diet group 1 15.1 ± 12.8 410.3 ± 15.2 Approx. 5 ± 19.2 319.6 ± 19.9 236.4 ± 23.8 Bunakaritake mixed diet group 132.0 ± 18.3 449.7 ± 26.7 420.4 ± 30.4 304.0 ± 40.0 208.2 ± 36.1

 18.5

Week 2 Basic food group 314.4 ± 39.9 620.4 ± 26.8 597.9 ± 19.9 538.6 ± 16.5 500.9 ±

 Beech agaric mixed diet group 238.2 ± 57.8 453.8 + 25.3 * 484.0 ± 25.8 * 41 1.7 ± 38.8 伞 370.4 ± 50.5 *

 8.2 608.9 Sat 17.5 572.3 ± 19.4

Week 4 Basic diet group 492.8 ± 43.4 801.4 + 43.1 727.0 ± 2

 Beech agaric mixed diet group 353.3 ± 69.8 * 626.3 ± 49.9 * 647.3 ± 48.4 521.6 ± 31.1 * 456.0 ± 39.6 *

*: There is a significant difference at risk of 5% Example 4 [Effect on Type 2 Diabetes]

Using KKA ^y diabetes model mice, which are a type 2 diabetes model animal, the effect of the fruit body of Buna hachitake was examined in the same manner as in type 1 diabetes as follows. The KKA ^y mouse is a type 2 diabetes model mouse in which the obesity gene A ^{y is} introduced into the KK mouse exhibiting hyperglycemia and the obesity and hyperglycemia is expressed. Widely used. Approximately 4 weeks old male KKA ^y ZT a Jc 1 mice were purchased from Clea Japan, and after one week of preliminary breeding, they were divided into 8 animals per group as in the type 1 diabetes test. The test was started in groups of mixed diets. Mice were housed individually in cages at room temperature of 23 ° (:, 12 hours light / dark cycle, with free access to food and water for 5 weeks. Blood was collected from the fundus of the mouse using a capillary blood collection tube, blood glucose was measured, and a glucose tolerance test was performed 5 weeks after the breeding was started, ie, after fasting for 20 hours, 2 g / kg of glucose Blood was collected from the fundus before glucose administration (0 hour), 0.5 hours, 1 hour, 1.5 hours, and 2 hours after glucose administration, and blood glucose levels were measured. The changes in the values are shown in Table 3. Compared to the basic diet group, the blood glucose level was significantly lower in the Bunahari Yuga mixed diet group, and the increase in the blood glucose level was suppressed. The results are shown in Table 4. Compared to the basic diet group, Bunach In the bamboo mixed diet group, it was confirmed that the increase in blood glucose level immediately after glucose administration was significantly suppressed, and that the blood glucose level was recovered quickly. It was shown.

Four Table 3

表 4

Table 4

実施例 5 [筋細胞に対する効果]

Example 5 [Effect on muscle cells]

Using the myoblast L6 derived from rat skeletal muscle, the effect of promoting glucose uptake on the myocytes of B. agaricus was demonstrated. L6 cells used for the test were purchased from Dainippon Pharmaceutical Co., Ltd. L 6 cells were cultured at 3 7 ° C, 5% C_〇 ₂ concentration under using carbon dioxide gas incubator at basal medium (D-MEM medium containing 1 0% FBS). Cells in the growth phase were prepared and suspended in a basal medium at ⁴ × 10 ⁴ cells / ml. This was added to a 24-multiwell cell culture plate (30464, manufactured by Falcon) in an amount of 1 ml each, and the cells were cultured until they became conferred. The medium of cells grown in the conference was replaced with a differentiation-inducing medium (D-MEM medium containing 2% FBS) to induce differentiation. Thereafter, the medium was replaced with a differentiation-inducing medium every 48 hours, and multinucleated myotube cells were obtained 45 days later.

The preparation of the Beechari Yuga extract was performed as follows. 0.25 of the dried powder of Beech agaricus described in Example 2 above was mixed with hexane, gethyruether, acetone, 10 ml of form, methanol, ethanol, and water respectively. Add 1 and shake overnight. For hot water extraction, 10 ml of water was added to 0.25 g of the dried powder of Bunakaritake, and the mixture was heated at 105 ° C for 5 minutes using an autoclave. These extracts were filtered with filter paper, and the filtrate was dried under reduced pressure to obtain an extract. Table 5 shows the yields. Hexane, getyl ether, acetone, chloroform, methanol, and ethanol extracts are dimethyl sulfoxide (DI MSO), and water and hot water extracts are water. And dissolved to obtain an extract.

Table 5

分化誘導して得られた多核筋管細胞の培地を、 新鮮な分化誘導培地に 交換した後、 1 1のブナハリタケ抽出液をそれぞれ培地に加え、 培養 した。 抽出液添加 48時間後に培地を取り出し、 培地中のグルコース量 を測定した。 培地中に残っているグルコース量を表 6に示す。 培地中に ブナハリタケ抽出物を添加していない無添加と比較して、 ジェチルェ一 テルとクロ口ホルム抽出液で、有意にグルコース量の低下が観察された。 従って、 ブナハリタケには筋組織においてグルコースの取り込みを増加 させ、 血糖値を低下させる可能性が示された。

After replacing the medium of the multinucleated myotube cells obtained by inducing differentiation with a fresh differentiation inducing medium, 11 B. agaricus extracts were added to each medium and cultured. The culture medium was removed 48 hours after the addition of the extract, and the amount of glucose in the culture medium was measured. Table 6 shows the amount of glucose remaining in the medium. Compared with the case where no Bunakaritake extract was added to the medium, a significant decrease in the amount of glucose was observed in the extract of Jetilether and in the extract of black-mouthed form. Thus, it was shown that B. agarita may increase glucose uptake in muscle tissue and lower blood glucose levels.

6 Table 6

 *: Significant difference at 5% risk Example 6 [Effect on fat cells]

 Preadipocytes 3 T 3 L 1 cells derived from mouse adipose tissue:

The effect of mushrooms on promoting glucose uptake on fat cells was examined. 3T3L1 cells used in the test were purchased from Dainippon Pharmaceutical Co., Ltd. The 3T3L1 cells were cultured in a basal medium (D-MEM medium containing 10% FBS) under a 37,5% C% ₂ concentration using a gas carbonate incubator. Cells in the growth phase were prepared and suspended in a basal medium to 5 × 10 ⁴ Zm 1. This was added to a 24 multi-well cell culture plate every 1 ml, and cultured until it became a confident. The culture medium of the cells grown in the conferent cells is transformed into a differentiation-inducing medium (basic medium contains 0.25 M dexamethasone, 0.5 mM 1-methyl-3-isobutylxanthine, and 10 Hg / m1 insulin). Medium) to induce differentiation. 48 After 8 hours, replace the medium with a maturation promoting medium (medium containing 5 g / m1 of insulin in the basal medium), and then replace the maturation promoting medium every 48 hours. After 4 to 5 days, mature adipocytes I got

After the medium of the mature adipocytes was replaced with a fresh basal medium, the fungus mushroom extract 1 I 1 prepared in Example 5 was added to each medium and cultured. The culture medium was taken out 48 hours after the addition of the extract, and the amount of glucose in the medium was measured. Table 7 shows the amount of Darcos remaining in the medium. As can be seen from Table 7, the amount of glucose was significantly lower in the hexane, getyl ether, acetone, chloroform, and methanol extracts compared to the case without the addition of the Buna mushroom extract in the medium. Was observed. Therefore, it was shown that B. agarita may increase glucose uptake in adipose tissue and decrease blood glucose level.

Table 7

 *: Significant difference with a risk factor of 5¾ Industrial applicability

 The functional food having the diabetes preventive and ameliorating agent and the diabetes preventive and ameliorating effect of the present invention is derived from a natural product, mushroom, and is highly safe, and has a blood sugar level suppressing effect on type 1 diabetes and type 2 diabetes. Since it has a glucose tolerance improving effect, ingestion of the antitumor agent of the present invention or a functional food having a tumor improving effect can be expected to have a preventive effect on diabetes and a therapeutic effect on diabetes. In addition, by using an artificially cultivated product of Buna hachitake as a fruiting body, it is possible to manufacture an inexpensive and stable agent for preventing and ameliorating diabetes mellitus with stable quality throughout the year.

Claims

The scope of the claims 1. An antidiabetic agent comprising a mushroom fruit body of Climacodontaceae or a processed product thereof as an active ingredient. 2. The diabetes preventive and ameliorating agent according to claim 1, wherein the mushroom of the family Climacodontaceae is a mushroom of the genus Mycoleptodonoides.  3. The preventive and ameliorating agent for diabetes according to claim 2, wherein the mushroom of the genus Mycoeptodonoides is Mycoeptodonoides aitchisonii.  4. The diabetes prevention / improvement agent according to any one of claims 1 to 3, wherein the processed product is a dry powder.  5. The diabetes prevention / improvement agent according to any one of claims 1 to 3, wherein the processed product is a solvent extract. 6. The preventive or ameliorating agent for diabetes according to any one of claims 3 to 5, wherein the fruit body of the fungus is a fruit body of a fungus obtained by artificial cultivation. 7. The preventive and ameliorating agent for diabetes according to claim 6, wherein the artificial cultivation is a fungal bed cultivation. 8. Bacterial bed cultivation is a sterilization step of sterilizing a medium in which water is mixed with a cultivation nutrient including at least one of a water retention body, dried okara and beer lees, and a seed culture of B. aeruginosa on the sterilized medium. An inoculation step of inoculation, a pre-culturing step of culturing a medium inoculated with a Bacillus inoculum inoculum and obtaining a bacterial bed on which the mycelia have grown, and a physical space in which the bacterial bed is cultured and can grow into fruiting bodies. A medium culturing step for obtaining unexposed B. agaricus primordium, and a primordia selected from among the unexposed B. agaricus primordia that have not been exposed to a physical space that can grow into the fruiting body. Under the conditions exposed to the physical space 8. The preventive and ameliorating agent for diabetes according to claim 7, which comprises a post-culturing step of culturing to obtain a fruit body of B. aeruginosa.  9. The preventive and ameliorating agent for diabetes according to any one of claims 3 to 8, wherein the fruiting body of Bunakaritake is a fruiting body of Bunakariyuke (FERMBP-66697).  10. A functional food having a diabetes prevention / ameliorating action, comprising the diabetes prevention / ameliorating agent according to any one of claims 1 to 9.