KR20080004515A - Hypotensive agent produced by cultivation of lactic acid bacterium - Google Patents

Abstract

A hypotensive agent comprising, as the active ingredient, a culture of a lactic acid bacterium which is produced by cultivating the lactic acid bacterium in a culture medium containing a microalga and the lactic acid bacterium.

Description

Blood pressure lowering agent obtained by lactic acid bacteria culture {HYPOTENSIVE AGENT PRODUCED BY CULTIVATION OF LACTIC ACID BACTERIUM}

The present invention relates to a blood pressure lowering agent comprising, as an active ingredient, a culture in which lactic acid bacteria are cultured in a specific culture solution.

Foods containing spirulina or chlorella have been used as foods that can easily consume nutritional ingredients that are easily lacking in normal diet, since they contain abundant nutritional ingredients and inherent nutritional ingredients unique to green and yellow vegetables. . In recent years, foods in which lactic acid bacteria are cultivated in spirulina or chlorella to improve the odor and flavor peculiar to spirulina and chlorella have been proposed (see Patent Documents 1 and 2).

Moreover, since a specific health food is effective for hypertension, the method of inoculating lactic acid bacteria to oil (patent document 3), the method of inoculating lactic acid bacteria to an enzyme followed by an enzyme (patent document 4), or bran, an embryo It is known by the method (patent document 5) which immerses rice bran etc. in water under fixed conditions.

However, although a culture obtained by maintaining a mixture of spirulina or chlorella and lactic acid in the presence of water and carrying out the culture of lactic acid bacteria is used as a health food, it is not known that it is effective for hypertension.

Patent Document 1: Japanese Patent Application Laid-Open No. 2004-081206

Patent Document 2: Japanese Patent Application Laid-Open No. 63-157963

Patent Document 3: Japanese Patent No. 3172150

Patent Document 4: Japanese Patent Application Laid-Open No. 2001-120179

Patent Document 5: Japanese Patent No. 2590423

(Initiation of invention)

(Tasks to be solved by the invention)

An object of the present invention is to provide a blood pressure reducing agent comprising, as an active ingredient, a lactic acid bacteria culture obtained by culturing lactic acid bacteria in a culture solution containing microalgae and lactic acid bacteria.

(Means to solve the task)

In order to solve the above problems, the present inventors have carried out a test regarding the blood pressure lowering action of the lactic acid bacteria culture obtained by maintaining the microalgae and lactic acid bacteria in the presence of water and culturing the lactic acid bacteria in the mixture under various conditions. It has been found that water has a very good action and has come to complete the present invention.

That is, the present invention is to provide a blood pressure reducing agent comprising a lactic acid bacteria culture obtained by culturing lactic acid bacteria in a culture solution containing microalgae and lactic acid bacteria as an active ingredient.

(Effects of the Invention)

According to the present invention, a blood pressure lowering agent comprising a lactic acid bacteria culture obtained by culturing lactic acid bacteria in a culture solution containing microalgae and lactic acid bacteria as an active ingredient can be obtained.

BRIEF DESCRIPTION OF THE DRAWINGS The figure which shows the comparison of GABA amount (mg / 100g) of Example 1 and Example 2 until the culture time of 24 hours.

Fig. 2 is a diagram showing the blood pressure increase rate at Week X based on the blood pressure at Week 0 (test example).

Hereinafter, the content of the present invention will be described in detail.

Examples of the microalgae used in the present invention include spirulina and chlorella.

The spirulina used in the present invention includes, for example.

Spirulina is a microscopic single-celled microorganism belonging to the genus Arthrospira and Spirulina, which is included in Cyanobacteria and has been collectively referred to as Spirulina. Arthrospira platensis, Arthrospira maxima, Arthrospira geitleri, Arthrospira siamese, Spirulina major, Spirulina subsalsa, Spirulina subsalsa, etc. Among them, arthrospyras pratensis, arspyras maxima, arsspira gate Larry and arsspira sham are preferable in view of the fact that they can be artificially cultured and are easily available.

As chlorella used by this invention, the following are mentioned, for example.

Chlorella is an algae of Chlorophyceae and Chlorella, which is easy to obtain and excellent in safety, such as C. Vulgaris, Chlorella regularis. , C. pyrenoidosa, C. ellipsoidea, and the like.

These microalgae have been widely used in conventional foods, and it has been confirmed that there is no problem about safety.

As these microalgae, the crude processed material etc. which were processed by the methods, such as a live granulation, a dry granulation, and a mechanical treatment, are mentioned.

The raw microorganisms are obtained by, for example, harvesting spirulina or chlorella cultured in water by a method such as centrifugation and filtration. The raw body may be used as it is after harvest from the culture tank, but it is preferable to wash with water or physiological saline.

Examples of the dry granulated product include those obtained by freeze-drying or spray drying the raw crude product obtained by the above method.

The processed product treated by the mechanical treatment method is obtained by, for example, performing a mechanical treatment such as ultrasonic irradiation treatment or homogenization of a living body. The mechanical processed material of the tank may be subjected to a drying treatment thereafter.

As the granules to be used in the present invention, the raw granules are also preferable in terms of taste and smell since they further retain the active ingredient of the microalgae.

Raw products are usually suspended or suspended in water depending on the degree of water removal at the time of harvest, or may be in the form of a paste having a lower water content than that of the suspended or a cake in which the water content is lower than that of the paste. Any state can be used. It is preferable to use spirulina and chlorella as suspension suspensions (sometimes referred to as suspensions). In addition, when using a dry granulated body and a coarse processed material, it may be in the dried state, and may be made into the suspension form, the paste form, or the cake form like water raw material by adding water.

In order not to damage the active ingredient which micro algae have, it is preferable not to heat-sterilize, but you may use what was heat-sterilized as needed.

Here, the microalgae may use spirulina or chlorella alone or may use a mixture of spirulina and chlorella.

Next, lactic acid bacteria will be described.

Lactic acid bacteria have long been used for the processing of many food products, such as fermented dairy products, cast products, pickled vegetables and fruits, for the purpose of storing and seasoning foods. As lactic acid bacteria used in the present invention, any lactic acid bacteria that can be used in food can be used without limitation. The lactic acid bacteria are classified into oil-based lactic acid bacteria, plant-based lactic acid bacteria, enteric lactic acid bacteria, and lactic acid bacteria derived from natural lakes in which algae grows depending on the growth environment derived. In addition, lactic acid bacteria are classified into mesophilic bacteria, thermophilic bacteria, flame resistant bacteria and the like according to their optimum growth conditions, and may be bacteria having any properties.

As lactic acid bacteria used in the present invention, Lactobacillus genus, Pediococcus genus, Teteragenococcus genus, Carnobacterium genus, Vagococcus genus, Genus Luconostoc, Genus Weissella, Genus Oenococcus, Genus Atopobium, Genus Streptococcus (formerly genus Enterococcus, herein) Belonging to the genus Cocous), Enterococcus genus, Lactococcus genus, Aerococcus genus, Alloiococcus genus, Melisos coccus genus, Bifidobacterium The genus Bifidobacterium, and the like, and further examples thereof include Lactobacillus delbrueckii, Lactobacillus plantarum, Lactobacillus acidophilus, and Lactobacillus acidophilus. And brevis (Lactobacillus brevis), Lactococcus lactis (Lactococcus lactis), Leuconostoc sp., Enterococcus casseliflavus and the like. As lactic acid bacteria used in the present invention, lactic acid bacteria belonging to the genus Lactobacillus, Lactococcus and Enterococcus are preferable.

Lactic acid bacteria may be used as a single species, or may mix and use two or more types of bacteria. In the culture step described later, the same type of bacteria may be cultured by dividing the bacteria into two or more stages, or different bacterial species may be divided into two or more stages and cultured.

The lactic acid bacteria may be cultured in an agar medium or a liquid medium, and then stored by a preservation method such as cold storage, cryopreservation, or dry preservation, but these stored lactic acid bacteria are cultured by incubating in a liquid medium (hereinafter, It is preferable to use a seed culture solution, which is omitted in view of the fact that the growth rate of the lactic acid bacteria is high and the activity of the production of flavors such as acetaldehyde, diacetyl, organic acid production ability and the like is high. The medium to be used for cultivating the species culture medium may be any medium that can be used to grow lactic acid bacteria, but there is no limitation. Generally, as a liquid medium for culturing lactic acid bacteria, for example, MRS medium (manufactured by Merck Co., Ltd.) designed by Man, Rogosa Sharpe And mediums such as whey medium and skim milk medium using milk components.

In order to prepare a seed culture liquid, the lactic acid bacteria preserve | saved in the said liquid medium are normally added, and it may make it culture | cultivate by standing or stirring, maintaining in aerobic state or a base state adapting to the lactic acid bacteria to culture | cultivate.

Next, the preparation method of the lactic acid bacteria culture used for this invention is demonstrated.

Lactic acid bacteria cultures are prepared by culturing lactic acid bacteria in a culture solution containing microalgae and lactic acid bacteria. Herein, the culture solution containing microalgae and lactic acid bacteria is not particularly limited as long as it contains microalgae and can grow lactic acid bacteria, but for example, a method of adding lactic acid bacteria to a suspension in which microalgae is suspended in water and microalgae in water It can be obtained by the method of adding lactic acid bacteria to the added wet liquid, paste, cake, etc., the method of adding the species culture solution to the suspension.

More specifically, it can obtain by the method illustrated below. (i) Lactic acid bacteria culture medium or dried lactic acid bacteria is added to the raw or dried suspension and paste. (ii) The lactic acid bacteria culture medium is added to the raw or dried cake. (iii) Add the lactic acid bacteria culture medium in the amount to be wetted to the dried granules.

Among these, (i) is preferable, and in (i), it is preferable to use a suspension of a living organism as a microalgae and to use a culture solution, especially a species culture solution, as lactic acid bacteria in view of high culture capacity and flavor production capacity of lactic acid bacteria.

The crude suspension, paste, cake or lactic acid bacteria culture medium contains water, but when the culture solution containing microalgae and lactic acid bacteria lacks water, water may be added and kept in a wet state or in water. It is preferable to use sterile water for water.

The content of the microalgae in the culture solution containing the microalgae and lactic acid bacteria is preferably from 0.1 to 30% by mass, and from 1 to 20, as the dry cells from the viewpoint of improving the efficiency in the harvesting step and the drying step after the lactic acid bacterium culture described later. Mass% is more preferable.

The lactic acid bacteria culture in the culture solution containing the microalgae and lactic acid bacteria may be a stationary culture, or if the culture solution is a liquid, a stirred culture by propeller stirring may be used. In addition, the system to be cultured so as to be suitable for the growth of lactic acid bacteria to be used may be in a base state or may be in an aerobic state.

The amount of lactic acid bacteria used is the number of lactic acid bacteria that grow, but the number of lactic acid bacteria at the start of cultivation of lactic acid bacteria from the viewpoint of improving the propagation of lactic acid bacteria is preferably 10 ⁵ to 10 ¹¹ per 1 g of solid-sized algae, and 10 ⁶ More preferably, 10 to ¹⁰ individuals.

The pH of the culture solution containing the microalgae and lactic acid bacteria is changed in the culture process by acids such as lactic acid produced by lactic acid bacteria culture. The pH at the start of the culture is preferably 4.0 to 9.0, more preferably 5.0 to 7.0. desirable.

The culture temperature is irrelevant as long as the lactic acid bacteria can grow, but 4 to 45 ° C is preferable, and 20 to 40 ° C is more preferable in that the lactic acid bacteria are suitable for the growth of the lactic acid bacteria and the active ingredients of the microalgae are not damaged.

The culture time is preferably 9 to 100 hours, more preferably 15 to 72 hours, and particularly preferably 18 to 50 hours in order to sufficiently propagate the lactic acid bacteria and exhibit an excellent blood pressure lowering effect.

The number of lactic acid bacteria at the start of the culture is sufficient to exert an excellent blood pressure lowering effect, to sufficiently suppress the increase of other microorganisms, to satisfactorily reduce the taste and smell peculiar to microalgae, and to exhibit a blood pressure lowering effect. It is preferable that it is increased by 10 to 1000 times.

In addition, in order to maintain an optimal pH for lactic acid bacteria growth, the pH may be adjusted by adding a basic compound such as potassium hydroxide or calcium hydroxide during the culturing, but the pH of the culture solution containing the microalgae and lactic acid bacteria may be changed by lactic acid produced by lactic acid bacteria. It is preferable to fall to around 5.0 because it lowers the bacteria and exhibits a blood pressure lowering action.

In the present invention, the addition of sugar to the culture medium containing the microalgae and lactic acid bacteria is preferable because it can suppress the growth of microorganisms and can further reduce the smell and taste peculiar to the microalgae. The growth inhibitory effect of these microorganisms is particularly remarkable when a dry coarse microorganism is easily reproduced as microalgae.

As said saccharide, monosaccharide, oligosaccharide, polysaccharide, etc. are mentioned, for example. As a monosaccharide, glucose, galactose, mannose, fructose, ribose, xylose, etc. are mentioned, for example. Examples of oligosaccharides include disaccharides such as sucrose and maltose, galactooligosaccharides, fructooligosaccharides, soy oligosaccharides, xylo oligosaccharides and raffinose. Examples of the polysaccharides include amylose, amylopectin, cellulose, glycogen, β-glucan, mucopolysaccharide and the like. As the saccharide, oligosaccharides are preferable, and galactooligosaccharide and xyloligosaccharide are particularly preferred.

There is no restriction | limiting in particular as a sugar addition method, For example, you may mix microalgae, lactic acid bacteria, and saccharides, You may mix the microalgae and lactic acid bacteria which added saccharides previously, Lactic acid bacteria and microalgae which added saccharides previously You may mix. In addition, although a solid thing may be used for a saccharide, it is preferable to use what was melt | dissolved in water etc. previously and made into aqueous solution.

As the usage-amount of a saccharide, 0.05-20 mass% is preferable with respect to the sum total of the culture liquid and microsaccharides containing microalgae and lactic acid bacteria, and 0.1-10 mass% is more preferable.

Due to the antibacterial action of lactic acid, organic acids such as acetic acid, and bacteriocin produced during the preparation of such lactic acid bacteria cultures, other contaminating bacteria are reduced, and lactic acid bacteria become superior species. Furthermore, flavors such as acetaldehyde and diacetyl produced by lactic acid bacteria lower the smell and taste peculiar to microalgae, making them easy to use in food.

The lactic acid bacteria culture obtained above can be used as a lactic acid bacteria beverage or a lactic acid bacteria added food as it is.

The lactic acid bacteria culture obtained above is characterized by having a different composition from the original microalgae before the culture.

For example, proline, cysteine, valine, leucine, isoleucine, γ-aminobutyric acid (hereinafter abbreviated as 'GABA'), lysine, histidine, and the like are related to the amount of free amino acid in the culture after 24 hours of culture. The content per 100 g is 10 times or more, and the total amount of free amino acids is also increased by 2 times or more.

In addition, the amount of protein contained in the raw material before the culture and the lactic acid bacteria cultures treated with a molecular weight 10,000 membrane was measured by the BCA (Bicinchoninic Acid) method, which is a general measurement method of protein amount. As a result, for example, when spirulina or raw material was used as a coarse substance, it was found that after 24 hours of culture, the component having a molecular weight of 10,000 or less increased about twice as much as before the start of culture, and the high molecular weight protein became low molecular weight and became an excellent absorbent component. It became.

In addition, the lactic acid culture obtained by the present invention is characterized by containing GABA at a high concentration as compared with the conventional lactic acid bacteria cultures of the present invention. For example, Patent Document 3 described above shows that when L. lactis YIT 2027 is used alone in oil, it is 15 mg / 100 mL in a three-day culture, and 38 mg / L is carried out by performing a mixed culture with L. casey YIT 9029. There is a description that increases to 100 mL.

However, according to the method of the present invention, it is evident that GABA can be produced at a concentration of 100 mg / 100 mL or more at a culture time of 24 hours, which is extremely excellent as a GABA preparation method.

The blood pressure reducing agent of this invention uses the lactic acid bacteria culture obtained above as an active ingredient. The blood pressure reducing agent of the present invention can be prepared in the form of tablets, granules, powders, solutions, and the like, such as uncoated tablets, film coated tablets, dragees, enteric tablets, and multilayer tablets. Preparation in these forms may be carried out by suspending, drying, pulverizing and molding the lactic acid bacteria culture in accordance with a general preparation method according to each form. In preparation in each form, the binder, surfactant, thickener, filler, disintegrating agent, excipient, etc. which are generally used for preparation in the form can be used. In addition, such a lactic acid bacteria culture may be used as a lactic acid bacteria beverage or a lactic acid bacterium additive as it is, and a lactic acid bacteria culture may be added to other food.

For example, when performing a drying process, although it is normally performed so that the moisture content of a crude body may be 4-7 mass%, the process which can maintain the bacteria count of lactic acid bacteria is preferable. As a preferable drying method, a freeze drying method, a spray drying method, etc. are mentioned, for example, A spray drying method is more preferable at an economic point. The drying temperature at the time of drying process improves production efficiency, so that the higher the ventilation wind temperature, but the quality of the microalgae is good and the number of lactic acid bacteria is not lowered, so that the drying temperature is preferably in the range of 30 to 70 ° C. More preferably, it is the range used as 40-60 degreeC. In addition, in this invention, product temperature shall mean the sample temperature after drying.

What is necessary is just to tablet the powder obtained above by the well-known tableting method, for example in preparation to a tablet.

In the case of a liquid preparation, for example, the powder obtained above may be dispersed in water, and the cultured lactic acid bacteria culture may be prepared as it is or diluted with water or the like.

The intake of the patient of the blood pressure lowering agent of the present invention is preferably determined in consideration of the sex, age, symptoms, etc. of the patient, but in general, 0.2 to 10 g / day, in particular 0.5 to 8 g / day in terms of lactic acid bacteria culture. These may be taken at once, or may be divided into several times a day.

Next, the blood pressure lowering effect of the blood pressure reducing agent of this invention is demonstrated.

The present inventors carried out the verification of the effect on hypertension, which is considered to be the main cause of various lifestyle diseases, with respect to the lactic acid bacteria culture obtained above, using experimental animals.

That is, spirulina lactic acid bacteria cultures and chlorella lactic acid cultures were mixed in a solid feed, and the blood pressure measurement was performed to check the blood pressure lowering effect in spontaneously hypertensive rats (SHR system). The experimental group is a group in which a solid feed is fed to the target group, and in addition, a group in which the test substance is mixed, and a group in which spirulina raw material (test substance 1) is mixed with the solid feed, spirulina lactobacillus culture (test substance 2) The group mixed with the solid feed, the group which mixed chlorella raw material (test material 3) with the solid feed, and the group which mixed the chlorella lactic acid culture (test material 4) with the solid feed were set. As a result, it was clear that the group in which the spirulina lactic acid culture or the chlorella lactic acid culture was mixed all had a strong blood pressure lowering effect as compared with the group in which the control, the spirulina original and the chlorella original were mixed (see Test Example).

In addition, acute toxicity was examined through the mouse to confirm the safety of the lactic acid bacteria culture. Mice were 10 male and female ddY-N mice (20-26 g body weight) 5 weeks old. In the administration method, the culture was pulverized and suspended in a CMC 1% solution, and a single oral forced administration of a 10 mass% suspension of the highest dose that could be administered by gastride. Cultures were observed for 1 week after administration. Both sexes LD ⁵⁰ is over 6,000㎎ / ㎏, were identified as food safety.

Example

Next, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to the following Examples.

( Example  One) Spirulina  Preparation of Lactic Acid Bacteria Culture (1) and Measurement of Lactic Acid Bacteria

25 kg of galactooligosaccharide and 825 kg of tap water were added to a 2500 L culture vessel, and after heat sterilization, 100 kg of spirulina was added thereto, and 50 kg of a seed culture solution of lactic acid bacteria Lactobacillus brevis was inoculated. Lactic acid bacteria were cultured by stirring the aeration propeller at 37 ° C for 72 hours. The culture solution was sampled at culture time 0, 9, 12, 15, 18, 24, 48 and 72 hours to determine the number of lactic acid bacteria.

In addition, the lactic acid bacteria number measurement was performed in the following procedures. The suspension was adjusted by suspending 1.0 ml of lactic acid bacteria culture solution in 19 ml of phosphate buffered saline. The suspension was further diluted 10 times, 10 ² times, 10 ³ times, 10 ⁴ times, 10 ⁵ times, 10 ⁶ times and 10 ⁷ times through phosphate buffered saline to obtain sample dilutions. 0.1 ml of dilution was smeared on MRS agar medium prepared by adding 15 g of agar to 1000 ml of MRS medium (Merck Cat. No. 10661), and incubated for 48 hours at 35 ° C. Using the agar medium in which 30-300 colonies of lactic acid bacteria were confirmed, the number of colonies of this agar medium was measured, and the number obtained by multiplying this number by the dilution factor was made into the colony number of lactic acid bacteria.

(Measurement of Amino Acid Amount)

A portion of each sampled culture was spray dried to provide for amino acid analysis. Each 200 mg of lactic acid bacteria culture of spirulina was added to 5 mL of water, suspended and left for 30 minutes. The mixture was separated into a supernatant and an insoluble matter by a centrifuge (3000 rpm, 10 minutes), and 1 mL of the supernatant was collected. 1 mL of 10% (w / v) trichloroacetic acid solution was added thereto, followed by centrifugation. The obtained supernatant was subjected to HPLC analysis through o-phthalaldehyde method (post column detection) to determine the amino acid amount.

(HPLC condition)

Column: Hitachi # 4619 (4㎜ × 15㎝)

Temperature: 60 ° C (same temperature even during post column reaction)

Flow rate: eluent; 0.4 mL / min. Reaction solution; 0.5 mL / min.

Detection: Fluorescence Detector

The amount of L-glutamic acid, GABA amount, and the number of lactic acid bacteria in the culture at each incubation time are shown in [Table 1].

Incubation time (hr) 0 9 12 15 24 36 48 72 L-Glutamic Acid (mg / 100g) 1240 1221 1180 995 812 295 85 87 L-Glutamic Acid (mg / 100mL) 155 152 148 124 102 37 11 11 GABA (mg / 100g) 4 6 6 72 205 678 875 758 GABA (mg / 100mL) One One One 9 26 85 110 95 Lactic acid bacteria count (× 10 ⁵ / mL) 6 1000 1100 1500 2050 2250 3150 2200

( Example  2) Spirulina  Preparation of Lactic Acid Bacteria Culture (2)

In this example, lactic acid bacteria were cultured in the same manner as in Example 1 except for maintaining the pH at 5.0 and the incubation temperature at 30 ° C. in a 50% (w / w) lactic acid solution. The culture solution was sampled at culture time 0, 9, 12, 15, 18, 21, 24 hours, and the GABA amount (mg / 100g) was measured.

( Example  3) Spirulina  Preparation of Lactic Acid Bacteria Culture (3)

In this example, the GABA amount (mg / 100g) was measured in the same manner as in Example 2 except that xyloligosaccharide was used instead of galactooligosaccharide.

A comparison between the GABA amounts (mg / 100g) of Example 1 and Example 2 up to 24 hours of incubation time is shown in FIG. 1 and the results of Example 2 and Example 3 are shown in [Table 2].

In Examples 1 to 3, more GABA production was possible in a short time by the proper selection of pH and incubation temperature.

GABA amount (mg / 100g) Incubation time (hr) 0 9 12 15 21 24 Example 2 5 20 105 210 510 900 Example 3 5 25 110 220 530 910

( Example  4) Preparation of Lactic Acid Bacteria Culture Using Chlorella and Measurement of Lactic Acid Bacteria

A lactic acid bacteria culture was prepared in the same manner as in Example 1 except that chlorella was used instead of spirulina in Example 1, and the number of lactic acid bacteria and amino acid amount were measured. The amount of L-glutamic acid, GABA amount, and the number of lactic acid bacteria in the culture at each incubation time are shown in [Table 3].

Incubation time (hr) 0 9 12 15 24 36 48 72 L-Glutamic Acid (mg / 100g) 672 665 660 594 448 181 48 49 L-Glutamic Acid (mg / 100mL) 84 83 83 74 56 23 6 6 GABA (mg / 100g) 6 7 7 61 180 401 515 505 GABA (mg / 100mL) One One One 8 23 50 64 63 Lactic acid bacteria count (× 10 ⁵ / mL) 6 1000 1050 1350 1850 2000 2800 1950

( Test Example A) blood pressure of the present invention On a lowering agent  Of blood pressure lowering action by

In order to clarify the blood pressure lowering action of the blood pressure lowering agent of the present invention, a test using the following experimental animals was performed.

(Test substance)

Target group: solid feed MF (Oriental Yeast Co., Ltd.)

Test substance 1: spirulina

Test substance 2: Lactic acid fermentation spirulina (lactic acid bacteria culture obtained in Example 1)

Test Substance 3: Chlorella

Test substance 4: Lactic acid fermentation chlorella (lactic acid bacteria culture obtained in Example 4)

(Adjustment and Administration of Test Substances)

Adjustment Method: The test materials 1 and 2 were mixed with 15% of the solid feed MF.

Route of administration: oral

Dosing method: free intake

Dosage: 15% of feed

Duration of administration: 8 weeks

Test animals

Animal species, lineage: Rat, SHR (8 per group)

Gender, age: 6 weeks

Fleating period: About one week after bringing in, fleas were reared together, and the general condition of the period was observed.

(Blood pressure measurement)

Blood pressure was measured by the tail-cuff method, which is a non-invasive measurement of blood pressure in the arteries. The measurement method was performed at a total of 8 points before administration of the test substance (week 7) and every week after administration.

The measurement at each point of blood pressure was performed 3 times, and the average value was made into the measured value. In addition, prior to conducting the test, the test was performed after confirming that the average blood pressure average of the animals was 160 mmHg or more.

The results are shown in [Table 4].

Test substance Week 0 after 2 weeks 4 weeks later 6 weeks later 8 weeks later Target group 165 170 195 215 220 Test substance 1 165 170 190 200 205 Test Substance 2 165 165 180 185 195 Test Substance 3 170 175 190 210 210 Test substance 4 165 170 185 190 200

Table 5 and FIG. 2 show the results of evaluating the blood pressure increase rate of the week X based on the blood pressure at week 0 by the following equation.

% Increase in blood pressure = (blood pressure at week X-blood pressure at week 0) / (blood pressure at week 0) × 100 (%)

Test substance Week 0 after 2 weeks 4 weeks later 6 weeks later 8 weeks later Target group 0% 3.0% 18.2% 30.3% 33.3% Test substance 1 0% 3.0% 15.2% 21.2% 24.2% Test Substance 2 0% 0% 9.1% 12.1% 18.2% Test Substance 3 0% 2.9% 11.8% 23.5% 23.5% Test substance 4 0% 3.0% 12.1% 15.2% 21.2%

As is clear from this test example, the spirulina lactic acid bacteria culture and the chlorella lactic acid culture were confirmed to have an effect of inhibiting blood pressure increase compared to the spirulina and chlorella originals before the culture, respectively.

For example, after 6 weeks, about 30% increase in blood pressure was observed in the subject group, whereas the rate of increase in spirulina and chlorella was suppressed to 20-25%, and in the lactic fermentation product of spirulina and chlorella of the present invention, 12 It became clear that it was suppressed to about 15%, and the blood pressure lowering effect of the lactic acid fermentation product which concerns on this invention was confirmed.

The present invention can be used in the pharmaceutical industry, health food, food and the like.

Claims (15)

A blood pressure reducing agent, comprising a lactic acid bacteria culture obtained by culturing lactic acid bacteria in a culture solution containing microalgae and lactic acid bacteria as an active ingredient. The method of claim 1, And said microalgae are spirulina, chlorella or a mixture of spirulina and chlorella. The method of claim 1, The lactic acid bacterium is a lactic acid bacterium belonging to the genus Lactobacillus. The method of claim 1, The lactic acid bacteria are lactic acid bacteria belonging to the genus Lactococcus. The method of claim 1, The lactic acid bacterium is a lactic acid bacterium belonging to the genus Enterococcus. The method of claim 1, Blood pressure lowering agent, characterized in that for 15 to 72 hours of lactic acid bacteria culture. The method of claim 1, A blood pressure reducing agent, characterized in that the culture is continued until the number of lactic acid bacteria at the start of the culture of lactic acid bacteria is 1 × 10 ⁵ to 10 ¹¹ per 1 g of the microalgae converted to solid content, and the lactic acid bacteria is 10 to 1000 times the number of lactic acid bacteria at the start of culture. The method of claim 1, Blood pressure lowering agent, characterized in that the culture solution further contains sugars. The method of claim 8, A blood pressure reducing agent, wherein the saccharide is galactooligosaccharide. The method of claim 8, Blood pressure lowering agent, characterized in that the saccharide is xyloligosaccharides. A method of producing a blood pressure reducing agent by performing a step of culturing a lactic acid bacterium in a culture solution containing fine algae and lactic acid bacteria, and using the lactic acid bacteria culture obtained by this step as an active ingredient. The method of claim 1, A blood pressure lowering agent, wherein the content of the microalgae contained in the culture solution is 0.1 to 30% by mass. The method of claim 1, The lactic acid bacterium is selected from the group consisting of Lactobacillus delbourg extract, Lactobacillus plantarum, Lactobacillus asidophilus, Lactobacillus brevis, Lactococcus lactis, Luconostoke sp., Enterococcus casselliflavus At least one or two or more kinds of blood pressure reducing agents, characterized in that. Use of the blood pressure lowering agent of Claim 1 to treat hypertension. A method of treating hypertension using the blood pressure reducing agent according to claim 1.

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