Antiallergic agent, intestinal immunoenhancer, intestinal adhesiveness improver of lactic acid bacteria

Abstract

To provide antiallergic agents, intestinal immunoenhancers, and intestinal adhesiveness improvers of lactic acid bacteria, which are safely ingestible, are inexpensively available, and are excellent in the intestinal adhesiveness improving action of lactic acid bacteria.SOLUTION: The agent of the present invention contains stems and / or leaves of plants which are green juice materials. The plants which are green juice materials preferably comprise one or more selected from (1) Hordeum vulgare, (2) Ipomoea batatas, (3) Brassica oleracea, (4) Morus, and (5) Peucedanum japonicum, more preferably comprise two or more of the (1) to (5), and further more preferably comprise the (1) and one or more selected from (2) to (5).SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to an antiallergic agent, an intestinal immunity enhancer, and an agent for improving the intestinal adhesion of lactic acid bacteria. [Background technology]

[0002] The human immune system is dominated by several types of T cells. There are two types of T cells: Th1 cells, which mainly stimulate IgA antibody production and activate cellular immunity, and Th2 cells, which stimulate IgE antibody production, which causes type I allergies, and activate humoral immunity. These Th1 and Th2 cells suppress each other, and allergic symptoms are more likely to occur when Th2 cells are dominant over Th1 cells.

[0003] It is said that 70% of the body's immune cells are concentrated in the intestinal tract, where they play a major role in the body's immune system. In recent years, it has become clear that lactic acid bacteria play an important role in intestinal immunity. For example, it is known that the bacterial components of lactic acid bacteria promote cytokine secretion from macrophages and dendritic cells in the intestinal tract, thereby activating the activity of Th1 cells and promoting IgA antibody production. This is thought to suppress the activity of Th2 cells, suppressing IgE antibody production, and improving allergies.

[0004] Adhesion of lactic acid bacteria (and their bacterial components) to the intestinal tract is considered to be very important for enhancing the intestinal immune stimulating effect and the associated anti-allergic effect. If oral agents that improve the intestinal adhesiveness of lactic acid bacteria can be used to promote intestinal adhesion, it is expected that allergies can be prevented or improved. Such oral agents must be highly safe and inexpensive to obtain, but their development has not progressed sufficiently and the diverse needs of patients and consumers have not been fully met.

[0005] On the other hand, green juice is a product that uses processed plant stems and / or leaves, such as dried powder or juice powder, and is used as a health food that allows vegetable ingredients to be easily ingested. The plant stems and / or leaves used as ingredients for green juice are known as safe and inexpensive food ingredients, and various effects have been investigated so far (e.g., Patent Document 1), but there has been no study on the anti-allergic effect focusing on the role of lactic acid bacteria in intestinal immunity. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] JP 2003-267880 A Summary of the Invention [Problem to be solved by the invention]

[0007] Therefore, an object of the present invention is to provide a novel agent for improving the intestinal adhesion of lactic acid bacteria, an agent for enhancing intestinal immunity, and an antiallergic agent. [Means for solving the problem]

[0008] The present inventors have conducted extensive research into substances capable of increasing the intestinal adhesiveness of lactic acid bacteria, and have found that the intestinal adhesiveness of lactic acid bacteria can be increased by using the stems and / or leaves of plants that are green juice ingredients, which are inexpensive and safe to obtain.

[0009] The present invention is based on the above findings, and provides an anti-allergic agent (excluding anti-allergic agents that inhibit interleukin 4 production) containing the stems and / or leaves of plants that are the raw materials for green juice.

[0010] The present invention also provides an intestinal immunity enhancer and an intestinal adhesion enhancer for lactic acid bacteria, which contain the stems and / or leaves of the plant that is the green juice ingredient. Effect of the Invention

[0011] According to the present invention, there are provided an antiallergic agent, an intestinal immunity enhancer, and an agent for improving the intestinal adhesion of lactic acid bacteria, which can be safely ingested, are available at low cost, and have an excellent effect of improving the intestinal adhesion of lactic acid bacteria. [Brief description of the drawings]

[0012] [Figure 1] FIG. 1 is a graph showing the results of evaluating the intestinal adhesion effect of the agents of the Examples and Comparative Examples. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] The present invention will be described below based on its preferred embodiments. The agent of the present invention contains the stems and / or leaves (hereinafter simply referred to as "stems and leaves") of a plant that is a raw material for green juice. Unless otherwise specified, the following description applies to the antiallergic agent, the intestinal immunity enhancer, and the agent for improving the intestinal adhesion of lactic acid bacteria of the present invention. The stems and leaves may be only stems or only leaves, or a mixture thereof, but are preferably only leaves or a mixture of stems and leaves. Furthermore, the agent of the present invention may contain flowers, roots, etc. in addition to stems and leaves as the plant that is a raw material for green juice, or may not contain any parts other than the stems and leaves.

[0014] Preferred examples of plants that are the raw materials for green juice include (1) barley, (2) sweet potato, (3) kale, (4) mulberry, and (5) Peony root. These are described below.

[0015] (1) Barley (scientific name: Hordeum vulgare) includes two-rowed barley, six-rowed barley, naked barley, etc., and these can be used alone or in combination of two or more. Barley stalks and leaves are preferably harvested before maturity, that is, from the start of tillering to the start of heading. Specifically, although it varies depending on the variety, it is generally preferable to harvest stalks and leaves from barley having a height of 10 cm or more, preferably about 10 to 90 cm, particularly preferably about 20 to 80 cm, and particularly preferably about 30 to 70 cm, but are not limited thereto. Barley stalks and leaves used in the present invention are not particularly limited as long as they are usually available, and those obtained by the processing method described below or commercially available products can be used.

[0016] (2) Sweet potato (scientific name Ipomoea batatas) includes Suio, Joy White, Koganesengan, Shiroyutaka, Satsuma Sachi, Ayamurasaki, etc., and these can be used alone or in combination of two or more. Among sweet potatoes, Suio, which has a high polyphenol content, is preferred. Suio has the characteristic that young sweet potato stems and leaves regenerate from the tip of the same stem even after the stems and leaves have been harvested once, so it is also suitable for use in terms of productivity. As sweet potato stems and leaves, it is preferable to use stems and / or leaves that are exposed above the ground during sweet potato cultivation. Sweet potato stems and leaves that have grown 5 cm or more, more preferably 10 cm or more, and even more preferably 20 cm or more out of the ground are preferred. In addition, the length from the position where the sweet potato stem is exposed above the ground to the tip of the sweet potato stem and leaves is preferably 300 cm or less, more preferably 200 cm or less, and even more preferably 150 cm or less. As sweet potato stalks and leaves, the tip part of sweet potato stalks and leaves ("young sweet potato stalks and leaves") is preferred, and young sweet potato stalks and leaves that retain a yellowish green color compared to other stalks and leaves are more preferred. As sweet potato stalks and leaves, stalks and leaves within 100 cm from the tip of sweet potato stalks and leaves are preferably used. Young sweet potato stalks and leaves are easy to process because the plant body itself is soft. Furthermore, when dried and powdered, young sweet potato stalks and leaves have a good texture and can be easily used in various foods. The sweet potato stalks and leaves used in the present invention are not particularly limited as long as they are normally available, and those obtained by the processing method described below or commercially available products can be used.

[0017] (3) Kale is Brassica oleracea L. (a crop of the genus Brassica). Specific examples of kale include kitchen kale, tree kale, bush kale, marrow kale, collard, green-leaf kale, drinking leaf, Portuguese kale, Chinese kale, thousand-headed kale, and Jersey kale, and these may be used alone or in combination of two or more. The kale stems and leaves used in the present invention are not particularly limited as long as they are usually available, and those obtained by the processing method described below or commercially available products may be used.

[0018] (4) Mulberry is a general term for the genus Morus (scientific name: Morus) of the family Moraceae. Examples of mulberry include Japanese mulberry, Japanese mulberry, log mulberry, striped mulberry, and Korean mulberry, as well as their hybrids and hybrids. These may be used alone or in combination of two or more. The mulberry stems and leaves used in the present invention are not particularly limited as long as they are normally available, and those obtained by the processing method described below or commercially available products may be used.

[0019] (5) Peucedanum japonicum (scientific name: Peucedanum japonicum) is a perennial plant of the Apiaceae family, Peucedanum genus. It is also known as Chou-mei-sou, and grows wild mainly from southern Kyushu to Okinawa. The stems and leaves of Peucedanum japonicum used in the present invention are not particularly limited as long as they are normally available, and those obtained by the processing method described below or commercially available products can be used.

[0020] In the present invention, various processed products obtained from the stems and leaves of plants that are green juice ingredients can be used. Examples of such processed products include dried powder (sometimes called pulverized powder) obtained by drying and pulverizing the stems and leaves, shredded stems and leaves and their dried powder, juice obtained by squeezing the stems and leaves and their dried powder, and extract obtained by extracting the stems and leaves with water, an organic solvent or a mixture thereof and their dried powder. These may be products obtained through a process of fermenting the stems and leaves (fermented products) or products obtained without a fermentation process (non-fermented products).

[0021] For example, a conventionally known method can be used to dry and powder the stems and leaves (to obtain a pulverized powder). As such a method, a method in which a drying process and a pulverization process are combined for the stems and leaves can be used. Either the drying process or the pulverization process may be performed first, but it is preferable to perform the drying process first. The drying and powderization process may be combined with one or more processes selected from a blanching process, a sterilization process, etc., as necessary. The pulverization process may be performed once or in combination of two or more processes, but it is preferable to combine a coarse pulverization process with a fine pulverization process for more fine pulverization.

[0022] The blanching treatment is a treatment for keeping the green color derived from the stems and leaves vivid, and examples of the blanching treatment include hot water treatment and steam treatment. The heating temperature in the blanching treatment is preferably higher than 80°C, more preferably 85°C or higher, and even more preferably 90°C or higher. The heating temperature is preferably 100°C or lower. The heating time in the heat treatment is preferably 10 minutes or less, more preferably 5 minutes or less, even more preferably 3 minutes or less, and particularly preferably 10 seconds to 1 minute. In addition, when hot water treatment is performed as the blanching treatment, it is preferable to dissolve carbonates such as magnesium carbonate or hydrogen carbonates such as sodium hydrogen carbonate in the hot water, since this can make the green color of the stems and leaves more vivid. In this case, the pH of the hot water is preferably 5.4 or more, more preferably 5.6 to 8.4, and even more preferably 5.6 to 8.0. In addition, as the steam treatment, an intermittent steam treatment in which the process of steaming the stems and leaves with water vapor and the process of cooling are repeated under normal pressure or pressure is preferable. In the intermittent steaming treatment, the steaming treatment with water vapor is preferably performed for 20 to 40 seconds, more preferably 30 seconds. The cooling treatment after the steaming treatment is preferably performed immediately, and the method is not particularly limited, but immersion in cold water, refrigeration, cooling with cold air, evaporative cooling with hot air, evaporative cooling by combining hot air and cold air, etc. are used. Among these, evaporative cooling by combining hot air and cold air is preferred. Such a cooling treatment is performed so that the product temperature of the stems and leaves is preferably 60°C or less, more preferably 50°C or less, and most preferably 40°C or less. In addition, in order to produce a powder of stems and leaves rich in nutritional components such as vitamins, minerals, and chlorophyll, it is preferable to repeat the intermittent steaming treatment 2 to 5 times.

[0023] Sterilization is a process that usually uses temperature, pressure, electromagnetic waves, chemicals, etc. to physically and chemically kill microbial cells. Among these methods, heat sterilization is a preferred method because it can improve the flavor of the stems and leaves. The heat treatment is preferably performed at a temperature of 110°C or higher, and specific equipment that can be used includes a high-pressure sterilizer, a heat sterilizer, and a pressurized steam sterilizer. For example, in the case of heat treatment by pressurized steam sterilization, coarsely crushed sweet potato stems and leaves are sterilized at a pressure of, for example, 0.5 kg / cm. 2 More than 10kg / cm 2 It is preferable that the pasteurized food is sterilized by heating with saturated steam at 110° C. to 200° C. for 2 to 10 seconds under the following pressure: After the pasteurization, the pasteurized food may be further dried to remove moisture contained in the pasteurized food during heating with saturated steam, if necessary.

[0024] In addition, the drying process is preferably a process in which the moisture content of the stems and leaves after drying is 10% or less, particularly 5% or less, since a dried powder of stems and leaves with good flavor and bright color can be obtained. This drying process can be performed by any method known to those skilled in the art, such as hot air drying, high-pressure steam drying, electromagnetic wave drying, and freeze drying. Drying by heating can be performed at a temperature and for a time period at which the stems and leaves do not discolor due to heating. In terms of obtaining a dried powder of stems and leaves with good flavor and bright color, it is preferable to perform the drying process at 60°C or more and 150°C or less, preferably 70°C or more and 100°C or less. When the stems and leaves are dried as they are, it is preferable to perform drying in two stages. The two-stage drying can be performed, for example, using a hot air dryer. In the two-stage drying, first, the stems and leaves are primarily dried at a temperature of 60°C or more and 80°C or less until the moisture content is 25% by mass or less. Then, the primary dried stems and leaves are secondary dried at a temperature higher than the primary drying temperature, preferably 70°C or more and 90°C or less, until the moisture content is 5% by mass or less.

[0025] The crushing treatment may be carried out by any method commonly used by those skilled in the art using a crusher, a mill, a blender, a stone mill, etc. The crushed stems and leaves may be sieved as necessary.

[0026] When blanching is performed in addition to drying and grinding, the blanching is preferably performed before drying. When sterilization is performed in addition to drying and grinding, the sterilization is preferably performed after drying or before or after grinding.

[0027] A specific method for drying and powdering includes, for example, cutting the stems and leaves, performing a blanching treatment, then drying so that the moisture content is 10% by mass or less, preferably 5% by mass or less, and then pulverizing (see JP 2004-000210 A). Another example includes cutting the stems and leaves, performing a blanching treatment, then rolling, then drying, and pulverizing (see JP 2002-065204 A). Another example includes drying the stems and leaves, coarsely pulverizing them, heating them at 110° C. or higher, and further finely pulverizing them (see JP 2003-033151 A). The particle size of the powder is preferably such that 50% by mass or more, particularly 70% by mass or more, passes through a 200 mesh.

[0028] As a method for shredding the stems and leaves, a method that is usually used by those skilled in the art when shredding a plant body, such as slicing, crushing, or chopping, can be used. As an example of shredding, the stems and leaves may be slurried. Slurrying is performed by subjecting the stems and leaves to a mixer, juicer, blender, mass colloider, or the like, and turning the stems and leaves into a mushy gruel (a suspension of liquid and solid). By slurried in this way, the stems and leaves are shredded so that 80% by mass or more of the shredded pieces preferably have an average diameter of 1 mm or less, more preferably 0.5 mm or less, even more preferably 0.1 mm or less, and most preferably 0.05 mm or less, and the shredded pieces become fluid.

[0029] Methods for squeezing the stems and leaves include squeezing the stems and leaves or shredded material thereof, or centrifuging or filtering the shredded material. A representative example is a method in which the juice is extracted by mechanical crushing means such as a mixer or juicer, and if necessary, the crude solid content is removed by means of sieving, filtration, or the like to obtain a squeezed juice, and specific examples of the method include those described in JP-A-08-245408 and JP-A-09-047252. In addition, the method for obtaining an extract of stems and leaves includes a method in which an extraction solvent commonly used by those skilled in the art, such as ethanol, water, or aqueous ethanol, is added to the stems and leaves or shredded material thereof, and extraction is performed by heating as necessary. The extract may be concentrated as necessary.

[0030] Among the processed products of stems and leaves, it is particularly preferable to use crushed products (crushed powder, etc.) or squeezed juice (squeezed juice powder, etc.) of stems and leaves, because the agent of the present invention can be made more vivid in color and flavor, and can be made rich in dietary fiber. Commercially available products can be used as processed products. For example, in the case of barley, commercially available crushed powder of barley stems and leaves (trade name "Barley Young Leaf Powder", etc.) or squeezed powder (trade name "Barley Young Leaf Extract", etc.) can be used, and in the case of sweet potato, commercially available crushed powder of sweet potato stems and leaves (trade name "Suio", etc.) can be used.

[0031] In terms of enhancing the intestinal adhesion activity of lactic acid bacteria, the agent of the present invention is more preferably such that the plant serving as the raw material for green juice contains two or more of the above-mentioned (1) barley, (2) sweet potato, (3) kale, (4) mulberry, and (5) Peony Bouhu. In particular, the plant serving as the raw material for green juice preferably contains (1) barley and at least one selected from (2) sweet potato, (3) kale, (4) mulberry, and (5) Peony Bouhu. More preferably, the agent contains (1) barley and two or more selected from (2) sweet potato, (3) kale, (4) mulberry, and (5) Peony Bouhu. When the agent contains (1) and two or more selected from (2) to (5), it is particularly preferable that the agent contains (1) barley, (3) kale, and (5) Peony Bouhu. Most preferably, the composition contains all of (1) barley, (2) sweet potato, (3) kale, (4) mulberry, and (5) Peony.

[0032] The total amount of stems and leaves of plants that are green juice ingredients in the agent of the present invention is preferably 0.01% by mass or more and 90% by mass or less, more preferably 0.05% by mass or more and 80% by mass or less, and particularly preferably 0.1% by mass or more and 70% by mass or less, in terms of dry mass in the agent of the present invention. The total amount referred to here is the content of only one plant that is a green juice ingredient when the agent of the present invention contains only one plant. When the agent of the present invention contains the stems and leaves of any of the plants (1) to (5) above, the preferred range of the total amount of stems and leaves of plants (1) to (5) in the agent of the present invention is the same as the total amount of stems and leaves of the plants that are green juice ingredients.

[0033] In particular, from the viewpoint of enhancing the intestinal adhesion effect of lactic acid bacteria by the agent of the present invention, when the agent contains the stems and leaves of any of the plants (1) to (5) above, the stems and leaves of each of the plants (1) to (5) are preferably contained in an amount of 0.01% by mass or more and 80% by mass or less, more preferably 0.05% by mass or more and 70% by mass or less, and particularly preferably 0.1% by mass or more and 50% by mass or less, on a dry mass basis, in the agent of the present invention.

[0034] When the agent of the present invention contains the above-mentioned (1) and one or more of (2) to (5), the total dry mass of (2) to (5) per 100 parts by mass of (1) is preferably 0.1 to 1,000 parts by mass, more preferably 0.5 to 800 parts by mass, and particularly preferably 1 to 500 parts by mass.

[0035] The agent of the present invention may consist essentially of only (1) to (5) as plant-derived components, or may contain other plant-derived components. For example, the plant-derived components other than (1) to (5) in the agent of the present invention are preferably 70% by mass or less, more preferably 50% by mass or less, in the agent of the present invention.

[0036] The agent of the present invention may not contain lactic acid bacteria, but it is preferable to contain lactic acid bacteria in order to further enhance the intestinal adhesion effect of lactic acid bacteria. The lactic acid bacteria may be live or dead. If it is live, it is preferable from the viewpoint of improving the intestinal environment, but it may be dead. This is based on the fact that not only live bacteria but also dead bacteria are considered to stimulate intestinal immunity. The lactic acid bacteria used in the present invention are not particularly limited as long as they produce lactic acid as a metabolic product, and include those that have been orally ingested in animals such as humans, for example, those of the genus Bifidobacterium, Lactbacillus, Enterococcus, Leuconostoc, Pediococcus, Staphylococcus, Tetragenococcus, and Bacillus. Bifidobacterium genus includes Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium lactis, Bifidobacterium longum, Bifidobacterium adolescentis, and Bifidobacterium mongoliense. Lactobacillus genus includes Lactobacillus brevis, Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus buchneri, Lactobacillus bulgaricus, Lactobacillus delburvecki, Lactobacillus casei, Lactobacillus crispatus, Lactobacillus curvatus, Lactobacillus halivaticus, Lactobacillus Examples include Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus salivarius, Lactobacillus sporogenes, Lactobacillus sakei, Lactobacillus fructivorans, Lactobacillus hilgardii, Lactobacillus reuteri, and Lactobacillus fermentum. Enterococcus includes Enterococcus faecalis (sometimes called Streptococcus faecalis), Enterococcus faesium (sometimes called Streptococcus faesium), Streptococcus thermophilus, and Lactococcus lactis (sometimes called Streptococcus lactis). Examples of the Leuconostoc genus include Leuconostoc mesenteroides and Leuconostoc oenos. The genus Pediococcus includes Pediococcus acidilactici and Pediococcus pentosaceus. The genus Staphylococcus includes Staphylococcus carnosus and Staphylococcus xylosus. The genus Tetragenococcus includes Tetragenococcus halophilus, and the genus Bacillus includes Bacillus coagulans and Bacillus mesentericus. Among these, Bacillus coagulans, Enterococcus faecalis, Bifidobacterium bifidum, Enterococcus faesium, and Lactobacillus acidophilus are preferable. These may be used alone or in combination of two or more.

[0037] When the agent of the present invention contains lactic acid bacteria, the content of the lactic acid bacteria is not particularly limited. For example, the number of bacteria in the agent of the present invention is 1×10 3 10 or more, preferably 1×10 5 pcs or more 1×10 20 10 or less, more preferably 1×10 7 pcs or more 1×10 15 A number of particles or less is preferable from the viewpoints of ease of formulation and enhancing the effect of the lactic acid bacteria adhering to the intestinal tract.

[0038] When the agent of the present invention contains lactic acid bacteria, the lactic acid bacteria and the stems and leaves of the plant may be contained in the same agent or in separate agents. When contained in separate agents, the agent of the present invention is a multi-drug type of two or more agents. When contained in the same agent, the agent of the present invention may be a single-drug type or a multi-drug type, but a single-drug type is preferable in terms of ease of intake, etc. The dosage form of the agent of the present invention may be a solid, liquid, paste, gel, etc., regardless of whether it is a single-drug type or a multi-drug type. For example, solid forms include powder, granules, tablets, rods, plates, blocks, solid forms, capsules such as hard capsules and soft capsules, caplets, tablets, chewable forms, sticks, etc. Examples of liquid forms include fluid forms and syrup forms. These can be formulated into various dosage forms by known formulation methods by adding pharmaceutically acceptable base materials or carriers.

[0039] For example, preferred dosage forms of the agent of the present invention containing lactic acid bacteria include a single dosage form containing lactic acid bacteria and the stems and leaves of the plant and being in any of powder, fine granules, granules, tablets, capsules such as soft capsules and hard capsules, and liquid, and a two-dosage form having a first agent containing the stems and leaves of the plant and being in any of powder, fine granules, granules, tablets, capsules such as soft capsules and hard capsules, and liquid, and a second agent containing lactic acid bacteria and being in any of powder, fine granules, granules, tablets, capsules such as soft capsules and hard capsules, and liquid. Note that in the present invention, the intestinal adhesiveness of lactic acid bacteria and bifidobacteria present in the intestine can also be improved by using the first agent containing the stems and leaves of the plant alone.

[0040] The oral intake method of the agent of the present invention is not limited, and may be taken as it is, or may be taken after dispersing or dissolving the agent in water, hot water, milk, yogurt, etc. In addition, when the agent of the present invention is a multi-drug type, the multiple agents may be taken simultaneously or at different times. When taken at different times, for example, either the lactic acid bacteria-containing agent or the plant stem and leaf-containing agent may be taken first, but the time difference between the intake of the two is preferably within 24 hours, more preferably within 18 hours, and particularly preferably within 15 hours. In addition, the intake methods of the two agents may be the same or different, and when they are different, for example, the lactic acid bacteria-containing agent is taken as it is, and the plant stem and leaf-containing agent is taken after dispersing in water, hot water, milk, yogurt, etc., or this may be reversed.

[0041] The agent of the present invention may contain other commonly used components in addition to the plant stems and leaves and lactic acid bacteria, as long as the effects of the present invention are not impaired. Such components include various excipients, binders, gloss agents, lubricants, stabilizers, diluents, bulking agents, thickeners, emulsifiers, antioxidants, pH adjusters, colorants, flavorings, additives, etc. The content of other components can be appropriately selected depending on the form of the agent of the present invention. In addition, when the agent of the present invention is a multi-form, which other components are contained in which agent can be appropriately selected depending on the form of the agent.

[0042] The daily oral dose of the agent of the present invention is preferably 10 mg or more as the dry mass of the stems and leaves of the plant. The agent of the present invention can be administered continuously, for example, every day, and can be administered continuously for a long period of time, for example, for one month or more.

[0043] Regardless of whether the agent of the present invention contains lactic acid bacteria or not, the daily oral dose of lactic acid bacteria is 1×10 3 More than 10, especially 1×10 5 pcs or more 1×10 20 It is preferable that the number is less than 1.

[0044] As will be apparent from the description of the examples below, the agent of the present invention can enhance the intestinal adhesiveness of lactic acid bacteria by the action of the stems and leaves of the plant. The intestinal adhesiveness referred to here preferably means the adhesiveness to the intestinal epithelium (intestinal epithelial cells). Therefore, the agent of the present invention can enhance the action of lactic acid bacteria in the intestinal tract by orally taking it, and can be used, for example, as an intestinal immunity enhancer, and can prevent or improve allergic symptoms such as atopic dermatitis, allergic rhinitis, hay fever, eczema, urticaria, and other rashes, diarrhea, and vomiting. The action of the intestinal immunity enhancer includes activating the activity of Th1 cells and promoting IgA antibody production. The agent of the present invention is preferably applied to humans, but can also be applied to animals other than humans (e.g., mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as each action and effect is exerted. The allergy agent of the present invention excludes those that exert anti-allergic action by suppressing interleukin 4 production.

[0045] Examples of lactic acid bacteria whose intestinal adhesion (attachment) is enhanced by the agent of the present invention include the same lactic acid bacteria as those listed above as lactic acid bacteria that may be contained in the agent of the present invention. The present invention provides foods, medicines, quasi-drugs, etc. that contain the agent of the present invention. The present invention provides a method (excluding medical procedures) for promoting lactic acid bacteria adhesion in the intestinal tract by ingesting the agent of the present invention together with lactic acid bacteria. EXAMPLES

[0046] The present invention will be described in more detail below with reference to examples. However, the scope of the present invention is not limited to such examples. In the following, unless otherwise specified, "%" means "mass %" and "parts" means "mass parts".

[0047] As barley stalks and leaves, crushed powder (manufactured by Toyo Shinyaku Co., Ltd.) was used, which was obtained by drying and crushing stalks and leaves cut before ear emergence. As sweet potato stalks and leaves, crushed powder (manufactured by the same company) was used, which was obtained by drying and crushing sweet potato stalks and leaves cut at a length of 30 to 80 cm from the tip of the stalk and leaves. As kale, crushed powder (manufactured by the same company) was used, which was obtained by drying and crushing kale stalks and leaves. As mulberry, crushed powder (manufactured by the same company) was used, which was obtained by drying and crushing mulberry stalks and leaves. As Peony peony, crushed powder (manufactured by the same company) was used, which was obtained by drying and crushing Peony peony stems and leaves. All of these crushed powders had a mass ratio of 90% or more passing through a 200 mesh section, and the moisture content was 5% or less by mass. These crushed powders were used as test substances in the following adhesion ability test.

[0048] (Adhesion test) (1) Preparation of monolayer using 96-well plate In a 37℃, 5% CO2 incubator, 75 cm 2 Intestinal epithelial cell line Caco-2 cells were cultured in a standard medium using a flask. This standard medium was prepared by adding 5 mL of FBS (Japan Bio Serum) and 0.5 mL of NEAA (Non-Essential Amino Acids, SIGMA) to 44.5 mL of DMEM (containing 1% penicillin-streptomycin). The cells were suspended by trypsinization and placed in a 75 cm 2 From flask to 96-well clear plate: 4.0 × 10 4 The cells were seeded at a density of 100 cells / well and cultured in a 5% CO2 incubator at 37°C for 72 hours to prepare a monolayer.

[0049] (2) Fluorescent labeling of B. coagulans Bacillus coagulans stocked in glycerol at -80°C was returned to room temperature (25°C) and inoculated onto MRS liquid medium warmed to 37°C. The cultured material was cultured at 37°C for 24 hours and used in the test. The pre-cultured B. coagulans was collected and centrifuged at room temperature and 1000g for 3 minutes. The supernatant was removed and the resulting pellet was suspended in 1mL of phosphate-buffered saline (PBS) and centrifuged at room temperature and 1000g for 3 minutes. The same suspension and centrifugation were repeated once more, the supernatant was removed, and the pellet was suspended in 1mL of PBS and 15μL of carboxyfluorescein diacetate (CFDA, Dojindo Laboratories) was added. After incubation at room temperature for 30 minutes in the dark, the cells were centrifuged at room temperature and 1000g for 3 minutes. The supernatant was removed and the cells were suspended in 1mL of PBS and centrifuged at room temperature and 1000g for 3 minutes twice. The supernatant was then removed and the cells were suspended in the test medium. A blank was prepared by carrying out the same procedure except that no CFDA was added. The test medium used was 45 mL RPMI1640 medium (containing 1% penicillin-streptomycin) to which 5 mL FBS (Japan Bio Serum) was added. 100 μL of the suspension was transferred to a black plate and the fluorescence intensity was measured. The excitation wavelength was 495 nm and the fluorescence wavelength was 515 nm. The blank value was subtracted from the measured fluorescence intensity, and the test medium was used to dilute the sample so that the fluorescence intensity was 200. This was used as the B. coagulans sample.

[0050] (3) Evaluation of adhesion ability of B. coagulans by measuring fluorescence intensity The medium of the monolayer of Caco-2 cells prepared in (1) was replaced from the normal medium to the test medium. The medium was removed, and the dispersion in which the test substance was dispersed in the test medium and the B. coagulans sample prepared in (2) were added to a 96-well plate in 100 μL each (final fluorescence intensity of B. coagulans: 100). After 2 hours of culture in a 5% by volume CO2 incubator at 37°C, 150 μL of formalin was added to each well and fixed for 30 minutes at 4°C in the dark. After washing three times with PBS and adding 100 μL of PBS, the fluorescence intensity was measured with a varioskan (excitation wavelength: 495 nm, fluorescence wavelength: 515 nm). Based on the measured value, the relative value (%) to the fluorescence intensity of the sample to which only lactic acid bacteria were added was calculated, and this was taken as the adhesion rate of lactic acid bacteria to the intestinal cell monolayer. The results are shown in Figure 1. The concentrations of the test substances in the Caco-2 cell culture medium during incubation are as shown in Table 1 below (the units of values ​​in Table 1 are μg / mL).

[0051] [Table 1]

[0052] The results in Figure 1 show that the stems and leaves of plants that are the raw materials for green juice increase the intestinal adhesion rate of lactic acid bacteria. Furthermore, it can be seen that the use of stems and leaves of two or more plants, including barley stems and leaves, further increases the intestinal adhesion rate of lactic acid bacteria.

Claims

[Claim 1] An intestinal immunity enhancing agent containing sweet potato stems and / or leaves.

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