Triglyceride metabolism enhancer

Abstract

To provide a lipid metabolism improver capable of obtaining a more reliable effect, by suppressing a burden on a subject.SOLUTION: In a soy milk culture medium, three genera of lactic acid bacteria, that is, lactic acid bacteria belonging to the genus Lactobacillus, lactic acid bacteria belonging to the genus Bifidobacterium, and lactic acid bacteria belonging to the genus Streptococcus are used, and a plurality of groups are formed using one or two or more kinds selected from the three genera of lactic acid bacteria, subculture each group to maintain a symbiotic state, and further subculture the lactic acid bacteria in each group to coexist with each other. A lactic acid bacteria culture solution obtained by culturing is used.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] This invention relates to neutral fat a metabolism enhancer.

Background Art

[0002] In recent years, various attempts have been made to improve obesity and dyslipidemia, which lead to serious lifestyle-related diseases. For example, improvements in diet such as exercise to consume ingested fat and the selection of low-fat foods have been proposed. In addition, various lipid metabolism promoters taken separately from diet have also been proposed.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0004] As described above, it is quite difficult for people with obesity and dyslipidemia to consume fat through exercise, and it imposes a great burden on them. On the other hand, it was also difficult to improve severe dyslipidemia only by diet. An object of this invention is to provide neutral fat a metabolism enhancer that imposes a small burden on the subject and can obtain a more reliable effect.

Means for Solving the Problems

[0007] BookThe invention classifies the above-mentioned lactic acid bacteria into the following groups: Group 1: E. faecium and L. helveticus; Group 2: E. faecium and L. acidophilus; Group 3: E. faecium and L. gasseri; Group 4: E. faecium, L. acidophilus and L. brevis; Group 5: E. faecium, L. acidophilus and L. brevis; Group 6: E. faecium, L. acidophilus, L. brevis, and L. paracasei; Group 7: B. adolescentis alone; Group 8: L. delbruickii and L. gasseri; Group 9: L. delbruickii alone; Group 10: E. faecium, L. gensenii, L. paracasei, and L. brevis alone; Group 1 Group 1 consists of E. faecium and L. gasseri, Group 12 consists of L. paracasei alone, Group 13 consists of L. gasseri, E. faecium and B. bifidum, Group 14 consists of B. longum, S. thermophilus and E. faecium, Group 15 consists of L. gasseri alone, Group 16 consists of L. bulgaricus and S. thermophilus, Group 17 consists of L. gasseri, L. lactis, L. gasseri and E. faecium, Group 18 consists of L. gasseri, S. thermophilus and L. bulgaricus, Group 19 consists of L. gasseri alone, Group 20 consists of L. lactis alone, Group 21 consists of L. gasseri and E. faecium, Group 22 consists of L. rhamnosus alone, Group 23 consists of L. casei alone, B.Longum was designated as the 24th group, and each group was subcultured to maintain a symbiotic state. Within these culture media, groups 1 and 2 were paired together, groups 3 and 4 together, groups 5 and 6 together, groups 7 and 8 together, groups 9 and 10 together, groups 11 and 12 together, groups 13 and 14 together, groups 15 and 16 together, groups 17 and 18 together, groups 19 and 20 together, groups 21 and 22 together, and 2 The lactic acid bacteria culture medium consists of a 3,24 primary culture, followed by secondary cultures with the following groups: groups 1 and 2 with groups 3 and 4, groups 5 and 6 with groups 7 and 8, groups 9 and 10 with groups 11 and 12, groups 13 and 14 with groups 15 and 16, groups 17 and 18 with groups 19 and 20, and groups 21 and 22 with groups 23 and 24. Finally, a tertiary culture is performed using a mixture of these secondary culture solutions. [Effects of the Invention]

[0009] Book The triglyceride metabolism-enhancing agent of this invention can easily improve triglyceride metabolism function. Therefore, it is expected to improve dyslipidemia while reducing the burden on subjects such as dietary restrictions and exercise. [Brief explanation of the drawing]

[0010] [Figure 1] This figure shows the culture procedure for producing the lactic acid bacteria culture solution of the embodiment. [Figure 2] This graph shows the effect of administered substances on changes in the body weight of mice. [Figure 3] This graph shows the effect of administered substances on the amount of free fatty acids in the blood of mice. [Figure 4] This graph shows the effect of administered substances on the amount of triglycerides in the blood of mice. [Figure 5] This graph shows the effect of administered substances on leptin levels in the blood of mice. [Figure 6] This graph shows the effect of administered substances on the daily food intake of mice. [Figure 7]This graph shows the effect of administered substances on the amount of lipids in mouse feces. [Figure 8] This graph shows the effect of administered substances on visceral fat percentage in mice. [Figure 9] This graph shows the effect of administered substances on the abdominal fat percentage of mice. [Figure 10] This graph shows the effect of administered substances on the ratio of liver to body weight in mice. [Figure 11] This graph shows the effect of administered substances on blood glucose levels in mice. [Modes for carrying out the invention]

[0011] [Embodiment] This invention neutral fat One embodiment of a metabolic enhancer will be described. In this embodiment, 24 groups were created by combining the following 16 types of lactic acid bacteria, and primary to tertiary co-culture was performed with these groups, and the final culture medium was sterilized. neutral fat It is intended to be used as a metabolic enhancer.

[0012] The 16 lactic acid bacteria are Lactobacillus acidophilus, Lactobacillus brevis, Lactobacillus jensenii, Lactobacillus paracasei subsp. paracasei, Lactobacillus gasseri, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus casei subsp. casei, Lactobacillus rhamnosus, Lactobacillus delbrueckii subsp. delbrueckii, Streptococcus thermophilus, Enterococcus faecium, Lactococcus lactis, Bifidobacterium longum, Bifidobacterium bifidum, and Bifidobacterium adolescentis.

[0013] In the group creation process, the above lactic acid bacteria are divided into 24 groups and subcultured for each group, and the grouping is as shown in Figure 1. Note that Figure 1 shows each process from the above group creation process to the third culture process. And "No." in the figure is the group number. That is, the first group is composed of Enterococcus faecium and Lactobacillus helveticus, the second group is composed of Enterococcus faecium and Lactobacillus acidophilus. The third group is composed of Enterococcus faecium and Lactobacillus gasseri, and the fourth group is composed of Enterococcus faecium, Lactobacillus acidophilus, and Lactobacillus brevis.

[0014] The fifth group consists of E. faecium, L. acidophilus, and L. brevis. The sixth group consists of E. faecium, L. acidophilus, L. brevis, and L. paracasei. The seventh group consists of B. adolescentis alone. The eighth group consists of L. delbrueckii and L. gasseri. The ninth group consists of L. delbrueckii alone. The tenth group consists of E. faecium, L. jensenii, L. paracasei, and L. brevis. The eleventh group consists of L. acidophilus alone.

[0015] The twelfth group consists of E. faecium and L. gasseri. The thirteenth group consists of L. paracasei alone. The fourteenth group consists of L. gasseri, E. faecium, and B. bifidum. The fifteenth group consists of B. longum, S. thermophilus, and E. faecium. The sixteenth group consists of L. gasseri alone. The seventeenth group consists of L. bulgaricus and S. thermophilus.

[0016] The eighteenth group consists of L. gasseri, L. lactis, L. gasseri, and E. faecium. The nineteenth group consists of L. gasseri, S. thermophilus, and L. bulgaricus. The twentieth group consists of L. lactis alone. The twenty-first group consists of L. gasseri and E. faecium. The twenty-second group consists of L. rhamnosus alone. The twenty-third group consists of L. casei alone. The twenty-fourth group consists of B. longum alone.

[0017] The above groups are subcultured, and among their culture solutions, the first and second groups, the third and fourth groups, the fifth and sixth groups, the seventh and eighth groups, the ninth and tenth groups, the eleventh and twelfth groups, the thirteenth and fourteenth groups, the fifteenth and sixteenth groups, the seventeenth and eighteenth groups, the nineteenth and twentieth groups, the twenty-first and twenty-second groups, and the twenty-third and twenty-fourth groups are co-cultured. This process is the primary culture process of this invention shown in (1) to (12) in FIG.

[0018] Furthermore, in the secondary culture process shown in Figure 1 (13) to (18), co-culture is performed with the above-mentioned groups 1 and 2 and groups 3 and 4, groups 5 and 6 and groups 7 and 8, groups 9 and 10 and groups 11 and 12, groups 13 and 14 and groups 15 and 16, groups 17 and 18 and groups 19 and 20, and groups 21 and 22 and groups 23 and 24. Furthermore, in the tertiary culture process shown in (19) of Figure 1, the secondary culture media of each group after the above-mentioned secondary culture are mixed, and this mixture is co-cultured.

[0019] Furthermore, the lactic acid bacteria from each group created in the above group creation process were subcultured in the following manner. That is, each group of lactic acid bacteria was selected with the expectation that they would be able to maintain a symbiotic state even after subculture. Maintaining a symbiotic state among multiple lactic acid bacteria means that multiple types of lactic acid bacteria coexist simultaneously, each maintaining its activity, and that this does not mean one type influences another, nor does it mean that only one type survives.

[0020] Each of the above lactic acid bacteria groups is formed by selecting symbiotic combinations from a variety of bacterial combinations. Furthermore, among the 24 groups of lactic acid bacteria mentioned above, some species belong to multiple groups simultaneously. This is because the species were combined considering that the symbiotic state would be maintained during the co-culturing process up to the third culture process. In addition, even within the same species of lactic acid bacteria, different strains are included.

[0021] For each lactic acid bacteria group, three types of culture media were used, selected according to the type of lactic acid bacteria: GAM semi-solid high-level medium, BL agar medium, or modified GAM agar medium, all manufactured by Nissui Pharmaceutical Co., Ltd. The cultures were incubated at 32°C for 12 hours, then at 37°C for 12 hours, and finally at 40°C for 24 hours. The grouped lactic acid bacteria were then subcultured as described above, and the culture solution was stored refrigerated at 5°C.

[0022] In this manner, the groups of lactic acid bacteria were subcultured and identified, with the identification tests being outsourced to the Japan Food Research Laboratories. The identification tests involved directly inoculating and culturing samples from each group on agar plates, picking colonies with different shapes that grew predominantly, and isolating the lactic acid bacteria from each group. These isolates were then subjected to morphological observation, physiological characteristic testing, and measurement of the GC content of intracellular DNA, and identified with reference to the following literature.

[0023] 1.Sneath, PHA, Mair, NS, Sharpe, ME and Holt, JG: “Bergey's Manual of Systematic Bacteriology” Vol. 2, (1986) Williams & Wilkins. 2. Holt, JG, Krieg, NR, Sneath, PHA, Staley, JT and Williams, ST: “Bergey's Manual of Determinative Bacteriology” Ninth Edition (1994) Williams & Wilkins. 3. Tomotari Mitsuoka: "The World of Intestinal Bacteria," (1984) Sobunsha. 4. Yoshimi Benno: Microorganisms 6, 3-14 (1990). 5. Supervised by the Ministry of Health and Welfare, Life Hygiene Bureau: "Guidelines for Food Hygiene Inspection - Microbiology Section -" (1990), Japan Food Hygiene Association. 6.Schleifer, KH and Kilpper-Balz, R.: Int.J.Syst.Bacteriol.,34,31-34(1984).

[0024] As a result of the above identification, 16 species of fungi and their strains, as shown in Figure 1, were identified, and it was confirmed that they maintain a symbiotic state within each group. Of the 24 groups that were subcultured as described above, lactic acid bacteria from adjacent groups in Figure 1 were combined and subjected to co-culture in the primary culture processes (1) to (12). In this primary culture process, three types of culture media, consisting of GAM semi-solid high-level medium, BL agar medium, or modified GAM agar medium manufactured by Nissui Pharmaceutical Co., Ltd., were used as primary culture media, and these primary culture media were selected according to the type of lactic acid bacteria.

[0025] Primary culture was performed by adding the grouped lactic acid bacteria to one of the three primary culture media described above. For example, if the culture medium for the grouped lactic acid bacteria was a highly fluid medium and also contained bacteria of the genus Bifidobacterium, 1 to 10 wt% of the lactic acid bacteria were added to the primary co-culture medium. Similarly, if the group did not contain bacteria of the genus Bifidobacterium, 1 to 10 wt% of the lactic acid bacteria were added to the primary culture medium.

[0026] When adding lactic acid bacteria to the primary culture medium described above, if the culture medium for lactic acid bacteria groups 1-24 in Figure 1 was a highly solid medium, a single amount lifted with a platinum loop was added to the primary culture medium. Then, as described above, the lactic acid bacteria were added to the primary culture medium and cultured at 37°C for approximately 6 to 12 hours until the pH reached 4.6. This is the primary culture process.

[0027] Of the 16 bacterial species listed above, those belonging to the Lactobacillus genus are L. acidophilus, L. brevis, L. gensenii, L. paracasei, L. gasseri, L. bulgaricus, L. helveticus, L. casei, L. rhamnosus, L. delbruickii, and L. lactis. Additionally, those belonging to the Bifidobacterium genus are B. longum, B. bifidum, and B. adolescentis, and those belonging to the Streptococcus genus are E. faecium and S. thermophilus.

[0028] The secondary culture process is the process shown in Figure 1, steps (13) to (18). In this secondary culture process, 0.5 wt% glucose and 0.4 wt% yeast extract were added to soy milk as the secondary culture medium.

[0029] The lactic acid bacteria, along with the primary culture medium, are added to this secondary culture medium along with the primary culture medium, and cultured at 37°C for approximately 6 to 10 hours until the pH reaches 4.55. The amount of primary culture medium added is 1 to 10 wt%. In each of the processes (13) to (18) in Figure 1, the secondary culture medium described above was used, but the secondary culture medium and added elements were the same for all lactic acid bacteria groups.

[0030] Furthermore, in the tertiary culture process shown in (19) in Figure 1, 0.5 wt% glucose and 0.4 wt% yeast extract were added to soy milk as the tertiary culture medium. All the lactic acid bacteria that had completed the secondary culture were added to this tertiary culture medium along with the secondary culture medium. At this time, the amount of secondary culture medium added was 1 to 10 wt% relative to the tertiary culture medium. The cells were then incubated at 32°C for 24 hours, then at 40°C for 48 hours, and finally at 37°C for 48 hours.

[0031] The final culture solution, consisting of lactic acid bacteria and culture medium after completing the tertiary culture process as described above, was sterilized by heating at 85°C or higher. This sterilized final culture solution is the stock solution of the lactic acid bacteria culture solution of the invention.

[0032] Then, the supernatant, residue, and ethanol extract were prepared from the stock solution. The supernatant was obtained by removing the solids from the stock solution by centrifugation (4°C, 10,000 × g, 15 min). The solids content was 11.6 [wt%] as a result of freeze-drying the stock solution.

[0033] Furthermore, the powder obtained by freeze-drying the above stock solution was mixed with ethanol and filtered through filter paper (ADVANTEC 5C), and concentrated under reduced pressure to obtain a yellow oily substance. This oily substance was redissolved in ether, sodium sulfate was added, and it was left overnight to remove water. After that, the sodium sulfate was filtered off, and the filtrate was concentrated to obtain an ethanol extract. 3.0 g of this ethanol extract was obtained from 100 g of the stock solution.

[0034] Furthermore, the residue consisted only of the insoluble components obtained by removing the water-soluble and lipid components from the stock solution. Specifically, the insoluble components obtained by removing the water-soluble and lipid components from the stock solution were washed with water, filtered, and freeze-dried. This residue was 5.6 wt% of the stock solution. Furthermore, the pH of the filtrate of the residue was 5.7. From this, it was determined that the water-soluble components had been removed, and that there were almost no leached substances from this residue. The above-mentioned stock solution, supernatant, residue, and ethanol extract are each of the following: neutral fat It is a metabolic enhancer.

[0035] [Lipid metabolism confirmation experiment] In this embodiment, mice were divided into six groups A to F, and the stock solution, supernatant, ethanol extract, and residue described above were administered to the mice. Experiments were then conducted to confirm various numerical values ​​related to the lipid metabolism function of the mice.

[0036] Each of the mouse groups A through F used in this experiment consisted of five mice. Of these, mouse group A was fed only standard diet. This mouse group A is referred to as normal group A. In addition, mouse group B was fed only high-fat diet throughout the experimental period, and this mouse group B is referred to as control group B.

[0037] Furthermore, mouse groups C to F were fed a high-fat diet for 4 weeks, and then, while continuing to be fed a high-fat diet, according to the embodiment... neutral fat The above-mentioned substance, which acts as a metabolic enhancer, was administered. 0.3 mL of the substance was administered daily in the morning via a tube. Changes in each mouse group A to F were then measured for 4 weeks. For the high-fat diet used to feed the mouse group, we used High Fat Diet 32 ​​for mice and rats manufactured by CLEA Japan Co., Ltd., and for the standard diet, we used CLEA Rodent CE-2 for mice, rats, and hamsters manufactured by CLEA Japan Co., Ltd.

[0038] The mouse group C was administered the undiluted solution described above, and this group of mice is referred to as the undiluted solution group C. The supernatant was administered to mouse group D, and this group of mice is referred to as supernatant group D. The above-mentioned residue was administered to mouse group E, and this group of mice is referred to as residue group E. The above ethanol extract was administered to mouse group F, and this mouse group is referred to as ethanol extract group F.

[0039] [body weight] The average body weight of all mice upon arrival was 25.8 ± 2.1 g. After four weeks of feeding with a high-fat diet, the average body weight of all mice increased to 32.9 ± 3.2 g, a 29.5 wt% increase. At this time, the average body weight of the normal group A, which was fed a standard diet, was 29.8 ± 2.4 g, which was less than the average body weight of all mice. Therefore, the mice fed with the high-fat diet were judged to be obese.

[0040] Figure 2 shows the changes in body weight of mice after administration of the above-mentioned substances. Each graph corresponds to the normal group A (Group A), control group B (Group B), undiluted solution group C (Group C), supernatant group D (Group D), residue group E (Group E), and ethanol extract group F (Group F).

[0041] As shown in Figure 2, the control group B gained weight, with a final weight of 36.3 ± 3.6 g, representing an increase of 3.0 g over four weeks. In contrast, the average body weight of group C (concentrated solution group) was 33.1 ± 0.6 [g], which was approximately the same as their body weight at the start of the experiment.

[0042] Furthermore, in the supernatant group D, a decrease in body weight was observed around the 7th day after administration, with the average body weight on the final day being 30.6 ± 3.4 [g], a decrease of approximately 3 [g] from the initial weight. The body weight of the residue group E was 37.0 ± 5.9 [g], which was almost the same as that of the control group B. Body weight after lipid administration was 34.0 ± 2.9 g, an increase of approximately 0.6 g during the experimental period.

[0043] These results confirm that the administration of substances other than the residue slowed or decreased weight gain. While each of the above-mentioned substances contains various substances from the lactic acid bacteria culture solution, the residue contains very few eluted components; therefore, it can be expected that the effect of the lactic acid bacteria culture solution is less pronounced in residue group E.

[0044] [Blood free fatty acids] Figure 3 shows the results of investigating the effect of the above-mentioned administered substances on free fatty acids in the blood of mice. It is known that blood free fatty acid levels increase mainly when there is a sugar deficiency or excessive lipid accumulation. In the experiment, the normal group A had a level of 1.50±0.22 [meq / L], the control group B had 1.54±0.21 [meq / L], the undiluted solution group C had 1.05±0.05 [meq / L], the supernatant group D had 0.92±0.07 [meq / L], the residue group E had 0.96±0.07 [meq / L], and the ethanol extract group F had 0.80±0.08 [meq / L]. Groups C to F, which were administered the above-mentioned substance consisting of lactic acid bacteria culture solution, showed a suppressed increase in blood free fatty acid levels compared to the control group B.

[0045] [Blood neutral fat] Figure 4 shows the results of an investigation into the effect of the above-mentioned administered substances on blood triglycerides. Blood triglycerides, like blood glucose levels, are known to increase with obesity. The results were 157.1 ± 5.5 [mg / dL] in the normal group A and 169.9 ± 16.1 [mg / dL] in the control group B.

[0046] Furthermore, the levels were 120.7±7.3 [mg / dL] in the undiluted solution group C, 135.1±9.8 [mg / dL] in the supernatant group D, 145.5±5.8 [mg / dL] in the residue group E, and 108.9±13.0 [mg / dL] in the ethanol extract group F. Compared to the control group B, triglycerides decreased in groups C to F that received the above-mentioned substances, with particularly large decreases in blood triglycerides in the undiluted solution group C and the ethanol extract group F.

[0047] [Blood leptin levels] Figure 5 shows the results of an investigation into the effects of the above-mentioned administered substances on blood leptin levels. Leptin is known as a hormone that suppresses appetite and is thought to be related to blood glucose levels and blood triglycerides.

[0048] The results were as follows: normal group A: 2280.5±90.0 [pg / mL], control group B: 21600±2470 [pg / mL], stock solution group C: 15600±3080 [pg / mL], supernatant group D: 10700±4120 [pg / mL], residue group E: 22400±5930 [pg / mL], and ethanol extract group F: 13700±2490 [pg / mL]. While a tendency toward decreased leptin levels was observed in stock solution group C, supernatant group D, and ethanol extract group F, statistical tests showed no significant difference compared to control group B. From this, we can infer that the slowdown in weight gain in the undiluted solution group C, the supernatant group D, and the ethanol extract group F, as shown in Figure 2, was not due to a decrease in appetite caused by leptin.

[0049] [Food intake] Figure 6 shows the results of investigating the effect of administered substances on the daily food intake of mice during the experimental period. The average daily food intake after administration was 6.6±0.1 g in the normal group A, 4.0±0.1 g in the control group B, 3.7±0.1 g in the undiluted solution group C, 3.3±0.1 g in the supernatant group D, 3.9±0.1 g in the residue group E, and 3.8±0.0 g in the ethanol extract group F. There was no significant difference in food intake among the groups fed high-fat diets, indicating no effect from the administered substance.

[0050] Thus, despite no difference in food intake between control group B and other high-fat diet groups C-F, differences in body weight, blood free fatty acid levels, and blood triglyceride levels were observed, which can be presumed to be due to the influence of the components of the administered substance consisting of lactic acid bacteria culture solution (see Figures 2, 3, and 4).

[0051] [Lipids in feces] Figure 7 shows the results of examining the percentage of lipids in feces excreted within 24 hours from the day before the final day of the experiment. In the normal group A, it was 2.8±0.1 [wt%], in the control group B, it was 1.5±0.1 [wt%], in the undiluted solution group C, it was 2.1±0.1 [wt%], in the supernatant group D, it was 1.4±0.1 [wt%], in the residue group E, it was 2.0±0.1 [wt%], and in the ethanol extract group F, it was 2.5±0.2 [wt%]. The results of the statistical test showed that the amount of lipids in the feces excreted was almost the same as in the control group, indicating no effect from the administered substance.

[0052] These results indicate that among the groups that ingested a large amount of fat through a high-fat diet, groups C-F, which were administered a substance consisting of lactic acid bacteria culture solution, showed a suppression of the increase in blood triglycerides and blood triglyceride levels compared to the control group B. This was not because the fat was excreted in the feces. In other words, it is thought that the components in the above-mentioned substance improved lipid metabolism function, and the ingested fat was metabolized within the mice's bodies.

[0053] [Visceral fat percentage] Figure 8 shows the visceral fat percentage quantified using image processing. The values ​​were 2.5±0.5 [vol%] for the normal group A, 13.1±0.8 [vol%] for the control group B, 10.6±1.0 [vol%] for the undiluted solution group C, 7.8±2.1 [vol%] for the supernatant group D, 16.2±1.2 [vol%] for the residue group E, and 12.1±1.5 [vol%] for the ethanol extract group F. In the supernatant group D, visceral fat was significantly reduced compared to the control group B, and in the undiluted solution group C, there was a trend toward reduction. Thus, it was confirmed that administration of both the undiluted solution and the supernatant reduced visceral fat.

[0054] [Abdominal fat percentage] Figure 9 shows the fat percentage, calculated by dividing the weight of excised abdominal lipids by body weight. The percentages were 1.6±0.3 [wt%] in the normal group A, 5.1±0.6 [wt%] in the control group B, 2.7±0.3 [wt%] in the undiluted solution group C, 2.8±0.4 [wt%] in the supernatant group D, 5.4±0.5 [wt%] in the residue group E, and 3.9±0.3 [wt%] in the ethanol extract group F. The amount of excised fat was significantly lower in the undiluted solution group C and the supernatant group D compared to the control group B.

[0055] [Liver weight] Figure 10 shows the effect of administered substances on the ratio of liver weight to body weight. A higher value indicates a larger liver and greater fat accumulation. The wt% levels were 1.9±0.2 in the normal group A, 2.8±0.4 in the control group B, 1.5±0.1 in the stock solution group C, 1.9±0.1 in the supernatant group D, 2.2±0.4 in the residue group E, and 1.8±0.2 in the ethanol extract group F.

[0056] Compared to control group B, groups C-F, which received the administered substance, showed lower values, suggesting that the administration of the lactic acid bacteria culture solution reduced fat accumulation in the liver. In particular, the liver of group C, which received the undiluted solution, was smaller, suggesting that the undiluted solution contains a large amount of components that suppress fat accumulation in the liver.

[0057] [Blood sugar levels] Figure 11 shows the results of an investigation into the effect of administered substances on blood glucose levels. Blood glucose levels increased with obesity, with normal group A at 158.0±6.8 mg / dL, control group B at 188.9±6.1 mg / dL, undiluted solution group C at 117.5±8.2 mg / dL, supernatant group D at 99.5±9.1 mg / dL, residue group E at 275.9±24.7 mg / dL, and ethanol extract group F at 161.2±14.0 mg / dL. Undiluted solution group C and supernatant group D significantly suppressed the rise in blood glucose levels. When consuming energy, the body first consumes sugar, and then breaks down lipids into fatty acids. Therefore, when the rise in blood glucose levels is suppressed, lipids are more easily consumed. In other words, it can be inferred that in the undiluted solution group C and the supernatant group D, the rise in blood glucose levels was suppressed, and lipid consumption was further promoted.

[0058] Furthermore, although specific figures are omitted, when the volume percentage of subcutaneous fat was measured using image analysis, no significant difference was observed among groups B to F, which were fed a high-fat diet. From the above results, it was confirmed that administering the undiluted lactic acid bacteria culture solution, supernatant, residue, and lipids of the embodiment resulted in a reduction of fat, particularly in internal organs and blood. The above reduction in fat is due to an improvement in fat metabolism function, and the above-mentioned undiluted solution, supernatant, residue, and lipids neutral fat This is because it functions as a metabolic enhancer. Therefore, the above embodiment neutral fat By taking metabolism-enhancing supplements, neutral fat Metabolic function improves, and improvement in dyslipidemia can be expected relatively easily.

[0059] For the measurement of triglycerides and free fatty acids, a laboratory assay kit from Fujifilm Wako Pure Chemical Corporation was used, and for leptin, a mouse / rat leptin kit from Biovendor was used. Blood samples were collected from the heart after an overnight fast. Furthermore, Tukey's test was used to verify the measured values.

[0060] Furthermore, when the ratio of spleen weight to body weight was measured in mice, no significant differences were observed in any of the groups. It is a known fact that ingesting poisons can lead to weight loss and spleen enlargement. However, as mentioned above, there were no significant differences between control group B and the undiluted solution group C, supernatant group D, residue group E, and ethanol extract group F, confirming that the administered substance consisting of lactic acid bacteria culture solution is not poisonous. [Industrial applicability]

[0061] Book wishInvention medium low fat Metabolism enhancers, when combined with exercise and dietary improvements, can be expected to further improve dyslipidemia.

Claims

[Claim 1] E. faecium and L. helveticus are in the first group, E. faecium and L. acidophilus are in the second group. E. faecium and L. gasseri are in the third group. E. faecium, L. acidophilus, and L. brevis are in Group 4. E. faecium, L. acidophilus, and L. brevis are in Group 5. E. faecium, L. acidophilus, L. brevis, and L. paracasei are in Group 6. B. Adolescentis alone is in Group 7. L. delbrickii and L. gasseri are in Group 8. L. delbrickii alone is in Group 9. E. faecium, L. gensenii, L. paracasei, and L. brevis are in Group 10. L. acidophilus alone is in Group 11, E. faecium and L. gasseri are in Group 12. L. paracasei alone is in Group 13. L. gasseri, E. faecium, and B. bifidum are in Group 14. B. longum, S. thermophilus, and E. faecium are in Group 15. L. Gasselly is placed in Group 16 on its own. L. bulgaricus and S. thermophilus are in Group 17. L. gasseri, L. lactis, L. gasseri, and E. faecium are in Group 18. L. gasseri, S. thermophilus, and L. bulgaricus are in Group 19. L. lactis alone is in group 20. L. gasseri and E. faecium are in Group 21. L. rhamnosus is listed as a separate species in Group 22. L. casei alone is in Group 23. B. Longham is placed in Group 24 on its own. In a soy milk culture medium, each of the above groups is subcultured to maintain a symbiotic state, and then, from these culture solutions, the groups 1 and 2 are cultured together, the groups 3 and 4 together, the groups 5 and 6 together, the groups 7 and 8 together, the groups 9 and 10 together, the groups 11 and 12 together, the groups 13 and 14 together, the groups 15 and 16 together, the groups 17 and 18 together, the groups 19 and 20 together, the groups 21 and 22 together, and the groups 23 and 24 together are cultured in a primary culture. Furthermore, secondary cultures are performed with the above groups 1 and 2 and 3 and 4, 5 and 6 and 7 and 8, 9 and 10 and 11 and 12, 13 and 14 and 15 and 16, 17 and 18 and 19 and 20, and 21 and 22 and 23 and 24 with each other. Furthermore, a neutral fat metabolism enhancer consisting of a lactic acid bacteria culture solution obtained by tertiary culture using a mixture of these secondary culture solutions.

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