Visceral fat reducing agent

Abstract

To provide a visceral fat reducing agent useful for reducing visceral fat.SOLUTION: A visceral fat reducing agent contains Blautia bacteria as an active ingredient.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to a visceral fat reducing agent.

Background Art

[0002] Visceral fat is adipose tissue distributed between and around the stomach and liver, and it has been clarified that its excessive accumulation poses a risk of atherosclerotic lesions. Currently, visceral fat accumulation is the first criterion for diagnosing metabolic syndrome. Managing visceral fat accumulation is extremely important for extending a healthy lifespan.

[0003] More than 1,000 species and over 100 trillion bacteria inhabit the human gastrointestinal tract, forming the gut microbiota. Recent research has revealed that the gut microbiota is involved in the pathogenesis of various diseases, not limited to digestive diseases, but also metabolic diseases, immune diseases, mental diseases, etc. In particular, it has been suggested that the composition of the gut microbiota may be closely related to obesity and metabolic diseases caused by obesity. Among the gut microbiota inhabiting the gastrointestinal tract, the two phyla Firmicutes and Bacteroidetes are the most dominant. Comparing the gut microbiota of obese mice and normal mice, it has been reported that in obese mice, there are more bacteria of the phylum Firmicutes and fewer bacteria of the phylum Bacteroidetes, and conversely, in normal mice, bacteria of the phylum Bacteroidetes predominate. In humans as well, it has been reported that the ratio of Firmicutes / Bacteroidetes is high in obese individuals. Furthermore, although no consistent correlation has been found between the presence of bacteria of the phyla Firmicutes and Bacteroidetes and visceral fat, when analyzing the bacterial species at a level smaller than the phylum, i.e., at the "genus" level, it has been reported that regardless of gender, individuals with a smaller visceral fat area have a higher abundance of Blautia bacteria in the gut, and among the gut bacteria, Blautia bacteria are related to visceral fat mass in both men and women (Non-Patent Document 1).

[0004] Bacteria of the genus Blautia are major bacteria that account for about 3 to 11% of all gut bacteria in humans and animals. Bacteria of the genus Blautia are known to produce short-chain fatty acids such as acetic acid, propionic acid, and butyric acid, which have the function of eliminating obesity in the body. In addition, it has been reported that they are decreased in patients with diabetes, cirrhosis, colorectal cancer, and rheumatoid arthritis. Also, it has been reported that bacterial strains of the genus Blautia, such as Blautia hydrogenotrophica, are effective in increasing or maintaining the diversity of the microbiota and are effective in treating brain damage (for example, Patent Documents 1 and 2). However, there is no report on the effect of the ingestion of bacteria of the genus Blautia on visceral fat accumulation.

Prior Art Documents

Patent Documents

[0005]

Patent Document 1

Patent Document 2

Non-Patent Documents

[0006]

Non-Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0007] The present invention relates to providing a visceral fat reducing agent useful for reducing visceral fat.

Means for Solving the Problems

[0008] In view of the above problems, the present inventor has intensively studied and found that when bacteria of the genus Blautia are orally ingested, visceral fat is significantly reduced.

[0009] That is, the present invention provides a visceral fat reducing agent containing bacteria belonging to the genus Blautia as an active ingredient.

Effects of the Invention

[0010] The visceral fat reducing agent of the present invention exhibits an excellent visceral fat reducing effect. Therefore, it can be expected to prevent, improve, or treat the onset or progression of health disorders or diseases closely related to visceral fat accumulation.

Brief Description of the Drawings

[0011]

Figure 1

Modes for Carrying Out the Invention

[0012] In the present invention, the bacteria belonging to the genus Blautia are bacteria belonging to the genus Blautia classified in the phylum Firmicutes. Examples of the bacteria belonging to the genus Blautia include Blautia hydrogenotrophica, Blautia stercoris, Blautia faecis, Blautia coccoides, Blautia glucerasea, Blautia hansenii, Blautia luti, Blautia producta, Blautia schinkii, Blautia wexlerae, etc. One or more of these can be used. Among these, from the viewpoint of improving the visceral fat reducing effect, Blautia hansenii is preferable. Braultea henseni can be exemplified by the strain JCM 14655 or bacteria identical thereto. Identical bacteria refer to bacteria of the same genus having a nucleotide sequence of the 16S rRNA gene with 98% or more, preferably 99% or more homology to the nucleotide sequence of the 16S rRNA gene of the cell strain and having the same mycological properties. The strain Brautia henseni JCM 14655 is generally available from the Microbial Material Development Unit of the RIKEN BioResource Center (RIKEN BRC) based on the JCM number of the strain described in the online catalog (for example, see https: / / www.jcm.riken.jp / cgi-bin / jcm / jcm_number?JCM=14655 for the strain JCM 14655).

[0013] The cells of Brautia bacteria can be obtained by culturing according to a method usually used for culturing Brautia bacteria. For example, using a GAM medium or the like, culture is carried out at a culture temperature of 30 to 45°C. The culture is preferably carried out under anaerobic conditions. After culture, the cells can be separated by means of cell collection such as centrifugation or membrane concentration.

[0014] The Brautia bacteria may be either live cells or dead cells, but from the viewpoints of quality stability and safety to the living body, it is preferable to use dead cells. Examples of the dead cells include heat-killed cells obtained by heat-sterilizing the cells after culture. The dead cells may be in the form of a suspension in water or the like, or in the form of a dried product. As the drying method, ordinary means such as spray drying and freeze drying can be used.

[0015] As shown in the following examples, ingestion of bacteria belonging to the genus Blautia suppresses the accumulation of visceral fat promoted by high-fat diet intake. That is, bacteria belonging to the genus Blautia have the effect of suppressing the accumulation of visceral fat and reducing visceral fat. By ingesting bacteria belonging to the genus Blautia, it is considered that the living conditions and the number of bacteria of the genus Blautia existing in the intestine are affected, and visceral fat is reduced. By reducing visceral fat, it becomes possible to prevent, improve or treat visceral fat type obesity and metabolic syndrome (visceral fat syndrome) based on visceral fat accumulation. Therefore, bacteria belonging to the genus Blautia can be a visceral fat reducing agent that reduces visceral fat, and can also be used for producing a visceral fat reducing agent. In addition, bacteria belonging to the genus Blautia can be used for reducing visceral fat. In this specification, "visceral fat" refers to fat accumulated in visceral tissues (for example, stomach, liver, kidney, etc.) in the abdominal cavity and around them. "Reduction of visceral fat" means reducing the amount of visceral fat and suppressing the accumulation of visceral fat. "Use" can be administration or ingestion to animals including humans, and can be either therapeutic use or non-therapeutic use. "Non-therapeutic" is a concept that does not include medical acts, that is, a concept that does not include methods of operating, treating or diagnosing humans. More specifically, it is a concept that does not include methods of operating, treating or diagnosing humans by a doctor or a person receiving instructions from a doctor.

[0016] The visceral fat reducing agent of the present invention becomes a pharmaceutical product, quasi-drug, food or feed for humans or animals that exhibits the effect of reducing visceral fat when administered or ingested to animals including humans, and can also be a material or preparation used by being formulated in the pharmaceutical product, quasi-drug, food or feed.

[0017] The pharmaceutical product (hereinafter including quasi-drugs) contains bacteria belonging to the genus Blautia as an active ingredient for reducing visceral fat. Furthermore, the pharmaceutical product may contain a pharmaceutically acceptable carrier, or other active ingredients, pharmacological components, etc. as necessary, as long as the function of the active ingredient is not lost. The pharmaceutical can be administered in any dosage form. Examples of dosage forms include oral solid preparations such as tablets (including chewable tablets, etc.), capsules, granules, powders, troches, etc., oral liquid preparations such as oral solutions, syrups, etc., parenteral administration preparations such as injections, suppositories, inhalants, transdermal absorbents, external preparations, etc. The preferred dosage form is oral administration. The form can be arbitrarily adjusted in size according to the purpose of use. Such preparations in various dosage forms can be prepared according to established methods by appropriately combining with pharmaceutically acceptable carriers, for example, excipients, binders, diluents, disintegrants, surfactants, lubricants, dispersants, buffers, osmotic pressure regulators, pH regulators, emulsifiers, preservatives, stabilizers, preservatives, thickeners, fluidity improvers, flavoring agents, foaming agents, fragrances, coating agents, diluents, etc., and other medicinal ingredients, etc.

[0018] The food contains bacteria of the genus Blautia as an active ingredient for reducing visceral fat. The food includes foods that claim to reduce visceral fat and for which the indication to that effect is permitted or notified as necessary (foods for specified health uses, foods with functional claims). Examples of indications include "reduces visceral fat", etc. Foods with permitted or notified functional claims can be distinguished from general foods. The form of the food can be solid, semi-solid or liquid (for example, beverages). Examples include various food compositions (breads, cakes, noodles, confectioneries, frozen foods, ice creams, candies, sprinkles, soups, dairy products, shakes, beverages, seasonings, etc.), and furthermore, nutritional supplement compositions in the same forms as the above-mentioned oral administration preparations (solid preparations such as granules, powders, tablets, capsules, microcapsules, troches, etc.). Foods in various forms can be prepared according to established methods by appropriately combining bacteria of the genus Blautia with any food material, or other active ingredients, or additives permitted in foods (for example, solvents, softeners, oils, emulsifiers, preservatives, acidulants, sweeteners, bittering agents, pH regulators, stabilizers, colorants, ultraviolet absorbers, antioxidants, humectants, thickeners, fixing agents, dispersants, fluidity improvers, wetting agents, fragrances, seasonings, flavor modifiers), etc.

[0019] The feed contains bacteria of the genus Blautia as an active ingredient for reducing visceral fat. Preferably, the form of the feed is pellet, flake, mash or liquid. Examples include livestock feed for cattle, pigs, chickens, sheep, horses, etc., small animal feed for rabbits, rats, mice, etc., and pet food for dogs, cats, birds, etc. The feed can be prepared according to a conventional method by appropriately combining bacteria of the genus Blautia with other feed materials such as meats, proteins, grains, bran, meal, sugars, vegetables, vitamins, minerals, gelling agents, shaping agents, pH adjusters, seasonings, preservatives, nutritional supplements, etc.

[0020] In the visceral fat reducing agent of the present invention, the content of bacteria of the genus Blautia may vary depending on their dosage form and form, but can be appropriately set in consideration of ease of administration or ingestion, etc.

[0021] The dosage or intake amount of the visceral fat reducing agent of the present invention can vary according to the species, body weight, sex, age, condition or other factors of the administration or intake subject. The dosage, route, interval of administration, and the amount and interval of intake can be appropriately determined by those skilled in the art. However, in the case of oral administration or oral intake, from the viewpoint of improving the visceral fat reducing effect, for an adult (body weight 60 kg), per day, as bacteria of the genus Blautia, preferably the number of bacteria is 1×10 4 or more, more preferably the number of bacteria is 1×10 5 or more, still more preferably the number of bacteria is 1×10 7 or more, and preferably the number of bacteria is 1×10 15 or less, more preferably the number of bacteria is 1×10 12 or less, still more preferably the number of bacteria is 1×10 9 or less. And the dosage or intake amount of the visceral fat reducing agent of the present invention, as bacteria of the genus Blautia, is preferably the number of bacteria 1×10 4 ~1×10 15 and more preferably the number of bacteria is 1×10 5 ~1×10 12 and still more preferably the number of bacteria is 1×10 7 ~1×10 9is a range. In the present invention, it is preferable to orally administer or orally ingest such an amount once, twice, or three or more times a day. In addition, in the examples described later, it is explained that the administered Blautia bacteria have an effect of reducing visceral fat in mice. However, from the viewpoints of ethics and safety considerations, instead of humans, mice established by public test methods or academic papers as a human assumption model are deliberately used. Therefore, the visceral fat-reducing effect in the present invention is a concept that shows activity not only for the visceral fat-reducing effect in mice but also for humans in general.

[0022] The visceral fat-reducing agent of the present invention can be administered or ingested according to any plan. The administration or ingestion period is not particularly limited, and it may be a single administration or ingestion, or it may be repeatedly and continuously administered or ingested. When repeatedly and continuously administering or ingesting, it is preferable to continuously administer or ingest for 2 weeks or more, further 4 weeks or more, and further 8 weeks or more.

[0023] The visceral fat-reducing agent of the present invention is preferably administered or ingested during, before, or after eating or feeding. When administering or ingesting before or after eating or feeding, it is preferably administered or ingested within 10 minutes before or after eating or feeding. The content of eating (feeding) is not particularly limited, and examples include a diet or feed containing protein, carbohydrates (including sugars), and lipids. The administration or ingestion subject is not particularly limited as long as it is a human or non-human animal that needs or desires it. Preferred examples of the subject include humans with a high accumulation amount of visceral fat (for example, humans with a visceral fat area of 100 cm 2 or more), humans who tend to consume a fatty diet, and the like.

Examples

[0024] Test Example 1 1. Test Feed and Method The cells of B. hansenii JCM 14655 strain (RIKEN BRC) were heat-sterilized in an autoclave to kill the cells. Then, the residue obtained by suspending the cells in distilled water and repeatedly washing the suspension by centrifugation was freeze-dried and powdered for use. Three-week-old male C57BL / 6J mice were pre-fed for one week and then fed a normal diet (AIN-93G, a standard diet defined by the National Institutes of Health, USA) and a high-fat diet (20% lipid by mass) prepared with reference to the composition of the standard diet ad libitum for nine weeks. The experimental groups were divided into three groups: a normal diet control group (n = 8), a high-fat diet control group (n = 8), and a high-fat diet (with B. hansenii added) group (n = 8) so that the initial body weights were approximately constant. The dose of B. hansenii administered was 1×10 9 cells / mouse / day. During the experimental feeding period, body weight, water intake were measured, and feces were collected. After the experimental feeding period, the mice were euthanized by cervical dislocation and then dissected. As visceral fat, the fat around the epididymis, mesenteric fat, and perirenal and retroperitoneal fat were excised, and the weights of various organs were measured. The total value of these was taken as the total weight of visceral fat.

[0025] 2. Method for Identifying B. hansenii in Foods Containing B. hansenii It is possible to identify B. hansenii using the real-time PCR method. Specific examples are shown below. Procedure 1. DNA Extraction · Extraction Method Pretreatment and crude extraction: Each sample was suspended in 4 M guanidine thiocyanate, 100 mM Tris-HCl (pH 9.0), and 40 mM EDTA, and the sample was pretreated using a cell disruption device (FastPrep FP100A (MP Biomedicals, USA), etc.). Further, DNA can be extracted from the bead-treated suspension using the Magtration System12GC and GC series MagDEADNA 200 (Precision System Science, Japan).

[0026] Procedure 2. PCR Conditions Amplify the DNA fragment. As a kit, the Blautia Hansenii Detection Kit (manufactured by Technosuga Lab Co., Ltd.) can be used.

[0027] Procedure 3. Real-time PCR method As the reagent for the real-time PCR method, for example, TB Green Premix Ex Taq 2 (Tli RNaseH Plus) (manufactured by Takara Bio Inc.) can be used. As the device, for example, Rotor-Gene Q (manufactured by QIAGEN) can be used. As the PCR conditions, for example, the methods described in Dig Dis Sci, 2015, 60(9)2654-61 etc. can be used.

[0028] For the test of significant difference, the t-test was used. The obtained numerical values were shown as the mean ± standard error.

[0029] 2. Results Figure 1 shows the total weight of visceral fat, the weight around the epididymis, the mesenteric weight, and the perirenal and retroperitoneal weight of each group. Different letters (a, b, c) in Figure 1 indicate a significance level of P < 0.05, and ♯ indicates P = 0.08 in comparison with the normal diet. As a result, in the high-fat diet (Blautia bacteria added) group, a decrease in visceral fat weight was observed compared to the high-fat diet control group, and it was confirmed that Blautia bacteria have an excellent inhibitory effect on visceral fat accumulation promoted by high-fat diet intake.

Claims

1. A visceral fat reducing agent containing Bratia hensenii as an active ingredient.

2. As Bracteolaria henseni, it is administered or ingested in an amount of 1×10 4 to 1×10 15 per day, and it is the visceral fat reducing agent according to Claim 1.

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