Composition for promoting sugar absorption

Abstract

To provide a novel sugar absorption promoting composition.SOLUTION: Cells of bacteria belonging to Lactobacillus, Lactococcus, Leuconostoc, or Bifidobacterium are used as active ingredients, to provide food and drink, and feed having enterically sugar absorption promoting action.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a composition for promoting sugar absorption in the intestinal tract, which contains lactic acid bacteria and / or bifidobacterial cells as active ingredients. [Background technology]

[0002] Carbohydrates are digested and absorbed, circulate in the blood, and become an energy source for tissues throughout the body. In particular, glucose in the blood is the main energy source for the brain, so an extreme lack of carbohydrates can lead to dizziness, lightheadedness, and a lack of concentration, and in severe cases, can cause loss of consciousness.

[0003] A lack of carbohydrates in the blood can occur when carbohydrate intake is constantly insufficient due to extreme dieting or eating disorders, or when carbohydrates are consumed excessively due to exercise on an empty stomach, etc. It has also been reported as a side effect of insulin preparations and oral hypoglycemic drugs used by diabetic patients.

[0004] Malabsorption of sugar in the intestine can also be a cause of carbohydrate deficiency. Sugar uptake in the intestine is mainly mediated by monosaccharide transporters expressed in intestinal epithelial cells. Three monosaccharide transporters, SGLT1, a sodium-dependent unidirectional transporter, and GLUT2 and GLUT5, facilitative bidirectional transporters, are expressed in the small intestine, and contribute to the uptake of sugar from the intestinal lumen into the body. When the expression of these monosaccharide transporters is inhibited, the uptake of sugar from the intestine is suppressed. For example, it has been reported that tomatoside A, a saponin derived from tomato seeds, inhibits the uptake of sugar by inhibiting the expression of SGLT1 in intestinal epithelial cells (Patent Document 1). In addition, it has been reported that quercetin contained in apples and onions, and apigenin contained in celery and parsley, all have the effect of significantly reducing the gene expression of GLUT2 and GLUT5, and inhibit the uptake of sugar (Non-Patent Document 1).

[0005] Promoting sugar uptake in the intestinal tract is thought to be one of the means to improve carbohydrate deficiency. However, while many foods and pharmaceutical compositions that suppress sugar uptake have been reported, there are few reports of foods and pharmaceutical compositions that promote sugar uptake.

[0006] Lactic acid bacteria and bifidobacteria have been used since ancient times to produce fermented foods, and recent research has revealed that they contribute to the promotion of various health functions. It has been suggested that lactic acid bacteria cultures also affect sugar absorption. Patent Document 2 shows that an extract of fermented milk kefir (molecular weight 1000 or less) made using various microorganisms including lactic acid bacteria contributes to promoting sugar uptake into myotube cells. However, neither document discloses nor suggests whether the bacterial cells of lactic acid bacteria or bifidobacteria themselves are involved in promoting sugar uptake in the intestinal tract. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] Patent Publication No. 2017-192312 [Patent Document 2] Patent No. 3813808 [Non-patent literature]

[0008] [Non-Patent Document 1] Journal of Functional Foods(2017), 36, 429-439 Summary of the Invention [Problem to be solved by the invention]

[0009] An object of the present invention is to provide a novel composition for promoting sugar absorption in the intestinal tract. [Means for solving the problem]

[0010] The present invention provides the following novel composition for promoting sugar absorption. <1> A composition for promoting sugar absorption in the intestinal tract, comprising as an active ingredient bacterial cells of a bacterium belonging to the genus Lactobacillus, Lactococcus, Leuconostoc or Bifidobacterium. <2> Bacteria belonging to the genus Lactobacillus, Lactococcus, Leuconostoc, or Bifidobacterium are, for example, Lactobacillus acidophilus, Lactobacillus amylovorus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii subsp. lactis, Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus paracasei subsp. paracasei, Lactobacillus Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. lactis, Leuconostoc mesenteroides subsp.mesenteroides, Leuconostoc pseudomesenteroides, Bifidobacterium longum, Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium pseudocatenulatum, and Bifidobacterium pseudolongum. <1> The composition for promoting sugar absorption in the intestinal tract according to claim 1. <3> Bacteria belonging to the genus Lactobacillus, Lactococcus, Leuconostoc or Bifidobacterium are Lactobacillus acidophilus JCM1132, Lactobacillus amylovorus JCM1126, Lactobacillus delbrueckii subsp. bulgaricus JCM1002, Lactobacillus delbrueckii subsp. lactis JCM1012, Lactobacillus helveticus JCM1120, Lactobacillus johnsonii JCM2012, Lactobacillus paracasei subsp. paracasei JCM8130, Lactobacillus plantarum JCM1149, Lactobacillus reuteri JCM1112, Lactobacillus rhamnosus JCM1136, Lactococcus lactis subsp. cremoris JCM16167, Lactococcus lactis subsp. lactis JCM5805, Leuconostoc mesenteroides subsp. mesenteroides JCM6124, Leuconostoc pseudomesenteroides JCM9696, Bifidobacterium longum JCM1217, Bifidobacterium adolescentis JCM1275, Bifidobacterium bifidum JCM1255, Bifidobacterium breve JCM1192, Bifidobacterium catenulatum JCM1194, Bifidobacterium pseudocatenulatum JCM1200, and Bifidobacterium pseudolongum JCM1205. <2> 2. A composition for promoting sugar absorption in the intestinal tract according to claim 1. <4> <1> ~ <3> A feed composition for promoting sugar absorption, a pharmaceutical composition for promoting sugar absorption, or a food composition for promoting sugar absorption, comprising any one of the compositions for promoting sugar absorption described above. Effect of the Invention

[0011] According to the present invention, there is provided a composition for promoting sugar absorption in the intestinal tract, which contains as an active ingredient bacterial cells of lactic acid bacteria belonging to the genera Lactobacillus, Lactococcus, and Leuconostoc, and bifidobacteria belonging to the genus Bifidobacterium. This can be done. [Brief description of the drawings]

[0012] [Figure 1] This graph compares the gene expression levels of various glucose transporters (SGLT1, GLUT2, GLUT5) after 24 hours of incubation when various heat-killed lactic acid bacteria or bifidobacteria were added to the human colon cancer-derived cell line Caco-2 cultured to resemble intestinal epithelium. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] (Lactobacillus and Bifidobacteria) The lactic acid bacteria and bifidobacteria of the present invention can be any lactic acid bacteria classified into the genera Lactobacillus, Lactococcus, and Leuconostoc, and any bifidobacteria classified into the genus Bifidobacterium. Specifically, Lactobacillus acidophilus, Lactobacillus amylovorus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii subsp. lactis, Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus paracasei subsp. paracasei, Lactobacillus plantarum, Lactobacillus reuteri reuteri, Lactobacillus rhamnosus, Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. lactis, Leuconostoc mesenteroides subsp.mesenteroides, Leuconostoc pseudomesenteroides, Bifidobacterium longum, Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium pseudolongum, and the like can be exemplified, but are not limited to these.

[0014] The lactic acid bacteria of the present invention include Lactobacillus acidophilus JCM1132, Lactobacillus amylovorus JCM1126, Lactobacillus delbrueckii subsp. bulgaricus JCM1002, Lactobacillus delbrueckii subsp. lactis JCM1012, Lactobacillus helveticus JCM1120, Lactobacillus johnsonii JCM2012, Lactobacillus paracasei subsp. paracasei JCM8130, Lactobacillus plantarum JCM1149, Lactobacillus reuteri JCM1112, Lactobacillus rhamnosus JCM1136, Lactobacillus lactis subsp. JCM5805, Leuconostoc mesenteroides subsp. mesenteroides JCM6124, Leuconostoc pseudomesenteroides JCM9696, Bifidobacterium longum JCM1217, Bifidobacterium adolescentis JCM1275, Bifidobacterium bifidum JCM1255, Bifidobacterium breve JCM1192, Bifidobacterium catenulatum Preferably, they are JCM1194, Bifidobacterium pseudocatenulatum JCM1200, and Bifidobacterium pseudolongum JCM1205. The above strains can be obtained from the RIKEN BioResource Center (Tsukuba, Ibaraki, Japan) and other sources.

[0015] (Preparation of Lactic Acid Bacteria and Bifidobacteria) Lactic acid bacteria and bifidobacteria may be cultured according to a conventional method for culturing each bacterium, and the desired amount may be prepared. An example of the preparation is shown below. Lactic acid bacteria belonging to the genus Lactobacillus are cultured using MRS medium (Difco), lactic acid bacteria belonging to the genus Lactococcus are cultured using M17 medium (Difco), and lactic acid bacteria belonging to the genus Leuconostoc and bifidobacteria belonging to the genus Bifidobacterium are cultured using 1% glucose-containing GAM bouillon (Nissui), and the resulting culture is collected by centrifugation to obtain bacterial cells. The obtained bacterial cells may be used as they are, or may be subjected to a concentration, drying, or freeze-drying process. The bacterial cells may be killed by heating and drying, etc., and used.

[0016] (How to use) As described above, the composition of the present invention can contain bacterial cells subjected to concentration, drying, or freeze-drying treatment, or killed bacterial cells obtained by heat drying, etc. as an active ingredient, and therefore can be widely used as a raw material for preparations, foods, beverages, and feeds. The bacterial cells may be cultured in a preparation, food, beverage, or feed as a medium, and the medium may be used as a preparation, food, beverage, or feed as it is, or the bacterial cells cultured in another medium may be added to the preparation, food, beverage, or feed.

[0017] The subject of administration of the composition of the present invention is not particularly limited, and the subject of administration can be humans, but the subject of administration can also be animals other than humans (e.g., dogs, cats, horses, rabbits, etc.). When the subject of administration is humans, the composition can be administered to minors under 20 years of age, adults, or elderly people over 65 years of age. The amount of the composition of the present invention to be ingested is determined individually for each subject taking into consideration the symptoms, age, etc. of the subject, but typically, for adults, it is sufficient to ingest about 1-5000 mg per day.

[0018] (Method for evaluating sugar absorption promoting ability) The evaluation can be performed by the method described in the Examples.

[0019] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples. EXAMPLES

[0020] (Example 1) Heat-killed lactic acid bacteria and bifidobacteria The test bacteria in the following (1) were inoculated into MRS medium (Difco) for Lactobacillus, M17 medium (Difco) for Lactococcus, and 1% glucose-containing GAM broth (Nissui) for Leuconostoc and Bifidobacterium, respectively, and were subjected to static culture at 37°C for 16 hours. The culture was washed twice with physiological saline and once with sterile water to obtain washed bacteria. The washed bacteria were freeze-dried to obtain a bacterial powder. The bacterial powder was diluted with sterile PBS (-) to 10 mg / ml, and heated at 80°C for 30 minutes to obtain heat-killed bacteria. The heat-killed bacteria were diluted with DMEM (Sigma) containing 10% FBS (Gibco), 1x Non-essential Amino Acids (Gibco), and 1% penicillin-streptomycin (Sigma) to 100 μg / ml.

[0021] (1) Test bacteria The following 21 strains of the genera Lactobacillus, Lactococcus, Leuconostoc, and Bifidobacterium were used as test bacteria. Lactobacillus acidophilus JCM1132, Lactobacillus amylovorus JCM1126, Lactobacillus delbrueckii subsp. bulgaricus JCM1002, Lactobacillus delbrueckii subsp. lactis JCM1012, Lactobacillus helveticus JCM1120, Lactobacillus johnsonii JCM2012, Lactobacillus paracasei subsp. paracasei JCM8130, Lactobacillus plantarum JCM1149, Lactobacillus reuteri JCM1112, Lactobacillus rhamnosus JCM1136, Lactococcus lactis subsp. cremoris JCM16167, Lactococcus lactis subsp. lactis JCM5805, Leuconostoc mesenteroides subsp. mesenteroides JCM6124, Leuconostoc pseudomesenteroides JCM9696, Bifidobacterium longum JCM1217, Bifidobacterium adolescentis JCM1275, Bifidobacterium bifidum JCM1255, Bifidobacterium breve JCM1192, Bifidobacterium catenulatum JCM1194, Bifidobacterium pseudocatenulatum JCM1200, Bifidobacterium pseudolongum JCM1205 The above strains can be obtained from the RIKEN BioResource Center (Tsukuba, Ibaraki, Japan) and other sources.

[0022] [Test Example 1] Test on the effect of lactic acid bacteria and bifidobacteria in promoting glucose transporter expression Example product 1 was subjected to the following tests. Human colon cancer cell line Caco-2 was cultured at 5 × 10 cells / well on a collagen-coated 24-well plate (BD) with Cell Matrix Type IC (Nitta Gelatin). 4 The cells were seeded at 100 μl / well and cultured in DMEM (Sigma) containing 10% FBS (Gibco), 1× Non-essential Amino Acids (Gibco), and 1% penicillin-streptomycin (Sigma). The cells were cultured for 24 days with medium changes every 2-3 days to differentiate into intestinal epithelium-like cells. After 24 days, the cells were washed once with PBS, and cultured for 24 hours with medium containing 100 μg / ml of various bacterial cells suspended in the test group and medium containing no bacterial cells in the control group. After 24 hours, the cells were washed once with cold PBS (-), and 300 μl of TRIzol Reagent (Invitrogen) was added to the washed cells, and the cells were lysed by pipetting. The cell lysate was collected in a 1.5 ml tube, incubated at room temperature for 5 minutes or more, and then stored at -80°C until RNA extraction was performed.

[0023] The cell lysate stored at -80°C was returned to room temperature, and 60 μl (one-fifth the volume) of chloroform was added. After vigorously shaking by hand, the mixture was left to stand at room temperature for 3 minutes. The mixture was centrifuged at 12,000×g and 4°C for 15 minutes, and the upper layer (aqueous layer) was transferred to another tube. An equal amount of isopropanol was added to the aqueous layer, mixed by pipetting, and left to stand at room temperature for 10 minutes. The mixture was centrifuged at 20,000×g and 4°C for 10 minutes, and the supernatant was removed, leaving the pellet. 300 μl of 75% ethanol was added, the pellet was washed by inversion, and the mixture was centrifuged at 20,000×g and 4°C for 5 minutes. The supernatant was completely removed, leaving the pellet, and the mixture was air-dried for 30 minutes. The pellet was suspended in 30 μl of Nuclease-Free Water, and incubated at 55°C for 10 minutes to dissolve the RNA. The dissolved RNA solution was cooled on ice, and the nucleic acid concentration was measured using BioSpec-nano (Shimadzu Corporation).

[0024] cDNA synthesis from RNA was performed according to a scaled-down version of the ReverTraAce (Toyobo) protocol. The total amount of RNA used for reverse transcription was 250 ng / μl. The synthesized cDNA was stored at -20°C until quantitative PCR (qPCR).

[0025] The qPCR reaction reagent used was KAPA SYBR FAST Master Mix (2x) Universal (Kapa Biosystems). The qPCR device used was StepOnePlus (Applied Biosystems). The qPCR reaction was carried out according to a scaled-down version of the "ABI device protocol (Fast program)" for KAPA SYBR FAST Master Mix (2x) Universal. 1 μl / well of 20-fold diluted cDNA was used as the template. The qPCR program used was the StepOnePlus system's Fast program with some modifications (reaction volume: 20 μl / well → 5 μl / well, reaction time of Denaturation Step: 3 seconds → 5 seconds, reaction temperature of Annealing Step: all 60°C).

[0026] The relative expression levels of SGLT1, GLUT2 and GLUT5 genes were calculated using the ΔΔCT method. The relative expression levels of each gene were corrected using the relative expression level of the housekeeping gene GAPDH. The base sequences of the primers used in the test are shown below. TIFF0007627369000001.tif51152

[0027] (Test Results) Figure 1 shows the gene expression levels of glucose transporters when 21 species of lactobacilli or bifidobacteria were added to Caco-2 cells differentiated into intestinal epithelium and cultured for 24 hours. All test strains increased gene expression of SGLT1 and GLUT5 (SGLT1 was about 1.5-2.5 times, and GLUT5 was about 1.2-2.7 times compared to the control group). In addition, four species of bacteria, Lactobacillus amylovorus, Lactobacillus paracasei subsp. paracasei, Lactobacillus reuteri, and Lactobacillus rhamnosus, had almost no effect on gene expression of GLUT2, but the other 17 species increased gene expression (about 1.2-2.3 times compared to the control group). Therefore, it was suggested that lactobacilli and bifidobacteria increase gene expression of monosaccharide transporters and increase sugar uptake in the intestinal tract regardless of the species. [Industrial Applicability]

[0028] According to the present invention, it is possible to provide a composition for promoting sugar absorption in the intestinal tract, which contains as an active ingredient bacterial cells of bacteria belonging to the genus Lactobacillus, Lactococcus, Leuconostoc, or Bifidobacterium.

Claims

1. A composition for promoting sugar absorption in the intestinal tract, comprising as an active ingredient cells of a bacterium belonging to the genus Leuconostoc.

2. The composition for promoting sugar absorption in the intestinal tract according to claim 1, characterized in that the bacterium belonging to the genus Leuconostoc is one or more selected from Leuconostoc mesenteroides subsp. mesenteroides and Leuconostoc pseudomesenteroides.

3. The composition for promoting sugar absorption in the intestine according to claim 2, characterized in that the bacterium belonging to the genus Leuconostoc is one or more selected from Leuconostoc mesenteroides subsp. mesenteroides JCM6124 and Leuconostoc pseudomesenteroides JCM9696.

4. A feed composition for promoting sugar absorption, a pharmaceutical composition for promoting sugar absorption, or a food composition for promoting sugar absorption, comprising the composition for promoting sugar absorption according to any one of claims 1 to 3.

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