JPH05201871A - Agent for inhibiting rise of blood pressure and for preventing cardiomegaly - Google Patents

Abstract

(57) [Summary] [Structure] An agent for suppressing hypertension and preventing cardiac hypertrophy, which comprises, as an active ingredient, bacterial cells of Enterococcus faecalis strain NF-1011 or a processed product thereof. [Effect] To provide a drug which has a remarkable blood pressure elevation inhibitory effect and a cardiac hypertrophy prevention effect and has no side effects at all.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an agent for suppressing hypertension and preventing cardiac hypertrophy, which contains bacterial cells of Enterococcus faecalis NF-1011 strain or a treated product thereof.

[0002]

2. Description of the Related Art There are 100 kinds and 100 trillion intestinal bacteria in the human intestinal tract. The existence of enterobacteria can be understood as one ecosystem. That is, the types and amounts of the constituent microorganisms of the intestinal flora exist while maintaining a complicated relationship such as mutual dependence between the microorganisms and the host organism, and interaction such as antagonism. This intestinal flora can be considered as one organ, and its importance in the host organism is becoming more and more widely recognized. It is recognized that the range of its role is directly or indirectly related to developmental differentiation, immune activity, nutrition (digestion and absorption), various diseases such as various adult diseases and digestive organ diseases, and eventually aging and life span. Has been. The microorganism of the genus Enterococcus is a typical microorganism existing in the intestinal tract.

[0003]

Of the so-called adult diseases, heart disease is one of the three major adult diseases, which together with cancer and brain disease, is the top cause of death. Various diseases are showing various aspects along with changes in the overall life including eating habits. That is, changes in food preferences, pathogenic food additives,
There are many factors such as excess and side effects of medicines, mental stress due to society becoming more complicated, obesity, lack of exercise, etc., but these broadly impaired immune resistance complicate the disease and prevent it. Making it difficult. Furthermore, for example, the number of cases of heart disease has increased remarkably due to westernization of diet such as an increase in fat intake.

[0004]

The present inventors have isolated a new strain of Enterococcus faecalis NF-1011 from the feces of healthy subjects.
The strain has been deposited in the Institute of Microbial Science and Technology of the Agency of Industrial Science and Technology as Microorganism Research Institute No. 12564.

Further, as a result of research, the present inventors have found that the enterococci Enterococcus faecalis strain NF-1011 strain, which is an enterococcus derived from human intestine (human intestinal resident), or a treated product thereof has increased blood pressure. It has been found to have a suppressive and cardiac hypertrophic effect. Since the present bacterium is one of the indigenous bacteria in the human intestinal tract, virtually no toxicity by ingestion was observed, which led to the present invention.

The following is Enterococcus faecalis NF
The isolation means of -1011 and the mycological and physiological properties of the strain are shown.

(1) Separation Means A 10-fold dilution of feces of a healthy person with heat-sterilized water is smeared on an appropriate selective medium (KMP agar plate and SF agar plate), and aerial conditions at 37 ° C for 48- After culturing for 72 hours, bacterial colonies appeared. This bacterial colony was streaked on another homogeneous plate medium and cultured in the same manner to reappear the bacterial colony. The same operation was repeated several times to separate a single colony consisting of a single bacterial species. This new isolate was examined for mycological (morphological, biochemical and serological) properties and classified and identified as belonging to Enterococcus faecalis .

(2) Bacteriological and physiological properties ───────────────────────── Judgment of properties ─────────── ─────────────── Gram stain + Bacterial morphology Spherical catalase − Hemolytic α serogroup D Proliferative 10 ℃ + 45 ℃ + 50 ℃ + Heat resistance 60 ℃ 30 minutes + Bile Esculin Growth in supplemented medium + Growth in pH 9.6 medium + Growth in 6.5% salt-supplemented medium + Methylene blue reducing + Gelatin liquefaction − Growth in 0.01% TTC-supplemented medium + Growth in tellurite-supplemented medium + Presence or absence of acid formation Glycerol + L-arabinose-D-ribose + D-xylose-D-glucose + D-galactose + D-fructose + D-mannose + maltose + mannitol + sucrose + L-sorbose S-D-sorbitol + L-rhamnose-lactose + amygdalin + esculin + cellobiose + melibiose-inulin-merezitose + ────────────────────────── + ; Positive,-; Negative TTC; 2,3,5-triphenyltetrazolium chloride

(Medium) The medium for culturing the microorganism used in the present invention is not particularly limited, but one example thereof is shown below. Glucose 20 g Yeast extract 20 g Meat extract 20 g Dipotassium hydrogen phosphate 40 g Distilled water 1,000 ml (pH adjusted to 7.0 with sodium hydroxide solution, heat sterilized at 115 ° C for 20 minutes)

(Culturing method) This strain was inoculated into 3 ml of the above liquid medium and aerobically statically cultivated (precultured) at 37 ° C. for 10 to 24 hours to give a culture solution of about 10 ⁹ cells / ml ( Seed). This is added to 1000 ml of the above sterilized liquid medium, and static culture is performed in the same manner.

(Collecting, Washing and Drying Method) The culture broth obtained as described above is subjected to centrifugation at 8,000 to 12,000 rpm to obtain viable cells (precipitate). Viable cells were washed 3 times with physiological saline (similar centrifugation operation), then suspended in distilled water to obtain 200 ml of bacterial solution (about 10 ¹¹ cells / ml).
To get

The resulting viable cell suspension is heated at 105 ° C. for 10 minutes to obtain a dead cell suspension. Next, the cells are dried by an appropriate method such as a hot air drying method or a freeze drying method to obtain dried dead cells.

As the agent of the present invention, live cells or dead cells obtained as described above, a ground product thereof, or a water extract can be used. To formulate these, tablets, tablets, etc. can be prepared by known methods using additives such as carriers such as starch, lactose, soybean protein, excipients, binders, disintegrants, lubricants, stabilizers, and flavoring agents. It is formulated into granules.

The amount used depends on symptoms, age, etc.
As an active ingredient, 0.002 to 0.1 g / kg body weight per day can be usually administered to an adult once a day or in several divided doses.

[0015]

INDUSTRIAL APPLICABILITY Since Enterococcus faecalis NF-1011 of the present invention is an intestinal bacterium of a healthy person, it is nontoxic, has no side effects, has an effect of suppressing an increase in blood pressure and preventing cardiac hypertrophy.

[0016]

【Example】

Example 1. (Preparation of bacterial cells) Enterococcus faecalis NF-1011 was inoculated into 1000 ml of the above liquid medium (the number of bacteria: 10 ⁶ cells / ml), and 37 ° C.
After static culture for 24 hours, a culture solution containing about 10 ⁹ viable cells / ml was obtained. The obtained culture solution was centrifuged at 12,000 rpm to collect the cells, which was washed 3 times with distilled water to obtain 200 ml.
The suspension was suspended in distilled water to obtain a bacterial solution. This bacterial solution is 1 at 105 ℃
After heating for 0 minutes, freeze-dried to obtain dead bacterial cell powder.

Embodiment 2. (Blood pressure suppression effect) 6-week-old, male, 1 group of 7 spontaneously hypertensive rats (SH
R) is allowed to freely ingest solid feed (CE-2: manufactured by CLEA Japan, Inc.) and tap water filtered with a PF filter model III (manufactured by Organo), room temperature 25.0 ± 1.0 ° C., humidity 55.0.
± 5.0%, light and dark were switched every 12 hours, and preliminary breeding was performed for 4 weeks.

After preliminary breeding, SH weighing 270-290 g
For the group R, the dead bacterial cell powder obtained in Example 1 was suspended in physiological saline (0.85% NaCl) to give 15, 30, 6
0, 120 mg / animal / day was orally administered for 28 consecutive days using a gastric sonde. The control SHR group was similarly administered with 1 ml of physiological saline. The systolic blood pressure was measured every 7 days from the start of administration for all animals in all groups. The results are shown in Fig. 1. By the oral administration of killed bacterial cell powder, a clear effect of suppressing blood pressure increase depending on the dose was observed.

Example 3. (Blood pressure suppression effect) 6-week-old, male, 1 group of 7 spontaneously hypertensive rats (SH
R) was bred in the same manner as in Example 1.

After preliminary breeding, SH weighing 270-290 g
In the R group, the dead bacterial cell powder obtained in Example 1 was suspended in physiological saline (0.85% NaCl) to give 120 mg / mouse of 2.
Oral administration was performed using a gastric sonde for 8 consecutive days. Control S
The HR group was similarly administered with 1 ml of physiological saline. 29
After the first day, administration of dead bacterial cell powder was stopped, and 1 ml of physiological saline was administered for 28 days in the same manner as in the control SHR group.
Changes in systolic blood pressure during this period are shown in FIG. Oral administration of killed bacterial cell powder suppressed the elevation of systolic blood pressure, and it was clarified that this inhibitory effect was maintained for a certain period even after the administration was discontinued.

Example 4. (Blood pressure suppression effect and cardiac hypertrophy prevention effect) 6-week-old, male, 1 group of 7 spontaneously hypertensive rats (SH
R) and normal control rats (WKY rats) were allowed to freely ingest tap water filtered with solid feed (CE-2: manufactured by CLEA Japan, Inc.) and PF filter model III (manufactured by Organo),
Room temperature 25.0 ° C ± 1.0 ° C, humidity 55.0 ± 5.0%,
Light and dark were switched every 12 hours, and preliminary breeding was performed for 4 weeks.

After preliminary breeding, SH of 270 to 290 g in weight
In the R group, the dead bacterial cell powder obtained in Example 1 was suspended in physiological saline (0.85% NaCl) to give 120 mg / mouse of 2.
For 8 days, 240 mg / mouse was orally administered every day from the 29th day to the 270th day using a gastric sonde. Control SH
The R group and the control WKY rat group were similarly administered with 1 ml of physiological saline.

FIG. 3 shows changes in systolic blood pressure in both SHR groups that were administered with dead bacterial cells and not administered. After the administration, the weight of each organ (heart, liver, kidney, spleen) in all three groups was measured, and as shown in Table 1, the heart showed a remarkable change in appearance, and the dead bacterial cell powder The administration group showed significant suppression in heart weight and body weight ratio between the non-administration group.

[0024]

[Table 1]

Example 5. (Preparation example) (1) 150 mg of dead bacterial cell powder obtained in Example 1 is mixed with 150 mg of purified starch powder and 700 mg of lactose to give tablets / granules.

(2) 300 mg of dead bacterial cell powder obtained in Example 1
Is mixed with 300 mg of soy protein and 400 mg of lactose to give tablets /
Or use granules.

As described above, as shown in Examples 1 to 5, the present invention has been obtained which has a remarkable blood pressure elevation inhibitory effect and a cardiac hypertrophy prevention effect, and has no side effects.

[Brief description of drawings]

FIG. 1 is a graph showing an effect of suppressing an increase in blood pressure.

FIG. 2 is a graph showing the blood pressure increase suppressing effect.

FIG. 3 is a graph showing an effect of suppressing an increase in blood pressure.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁵ Identification code Office reference number FI technical display location C12R 1:01) 7804-4B

Claims (1)

[Claims] 1. An agent for suppressing hypertension and preventing cardiac hypertrophy, which comprises, as an active ingredient, bacterial cells of Enterococcus faecalis strain NF-1011 or a processed product thereof.