JP2003252772A - Agent for prevention, improvement and treatment of age-related metabolic disorder - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an agent for the prevention, improvement and treatment of the age-related metabolic disorder and/or foods and drinks having preventing, improving and treating effect on the age-related metabolic disorder. <P>SOLUTION: The agent for the prevention, improvement and treatment of the metabolic disorder contains cultured product of lactobacillus belonging to Lactobacillus gasseri and/or the bacterial cell as an active component. The food and drink effective for the prevention, improvement and treatment of the metabolic disorder contains the active component. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention has a preventive / ameliorating / treating agent for metabolic disorders associated with aging and / or a preventive / ameliorating / therapeutic action for metabolic disorders associated with aging. Regarding food and drink.

[0002]

Background Art Aging is an unavoidable life phenomenon for living things. Various factors have been hypothesized as causes of aging. For example, at the cell level, there are a program theory and an error failure theory. Whichever theory is used, it is unclear how changes in the cells caused by the aging mechanism are related to the aging phenomenon of the whole body. However, there is also the idea that those changes occur in central cell groups and have a general spillover effect, and that changes that occur peripherally in individual cells affect each other and are generally reflected in the aging phenomenon. is there. Various indicators have been proposed to use these changes as indicators of the phenomenon of aging, but there is no definite indicator yet. In recent years, it has been said that by improving various metabolic disorders, metabolic disorders, or functional disorders that occur with aging, lifespan extension is realized, and various aging indices decrease. With the establishment of westernized dietary habits, metabolic disorders in Japanese are changing with age.

[0003] Various diseases which are considered to be caused by metabolic abnormality occurring with aging, which has been particularly problematic in recent years, include hyperlipidemia due to obesity, senile dementia with brain amyloid accumulation, immune abnormality, and intestinal tract. These include renal dysfunction such as colon cancer and uremia caused by functional deterioration and alternation of intestinal microorganisms. There are various causes for obesity, but no radical therapeutic method that is more effective than diet and exercise has been found. Heretofore, as a body fat accumulation inhibitor with aging other than dietary treatment, a method of administering fruits or essences of Rosaceae plants (JP-A-8-198768), a method of administering a composition derived from buckwheat flour (JP-A-9-183735), a method of administering procyanidins (JP-A-9-291039), and a method of administering winter melon peel and eggplant skin (JP-A-11-1).
64668), a method of changing energy metabolism efficiency by administering a specific fatty acid (JP 2001-286268 A), various methods and agents have been proposed. However, there is no proposal for suppressing fat accumulation due to Lactobacillus gasseri having intestinal colonization properties described below in the present invention. There are two types of senile dementia, cerebral vascular dementia and Alzheimer's dementia, and particularly in recent years, the occurrence of Alzheimer's dementia accompanied by accumulation of amyloid in the brain (amyloidosis) has been increasing. As a substance that suppresses amyloidosis (amyloidosis), a method of administering amyloid β protein (Japanese Patent Publication No. 6-507585) or a fragment of amyloid protein (Japanese Patent Publication No. 11-500462) , A method of administering a sugar composition having a specific structure substituted with anions (Japanese Patent Publication No. 2001-513569) has been proposed. However, there is no proposal to suppress the intracerebral amyloid storage disease due to Lactobacillus gasseri having intestinal colonization properties described below in the present invention.

[0004] A decrease in renal function due to aging has been cited as various causes such as a decrease in glomerular function due to aging, a decrease in function of mesangium cells, and accumulation of waste products / IgA. In order to suppress the deterioration of renal function with age, IGF-
There have been proposed novel therapeutic agents and therapeutic methods such as a method of administering 1 or IGF-3 and a method of administering an IP receptor agonist (JP 2000-191523 A). However, there is no proposal for suppressing a decrease in renal function or metabolism due to Lactobacillus gasseri having an intestinal colonization property described below in the present invention. All of the above diseases severely limit the thinking and behavior of the elderly, and as a result, reduce the quality of life (QOL) of the elderly. Various therapeutic agents for improving the symptoms of these diseases have been proposed and put into practical use, but they do not drastically improve the QOL of the elderly.

[0005]

The present inventors have [0006] is, where has been conducting research on lactic acid bacteria with fixing ability within the human intestinal tract, belong lactic acid bacteria, in particular Lactobacillus gasseri (Lactobacillusgasseri) with intestinal fixing property When a culture obtained by culturing lactic acid bacteria and / or cells is administered,
It has been found that these microorganisms act on the host by colonizing the intestinal tract, resulting in a significant prolongation of life in experimental animals. As a result of further investigation, when administered Lactobacillus gasseri the (Lactobacillus gasseri) aging animals, Lactobacillus gasseri that is fixed to the intestinal tract (Lactobacillus gasseri) acts on the host, the metabolic abnormalities associated with aging Improve, suppress the decline and deterioration of kidney function,
It was found that it has a surprising effect of improving lipid metabolism that causes obesity and further suppressing the accumulation of amyloid in the brain. As a result, they have found that the quality of life associated with aging clearly improves, and have completed the present invention. Accordingly, the present invention is directed to Lactobacillus ga.
a culture obtained by culturing a lactic acid bacterium belonging to
Or prevent / ameliorate metabolic disorders with bacterial cells as active ingredients
An object is to provide a therapeutic agent. Furthermore, the present invention relates to Lactobacillus gasseri .
It is an object of the present invention to provide a culture product obtained by culturing a lactic acid bacterium belonging to and belonging to, and / or a food or drink, which has a preventive / ameliorating / therapeutic effect on dysmetabolism associated with aging, to which cells are added.

[0006] The inventors of the present invention have been conducting various studies on fermented milk, and among them, among lactic acid bacteria and human-derived lactic acid bacteria isolated from these fermented milks, particularly lactobacillus
It was found that the gasseri ( Lactobacillus gasseri ) has a high intestinal colonization property in humans that has never been seen, and further, this Lactobacillus gasseri ( Lactobacillus gasseri )
However, by improving the metabolic abnormality with age by intestinal colonization that has not been known, as a result, suppress the deterioration or deterioration of renal function, reduce the accumulation of lipids that cause obesity,
Furthermore, they found that the accumulation of amyloid in the brain was suppressed and the life span was extended, and they succeeded in solving the above problems.

[0007]

Means for Solving the Invention The present invention relates to age-related metabolic disorders containing a culture and / or cells obtained by culturing a lactic acid bacterium belonging to Lactobacillus gasseri as an active ingredient. Related to preventive / improvement / therapeutic agents. The present invention also relates to foods and drinks to which such active ingredients are added, which have the preventive, ameliorating, and treating effects on metabolic disorders associated with aging. That is, the present invention is an invention having the following configurations described in the claims.

[0008] (1) Lactobacillus gasseri (Lactobac
illus gasseri ) culturing lactic acid bacteria belonging to illus gasseri ) and / or a preventive / ameliorating / therapeutic agent for dysmetabolic disease containing a bacterial cell as an active ingredient. (2) Lactobacillus gasser
The lactic acid bacterium belonging to i ) is Lactobacillus gasseri , which has human intestinal colonization (1)
The preventive / ameliorating / therapeutic agent for the described metabolic disorders. (3) The preventive / ameliorating / therapeutic agent for metabolic disorders according to (1) or (2), wherein the metabolic disorders are renal dysfunction, brain amyloidosis, and dyslipidemia. (4) Lactobacillus gasser
The preventive / ameliorating / therapeutic agent for metabolic disorders according to any one of (1) to (3), wherein the culture obtained by culturing the lactic acid bacterium belonging to i ) is fermented milk. (5) Lactobacillus gasser
Lactic acid bacteria belonging to i ) are Lactobacillus gasseri ( Lactobac
illus gasseri ) SBT2055 (FERM P-15535) (1) to (4) prevention of metabolic disorders according to any one of
Improvement / therapeutic agent. (6) Lactobacillus gas ser
A food and drink having a preventive, ameliorating, and therapeutic action for metabolic disorders, which is obtained by culturing lactic acid bacteria belonging to i) and / or cells.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention has been made in order to solve the above-mentioned problems, and the present inventors newly set the following criteria when screening a target lactic acid bacterium. Then, a strain that matches the purpose was selected. That is, the present inventors show that among many Lactobacillus gasseri derived from fermented milk and humans, gastric acid resistance is high, growth is good under low pH conditions, and high colonization in the human intestine is shown. Shows intestinal cell affinity, is resistant to bile acids, has the effect of preventing, ameliorating, and treating various metabolic disorders associated with aging by establishing in the intestine, and has high survival potential when applied to foods. By setting conditions such as excellent flavor, physical properties, etc., we have conducted intensive studies on selection of strains. As a result of screening under these conditions, the following strains could be selected as strains that meet these conditions. The strain has been deposited at the Patent Microorganism Depositary Center, National Institute of Advanced Industrial Science and Technology, with the following deposit number.

Strain Lactobacillus gasseri SBT2055 FERM
P-15535 This strain has a high affinity for human intestinal cells, is capable of surviving and reaching the intestinal tract when orally administered, and can remain resident in the intestinal tract for a long time. By growing in-house, it acts on the host and prevents / improves metabolic disorders associated with aging.
treat. Lactobacillus gasseri ( Lactobacillus gasseri ) lactic acid bacteria that have been administered from the outside of the body are colonized in the intestine, and it is not known at all that such a physiological effect is exhibited, and it was first revealed by the present inventors.

Further, in the present invention, not only the above-mentioned deposited strain but also Lactobacillus gasseri which is isolated from human or fermented milk and has the above-mentioned action can be used. . Next, the method for culturing these lactic acid bacteria will be described. As the medium of Lactobacillus gasseri of the present invention, various media such as a milk medium, a medium containing milk components, a semi-synthetic medium containing no milk components, and the like can be used. Examples of such a medium include a reduced skim milk medium obtained by reducing skim milk and sterilizing by heating. The culture method is static culture or neutralization culture in which the pH is controlled to be constant, but the culture method is not particularly limited as long as the bacteria grow well.

The present invention uses the culture and / or cells obtained as described above as an active ingredient. Further, dried powder may be used as an active ingredient. It is preferable that these are dried by freeze-drying because they can be dried without degrading the cells. It is desirable to ingest these active ingredients orally. Further, these powders can be orally administered as powders, tablets, pills, capsules or granules by mixing with a suitable excipient such as lactose. The dose is appropriately determined individually in consideration of the subject's symptoms, age, etc., but is usually 0.5 to 1 as a dry matter per adult per day.
It is 0 g, and this may be administered in several divided doses per day.
Particularly preferably, each strain can be administered as a live bacterium and administered at 10 ⁸ to 10 ¹² cfu / day per adult, whereby the desired effects of the present invention can be exhibited. By ingesting in this way, it colonizes the intestinal tract and exerts the desired effect.

Further, the active ingredient of the present invention may be added to the raw material during the manufacturing process of food or drink. As the food and drink, any food and drink may be used, and examples thereof include milk drinks, fermented milk,
Examples thereof include fruit juices, jellies, candies, dairy products, processed egg products such as mayonnaise, and food products such as confectionery breads such as butter cake. However, as a characteristic of the present invention, it is necessary for lactic acid bacteria to settle in the intestinal tract in a living state, and it is preferable to avoid excessive heating. Further, a conventional technique such as microcapsules may be adopted to take measures to avoid heating. Furthermore, the food or drink according to the present invention is the above-mentioned Lactobacillus gasseri .
It may be yogurt or the like produced by lactic acid fermentation using the above strain. Lactobacillus gasseri ( Lactobacillus gass)
eri ) .SBT2055 (LG2055) is used to specifically describe the mycological properties and in vitro and in vivo effects. However, the present invention is not limited to this description.

[0014]

[Test Example] Properties of LG2055 strain 1. Bacteriological properties (1) Shows the results after anaerobic culture at 37 ° C. for 48 hours using a microbial LBS agar plate medium. Shape: Bacillus Size: 0.5-1 × 3-4 μm Many linked (2) Gram stainability Positive (3) Colony morphology: Circular rim: Wavy size: 2-3 mm diameter Color: White Surface: Smooth (4 ) Spore formation Negative (5) Gas production None (6) Motility None (7) Catalase activity negative (8) Skim milk coagulation Coagulation (9) Gelatin liquefaction None (10) Nitrate reduction None (11) Indole production None (12) Hydrogen sulfide productivity None

2. Fermentability of sugar Commercially available bacterial identification kit (Api 50CH, Biomerieu)
The results of examining the fermentability of sugar by others are described below. Glycerol-erythritol-D-arabinose-L-arabinose + ribose-D-xylose-L-xylose-adnitol-β-methyl-D-xyloside-galactose + D-glucose + L-fructose + D- Mannose + L-sorbose-rhamnose-dulcitol-inositol-mannitol-sorbitol-α? Methyl-D-mannoside-α? Methyl-D-glucoside-N-acetyl-glucosamine + amygdalin + arbutin + esculin + salicin + cellobiose + maltose + lactose + melibiose-saccharose + trehalose + inulin-merezitose-D-raffinose-β-amiditol-glycologen-glycologen-glycologen Gentiobiose + D-tulanose-D-lyxose-D-tagatose + D-arabitol-L-arabitol-gluconate-2-ketogluconate-5-ketogluconate-(+ indicates fermentable, -not fermentable) Is shown.)

The above-mentioned taxonomical characteristics and fermentability of sugar are determined by the typical Lactobacillus acidophilus complex strain (Lactob).
acillus acidophilus complex).

Then, a confirmation test by a DNA homology test was carried out as follows. DNA homology test Lactobacillus reference strains listed below, test strain LG2055
Strain, and as a control, Escherichia coli (Escherichi
a. coli) DNA was extracted and purified. The DNA homology between the LG2055 strain and each reference strain was examined by the DNA hybridization method when the homology between the DNAs of the LG2055 strain was 100% and the DNA homology between the LG2055 strain and E. coli was 0%. As a result, the LG2055 strain became Lactobacillus gasseri.
LG205 has more than 90% homology with JCM1131 strain.
Five strains were identified as Lactobacillus gasseri.

Gastric acid resistance The gastric acid resistance test was performed by Takiguchi et al. (Journal of Enterobacteriaceae 14.11-1).
8.2000), an artificial gastric juice having a pH of 2.0 was prepared and a gastric acid resistance test was conducted. The LG2055 strain survived for 3 hours or more.

Artificial intestinal fluid resistance An artificial intestinal fluid containing bile powder was prepared according to the method of Takiguchi et al. (Journal of Intestinal Bacteriology 14.11-18.2000), and the LG2055 strain treated with the above-mentioned artificial gastric juice was added thereto to improve resistance. When tested, it showed a viability of 20 hours or more. It was confirmed that the LG2055 strain passed through the digestive tract and survived and reached the large intestine.

Human intestinal transit ability and intestinal colonization Nonfat milk solid 9.5%, milk fat 3.0% milk 4% LG2055 starter inoculated and fermented for 4 hours at 39 degrees fermented milk healthy adult 42 volunteers get 100g daily for 4 weeks 1 day
It was swallowed and observed for changes in intestinal bacteria. During the test period, evaluation was conducted by freely eating except for prohibiting the intake of foods, oligosaccharides, and drugs that affect the intestinal bacteria. The LG2055 strain, which was not detected before the test, was detected in all the subjects after 4 weeks, and both strains were found to have high intestinal colonization.

Animal test A. In vivo effect test on the longevity of experimental animals 1. Preparation of lactic acid bacteria skim milk culture LG2055 strain was used. The lactic acid bacterium was cultivated at 37 ° C. for 16 hours in an 11.55% skim milk medium containing 0.5% yeast extract (manufactured by Asahi Breweries) sterilized at 115 ° C. for 20 minutes. The obtained culture was freeze-dried and then ground in a mortar.

2. Preparation of test feed Table 1 shows the diet composition based on AIN-93M (manufactured by Oriental Yeast Co., Ltd.). 5% skim milk or the above skim milk lactic acid bacteria culture was added to this feed.

[0023]

[Table 1]

3. Senescence-Accelerated, which is a model for accelerated aging of 5-week-old test animals
Mouse (SAM) P8 strain female mice (obtained from Japan SLC) were divided into 2 groups and 30 in each group. After preliminarily breeding in AIN-93M for 1 week,
The animals were raised on the above test diet until they died. During this period, ion-exchanged water and feed (within 7 g to 8 g) were freely taken.

4. Body weight measurement The body weight was measured once a week during the breeding period. Analysis of variance was performed by the division method for body weights from 5 weeks to 60 weeks of age.

5. Method for measuring food intake Food intake was measured about every 4 weeks. A pre-weighed feed was given, the amount of food remaining in the container was measured at the same time on the next day, and the food intake per day was calculated.

6. Flora analysis Fresh feces were collected, and according to the method of Mitsuoka et al. Described in "Isolation and identification of anaerobic bacteria" (Nane Shuppan), 5 weeks old (initial value), 3 months old, 5 months old Intestinal flora analysis was performed at ages of 7 months, 10 months, and 12 months. The obtained data was subjected to analysis of variance by the division method.

B. Test for effects on aging index (biological function at 40 weeks of age) 1. Preparation of lactic acid bacterium skim milk culture LG2055 strain was used in the same manner as in the above-mentioned A. Lactic acid bacteria, 115 ℃, 2
It was cultivated at 37 ° C. for 16 hours in a 11.55% skim milk medium containing 0.5% yeast extract (manufactured by Asahi Breweries, Ltd.) sterilized for 0 minutes. The obtained culture was freeze-dried and then ground in a mortar.

2. Test feed In the same manner as above, a diet based on AIN-93M (manufactured by Oriental Yeast Co., Ltd.) was prepared, and 5% skim milk or the above skimmed milk lactic acid bacterium culture was added to this diet.

3. Experimental animals Two 5-week-old SAM P8 female mice (obtained from Japan SLC)
Group, 10 animals were used for each group. After preliminarily breeding with AIN-93M for 1 week, it was fed with the above test feed for 36 weeks. During this period, ion-exchanged water and feed were freely taken. After breeding up to 40 weeks of age, they were fasted for about 16 hours, opened under anesthesia with Nembutal (Dainippon Pharmaceutical Co., Ltd.), and sacrificed by collecting blood from the inferior vena cava.

4. Body weight measurement The body weight was measured once a week during the breeding period. The obtained data was subjected to analysis of variance by the division method.

5. Food intake measurement method Food intake should be measured weekly at an early age and about 1 afterwards.
The measurement was performed once a month (one week). A pre-weighed feed was given, the amount of food remaining in the container was measured at the same time on the next day, and the food intake per day was calculated.

6. During the period of breeding in the metabolic cage, the animals were raised in the metabolic cage four times, and urine and feces were collected. Breeding in metabolic cages was performed at 14 weeks, 22 weeks, 29 weeks and 38 weeks of age. After the acclimatization for 2 days, feces and urine were collected for about 24 hours from the 3rd day to the 4th day. Drinking water, feed intake and body weight were also monitored during the period.

7. Measurement of urine 8-hydroxydeoxyguanosine For measurement of 8-OHDG of urine obtained by breeding metabolic cages
It measured using the ELISA kit (made by Japan Aging Control Research Institute).

8. Measurement of the amount of accumulated amyloid in the brain Immediately after sacrifice, dissection was performed and the cerebrum was excised. Obtained cerebrum 0.5% protein inhibitor mixture (manufactured by SIGMA)
It homogenized with the Teflon (trademark) homogenizer using the addition PBS, and measured using the amyloid beta (1-42) measurement kit (made by the Institute for Immunological Biology).

9. The abdominal wall fat weight was dissected immediately after slaughter, the abdominal abdominal fat was excised from the left and right, and the weight was measured with a precision balance METTLER AE240 (manufactured by Japan Siber-Hegner).

10. Standing movement and locomotor activity measurement method At 40 weeks of age, the spontaneous movement and standing movement of mice were measured. The number of movements between the coils and the number of standings in 30 minutes were measured. An experimental animal momentum measuring device (manufactured by Muromachi Kikai Co., Ltd.) was used for the measurement.

11. Plasma analysis After fasting for about 16 hours, under Nembutal anesthesia, whole blood was collected from the inferior vena cava and treated with an anticoagulant (heparin 10 units / m
After l), plasma was obtained by centrifugation. Then, it measured using the biochemical automatic analyzer FDC5500 (made by Fuji Film Medical).

12. Measurement of IgA in stool Feces obtained by breeding in a metabolic cage were dried with a centrifugal evaporator CVE-200D (manufactured by EYELA), then weighed, and protein inhibitor (manufactured by SIGMA) 0.5%
Added PBS was added, the feces were dispersed to a concentration of 0.05 g / ml, and homogenized with a homogenizer (manufactured by Ikemoto Rika Kogyo Co., Ltd.). This dispersion was centrifuged at 4 ° C. and 15,000 rpm for 10 minutes, and the supernatant was subjected to mouse IgA measurement. ELISA kit for mouse IgA measurement (B
(Manufactured by ETHYL).

13. Blood cell analysis After fasting the mouse for about 16 hours, under Nembutal anesthesia,
Using a syringe treated with anticoagulant (heparin 10 units / ml), after collecting whole blood from the inferior vena cava, fully automatic blood cell counter MEK-6
It was measured using 158 (manufactured by Nihon Kohden).

14. Measurement of Th1 / Th2 cell ratio After removing the spleen, it is crushed aseptically, and a spleen cell suspension is prepared while adding 25 mM HEPES-RPMI. This spleen cell suspension was passed through a cell strainer into a single cell suspension according to a standard method. In accordance with the method of EPICS Application Note8, cell surface CD3 of spleen lymphocytes, intracellular IL4, intracellular
INF-gamma was fluorescently stained. After this, Coulter EPICS
Analysis was performed with XL, and the CD3 positive cell ratio and Th1 / Th2 cell ratio were calculated.

Test Results A. Results of test on effects on lifespan of experimental animals in vivo Fig. 1 shows the lifespan of mice used in the experiment. It was revealed that the fermented milk (LG2055) -administered group of the present invention was more effective in prolonging life than the control. Also, when comparing the weight gain curves (Fig. 2), no significant difference was observed.

B. Test results on effects on aging index Table 2 shows the results of analysis of changes in intestinal flora up to the age of 12 months. The fermented milk (LG2055) administration group of the present invention showed a significant increase in fecal Bifidobacterium and a marked decrease in lecithinase-negative Clostridium. A time-dependent change was observed in both bacterial groups.

[0044]

[Table 2]

In the fermented milk (LG2055) -administered group of the present invention, it was revealed that the number of Bifidobacterium bacteria increases with the age of life (FIG. 3). On the other hand, it was confirmed that the number of Clostridium bacteria was remarkably reduced in the fermented milk (LG2055) -administered group of the present invention (Fig. 4).

FIG. 5 shows the results of analysis of the amount of excreted urine observed throughout the experimental period. Fermented milk of the present invention (LG2055 strain)
Urine excretion increased in the treated group.

FIG. 6 shows the excretion amount of the oxidative marker (8-OHDG) excreted in urine. Excretion is active in both groups during periods of high metabolism, but it was revealed that the fermented milk of the present invention (LG2055 strain) was highly excreted throughout the entire test period. From the above measurement results of the urinary excretion amount and the 8-OHDG excretion amount, it became clear that the fermented milk of the present invention (containing the LG2055 strain) prevents / improves the decline in renal function with aging.

FIG. 7 shows the measurement results of the amount of cerebral amyloid accumulation. In the fermented milk (LG2055 strain) administration group of the present invention, suppression of amyloid accumulation with aging was confirmed, and a preventive / ameliorating effect on amyloidosis was confirmed. In addition, in combination with the results of the standing exercise test shown below, prevention of dementia
The improvement effect was confirmed.

FIG. 8 shows the comparison results of the amount of fat accumulated in the abdominal wall. In the fermented milk (LG2055 strain) -administered group of the present invention, it was confirmed that it was significantly decreased, and it was revealed that the lipid metabolism abnormality associated with aging is improved.

FIG. 9 shows the results of observation of spontaneous locomotion of mice. Abnormal movement was observed in the control group, and such abnormal behavior was small in the fermented milk (LG2055 strain) administration group of the present invention. With the fermented milk of the present invention (containing LG2055 strain),
It was found that the accumulation of amyloid in the brain was reduced and that it was effective in suppressing dementia with aging.

FIG. 10 shows the measurement result of blood glucose level. The fermented milk (LG2055 strain) administration group of the present invention was able to suppress the occurrence of hyperglycemia with age.

FIG. 11 shows the analysis results of the splenic helper T cell repertoire. It was confirmed that activation of cell-mediated immune function occurred in the fermented milk (LG2055 strain) administration group of the present invention. At the same time, CD3 positive cell ratio (T cell) in spleen
It was confirmed that it was also higher.

FIG. 12 shows the results of cecal content pH measurement. It was confirmed that the fermented milk of the present invention (LG2055 administration group) lowers intestinal pH and improves intestinal metabolism.

FIG. 13 shows the excretion amount of fecal IgA. Fecal IgA
Was suppressed in the fermented milk (LG2055 strain) administration group of the present invention.

FIG. 14 shows the feed efficiency throughout the test period. The feed efficiency throughout the test period is determined by the fermented milk of the present invention (LG
It was confirmed that the 2055 strain) administration group was significantly higher. on the other hand,
From Fig. 14, it was found that the negative effects on feed efficiency due to aging were suppressed. This result proves that "you can live well and lose weight," and you can confirm the effect of maintaining and improving QOL.

[0056]

[Example] 1. The dry powder LG2055 strain was cultured in a 10% reduced skim milk medium (121 ° C., heated for 10 minutes), and the main culture was freeze-dried and powdered to prepare the preventive and ameliorating agent of the present invention (A).

2. The fermented milk LG2055 strain was cultured in an MRS liquid medium. 1% of each culture solution in the logarithmic growth phase was inoculated into 10% reduced skim milk (sterilized at 115 ° C. for 20 minutes) containing 0.35% yeast extract to prepare individual mother cultures. This was prepared by adding 2.5% of each to a yogurt mix (a mixture of 10% reduced skim milk and heating at 100 ° C. for 10 minutes). Fermentation was carried out at 37 ° C., and when the lactic acid degree reached 0.85, the fermentation was terminated and the fermentation was terminated to prepare fermented milk having the preventive, ameliorating and treating effect of the present invention (B).

3. Formulation Example 1 4 cells in the logarithmic growth phase from the liquid culture of LG2055 strain
The cells were centrifuged at 7,000 rpm for 15 minutes at 7,000 rpm and washed with sterilized water. This was repeated 3 times to obtain washed bacterial cells. This was freeze-dried to obtain bacterial cell powder. 1 part of this bacterial cell powder was mixed with 4 parts of skim milk, and 1 g of this powder was tabletted by a tableting machine according to a standard method to prepare a tablet containing 200 mg of bacterial cells. Further, the above-mentioned fermented milk containing the LG2055 strain was freeze-dried, and the obtained powder was directly tableted.

4. Encapsulating agent The freeze-dried powder was made into a powder, and then granulated by granulation, and then 10 mg each was filled in empty capsules to give a capsule.

5. Formulation Example 2 LG2055 strain was inoculated into 5 L of MRS liquid medium (manufactured by Difco), and then 37
C., static culture was performed for 18 hours. After inoculating 5 L of skim milk with the LG2055 strain, static culture was performed at 37 ° C. for 18 hours. After culturing,
Centrifugation was performed at 7,000 rpm for 15 minutes to obtain 1/50 volume of each concentrated bacterial cell of the culture solution. Next, the concentrated bacterial cells were mixed with the same amount of a dispersion medium containing 10% by weight of skim milk powder and 1% by weight of sodium glutamate, adjusted to pH 7, and then freeze-dried. The obtained lyophilized product was sized with a 60-mesh sieve to obtain a lyophilized bacterial powder. 13th Revised Japanese Pharmacopoeia Manual According to the general rules for formulation, "Powder", add 1 g of freeze-dried bacterial powder obtained in 5 above to 400 g of lactose (JP) and 600 g of potato starch (JP) Mix to prepare a powder.

6. Formulation Example 3 A tablet was produced according to the following formulation. (1) 50 g of freeze-dried product of culture of non-fat dry milk medium of LG2055 strain, (2) 90 g of lactose, (3) 29 g of corn starch, (4) 1 g of magnesium stearate, the mixture was compressed with a compression tablet machine to give 1 100 tablets were prepared containing 40 mg of active ingredient per tablet.

7. Formulation Example 4 The LG2055 strain was cultured in a whey medium (0.5% yeast extract, 0.1% trypticase peptone was added), and then the cells were recovered by centrifugation. 1 g of this culture was mixed with 5 g of lactose and molded into granules to obtain granules.

8. LG2055 mixed with 200 ml of milk as food (1) LG2055 strain lyophilized powder beverage washed cells are included, each 10 ⁸ or more, including to obtain a prophylactic improving therapeutic agent beverage of the present invention. It had a good flavor. (2) Fermented milk LG2055 strain was inoculated into yogurt mix (2% skim milk was added to raw milk and heated at 100 ° C for 10 minutes) and cultured at 20 ° C for 24 hours. After being filled in a paper cup and cooled, the viable cell concentration of the LG2055 strain in the yogurt product was 10 ⁸ or more per 100 g.

(3) Fermented butter Fermented butter (wt / wt) Milk fat 96.8% Salt 1.2 1. Sample (A) 2 obtained in

(4) Butter cake Butter cake (wt / wt) 24% butter Soft flour 24 Sugar 24 Whole egg 24 2. Sample (B) 4 obtained in A little spice

(5) Mayonnaise Mayonnaise (wt / wt) Salad oil 65% Egg yolk 17 Vinegar 10 1. Sample (A) 3 obtained in Spice 4.4 Monosodium glutamate 0.6

[0067]

INDUSTRIAL APPLICABILITY According to the present invention, a prophylactic / ameliorating / therapeutic agent for metabolic disorders containing a culture and / or a bacterial cell obtained by culturing a lactic acid bacterium belonging to Lactobacillus gasseri as an active ingredient It is possible to provide foods and drinks having the preventive, ameliorating, and treating effects on metabolic disorders. The preventive / ameliorating / therapeutic agent for metabolic disorders provided by the present invention has extremely little toxicity and side effects, and is also useful as a food material.

[Brief description of drawings]

1 shows the lifespan of experimental animals.

FIG. 2 shows a weight gain curve.

FIG. 3 shows the number of Bifidobacterium in feces.

FIG. 4 shows the number of Clostridium in feces.

FIG. 5 shows changes in urinary excretion.

FIG. 6 shows the amount of 8-OHDG excreted in urine.

FIG. 7 shows the accumulated amount of amyloid in the brain.

FIG. 8 shows a correlation analysis diagram of body weight and posterior abdominal wall fat weight.

FIG. 9 shows the measurement results of the self-starting movement of the animal.

FIG. 10 shows the measurement results of blood glucose level.

FIG. 11 shows a helper T cell composition ratio.

FIG. 12 shows cecal pH.

FIG. 13 shows the fluctuation values of fecal IgA.

FIG. 14 shows feed efficiency.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A23L 2/52 A61P 3/00 A61P 3/00 13/12 13/12 25/28 25/28 A23L 2 / 00 F (72) Inventor Shigeru Fujiwara 1385-34 Tsurugashima City, Saitama Prefecture (72) Inventor Toshiro Suzuki 1187 Mikatagahara Town, Hamamatsu City, Shizuoka Prefecture (72) Inventor Masanori Hosokawa Shogoin Kawahara Town, Sakyo Ward, Kyoto City, Kyoto Prefecture 53 F term (reference) 4B017 LC03 LK21 LL09 LP05 4B018 LB08 LE01 MD86 ME14 MF13 4C087 AA01 AA02 AA03 BB39 BC56 MA02 MA52 NA14 ZA15 ZA81 ZC21 ZC33

Claims (6)

[Claims] 1. Lactobacillus Lactobacillus
A preventive / ameliorating / therapeutic agent for metabolic disorders, which comprises a culture obtained by culturing lactic acid bacteria belonging to gasseri ) and / or bacterial cells as an active ingredient. 2. Lactobacillus
prevention, improvement and therapeutic agent for metabolic disorders of claim 1 wherein the Lactobacillus gasseri (Lactobacillus gasseri) lactic acid bacteria having a human intestinal fixability belonging to gasseri). 3. The preventive / ameliorating / therapeutic agent according to claim 1 or 2, wherein the dysbolism is renal dysfunction, cerebral amyloidosis, or dyslipidemia. 4. Lactobacillus Lactobacillus
The preventive / ameliorating / therapeutic agent for metabolic disorders according to any one of claims 1 to 3, wherein a culture obtained by culturing a lactic acid bacterium belonging to gasseri ) is fermented milk. 5. Lactobacillus
Lactobacillus belonging to gasseri is Lactobacillus gasseri ( La
ctobacillus gasseri ) ・ SBT2055 (hereinafter referred to as LG2055)
(FERM P-15535), The preventive / ameliorating / therapeutic agent for metabolic disorders according to any one of claims 1 to 4. 6. Lactobacillus Lactobacillus
Foods and drinks having a preventive, ameliorating, or therapeutic action on metabolic disorders by adding a culture obtained by culturing lactic acid bacteria belonging to gasseri ) and / or cells.