WO2006112364A1

WO2006112364A1 - Hypotensive agent produced by cultivation of lactic acid bacterium

Info

Publication number: WO2006112364A1
Application number: PCT/JP2006/307856
Authority: WO
Inventors: Keiichi Sadoyama; Takeshi Miyagi; Keiichiro Inafuku; Masaki Sakakibara; Tomohiro Hirahashi; Noritaka Yoshikawa
Original assignee: Dainippon Ink And Chemicals, Inc.
Priority date: 2005-04-15
Filing date: 2006-04-13
Publication date: 2006-10-26
Also published as: KR20080004515A; JPWO2006112364A1; CN101160136A; JP4762987B2

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

Specification

Antihypertensive agent obtained by lactic acid bacteria culture

Technical field

[0001] The present invention relates to an antihypertensive agent comprising a culture obtained by culturing lactic acid bacteria in a specific culture solution as an active ingredient.

Background art

[0002] Foods containing spirulina or chlorella have traditionally been rich in nutrients that are unique to green-yellow vegetables and are inherently rich in nutrients, making it easy to ingest nutrients that tend to be deficient in normal eating habits. It is used as food. Recently, foods have been proposed in which lactobacillus is cultured in Spirulina or Chlorella to improve the peculiar smell and flavor of Spirulina and Chlorella (see Patent Documents 1 and 2).

By the way, it is said that specific health foods are effective for hypertension, and a method of inoculating milk with lactobacilli (Patent Document 3), a method of inoculating milk products with enzymes and then lactic acid bacteria (Patent Document 4). ), Or a method of immersing rice bran or rice bran containing germ under certain conditions in water (Patent Document 5).

However, although a culture obtained by maintaining a mixture of spirulina or chlorella and lactic acid bacteria in the presence of water and culturing lactic acid bacteria is used as a health food, it is effective for hypertension. ! / That is completely known! /.

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

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

Patent Document 3: PR No. 3172150

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

Patent Document 5: Patent 2590423

Disclosure of the invention

Problems to be solved by the invention

[0003] An object of the present invention is to provide an antihypertensive 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 for solving the problem

[0004] In order to solve the above-mentioned problems, the present inventors maintain a microalgae and lactic acid bacteria in the presence of water, and obtain a lactic acid bacteria culture obtained by culturing the lactic acid bacteria in the mixture under various conditions. On the other hand, when a blood pressure lowering effect was tested, it was found that the culture had a very excellent action, and the present invention was completed.

That is, the present invention provides an antihypertensive agent comprising a lactic acid bacterium culture obtained by culturing lactic acid bacteria in a culture solution containing microalgae and lactic acid bacteria as an active ingredient. The invention's effect

[0006] According to the present invention, it is possible to obtain a blood pressure-lowering agent containing 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.

Brief Description of Drawings

[0007] FIG. 1 is a graph showing a comparison of GABA amounts (mg / 100 g) of Example 1 and Example 2 up to a culture time of 24 hours.

FIG. 2 is a graph showing the rate of blood pressure increase at week X with reference to blood pressure at week 0. (Examination example)

BEST MODE FOR CARRYING OUT THE INVENTION

[0008] The contents of the present invention are described in detail below.

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

Examples of the spirulina used in the present invention include the following. Spirulina is a fine single-celled microorganism belonging to the genus Arthrospira and Spirulina, which has been included in cyanobacteria and has been collectively referred to as the genus Spirulina. For example Arusurosubira 'platensis (Ar throspira platensis), Anoresurosuhifu' Ma ³ r Shima (Arthrospira maxima, Anoresu Rosupira-Geitoreri (Arthrospira geitleri), Arusurosupira-Saiamize (Arthros pira Siamese), Spirulina 'Mayer (Spirulina major), Spirulina Sabusarusa ( Spirulina subsalsa), among others, can be cultured artificially and are easily available. Gatorelli, Alsrosvilla Siamese are preferred.

[0009] Examples of the chlorella used in the present invention include the following.

Chlorella is a green algae (Chlorophyceae), Chlorella genus (Chlorella) algae (Chlorella) (Chlorella), is easy to obtain and excellent in safety. For example, Chlorella 'Bulgaris (C. Examples include 'Chlorella regularis', chlorella 'Pyrenoidosa', and chlorella 'C. ellipsidea'.

[0010] These microalgae have been widely used as foods in the past, and it has been confirmed that there are no problems with regard to safety.

Examples of these microalgae include raw alga bodies, dried alga bodies, and processed alga bodies processed by a method such as mechanical treatment.

The raw algal bodies can be obtained, for example, by harvesting spirulina or chlorella cultured in water by a method such as centrifugation or filtration. Raw algal cells can be used as they are after harvesting, but it is preferable to wash them with water or saline!

Examples of the dried alga body include those obtained by freeze-drying or spray-drying the raw alga body obtained by the above method.

The processed alga body treated by the mechanical treatment method can be obtained, for example, by subjecting raw alga bodies to ultrasonic treatment or mechanical treatment such as homogenization. The mechanically processed alga body may be subjected to a drying treatment thereafter.

[0011] The algal bodies used in the present invention are preferably raw algal bodies because they retain more active ingredients of microalgae, and are also preferable in terms of taste and smell.

Raw alga bodies are usually suspended in water depending on the degree of water removal during harvesting, pastes with less water content than suspended ones, There is a cake-like state in which the water content is less than that in the pasty state. Spirulina and chlorella are preferably used in the form of suspended alga bodies (hereinafter sometimes referred to as suspension). In addition, when using dried algae or processed algae, it may be in a dry state, or it may be a suspension, paste or cake like raw algae by adding water. .

[0012] Microalgae should not be heat sterilized so as not to impair their active ingredients. Although it is preferable, what was heat-sterilized as needed can also be used.

Here, as the microalgae, either spirulina or chlorella may be used alone, or a mixture of spirulina and chlorella may be used.

Next, lactic acid bacteria will be described.

Lactic acid bacteria have long been used to process many foods, including fermented dairy products, brewed products, vegetables, and pickles of fruits, for the purpose of storing and seasoning foods. As the lactic acid bacteria used in the present invention, any lactic acid bacteria that can be used for food can be used without limitation. Lactic acid bacteria are classified into milk-based lactic acid bacteria, plant-based lactic acid bacteria, intestinal lactic acid bacteria, and lactic acid bacteria derived from natural lakes where algae grow, depending on the growth environment. Lactic acid bacteria are classified into mesophilic bacteria, thermophilic bacteria, salt-tolerant bacteria, and the like depending on the optimal growth conditions, but bacteria having any property may be used.

[0014] The lactic acid bacteria used in the present invention are taxonomically classified as Lactobacillus, Pediococcus, Tetragenococcus, Carnobacterium, and Vagococcus. Genus, Leuconostoc genus, Weissella genus, Oenococcus genus, Atopobium genus, Streptococcus genus (official name is Enterococcus genus, and in this document, Enterococcus genus) ), Enterococcus

(Enterococcus), Factococcus, Aerococcus, Alloiococcus, Melissococcus, Bifidobacterium, and the like. delbrueckii), Lacto bacillus plantarum, Lactobacillus acidophil us, Lactobacillus brevis, Lactococcus lactis, sp Species such as casserole flavus (Enterococcus casseliflavus) can be mentioned. The lactic acid bacteria used in the present invention are preferably lactic acid bacteria belonging to the genus Lactobacillus, Ratatococcus, or Enterococcus.

[0015] Lactic acid bacteria may be used alone, or two or more bacteria may be mixed and used. . In the culture process described later, the same type of bacteria may be inoculated and cultured in two or more stages, or different bacterial species may be inoculated and cultured in two or more stages. Lactic acid bacteria can be stored after culturing in an agar medium or liquid medium, and then stored by refrigeration storage, frozen storage, dry storage, or other storage methods. It is used to inoculate and culture in a liquid medium (hereinafter abbreviated as “seed culture medium”). The ability to produce flavors such as acetaldehyde and diacetyl, which have a fast growth rate of lactic acid bacteria, and the ability to produce organic acids. It is preferable because of its high activity. The medium used for cultivating the seed culture is not particularly limited as long as the lactic acid bacterium to be used can grow, but in general, as a liquid medium for culturing lactic acid bacteria, for example, MRS medium devised by Man, Rogosa, Sharpe ( And a whey medium and a skim milk medium using milk components.

[0016] In order to prepare a seed culture solution, usually, a stored lactic acid bacterium is added to the liquid medium, and maintained in an aerobic state or an anaerobic state suitable for the lactic acid bacterium to be cultured, and left or stirred. Cultivate.

[0017] Next, a method for preparing a culture of lactic acid bacteria used in the present invention will be described.

The lactic acid bacteria culture is prepared by culturing lactic acid bacteria in a culture solution containing microalgae and lactic acid bacteria. Here, 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. For example, a suspension in which microalgae is suspended in water. It can be obtained by a method of collecting lactic acid bacteria in the liquid, a moist liquid containing fine algae in water, a method of adding lactic acid bacteria to a paste, cake, etc., a method of adding the seed culture solution to the suspension, etc. Monkey.

More specifically, it can be obtained by the method exemplified below. (I) Add a culture solution of lactic acid bacteria or a dried lactic acid bacterium to a suspension or paste of raw algae or dried algae. (ii) A culture solution of lactic acid bacteria is added to a raw algal body or a dried algal body cake. (Iii) Add a culture solution of lactic acid bacteria in a wet state to the dried algae.

[0018] Among these, (i) is preferred. Further, in (i), using a suspension of raw algal bodies as microalgae and using a culture solution, particularly a seed culture solution as lactic acid bacteria, is a culture of lactic acid bacteria. High performance and flavor production ability! The algae suspension, paste, cake, and lactic acid bacteria culture solution contain water. If the culture solution containing microalgae and lactic acid bacteria is insufficient, add water to You may make it the state maintained underwater. The water is preferably sterilized water.

[0019] The content of the microalgae in the culture solution containing the microalgae and lactic acid bacteria is 0.1 as dry cells from the viewpoint of improving efficiency in the harvesting and drying steps after lactic acid bacterium culture described below. -30% by mass is preferred 1-20% by mass is more preferred.

Cultivation of lactic acid bacteria in a culture solution containing microalgae and lactic acid bacteria may be stationary culture, or may be agitation culture with propeller stirring if the culture solution is liquid. Further, the culture system may be anaerobic or aerobic so as to be suitable for the growth of the lactobacillus used.

[0020] The amount of lactic acid bacteria used may be the number of bacteria in which lactic acid bacteria grow, but from the viewpoint of improving the suppression of the propagation of contaminating bacteria, the number of lactic acid bacteria at the start of cultivation of lactic acid bacteria is the algae in terms of solid content. It is preferred that there are ¹¹ lC lO per class lg, more preferably 10 ⁶ to 10 ¹ G. The pH of the culture solution containing microalgae and lactic acid bacteria is a force that changes during the cultivation process due to acids such as lactic acid produced by culturing lactic acid bacteria. The pH force at the start of culture is preferably 4.0 to 9.0. Mashima 5.0 to 7.0 is more preferable.

The culture temperature may be any temperature as long as the lactic acid bacteria can grow, but 4-45 ° C is preferred because it is suitable for the growth of lactic acid bacteria and the active ingredients of microalgae are not impaired. ° C is more preferred.

[0021] The culture time is sufficient to proliferate lactic acid bacteria and exert an excellent blood pressure lowering effect. For example, 9 to: L00 time is preferred 15 to 72 hours is more preferred 18 to 50 hours is particularly preferred It is preferable.

The number of lactic acid bacteria after culturing exerts an excellent blood pressure lowering effect, is sufficient to suppress the increase of other contaminants, reduces the taste and odor peculiar to microalgae, and exerts a blood pressure lowering effect. It is preferable that the number of lactic acid bacteria at the beginning is 10 to: L000 times.

In addition, in order to maintain the optimum pH for the growth of lactic acid bacteria, a basic compound such as potassium hydroxide or potassium hydroxide may be added during the cultivation to adjust the pH. Milk to produce It is preferable that the pH of the culture solution containing microalgae and lactic acid bacteria is lowered to around 5.0 due to acid or the like because it reduces impurities and exerts a blood pressure lowering effect.

[0022] In the present invention, the addition of sugar to a culture solution containing microalgae and lactic acid bacteria can suppress the growth of contaminating bacteria, and the smell and taste peculiar to microalgae. Is preferable because it can be further reduced. This inhibitory effect on the growth of germs is particularly prominent when the germs are easy to propagate as microalgae / and dried algae are used.

Examples of the saccharide include monosaccharides, oligosaccharides, and polysaccharides. Examples of monosaccharides include glucose, galactose, mannose, fructose, ribose, xylose and the like. Examples of the oligosaccharide include disaccharides such as sucrose and maltose, galatato-oligosaccharide, furato-oligosaccharide, soybean oligosaccharide, xylo-oligosaccharide, and raffinose. Examples of the polysaccharide include amylose, amylopectin, cellulose, dalicogen, j8-glucan, mucopolysaccharide and the like. As the saccharide, an oligosaccharide is preferable, and among them, a galato-oligosaccharide and a xylo-oligosaccharide are preferable.

[0023] There are no particular restrictions on the method of adding saccharides. For example, microalgae, lactic acid bacteria, and saccharides may be mixed, or microalgae added with saccharides in advance and lactic acid bacteria may be mixed. Lactic acid bacteria added with sugars and microalgae may be mixed. The saccharide may be solid, but it is preferable to use a saccharide previously dissolved in water or the like to form an aqueous solution.

The amount of saccharide used is preferably 0.05 to 20% by mass, more preferably 0.1 to 10% by mass, based on the total of the culture solution containing microalgae and lactic acid bacteria and the saccharide.

[0024] Due to the antibacterial action of organic acids such as lactic acid and acetic acid, bacteriocins and the like produced in the process of preparing the lactic acid bacteria culture, other contaminating bacteria are reduced and lactic acid bacteria become the dominant species. Furthermore, flavors such as acetaldehyde and diacetyl produced by lactic acid bacteria reduce the odor and taste peculiar to microalgae, making it easy to use in food and flavor.

[0025] The lactic acid bacteria culture obtained as described above can be used as it is as a lactic acid bacteria beverage or a food supplemented with lactic acid bacteria.

The lactic acid bacteria culture obtained above has a characteristic that it has a component composition different from that of the raw powder of microalgae before cultivation.

For example, regarding the amount of free amino acids in the culture after 24 hours of culture, proline, cystine , Lysine, leucine, isoleucine, γ-aminobutyric acid (hereinafter abbreviated as “GABA”), lysine, histidine, etc., the content per algal body lOOg is 10 times or more compared to that before culturing, and free amino acids The total amount is also more than doubled.

In addition, the amount of protein contained in the bulk powder before culturing treated with a separation membrane with a molecular weight of 10,000 and lactic acid bacteria culture is measured by the BCA (Bicincho ninic Acid) method, which is a general method for measuring protein amount. did. As a result, for example, when Spirulina bulk powder is used as an algal body, the component with a molecular weight of 10,000 or less increases about twice as much after 24 hours of culture compared to before the start of culture, and the high molecular weight protein is low. It was turned into a molecule and found to be a component with excellent absorbability.

Further, 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 culture similar to the present invention. For example, in the above-mentioned (Patent Document 3), when L. lactis YIT 2027 is used alone in milk, it is 15 mg Zl00 mL after 3 days of culture, and mixed culture with L. casei YIT 9029 There is a description that increases to 38 mgZ 1 OOmL.

However, according to the method of the present invention, it is apparent that GABA can be produced at a concentration of lOOmgZlOOmL or more in a culture time of 24 hours, which is extremely excellent as a GABA production method.

[0026] The antihypertensive agent of the present invention comprises the lactic acid bacteria culture obtained above as an active ingredient. The blood pressure lowering agent of the present invention can be prepared in the form of tablets such as naked tablets, film-coated tablets, sugar-coated tablets, enteric tablets, multilayer tablets, granules, powders, liquids and the like. Preparation into these forms may be carried out by suspending, drying, pulverizing, molding, etc. in the lactic acid bacteria culture according to a conventional method according to each form. In preparation for each form, binders, surfactants, thickeners, fillers, disintegrants, excipients and the like that are generally used for the preparation can be used. In addition, a strong lactic acid bacteria culture can be used as it is as a lactic acid bacteria beverage or a lactic acid bacteria additive, or the lactic acid bacteria culture can be added to other foods.

[0027] For example, when the drying treatment is performed, it is usually performed so that the moisture content of the algal bodies is 4 to 7% by mass, but a treatment capable of maintaining the number of lactic acid bacteria is preferable. Preferable drying methods include, for example, freeze-drying method and spray-drying method. A drying method is more preferable. The drying temperature during drying treatment increases the production efficiency as the exhaust air temperature is higher, but the quality of microalgae is good and the number of lactic acid bacteria does not decrease, so the product temperature is 30-70 ° C. It is preferable that the drying treatment is performed in a range of 40 to 60 ° C. In the present invention, the product temperature refers to the sample temperature after drying.

[0028] For preparation into tablets, for example, the powder obtained above may be tableted by a known tableting method.

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

[0029] The intake of the blood pressure lowering agent of the present invention to the patient is preferably determined in consideration of the patient's sex, age, symptom, etc., but is generally 0.2 to : LOg / day, especially 0.5 to 8 g / day is preferred. These may be taken together at one time, or the daily intake may be taken in several parts.

[0030] Next, the blood pressure lowering action of the blood pressure lowering agent of the present invention will be described.

The inventors of the present invention confirmed the effect on hypertension, which is considered to be a major cause of various lifestyle-related diseases, using experimental animals. That is, spontaneously hypertensive rats (SHR strain) were mixed with spirulina lactic acid bacteria culture and chlorella lactic acid bacteria culture in solid feed, and after free intake, blood pressure was measured to confirm blood pressure lowering action. In the experimental group, a group fed with solid feed was set as the target group, and in addition, as a group fed with the test substance, a group of spirulina powder (test substance 1) mixed with the solid feed, spirulina lactic acid bacteria culture The group in which the product (test substance 2) was mixed with the solid feed, the group in which the chlorella bulk powder (test substance 3) was mixed in the solid feed, and the group in which the chlorella lactic acid bacteria culture (test substance 4) was mixed in the solid feed were set. As a result, it is clear that the group fed with the spirulina lactic acid bacteria culture or the chlorella lactic acid bacteria culture has a stronger blood pressure lowering effect than the control group, the spirulina bulk powder, and the group fed with the chlorella bulk powder. (See test example).

[0031] To confirm the safety of lactic acid bacteria cultures, the acute toxicity of mice was examined. As mice, 10 male and 10 female ddY-N mice (body weight 20-26 g) were used. The administration method is that the culture can be finely pulverized and suspended in a CMC 1% solution. A suspension with a concentration of 10% by mass was orally gavaged once. The culture was observed for 1 week after administration. LD for both males and females is over 6, OOOmg / kg, confirming food safety.

50

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 1 Preparation of Lactic Acid Bacteria Culture Using Spirulina (1) and Measurement of Lactic Acid Bacteria Count

A 2500 L culture tank was charged with 25 kg of galactooligosaccharide and 825 kg of tap water. After heat sterilization, 100 kg of spirulina powder was added and inoculated with 50 kg of a seed culture solution of lactic acid bacteria Lactobacillus brevis. Lactic acid bacteria were cultured with aeration propeller stirring at 37 ° C for 72 hours. The culture solution was sampled at culture times of 0, 9, 12, 15, 18, 24, 48, and 72 hours, and the number of lactic acid bacteria was measured.

[0034] The number of lactic acid bacteria was measured by the following procedure. Lactic acid bacteria culture solution 1. Oml was suspended in 19ml of phosphate buffered saline to prepare a suspension. The suspension further with phosphate-buffered saline, 10x, 10 ^twice, 10 ^three times, 10 ⁴ times, 10 ⁵ times, 10 ⁶ times, and sample diluent was diluted to 10 ⁷ times Got. MRS agar medium prepared by adding 15 g of agar to 1000 ml of MRS medium (Merck Cat. No. 10661) is smeared with 0.1 ml of diluent and cultured at 35 ° C for 48 hours. did. Using an agar medium in which 30 to 300 colonies of lactic acid bacteria were confirmed, the number of colonies in the agar medium was measured, and the number obtained by multiplying this number by the dilution factor was taken as the number of lactic acid bacteria.

[0035] (Measurement of amino acid content)

A part of each sampled culture solution was spray-dried and subjected to amino acid analysis. Each 200 mg of Spirulina lactic acid bacteria culture was added to 5 mL of water, suspended, and left for 30 minutes. The supernatant and insoluble material were separated by a centrifuge (3000 rpm, 10 minutes), and 1 mL of the supernatant was collected. To this, 1 mL of 10% (w / v) triclonal acetic acid solution was added and centrifuged. The obtained supernatant was subjected to HPLC analysis by the 0-phthalaldehyde method (post-column detection) to measure the amount of amino acids.

(HPLC conditions) Column: Hitachi # 4619 (4mm X 15cm)

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

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

Detection: Fluorescence detector

Table 1 shows the amounts of L-glutamic acid, GABA, and lactic acid bacteria in the culture at each culture time.

[table 1]

(Example 2) Preparation of lactic acid bacteria culture using Spirulina (2)

In this example, lactic acid bacteria were cultured in the same manner as in Example 1 except that a 50% (w / w) lactic acid solution was used to maintain the pH at 5.0 and the culture temperature at 30 ° C. The culture solution was sampled at culture times of 0, 9, 12, 15, 18, 21, and 24 hours, and the amount of GABA (mg / lOOg) was measured.

(Example 3) Preparation of lactic acid bacteria culture using Spirulina (3)

In this example, GABA amount (mg / lOOg) was measured in the same manner as in Example 2 except that xylo-oligosaccharide was used instead of galatato-oligosaccharide.

A comparison of GABA amounts (mg / lOOg) between Example 1 and Example 2 up to a culture time of 24 hours is shown in FIG. 1, and the results of Example 2 and Example 3 are shown in Table 2.

From Examples 1 to 3, it was possible to produce more GABA in a short time by appropriately selecting pH and culture temperature.

[Table 2] GABA amount (mg / l OO g)

(Example 4) Preparation of lactic acid bacteria culture using chlorella and measurement of the number of lactic acid bacteria

In Example 1, except that chlorella was used instead of Spirulina, a lactic acid bacteria culture was prepared in the same manner as in Example 1, and the number of lactic acid bacteria and the amount of amino acids were measured. Table 3 shows the amounts of L-glutamic acid, GABA, and lactic acid bacteria in the culture at each culture time.

[0038] [Table 3]

(Test example) Confirmation of blood pressure lowering effect of the blood pressure lowering agent of the present invention

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 conducted.

(Test substance)

Target group: Solid feed MF (Made by Oriental Yeast Co., Ltd.)

Test substance 1: Spirulina bulk powder

Test substance 2: Lactic acid fermented spirulina (Lactic acid bacteria culture obtained in Example 1) Test substance 3: Chlorella bulk powder

Test substance 4: Lactic acid fermentation chlorella (Lactic acid bacteria culture obtained in Example 4) (Test substance preparation and administration) Preparation method: Solid feed MF was mixed with 15% each of test substances 1-2.

Route of administration: Oral

Administration method: Free intake

Dosing volume: 15% of feed

Administration period: 8 weeks

Test animal

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

Sex, age at import: Male, 6 weeks old

Acclimatization period: About 1 week after introduction, the animals were bred for acclimatization, and the general condition during that period was observed (blood pressure measurement method)

Blood pressure was measured by the tail-cuff method, which measures blood pressure in the tail artery noninvasively. The measurement was performed before the test substance administration (week 7) and every week after administration. The blood pressure was measured at 8 points in total, 3 times each, and the average value was taken as the measurement value. Prior to the test, it was confirmed that the average value of the maximum blood pressure of the test animals was ^{160 mmHg} or more on average for each animal.

The results are shown in Table 4.

[Table 4] Test substance 0 weeks 2 weeks 4 weeks 6 weeks 8 weeks later Target group 1 6 5 1 70 1 9 5 2 1 5 2 2 0 Test substance 1 6 5 1 7 0 1 9 0 2 0 0 2 0 5

1

Test substance 1 6 5 1 6 5 1 8 0 1 8 5 1 9 5

2

Test substance 1 7 0 1 7 5 1 9 0 2 1 0 2 1 0

Three

Test substance 1 6 5 1 7 0 1 8 5 1 9 0 2 0 0

Four Table 5 and Fig. 2 show the results of evaluating the blood pressure increase rate at week X based on blood pressure at week 0 using the following formula. Rate of increase in blood pressure (%) = (blood pressure at week X, blood pressure at week 0) I (blood pressure at week 0) X 100 (%) [¾5]

[0042] As is clear from this test example, the spirulina lactic acid bacteria culture and the chlorella lactic acid bacteria culture were confirmed to have an effect of suppressing an increase in blood pressure as compared with the spirulina bulk powder and the chlorella bulk powder before culture, respectively.

For example, after 6 weeks, the blood pressure increase of about 30% is observed in the subject group, whereas in spirulina and chlorella, the increase rate is suppressed to 20 to 25%. In the lactic acid fermented product of Chlorella and chlorella, it is apparent that the lactic acid fermented product according to the present invention is suppressed to about 12 to 15%. Industrial applicability

[0043] The present invention can be used in fields such as the pharmaceutical industry, health food, and food.

Claims

The scope of the claims

[I] An antihypertensive 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.

[2] The blood pressure lowering agent according to claim 1, wherein the microalga is spirulina, chlorella, or a mixture of spirulina and chlorella.

[3] The blood pressure lowering agent according to claim 1, wherein the lactic acid bacterium is a lactic acid bacterium belonging to the genus Lactobacillus.

[4] The blood pressure lowering agent according to claim 1, wherein the lactic acid bacterium is a lactic acid bacterium belonging to the genus Ratatococcus.

5. The blood pressure lowering agent according to claim 1, wherein the lactic acid bacterium is a lactic acid bacterium belonging to the genus Enterococcus.

6. The blood pressure lowering agent according to claim 1, wherein the lactic acid bacteria are cultured for 15 to 72 hours.

[7] Lactobacillus number at the start of cultivation of lactic acid bacteria is a ^¹ X ΙΟ ^ ΙΟ 1 1 per solid equivalent microalgae lg, lactic acid bacteria 10 to the number of lactic acid bacteria at the initiation of culture: in until L000 times, cultured 2. The blood pressure lowering agent according to claim 1, wherein

8. The blood pressure lowering agent according to claim 1, wherein the culture solution further contains a saccharide.

9. The blood pressure lowering agent according to claim 8, wherein the saccharide is galata oligosaccharide.

10. The blood pressure lowering agent according to claim 8, wherein the saccharide is xylo-oligosaccharide.

[II] A method for producing a blood pressure-lowering agent by performing a step of culturing lactic acid bacteria in a culture solution containing microalgae and lactic acid bacteria, and using the lactic acid bacteria culture obtained by this step as an active ingredient.

[12] The blood pressure-lowering agent according to claim 1, wherein the content of microalgae contained in the culture solution is 0.1 to 30% by mass.

[13] Lactobacillus power Lactobacillus delbruecki, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus brevis, Ratatococcus lacteis, Leucostock sp, Group power of at least one selected from two or more kinds The blood pressure lowering agent according to claim 1.

[14] Use of the blood pressure lowering agent according to claim 1 for the treatment of hypertension. [15] A method for treating hypertension using the blood pressure lowering agent according to claim 1.