JP5155960B2 - Preventive, ameliorating, and therapeutic agents for metabolic disorders associated with aging - Google Patents

Description

  The present invention relates to a preventive / ameliorating / treating agent for metabolic disorders occurring with aging and / or a food / beverage product having an effect of preventing / ameliorating / treating metabolic disorders occurring with aging.

Aging is an unavoidable life phenomenon for organisms. Various factors have been cited as hypotheses as causes of aging. For example, there are a program theory and an error failure theory at the cell level. Whichever theories are taken, it is unclear how changes caused by the aging mechanism in the cells are related to the aging phenomenon in the whole organism. However, there is also an idea that these changes occur in central cell groups and have a ripple effect in general, and changes that occur in individual cells in the periphery influence each other and reflect them in the aging phenomenon in general. 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 dysfunctions that occur with aging, life extension is realized, and various aging indices are lowered. With the establishment of Westernized dietary habits, the nature of metabolic abnormalities in Japanese with age has changed.

In recent years, various diseases that are considered to be caused by metabolic abnormalities that occur with aging include hyperlipidemia due to obesity, senile dementia with amyloid accumulation in the brain, immune abnormalities, and decreased bowel function. -Kidney dysfunction such as colorectal cancer and uremia caused by changes in intestinal microorganisms.
There are various causes of obesity, but no radical treatment has been found that is more effective than diet or exercise. Conventionally, as a body fat accumulation inhibitor other than diet therapy, a method of administering a fruit or essence of a rose family plant (Patent Document 1), a method of administering a composition derived from buckwheat (Patent Document 2) ), A method of administering procyanidins (Patent Document 3), a method of administering winter pericarp and coconut pericarp (Patent Document 4), a method of changing the energy metabolism efficiency by administering a specific fatty acid (Patent Document 5) And agents have been proposed. However, there is no proposal for suppressing fat accumulation by Bifidobacterium longum having intestinal colonization described below in the present invention.
There are two types of senile dementia, cerebrovascular dementia and Alzheimer-type dementia. In particular, the occurrence of Alzheimer-type dementia accompanied by accumulation of amyloid in the brain (amyloidosis) has increased in recent years. Examples of substances that suppress amyloid accumulation (amyloidosis) include a method of administering amyloid β protein (Patent Document 6), a method of administering fragments of amyloid protein (Patent Document 7), and a sugar having a specific structure substituted with an anion. A method of administering the composition (Patent Document 8) and the like have been proposed. However, there is no proposal to suppress the accumulation of amyloid in the brain by Bifidobacterium longum having intestinal colonization described below in the present invention.

The decline in kidney function with aging has been attributed to various causes such as a decrease in glomerular function due to aging, a decrease in mesangial cell function, and accumulation of waste products / IgA. Proposed new therapeutic agents and methods, such as a method of administering IGF-1 or IGF-3, a method of administering an IP receptor agonist (Patent Document 9), etc., in order to suppress a decrease in kidney function associated with aging Has been. However, there is no proposal to suppress the decrease in kidney function and metabolism due to Bifidobacterium longum having intestinal colonization described below in the present invention.
All of the above-mentioned diseases greatly limit the thinking and behavior of the elderly, and as a result, the quality of life (QOL) of the elderly is reduced.
Various therapeutic agents for improving the symptoms of these diseases have been proposed and put into practical use, but do not drastically improve the QOL of the elderly.

JP-A-8-198768 JP 9-183735 A JP-A-9-291039 Japanese Patent Laid-Open No. 11-164668 JP 2001-286268 A Japanese Patent Publication No. 6-502387 Japanese National Patent Publication No. 11-500462 Special table 2001-513569 JP 2000-191523 A

The present inventors have been studying lactic acid bacteria having colonization in the human intestinal tract, and obtained by culturing lactic acid bacteria having colonization in the intestinal tract, particularly lactic acid bacteria belonging to Bifidobacterium longum. It has been found that upon administration of the resulting culture and / or cells, these microorganisms act on the host by colonizing in the intestinal tract, leading to a significant increase in life in laboratory animals. As a result of further investigation, when administered Bifidobacterium longum a (Bifidobacterium longum) aging animals, bi and fixed to the intestinal tract Bifidobacterium longum (Bifidobacterium longum) acts on the host, the age The present inventors have found that it has surprising effects of improving accompanying metabolic abnormalities, suppressing decline and deterioration of kidney function, improving lipid metabolism causing obesity, and further suppressing accumulation of amyloid in the brain. As a result, it was found that QOL accompanying aging was clearly improved, and the present invention was completed.
Accordingly, the present invention provides a culture obtained by culturing lactic acid bacteria belonging to Bifidobacterium longum and / or a preventive, ameliorating, or therapeutic agent for metabolic disorders containing the bacterial cells as active ingredients. Is an issue.
Furthermore, the present invention provides a preventive, ameliorating, and therapeutic action for metabolic disorders associated with aging by containing a culture obtained by culturing lactic acid bacteria belonging to Bifidobacterium longum and / or bacterial cells. It is an object of the present invention to provide a food / beverage product having the following.

The inventors of the present invention have been researching various fermented milks, and Bifidobacterium longum ( Bifidobacterium longum ) has been conventionally used among lactic acid bacteria isolated from these fermented milks and human-derived lactic acid bacteria. It has been found that it has a high intestinal colonization that is not found in humans, and this Bifidobacterium longum has a metabolic disorder associated with aging due to the intestinal colonization that was not previously known. The present inventors have found that the present invention has improved, suppressed kidney function deterioration and deterioration, improved lipid metabolism that causes obesity, and further suppressed the accumulation of amyloid in the brain.

The present invention relates to a culture obtained by culturing lactic acid bacteria belonging to Bifidobacterium longum and / or a preventive, ameliorating, and treating agent for metabolic disorders associated with aging, which comprises microbial cells as active ingredients. .
Moreover, this invention relates to the food / beverage products which have the preventive / improving / therapeutic effect of the metabolic disorders accompanying aging which contain such an active ingredient. That is, this invention is invention which consists of the following structure described in the claim.

(1) A preventive / ameliorating / treating agent for metabolic disorders comprising as an active ingredient a culture obtained by culturing lactic acid bacteria belonging to Bifidobacterium longum and / or bacterial cells.
(2) Bifidobacterium longum (Bifidobacterium longum) belonging to lactic acid bacteria, a bi having a human intestinal fixability Bifidobacterium longum (Bifidobacterium longum) (1) prevention, improvement and metabolic disorders described Therapeutic agent.
(3) The preventive / improving / treating agent for metabolic disorders according to (1) or (2), wherein the metabolic disorder is any of renal dysfunction, cerebral amyloidosis, and lipid metabolism disorder.
(4) Prevention / amelioration of metabolic disorders according to any one of (1) to (3), wherein the culture obtained by culturing lactic acid bacteria belonging to Bifidobacterium longum is fermented milk. Therapeutic agent.
(5) The lactic acid bacterium belonging to Bifidobacterium longum is Bifidobacterium longum SBT2928 (FERM P-10657) according to any one of (1) to (4) A preventive, ameliorating, or therapeutic agent for metabolic disorders.
(6) A culture obtained by culturing lactic acid bacteria belonging to Bifidobacterium longum and / or a food or drink having a preventive, ameliorating, or therapeutic effect on metabolic disorders with the addition of bacterial cells.

According to the present invention, a prophylactic / improving / therapeutic agent for metabolic disorders and metabolic abnormalities comprising a culture obtained by culturing lactic acid bacteria belonging to Bifidobacterium longum and / or bacterial cells as active ingredients It is possible to provide foods and drinks that have preventive, ameliorating, and therapeutic effects on symptoms.
The agent for preventing / ameliorating / treating metabolic disorders provided by the present invention has extremely low toxicity and side effects, and is also useful as a food material.

The life span of experimental animals is shown. Shows the number of Bifidobacterium in feces. The number of clostridium in feces is shown. Shows changes in urinary excretion. Shows changes in urinary excretion. Shows urinary 8-OHDG excretion. The accumulation amount of amyloid in the brain is shown. The correlation between body weight and retroabdominal wall fat weight is shown. The self-standing standing movement measurement result of an animal is shown. The helper T cell component ratio is shown. Shows pH in the cecum. The fluctuation value of fecal IgA is shown. Shows feed efficiency.

  The present invention has been made in order to solve the above-mentioned problems, and the present inventors newly set the following criteria and screened strains that meet the purpose when screening the target lactic acid bacteria. Selected. That is, the present inventors show high fixability to the human intestinal tract among fermented milk and human-derived Bifidobacterium longum having high gastric acid resistance and good growth under low pH conditions. , Human gut cell affinity, bile acid tolerance, colonization in the intestine has the effect of preventing, ameliorating and treating various metabolic abnormalities associated with aging, and survival when applied to food However, we set the conditions such as high taste, excellent flavor and physical properties. As a result of screening under these conditions, the following strains could be selected as strains meeting these conditions. This strain has been deposited with the Patent Microorganism Deposit Center of the National Institute of Advanced Industrial Science and Technology under the following deposit number.

Strain Bifidobacterium longum
SBT2928 FERM P-10657
This strain has a high affinity for human intestinal cells, and when orally administered, it can survive and reach the intestinal tract and can be resident in the intestinal tract for a long period of time, and grows in the intestinal tract It acts on the host to prevent, ameliorate, and treat metabolic disorders associated with aging. Lactic acid bacteria belonging to Bifidobacterium longum administered from outside the body have settled in the intestine, and it is not known at all that it was revealed for the first time by the present inventors. .

Furthermore, in the present invention, not only the above-mentioned deposited strain, but any Bifidobacterium longum isolated from humans or fermented milk can be used as long as it exhibits the above-mentioned action. .
Next, the culture method of these lactic acid bacteria will be described.
As the Bifidobacterium longum medium of the present invention, various media such as a milk medium or a medium containing milk components, a semi-synthetic medium not containing this, or the like can be used. An example of such a medium is a reduced skim milk medium obtained by reducing skim milk and then heat sterilizing.
The culture method is a static culture or a neutralization culture in which pH is controlled to be constant. However, the culture method is not particularly limited as long as the bacteria grow well.

In the present invention, the culture and / or cells obtained as described above are used as active ingredients. Moreover, it is good also considering the dried powder as an active ingredient. Such drying is preferably performed by freeze-drying because the cells can be dried without deteriorating the cells.
These active ingredients are preferably taken orally. These powders can be mixed with an appropriate excipient such as lactose and orally administered as a powder, tablet, pill, capsule, granule or the like. The dosage is appropriately determined individually in consideration of the symptom, age, etc. of the administration subject, but is usually 0.5 to 10 g as a dry product per day for an adult, and this may be divided into several times a day. Particularly preferably, administration of 10 ⁸ to 10 ¹² cuf / day per adult is possible with each strain as a living bacterium, and the intended effect of the present invention can be exhibited. By taking in this way, it settles in the intestinal tract and exhibits the desired effect.

Moreover, you may add the active ingredient of this invention to a raw material during the manufacturing process of food-drinks. As the food and drink, any food and drink may be used. Examples thereof include milk drinks, fermented milk, fruit juice drinks, jelly, candy, dairy products, processed eggs such as mayonnaise, and foods such as butter cakes and other confectionery breads. be able to. However, as a characteristic of the present invention, it is necessary to settle in the intestinal tract while the lactic acid bacteria are alive, and it is preferable to avoid excessive heating. In addition, conventional techniques such as microcapsules may be employed to take measures to avoid heating.
Furthermore, the food and drink in the present invention may be yogurt produced by lactic acid fermentation using the aforementioned strain of Bifidobacterium longum .
In the following, test examples using Bifidobacterium longum and SBT2928 (BL2928) strains as lactic acid strains used in the present invention are shown, and the bacteriological properties and in vitro and in vivo effects are specifically explained. To do. However, the present invention is not limited to this description.

[Test example]
Characteristics of BL2928 strain Mycological properties (1) Fungal form
The results after anaerobic culture at 37 ° C. for 48 hours using BL blood agar plates are shown.
Shape: Aspergillus Size: 0.5-1 × 3-4μm
Many linked (2) Gram stain positive (3) Colony shape Shape: Circular Perimeter: Sliding Size: Diameter 2-3 mm
Color: Brown Surface: Smooth (4) Spore formation Negative (5) Gas production None (6) Motility None (7) Catalase activity Negative (8) Nonfat milk coagulability Coagulation (9) Gelatin liquefaction None (10) Nitrate reduction None (11) Indole productivity None (12) Hydrogen sulfide productivity None

2. Fermentability of sugars The results of examining the fermentability of sugars with a commercially available bacterial identification kit (Api 50CH, Biomelieu) are described below.
L-arabinose +
L-ribose +
Sorbitol −
Cellobiose −
Lactose +
Merezitose +
Raffinose +
Starch −
Gluconate −
(+ Indicates that there is fermentability, − indicates that there is no fermentability.)

The above taxonomic properties and sugar fermentability showed typical Bifidobacterium longum properties.

Subsequently, the confirmation test by a DNA homology test was implemented as follows.
DNA homology test The DNA of Bifidobacterium described below, the test strain BL2928, and Escherichia coli as a control were extracted and purified.
Test strain: BL2928 strain Reference strain: Bifidobacterium longum JCM1217 Bifidobacterium infantis JCM1222 Bifidobacterium bifidum JCM1255 Bifidobacterium brebe JCM1192 Bifidobacterium adrescentis JCM1275 strain Bifidobacterium animalis JCM1190 strain Escherichia coli
DNA homology between BL2928 strain and each reference strain was examined by DNA hybridization method, assuming that the homology between DNA of BL2928 strain was 100% and the DNA homology between BL2928 strain and E. coli was 0%. As a result, BL 2928 strain had 90% or more homology with the Bifidobacterium longum reference strain, and therefore BL2928 strain was identified as Bifidobacterium longum.

Gastric acid resistance The gastric acid resistance test was carried out according to the method of Higuchi et al. (Intestinal Bacteriology Journal 14.11-18.2000), and a gastric acid resistance test was carried out by preparing an artificial gastric juice having a pH of 2.0. The BL2928 strain survived for 3 hours or more.

Artificial intestinal juice resistance Artificial intestinal fluid containing bile powder was prepared according to the method of Higuchi et al. (Intestinal Bacteriology Journal 14.11-18.2000), and the above-described artificial gastric juice treated BL2928 strain was added to test the resistance. , Showed viability of more than 20 hours. The BL2928 strain was confirmed to pass through the gastrointestinal tract and survive to reach the large intestine.

Human intestinal transit ability and intestinal colonization Healthy adult volunteers using fermented milk that was fermented at 39 ° C for 4 hours after inoculating 4% of BL2928 starter into non-fat milk solids 9.5% and milk fat 3.0% Names were fed 100 g once a day for 4 weeks and observed changes in intestinal bacteria. During the test period, evaluation was performed by eating freely except forbidding foods, oligosaccharides, and drugs that affect enterobacteria. The BL2928 strain, which was not detected before the test, was detected in all subjects after 4 weeks, indicating that the strain has high intestinal colonization.

Animal test A. Effect test on the life span of experimental animals in vivo Preparation of lactic acid bacteria skim milk culture
BL2928 strain was used. Lactic acid bacteria were cultured at 37 ° C. for 16 hours in an 11.55% skim milk medium supplemented with 0.5% yeast extract (manufactured by Asahi Beer Foods) 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 (Oriental Yeast Co., Ltd.). To this feed, 5% skim milk or the above skim milk lactic acid bacteria culture was added.

3. Test animals Two groups of Senescence-Accelerated Mouse (SAM) P8 female mice (obtained from SLC, Japan), a 5-week-old aging promotion model, were divided into 30 groups. After pre-breeding for 1 week in AIN-93M, the breeding was continued until the above test feed died. During this time, ion-exchanged water and feed (within 7 to 8 g) were freely ingested.

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

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

6). Flora analysis Fresh stool was collected, and 5 weeks old (initial value), 3 months old, 5 months in accordance with the method of Mitsuoka et al. Intestinal flora analysis was performed at age 7, 7 months, 10 months, and 12 months. About the obtained data, analysis of variance by the division method was performed.

B. Test on effects on aging index (biological function at 40 weeks of age)
1. Preparation of lactic acid bacteria skim milk culture As in the case of A, BL2928 strain was used. Lactic acid bacteria were cultured at 37 ° C. for 16 hours in a 11.55% skim milk medium supplemented with 0.5% yeast extract (manufactured by Asahi Beer Foods) sterilized at 115 ° C. for 20 minutes. The obtained culture was freeze-dried and then ground in a mortar.

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

3. Experimental animals
Two groups of 5-week-old SAM P8 female mice (obtained from SLC, Japan) were used as 10 groups. After preliminarily raised in AIN-93M for 1 week, it was raised for 36 weeks with the above test feed. During this time, ion-exchanged water and feed were ingested freely. After raising to 40 weeks of age, the animals were fasted for about 16 hours, opened under anesthesia with Nembutal (Dainippon Pharmaceutical Co., Ltd.), and sacrificed by inferior vena cava blood sampling.

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

5. Dietary intake measurement method Dietary intake was measured at a rate of about once a week (1 week), weekly after childhood. A pre-weighed feed was given, and the remaining amount of food in the container was measured at the same time zone the next day, and the amount of food intake per day was calculated.

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

7). Measurement of urinary 8-hydroxydeoxyguanosine The urine obtained in metabolic cage breeding was measured using an ELISA kit for 8-OHDG measurement (manufactured by Japan Aging Control Laboratory).

8). Measurement of amyloid accumulation in the brain Dissection was performed immediately after sacrifice and the cerebrum was removed. The obtained cerebrum was homogenized with a Teflon (registered trademark) homogenizer with PBS containing 0.5% protein inhibitor mixture (manufactured by SIGMA), and an amyloid beta (1-42) measurement kit (manufactured by Immunobiological Research Institute) was used. Measured.

9. The weight of the abdominal wall fat was dissected immediately after slaughtering, and the left and right abdominal wall fats were excised respectively, and the weight was measured with a precision balance METTLER AE240 (manufactured by Siebel Hegner Japan).

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

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

12 Measurement of fecal IgA The feces obtained from metabolic cages were dried with a centrifugal evaporator CVE-200D (EYELA), weighed, and added with PBS containing 0.5% protein inhibitor (SIGMA) Feces were dispersed at a concentration of 0.05 g / ml and homogenized with a homogenizer (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. Measurement was performed using an ELISA kit for mouse IgA measurement (manufactured by BETHYL).

13. Blood cell analysis After mice were fasted for about 16 hours, a whole blood sample was collected from the inferior vena cava using an anticoagulant-treated (heparin 10 units / ml) syringe under Nembutal anesthesia, and then a fully automatic hemocytometer MEK-6158 ( Nihon Kohden Co., Ltd.).

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 MHEPES-RPMI. This spleen cell suspension was made into a single cell suspension through a cell strainer according to a conventional method. According to the method of EPICS Application Note 8, spleen lymphocytes were stained for cell surface CD3, intracellular IL4, and intracellular INF-gamma. Then, it analyzed by flow cytometer Coulter EPICS XL, and calculated CD3-positive cell ratio and Th1 / Th2 cell ratio.

Test results A. Test results on effects on the lifespan of experimental animals in vivo FIG. 1 shows the lifespan of mice used in the experiments. Compared with the control, the fermented milk (BL2928 strain) administration group of the present invention markedly prolongs the survival time, and it was revealed that the present invention has a remarkable effect on the life extension.
FIG. 2 shows the change in body weight of the mice used in the experiment from 5 weeks of age to 60 weeks. It was confirmed that the weight gain of the fermented milk (BL2928 strain) administration group of the present invention was small compared to the control, and the weight gain was small in the vicinity of 30 weeks.

B. Test results on effects on aging index Table 2 shows the analysis results of changes in intestinal flora up to 12 months of age. The administration of the fermented milk of the present invention (BL2928 strain) showed a significant increase in the genus Bifidobacterium and a significant decrease in the lecithinase-negative Clostridium genus. Time-dependent changes were observed in both fungal groups. It was confirmed that there was a difference in the change in the number of Bifidobacterium in each group.

FIG. 3 shows changes in the number of Bifidobacterium in the stool. In the fermented milk (BL2928 strain) administration group of the present invention, it became clear that the number of Bifidobacterium spp. Increases with the age of life.
FIG. 4 shows the change in the number of bacteria of the genus Clostridium. In the fermented milk (BL2928 strain) administration group of the present invention, it was revealed that the number of Clostridium bacteria was significantly reduced.

  FIG. 5 shows the analysis results of the urinary excretion observed throughout the experimental period. In the fermented milk (BL2928 strain) administration group of the present invention, urinary excretion increased.

FIG. 6 shows the excretion amount of the oxidation marker (8-OHDG) excreted in urine. Although excretion was active in both groups during the period of high metabolism, it was found that the excretion amount of the fermented milk (BL2928 strain) administration group of the present invention was high throughout the entire test period.
From the above measurement results of urinary excretion and 8-OHDG excretion, it became clear that fermented milk containing fermented milk of the present invention (containing BL2928 strain) prevents and improves kidney function decline with age. .

  FIG. 7 shows the measurement results of the amount of amyloid accumulated in the brain. In the fermented milk (BL2928 strain) administration group of the present invention, inhibition of amyloid accumulation accompanying aging was confirmed, and the prevention / amelioration effect of amyloidosis could be confirmed. In addition, the preventive / improving effect of dementia could be confirmed together with the results of the standing exercise test shown below.

  FIG. 8 shows a comparison result of the abdominal wall fat accumulation amount. In the fermented milk (BL2928 strain) administration group of the present invention, it was confirmed that it decreased significantly, and it became clear that lipid metabolism abnormality accompanying aging was improved.

  FIG. 9 shows the observation result of the spontaneous movement of the mouse. Abnormal movement was confirmed in the control group, and such abnormal behavior was small in the fermented milk (BL2928 strain) administration group of the present invention. It was revealed that administration of the fermented milk of the present invention (containing BL2928 strain) reduces the accumulation of amyloid in the brain and is effective in suppressing dementia associated with aging.

  Table 3 summarizes the plasma analysis values. In the fermented milk (BL2928 strain) administration group of the present invention, a tendency to decrease plasma cholesterol levels was confirmed. It was confirmed that lipid metabolism abnormalities associated with aging were improved by fermented milk of the present invention (containing BL2928 strain) together with a decrease in accumulated fat weight.

  FIG. 10 shows the analysis result of the spleen helper T cell repertoire. In the fermented milk (BL2928 strain) administration group of the present invention, it was confirmed that cellular immune function was activated. At the same time, it was confirmed that the ratio of CD3-positive cells (T cells) in the spleen was also high (P = 0.0842).

  FIG. 11 shows the measurement results of the pH of the cecum contents. In the fermented milk (BL2928 strain) administration group of the present invention, it was confirmed that the pH of the cecal contents was significantly reduced.

  FIG. 12 shows the measurement results of fecal IgA. The excretion amount of fecal IgA tended to be high in the fermented milk (BL2928 strain) administration group of the present invention.

  FIG. 13 shows the feed efficiency throughout the test period. It was confirmed that the feed efficiency of the fermented milk (BL2928 strain) administration group of the present invention was significantly high. On the other hand, it was found from FIG. 13 that the negative feed efficiency due to aging was suppressed. This result proves that “I live well and get lost”, and I was able to confirm the effect of maintaining and improving QOL.

Examples are shown below.

1. Dry powder
BL2928 strain was cultured in a 10% reduced skim milk medium (heated at 121 ° C. for 10 minutes), and this culture was freeze-dried and powdered to prepare the preventive / ameliorating therapeutic agent of the present invention (A).

2. Fermented milk
BL2928 strain was cultured in Briggs liver medium. Each culture solution in the logarithmic growth phase was inoculated with 1% of 10% reduced skim milk (115 ° C., sterilized for 20 minutes) supplemented with 0.3% yeast extract to prepare individual mother cultures.
This was prepared by adding 2.5% each to yogurt mix (10% reduced skim milk was added and heated at 100 ° C. for 10 minutes). Fermentation was performed at 37 ° C., and when the lactic acid acidity reached 0.85, the mixture was cooled, the fermentation was terminated, and fermented milk having the preventive improvement therapeutic action of the present invention was prepared (B).

2. Formulation Example 1
The cells in the logarithmic growth phase were centrifuged at 4 ° C. and 7,000 rpm for 15 minutes from the BL2928 strain liquid culture, washed with sterilized water, and this was repeated three times to obtain washed cells. This was freeze-dried to obtain cell powder. 4 parts of skim milk was mixed with 1 part of this bacterial cell powder, and 1 g of this powder was tableted by a conventional method with a tableting machine to prepare a tablet containing 200 mg of bacterial cell. Further, the fermented milk containing the above BL2928 strain was freeze-dried and directly compressed using the obtained powder.

4). Encapsulating agent After freeze-dried powder was powdered, it was granulated by granulation, and 10 mg each was filled into empty capsules to form capsules.

5. Formulation Example 2
BL2928 strain was inoculated into 5 L of MRS liquid medium (Difco), followed by stationary culture at 37 ° C. for 18 hours. BL2928 strain was inoculated into 5 L of skim milk, and then static culture was performed at 37 ° C. for 18 hours. After completion of the culture, centrifugation was performed at 7,000 rp for 15 minutes to obtain 1/50 amount of each concentrated bacterial cell of the culture solution. Next, the same amount of the concentrated cells was mixed with a dispersion medium containing 10% (weight) of skim milk powder and 1% (weight) of sodium glutamate, adjusted to pH 7, and then lyophilized. The obtained freeze-dried product was sized with a 60-mesh sieve to obtain a freeze-dried bacterial powder.
In accordance with the provisions of the 13th revised Japanese Pharmacopoeia General Rules for Preparations, “Powder”, add 1 g of freeze-dried bacteria obtained in 5 above to 400 g of lactose (Japan Pharmacopoeia) and 600 g of potato starch (Japan Pharmacopoeia) to evenly Mixed to produce a powder.

6). Formulation Example 3
An anti-ulcer agent was produced by the following composition (1) 50 g of freeze-dried culture of BL2928 strain nonfat dry milk medium,
(2) 90 g lactose,
(3) Corn starch 29g,
(4) Magnesium stearate 1 g,
This mixture was compressed by a compression tablet machine to produce 100 tablets containing 40 mg of active ingredient per tablet.

7). Formulation Example 4
BL2928 strain was cultured in whey medium (0.5% yeast extract and 0.1% trypticase peptone added), and the cells were collected by centrifugation. 1 g of this culture was mixed with 5 g of lactose and formed into granules to obtain granules.

8). The freeze-dried powder (1) Preparation of beverage washed cells of foods containing BL2928 strain was mixed with 200 ml of milk so BL2928 strain contained respectively 10 ⁸ or more, the prophylactic improving therapeutic agent mixed beverage of the present invention Obtained. It had a good flavor.

(2) Fermented milk
BL2928 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 filling in a paper cup and cooling, the viable cell count concentration of BL2928 strain in the product made into yogurt 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)
Butter 24%
Light flour 24
Sugar 24
Whole egg 24
2. Sample (B) obtained in 4
Fragrance a little

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

Claims (3)

An agent for the prevention, amelioration, and treatment of Alzheimer-type dementia, comprising as an active ingredient a culture obtained by culturing lactic acid bacteria belonging to Bifidobacterium longum and / or bacterial cells. The preventive / ameliorating / treating agent for Alzheimer-type dementia according to claim 1, wherein the culture obtained by culturing lactic acid bacteria belonging to Bifidobacterium longum is fermented milk. Bifidobacterium longum (hereinafter referred to BL2928) (Bifidobacterium longum) belonging to lactic acid bacteria Bifidobacterium longum (Bifidobacteriumlongum) · SBT2928 of (FERMP-10657) in Alzheimer's disease according to claim 1 or 2, Prevention / improvement / therapeutic agent.