An agent for improving the absorption of propionic acid and / or butyrate in the small intestine, and a method for improving the absorption of propionic acid and / or butyrate in the small intestine.

Abstract

To provide new functions of dried young barley grass. [Solution] An agent for improving the absorption of propionic acid and / or butyrate in the small intestine, comprising dried young barley leaves as the active ingredient.

Description

Technical Field

[0001] The present invention relates to an agent for improving the absorption of propionic acid and / or butyric acid in the small intestine and a method for improving the absorption of propionic acid and / or butyric acid in the small intestine.

Background Art

[0002] Both butyric acid and propionic acid are short-chain fatty acids, which are absorbed from the digestive tract and exhibit various physiological effects in the body, such as blood cholesterol reduction, allergy reaction reduction, antidepressant effect, mood disorder improvement, stress reaction alleviation, immune function regulation, inflammation suppression, obesity suppression, etc. Butyric acid (or butyrate and other derivatives of butyric acid) is a useful substance that can provide an energy source that is rapidly and easily absorbed for animals and tissue cells. In in vitro studies, when glucose, ketone bodies, glutamine, etc. are used as respiratory energy sources for animals, it has been shown that colon epithelial cells first utilize butyric acid, and it has been reported that almost all butyric acid is absorbed and utilized by intestinal epithelial cells. Therefore, it is presumed that most of the butyric acid present in the intestinal tract is consumed by intestinal epithelial cells before being absorbed into the body, and the absorption efficiency into the body is not good. Isotope tracer tests have further shown that butyric acid can enter the human blood circulation and be metabolized by body tissues. In addition, in peripheral tissues, butyric acid is rapidly oxidized for use in fat synthesis and is also reported to be taken up by the mammary gland for synthesis of milk fat. Propionic acid (or propionate and other derivatives of propionic acid) is metabolized in the liver when absorbed into the body and has the function of reducing cholesterol in the blood. In addition, when propionic acid is administered to lactating mother mice, allergic airway inflammation, which is one of the pathological conditions of infantile bronchial asthma, is found to be suppressed (Non-Patent Document 1). Propionic acid is deeply involved in extraintestinal diseases such as obesity suppression, cholesterol reduction, and allergic diseases, and is expected to contribute to the development of new treatment methods for obesity suppression, cholesterol reduction, and allergic diseases including bronchial asthma in the future. Furthermore, monocarboxylic acid transporters are involved in the absorption of butyrate and propionic acid in the small intestine. It is known that there are several types of monocarboxylic acid transporters, and among them, SMCT1, a sodium-dependent transporter, is known to have a high affinity for butyrate and propionic acid.

[0003] On the other hand, as a beverage that easily addresses the problem of insufficient vegetable intake, so-called "green juice" is known along with vegetable juices. The raw materials for this "green juice" include chlorophyll-containing green leaf plants such as kale, barley grass, wheatgrass, angelica tree, and mulberry leaves. Among these, barley grass is a representative ingredient of green juice and forms a large market for health food ingredients as one of the green juice products. It is known that consuming barley grass can improve the balance of the gut microbiota in the large intestine and promote the growth of gut bacteria that produce short-chain fatty acids such as acetic acid, propionic acid, and butyric acid (Patent Documents 1 and 2). On the other hand, there is no knowledge about what effect barley grass intake has on the small intestine, and since it is known that the gut microbiota of the small intestine and the large intestine are different, it is not possible to apply the knowledge about the effects of dried barley grass on the gut microbiota of the large intestine to the gut microbiota of the small intestine. Furthermore, because the large intestine is located behind the small intestine, short-chain fatty acids produced by the gut microbiota of the large intestine are utilized within the large intestine and are unlikely to reflux and affect the absorption of short-chain fatty acids in the small intestine. [Prior art documents] [Patent Documents]

[0004] [Patent Document 1] Japanese Patent Publication No. 2021-35346 [Patent Document 2] Japanese Patent Publication No. 2023-51312 [Non-patent literature]

[0005] [Non-Patent Document 1] Gut Microbes, 2023, Vol. 15, No. 1, https: / / doi.org / 10.1080 / 19490976.2023.2206507 [Overview of the Initiative] [Problems that the invention aims to solve]

[0006] This invention was made in view of the above circumstances and aims to provide new functions of dried barley grass. [Means for solving the problem]

[0007] While researching the novel functions of dried barley grass, the inventors discovered that dried barley grass can contribute to improving the absorption of propionic acid and / or butyrate in the small intestine of mammals such as humans, and thus completed the present invention.

[0008] 1. An agent for improving the absorption of propionic acid and / or butyrate in the small intestine, comprising dried barley grass as the active ingredient. 2. A method for improving the absorption of propionic acid and / or butyric acid in the small intestine, characterized by ingesting dried barley grass. [Effects of the Invention]

[0009] According to the present invention, ingesting dried barley grass effectively improves the absorption of propionic acid and / or butyric acid in the small intestine. As a result, the improved absorption of propionic acid and / or butyric acid in the small intestine is expected to more strongly express the physiological effects that propionic acid and / or butyric acid itself exhibits in the body, such as blood cholesterol reduction, allergic reaction reduction, depressive effect, mood disorder improvement, stress response mitigation, immune function regulation, inflammation suppression, and obesity suppression. Furthermore, it is useful as an energy source for mammals such as humans. Furthermore, a key difference between this invention and the prior art is that ingesting dried barley grass does not improve the absorption of acetic acid in the small intestine. [Brief explanation of the drawing]

[0010] [Figure 1] This graph shows the concentration of acetate in portal vein blood in the examples. [Figure 2] This graph shows the propionic acid concentration in portal vein blood in the examples. [Figure 3] This graph shows the isobutyric acid concentration in portal vein blood in the examples. [Figure 4] This graph shows the butyrate concentration in portal vein blood in the example. [Figure 5] This graph shows the valeric acid concentration in portal vein blood in the examples. [Modes for carrying out the invention]

[0011] <About dried barley grass> The dried barley grass product of the present invention will be described in detail below. In this specification, "barley grass" refers to the leaves of barley and the stems to which those leaves are attached. In this invention, barley can be specifically referred to as two-row barley, six-row barley, or hulless barley, depending on the shape of the ear and seeds, but these are not particularly limited and any of them can be used. Examples of barley varieties include Aominori, Agurimochi, Kusumochi Nijo, Asakagold, Kinuyutaka, Oumiyutaka, Kawahonami, Kawamizuki, Kirinijo, Haruna Nijo, Amagi Nijo, Fuji Nijo, Ishukushirazu, Kinuka Nijo, Komaki Nijo, Sakitama Nijo, Tone Nijo, Sachihogolden, Satsukibare, Shunrei, Skygolden, and Takahogo. Golden, Tsuyushirazu, Nasu Nijo, Nishino Gold, Nishino Chikara, Nishino Hoshi, Haruka Nijo, Nirasaki Nijo, Nira Nijo, Harushizuku, Satsuki Nijo, Hoshun, Mikamo Golden, Myogi Nijo, Misato Golden, Miharu Gold, Miho Golden, Yashio Golden, Yachiho Golden, Kitaiku No. 41, Ayano Hoshi, Ryofu, and other two-row hulled wheat, View Fiber -, two-row hulless barley such as Kirarimochi, Yumesakiboshi, Waxy Fiber, Asamamugi, Kashimagol, Minorimugi, Katorimugi, Musashinomugi, Drillmugi, Sanadamugi, Haganemugi, Sayakaze, Suzukaze, Shunrai, Silky Snow, Shinjuboshi, Setsugenmochi, Natorioomugi, Hamayutaka, Hayamioomugi, Fiber Snow, Hanemamochi, White Examples of hulless barley include six-row barley such as Fiber, Baitori, and Miyuki barley, as well as hulless barley such as Ichibanboshi, Senbonhadaka, Sanukihadaka, Kikaihadaka, Sanshu, Nanpuhadaka, Shiratamahadaka, Yuunagihadaka, Daishimochi, Toyonokaze, Hayatehadaka, Ichiwase, Biwairohadaka, Benihadaka, Hashirihadaka, and Mannenboshi (formerly Mantenboshi), but are not particularly limited. These barleys may be used individually or in combination of two or more. In particular, among the dried barley grass products used in this invention, those derived from two-row barley are preferred, and those derived from two-row hulless barley are more preferred. Furthermore, those derived from glutinous barley are also preferred.

[0012] The dried barley grass product of the present invention may be manufactured by a conventionally known manufacturing method, and the manufacturing method is not limited. Conventional known manufacturing methods include processes such as "blanching," "juicing and powdering," "drying," "shredding," "sterilization," and "grinding," using fresh young barley grass as a raw material. Let me briefly explain one example. The harvested raw materials are preferably processed immediately after harvesting. However, if it takes time until processing, they are stored by means such as low-temperature storage to prevent deterioration of the raw materials. After removing defective materials such as foreign substances and dead leaves from the raw materials, they are cut to about 3 to 5 cm and washed with water, followed by dehydration treatment by centrifugation or the like. · Blanching treatment Subsequently, blanching treatment of the dehydrated raw materials is carried out. This blanching treatment is performed to inactivate the enzymes in the leaves that cause discoloration and deterioration of the young barley leaves and stems, and to reduce the soil bacteria adhering to the young barley leaves and stems. Specific examples of the blanching treatment include hot water treatment and steam treatment. The hot water treatment is carried out, for example, by immersing the young barley leaves and stems in boiling water for about several tens of seconds to 5 minutes. The steam treatment is carried out, for example, by using a belt conveyor type or rotary wire mesh cylindrical steamer and heating the young barley leaves and stems with steam for 20 to 100 seconds. Furthermore, superheated steam treatment can also be mentioned as a new technique, which is carried out with superheated steam heated to 200°C or higher and 600°C or lower. In this blanching treatment, the enzymes are inactivated and the soil bacteria and the like are reduced, and at the same time, flexibility is imparted to the young barley leaves and stems. In order to impart appropriate flexibility to the young barley leaves and stems, the hot water treatment is preferably carried out by immersing them in boiling water for 40 seconds to 3 minutes, and the superheated steam treatment is preferably carried out by heating them with superheated steam heated to 200°C or higher and 600°C or lower for 5 to 120 seconds.

[0013] · Juice extraction and powderization After cooling, the young barley leaves that have undergone blanching treatment are preferably finely chopped if necessary, and juice extraction is carried out using a device capable of applying pressure, such as a screw press or a cold press, according to a known method. The juice extract may be used in a low-temperature liquid state or frozen, or if necessary, it can also be powdered by spray drying or freeze drying after adding excipients such as dextrin, cyclodextrin, starch, maltose, etc. [[ID=十六]] [[ID=十七]]

[0014] [[ID=十八]] [[ID=十九]] [[ID=二十]] It is also preferable to dry the blanched and cooled barley grass. This drying process is carried out, for example, by placing the blanched barley grass on a ventilated belt conveyor and blowing hot air at about 70-100°C to achieve a product temperature of 30°C to 40°C. The final moisture content is preferably about 10%, and more preferably less than 10%. The final drying may be carried out, for example, by placing the barley grass on a belt conveyor and drying it continuously with hot air, or by static hot air drying on shelves or the like.

[0015] • Shredding and sterilization The shredding process can be carried out using methods commonly used by those skilled in the art when shredding plants, and it is preferable to shred the material into pieces smaller than 1 cm in length. It is preferable to sterilize the shredded barley grass. The sterilization treatment is not particularly limited as long as it is a treatment that is commonly known to those skilled in the art, but for example, it can be a treatment that physically or chemically kills microorganisms using temperature, pressure, electromagnetic waves, chemicals, etc. Among these, sterilization using an airflow sterilizer is preferred.

[0016] • Grinding process The dried barley grass obtained through shredding and sterilization processes can be pulverized to produce a fine powder. The pulverization process can be carried out using, for example, an impact pulverizer with a grinding effect (a grinding-type impact pulverizer), which is an example of an impact pulverizer, but is not limited to this. It may also be carried out using other known pulverizers such as impact pulverizers like pin mills and jet mills, shear pulverizers, grinding pulverizers like mascolloiders, grinders, millstones, or composite pulverizers such as low-temperature freeze pulverizers. In the grinding process, it is preferable to have a particle size of barley grass powder that passes through 30 mesh to 250 mesh remaining as powder. If the particle size is small enough to pass through 250 mesh, it may be difficult to handle when processing as a food ingredient or pharmaceutical raw material. On the other hand, if the particle size is large enough to remain in 30 mesh, uniform mixing with other food ingredients may be hindered. However, this does not prevent using other particle sizes depending on the application and preference. The equipment used in the above process is not particularly limited; an appropriate piece of equipment should be selected considering the hardness of the barley grass, the directionality of the fibers, the leaf tissue, etc.

[0017] <Confirmation of the effects of consuming dried barley grass> The in vivo effects of ingesting dried barley grass were confirmed by rat portal vein catheterization, as will be explained in detail in the examples described later. The rat portal vein catheterization method is known as an animal experimental model that allows for rapid screening of functional components using a small number of experimental animals (Digestion and Absorption, Vol. 19, No. 2, pp. 56-60, 1996, etc.). The rat portal vein catheterization method is an experimental model in which a catheter is placed in the portal vein of a rat in a manner that does not obstruct blood flow, and a harness with a protective coil is attached, allowing for the collection of portal vein blood after oral administration of a test feed under unanesthetized and unrestrained conditions. It has the advantage of allowing for the collection of portal vein blood at short intervals or over long periods without stressing the animals. In this experimental model, components absorbed from the small intestine and released into the portal vein can be directly collected and measured, offering the advantage of high sensitivity detection without undergoing metabolism in the liver. Furthermore, it allows for long-term rearing under unanesthetized and unrestrained conditions. In short, the rat portal vein catheterization method is an experimental model suitable for evaluating components absorbed from the small intestine and released into the portal vein. Using this rat portal vein catheterization method, rats were orally administered a test diet containing dried barley grass. Portal vein blood was collected over three days: on the day of administration (day 0), day 1, and day 2. The short-chain fatty acid concentrations in the plasma of the collected blood were measured. As shown in the examples described later, a significant increase in propionic acid and butyrate concentrations in the portal vein blood was confirmed. On the other hand, there was no change in acetic acid concentration in the portal vein blood, and no increase in the concentrations of short-chain fatty acids other than propionic acid and butyrate was observed. The above results are based on portal vein blood analysis and are attributed to a significant improvement in propionic acid absorption and butyrate absorption in the small intestine. Therefore, it can be said that the intake of dried barley grass improved the absorption of propionic acid and butyrate in the small intestine. Furthermore, since the intake of dried barley grass did not improve the absorption of short-chain fatty acids other than propionic acid and butyric acid, such as acetic acid, in the small intestine, it is possible that there is another mechanism that affects only the absorption of propionic acid and / or butyric acid, rather than the increase in the overall production of short-chain fatty acids in the small intestine due to the promotion of the growth of intestinal bacteria that produce short-chain fatty acids in general, and the resulting increase in the absorption of short-chain fatty acids in the small intestine. For example, it is possible that there is another mechanism that affects only the absorption of propionic acid and / or butyric acid, such as an increase in the expression of monocarboxylic acid transporters that do not involve the growth of intestinal bacteria, or an enhancement of the affinity between monocarboxylic acid transporters and propionic acid and butyric acid. Therefore, the significant improvement in propionic acid and butyric acid absorption in the small intestine due to the intake of dried barley grass is considered to be an extremely selective event. The propionic acid and / or butyrate absorption enhancer in the small intestine according to the present invention exhibits the function of improving the absorption of propionic acid, the function of improving the absorption of butyrate, and the function of improving the absorption of propionic acid and butyrate, and among these, it is preferable that it exhibits the function of improving the absorption of propionic acid and butyrate, and the function of improving the absorption of propionic acid.

[0018] <Regarding agents that enhance propionic acid absorption and butyrate absorption in the small intestine> In the present invention's small intestine propionic acid absorption and butyrate absorption enhancer, when using "dried barley grass," the amount is not particularly limited as it varies depending on age, sex, weight, symptoms, or intended use. However, for example, the daily intake for an adult human is preferably in the range of 0.1 to 20 g, more preferably in the range of 1.5 to 10 g, even more preferably in the range of 1 to 5 g, and particularly preferably in the range of 2 to 3 g.

[0019] The small intestine propionic acid and / or butyrate absorption enhancer of the present invention does not necessarily need to be formulated, but it is preferable to use it in a formulation. When the small intestine propionic acid and / or butyrate absorption enhancer of the present invention is used in a formulation, preferred dosage forms include powders, granules, tablets, capsules such as soft capsules and hard capsules, suspensions, liquids, and emulsions, all containing dried barley grass. The small intestine propionic acid and / or butyrate absorption enhancer of the present invention may contain other commonly used components in addition to dried barley grass during formulation, to the extent that they do not impair the effects of the present invention. Examples of such components include various excipients, binders, glazing agents, lubricants, stabilizers, diluents, bulking agents, thickening agents, emulsifiers, antioxidants, pH adjusters, colorants, flavorings, and additives. The content of other components can be appropriately selected depending on the form of the agent of the present invention. Furthermore, if the agent of the present invention is in multiple dosage forms, the inclusion of each other component in each dosage form can also be appropriately selected depending on the form of the dosage form. Furthermore, the small intestine absorption enhancer of propionic acid and / or butyrate according to the present invention may also include foods, tablets, capsules, and other supplements that can claim health functional foods as defined by food-related laws and regulations such as the Food Sanitation Act, the Food Labeling Act, and the Health Promotion Act, that is, foods that can claim efficacy and health maintenance and promotion effects based on the action of the present invention. [Examples]

[0020] The present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples. In the examples, unless otherwise specified, "parts" means parts by weight.

[0021] (1) Test feed and comparative feed Test feeds and comparison feeds were prepared according to the contents shown in Table 1 below. Young barley grass of the Kirarimochi variety was used. The basis for calculating the dried barley grass content in the test feed is as follows: The recommended daily intake of dried barley grass for humans is 3g (=0.06g / kg of body weight). For this experiment, the experimental animals, rats (approximately 300g of body weight), were given 10 times the amount of dried barley grass for humans (0.67g / kg of body weight), resulting in a dosage of 10g / kg of feed. The amount of dried barley grass was determined considering the intake amount and duration for humans, as well as differences in sensitivity between rats and rodents. [Table 1]

[0022] (2) Laboratory animals Four-week-old male Sprague-Dawley rats (Slc:SD, SLC Co., Ltd.) were reared under conditions of room temperature 23±1℃, humidity 55±7%, and a 12-hour light-dark cycle (light period 8:00-20:00). Solid feed (MF; Oriental Yeast Co., Ltd.) and water were freely provided, and after a three-week pre-maturation period, the rats were used in the experiment. Catheter placement into the portal vein was performed according to previously reported procedures (e.g., Digestion and Absorption, Vol. 19, No. 2, pp. 56-60, 1996). Specifically, rats were subjected to laparotomy under nitrous oxide, oxygen, and phlosen mixed gas anesthesia (2.5% induction phase, 1.5% maintenance phase), and a blood collection catheter (polyethylene tube: inner diameter 0.28 mm, outer diameter 1.0 mm) was placed in the portal vein. The other end of this catheter was brought out through a subcutaneous tunnel to the dorsum. The blood collection catheter was filled with an anticoagulant to prevent occlusion due to blood coagulation. The anticoagulant used was heparin sodium injection (Ajinomoto Pharmaceutical Co., Ltd.) diluted with sterile physiological saline, resulting in 30% heparinized saline. Rats in which a catheter was placed in the portal vein were randomly selected and divided into groups of six, and each group was individually housed in a stainless steel metabolic cage.

[0023] (3) Experimental method Six rats each, with a catheter placed in the portal vein, were given either the test diet or the control diet described in Table 1 for three days. Portal vein blood (0.5 mL) was collected on the day of ingestion (day 0), day 1, day 2, and day 3. The collected blood was centrifuged to obtain plasma, and the concentrations of short-chain fatty acids (acetic acid, propionic acid, isobutyric acid, butyric acid, and valeric acid) in this plasma were measured using high-performance liquid chromatography (HPLC). The analysis of short-chain fatty acid concentrations in plasma was performed using labeling reagents for long-chain and short-chain fatty acid analysis for high-performance liquid chromatography (manufactured by YMC Corporation). All experimental data are presented as mean values ​​and standard errors. Statistical analysis was performed using the 4steps Excel statistical add-in software Statcel3 (OMS Publishing). Significant difference testing for portal blood concentrations was performed using one-way ANOVA followed by multiple comparison tests (Scheffe's F test). Figures 1-5 show graphs plotting the concentrations of acetate, propionic acid, isobutyric acid, butyric acid, and valeric acid in portal blood on the day of ingestion (day 0), day 1, day 2, and day 3, respectively. In Figures 2 and 4, the asterisk (*) indicates a significant difference between the test feed containing dried barley grass and the comparative feed without dried barley grass.

[0024] As shown in Figures 2 and 4, the propionic acid and butyric acid concentrations in the portal blood of rats that ingested a test diet containing dried barley grass were significantly higher than those in rats that ingested a control diet without dried barley grass. These results indicate that the intake of dried barley grass was a factor in the improved absorption of propionic acid and butyrate in the small intestine, which draws blood into the portal vein. Therefore, it is clear that the intake of dried barley grass can effectively improve the absorption of propionic acid and butyrate in the small intestine. On the other hand, as shown in Figures 1, 3, and 5, it was confirmed that the concentrations of acetate, isobutyric acid, and valeric acid in the portal blood of rats that ingested a test diet containing dried barley grass were no different from the concentrations in the portal blood of rats that ingested a control diet that did not contain dried barley grass.

Claims

1. An agent for improving the absorption of propionic acid and / or butyrate in the small intestine, with dried young barley grass as the active ingredient.

2. A method for improving the absorption of propionic acid and / or butyric acid in the small intestine, characterized by ingesting dried barley grass.

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