JP6145352B2 - Antihypertensive agent of fermented makonbu using Bacillus natto - Google Patents

Description

The present invention relates to an antihypertensive action comprising a fermented makonbu product using Bacillus subtilis NATTO as an active ingredient. The present invention relates to a blood pressure increase-suppressing functional food or food material, or an internal preparation for pharmaceuticals (including animal therapeutics) given to humans or animals other than humans, for foods and drinks, and for feeds or pet foods. Means a thing.

Hypertension is one of lifestyle-related diseases. There are two types of hypertension: secondary hypertension whose cause is known and essential hypertension whose cause is unknown. About 90% of hypertensive patients in Japan have essential hypertension. Blood pressure is regulated by various factors such as cardiac output, peripheral vascular resistance, and the renin-angiotensin system plays a major role in controlling blood pressure. Angiotensinogen is synthesized in the liver, and angiotensinogen is converted to angiotensin I by renin synthesized in the kidney. Furthermore, it is converted into angiotensin II by angiotensin converting enzyme (ACE). By binding angiotensin II to the angiotensin II receptor, vascular smooth muscle contracts, resulting in an increase in blood pressure. Hypertension has no subjective symptoms and is often left unattended, which can cause arteriosclerosis, heart disease, and stroke.

Regarding the above-mentioned hypertension, various medical products for solving this problem have been proposed and many are already on the market. Therapeutic agents are diuretics that suppress the reabsorption of sodium and water in the kidneys and reduce the blood circulation to lower blood pressure, block the vasoconstrictive action of sympathetic alpha receptors and dilate blood vessels to lower blood pressure. α-blocker, β-blocker that lowers blood pressure by blocking the effects of sympathetic β-receptor enhancement of cardiac contraction and production of pressor hormones, suppresses calcium ion uptake into cells and dilates blood vessels Calcium antagonist that lowers blood pressure, ACE inhibitor that decreases the production of angiotensin II, a powerful pressor hormone, and angiotensin that lowers blood pressure by inhibiting the action of the receptor that angiotensin II, a pressor hormone acts on II receptor inhibitors (ARBs are used, but these synthetic drugs are not easily available because they require prescriptions. Various side effects may accompany the administration or administration of the preparation, and therefore, anti-hypertensive agents used for the treatment of hypertension are required to be easily obtained and have a natural product origin with as few side effects as possible. Antihypertensive agents such as glue, bonito, seaweed, etc. that inhibit ACE, parasympathetic nerves are stimulated, vascular smooth muscle is relaxed, blood pressure is lowered, and noradrenaline secretion is suppressed. Specific health foods containing γ-aminobutyric acid exhibiting a blood pressure lowering action are known, and these have been approved as foods for specific health.

On the other hand, the seaweed mambu is known as a seaweed belonging to the genus Kombu in the order of brown algae, and is a thick and high quality kelp among the kombu. This seaweed is a very familiar seaweed produced mainly in Hokkaido's southern Hokkaido region, with a large amount of aquaculture. It has been reported that a component extracted from macomb has a blood glucose level increase inhibitory effect and an antioxidant effect. Moreover, it is known that the chemical | medical agent containing the component water-extracted from specific brown algae and red algae, such as a macomb, reduces drowning by the bacterial infection in the fish farm.

An object of the present invention is to provide a functional food or food material for prevention and improvement, which can be easily obtained and ingested in daily eating habits, and can suppress an increase in blood pressure, or a feed material, and feed.

In order to achieve the above-mentioned object, the present inventor has paid attention to the functional components of Macombu and the functionality of Bacillus natto, and as a result of intensive studies, the fermented Macombib 10 mM Tris-HCl buffer using Bacillus natto (PH 8.5) It has been found that the extract has an antihypertensive effect.

The present invention has been completed based on such findings. That is, the antihypertensive action of the present invention is characterized in that the fermented makonbu is an active ingredient.

Although it is still unclear what kind of compound the substance having an inhibitory effect on blood pressure increase contained in the fermented macomb using Bacillus natto is, the 10 mM Tris-HCl buffer (pH 8.5) is used. ) It has been experimentally confirmed that the components eluted by extraction exert a blood pressure increase inhibitory effect. Therefore, the fermented makonbu using Bacillus natto exhibits prevention and improvement as it is as a constituent component of the blood pressure increase inhibitory action of the present invention.

That is, the present invention provides the following.

[1] It contains a fermented makonbu using natto bacteria as an active ingredient.

[2] An antihypertensive agent comprising, as an active ingredient, a composition containing a fermented makonbu product using Bacillus natto.

[3] A functional food comprising the antihypertensive agent according to [1] or [2].

[4] A food material comprising the antihypertensive agent according to [1] or [2].

[5] A pet food comprising the antihypertensive agent according to [1] or [2].

[6] A food and drink composition containing the blood pressure elevation inhibitor according to [1] to [4].

[7] A pharmaceutical composition comprising the antihypertensive agent according to [1] to [4].

The antihypertensive agent of the present invention or a composition containing the antihypertensive agent suppresses the activity of ACE that converts angiotensin I into angiotensin II having an antihypertensive action in the body, and therefore an antihypertensive action can be expected. Therefore, it is useful for suppressing blood pressure increase. Furthermore, since it is possible to produce the agent or composition of the present invention from ingredients with food experience, it is safe to take.

The antihypertensive agent of the present specification is an agent and a composition having an ACE harmful action containing a fermented makonbu product using Bacillus natto as an active ingredient. Although content of said compound is not limited, it should just be contained in the range which can exhibit the blood pressure rise inhibitory effect.

The composition for eating and drinking of the present invention is a composition containing the above-mentioned blood pressure increase inhibitor, and these are mixed with general foods. Further, capsules, tablets, granules and the like that are easy to take may be used by using appropriate carriers and auxiliaries as known foods. The food and drink here refers to, for example, general foods, health functional foods (special health foods, nutritional functional foods), health foods, nutritional supplements, pet foods, and the like. General food as used herein includes, but is not limited to, beverages, dairy products, fermented milk, lactic acid bacteria beverages, processed milk, coffee beverages, juices, ice creams, strawberries, biscuits, wafers, jellies, soups, and noodles. is not. Preferably, beverages, dairy products, processed milk, fermented milk, lactic acid bacteria beverages, wafers, and jelly are included.

The pharmaceutical composition of the present invention is a composition containing the above antihypertensive agent, may be the above agent itself, or a composition containing a pharmaceutically acceptable carrier as desired. Also good. The use is not limited, and examples thereof include readily available drugs such as over-the-counter drugs (OTC) or quasi drugs. The form of the pharmaceutical composition is not limited, and examples thereof include pills, solutions, powders, granules, tablets, capsule tablets, troches, syrups, and dry syrups. Preferably, capsules, liquids, elixirs, tablets, cachets, suppositories and the like are used. A pharmaceutically acceptable carrier is any material suitable for oral, enteral, transdermal, and subcutaneous administration, such as water, gelatin, gum arabic, lactose, microcrystalline cellulose, starch. Sodium starch glycolate, calcium hydrogen phosphate, magnesium stearate, talc, colloidal silicon dioxide, and the like.

The fermented makonbu using natto and the composition are blended as an active ingredient of an antihypertensive agent, but when formulated as an antihypertensive agent, the dosage form is powder, granule, tablet Any dosage form can be employed. At that time, in order to facilitate storage and handling, carriers such as dextrin and cyclodextrin, preservatives and other optional auxiliaries can be blended as necessary. Moreover, the form is not limited, The composition for food and drink and the composition for pharmaceuticals are included. Although these content is not limited, it should just be contained in the range which can exhibit an effect with respect to a blood pressure rise inhibitory effect.

The antihypertensive agent inhibits the activity of ACE so that angiotensin II is not generated.

The functional food or food material for preventing or improving blood pressure elevation according to the present invention, which contains a fermented makonbu product using Bacillus natto as an active ingredient, is a part of the raw material for foods and beverages for preventing or improving blood pressure elevation according to the present invention. Or can be obtained by adding and blending after the production process or production. Such functional foods are not particularly limited, and specific examples include seasonings, sprinkles, confectionery, snacks, noodles, fish paste products, dairy products, beverages, and the like.

Examples are given below to describe the present invention in more detail, but the present invention is not limited by these.

The sample was sterilized by autoclaving after adding 2.5 g of macomb powder (produced in Hokkaido, Hokkaido) pulverized to about 500 μm to 50 ml of distilled water and adjusting the pH to 8.0. 500 μl of the Bacillus natto preculture was inoculated and cultured at 30 ° C. and 160 rpm. After culturing for 5 days, lyophilization was carried out, 100 ml of 10 mM Tris-HCl buffer (pH 8.5) was added to 1 g of the macomb fermented product, shaken at 50 ° C. for 1 hour, and extracted with warm water as a sample. .

Examination of ACE activity inhibitory action of fermented macabon using Bacillus natto The influence of fermented macomb using natto bacillus on ACE activity was examined using an enzyme reaction system in a test tube. 10 μl of sample solution of each concentration diluted with distilled water was added to make a total volume of 130 μl / reaction well.

This test used ACE Kit-WST of Dojindo Laboratories. The solutions (A to C solutions) used for the reaction in this test were prepared as follows. A liquid is Substrate buffer attached to the kit. For solution B, Enzyme B attached to the kit was dissolved in 2 ml of distilled water, and 1.5 ml of the dissolved enzyme solution was dissolved in Enzyme A attached to the kit. The solution C was obtained by dissolving Enzyme C attached to the kit with 3 ml of distilled water. Similarly, Coenzyme included in the kit was dissolved in 3 ml of distilled water. Each 2.8 ml of dissolved Enzyme C and Coenzyme was added to the Indicator solution included in the kit.

In the ACE reaction in the plate, 10 μl of sample diluted to each concentration, 10 μl of solution A and 10 μl of solution B were mixed and preincubated at 37 ° C. for 1 hour. Next, 100 μl of solution C was added and incubated at room temperature for 10 minutes. Thereafter, the ACE inhibitory activity was evaluated by measuring the absorbance at 450 nm. As a control, 10 mM Tris-HCl buffer (pH 8.5) equivalent to each sample was added instead of the sample. In each test group, a blank in which solution A was added instead of solution B was set.
The inhibition rate was calculated from the following calculation formula. Inhibition rate = 1- (absorbance of sample-absorbance of sample blank) / (absorbance of control-absorbance of control blank). The relative ACE inhibition rate in relation to the solvent and concentration is shown in Table 2 with respect to 100 as a control.

As is clear from FIG. 1, ACE activity inhibition was observed in the fermented macomb using Bacillus natto. The IC ₅₀ value for inhibition was 70 μg / ml. Therefore, it was demonstrated that the fermented macomb using Bacillus natto is effective in inhibiting ACE activity.

Study on suppression of blood pressure increase in fermented macomb using Bacillus natto The effect of fermented macomb on ACE using Bacillus natto was examined using the tail-cuff method in animal experiments.

In this test, the hot water extract was dried under reduced pressure and used as a sample. In this test, the solutions (A to B solutions) were prepared as follows. As the liquid A, 300 mg of a sample was dissolved in 2 ml of distilled water. For solution B, 4.5 mg of angiotensin I (Wako Pure Chemical Industries, Ltd.) was dissolved in 10 ml of distilled water.

  A ddY mouse, male, 5 weeks old (Japan SLC) was preliminarily raised in a breeding room (25 ° C. ± 2) in a solid feed (MR stock, Nippon Nosan Kogyo) for 1 week, and then divided into 8 groups per group. . The mice were orally administered with 0.4 ml of distilled water or solution A, fitted with an intraperitoneal cannula, and then placed in a measurement cage. Thirty minutes after administration, wear a cuff and use an unheated non-invasive sphygmomanometer for mice and rats (BLOOD PRESURE MONITOR FOR MICE & RAT Model MK-2000, Muromachi Kikai) to automatically control the caudal artery blood pressure 6 times The average value was measured as the systolic blood pressure before angiotensin I administration. 100 μl of solution B was administered using an intraperitoneal cannula. Two minutes after the administration, the blood pressure was measured in the same manner as the systolic blood pressure after angiotensin I administration. The difference in systolic blood pressure before and after angiotensin I administration was taken as the blood pressure increase value.

As apparent from FIG. 2 and FIG. 3, the ACE inhibitory activity was recognized in the animal body in the fermented macomb using natto. Therefore, it was demonstrated that the fermented macomb using Bacillus natto is effective in inhibiting ACE activity.

It is a graph which shows the effect by the macomb fermented material using Bacillus natto, and showed the relative ACE inhibition rate which makes control 100. It is a graph which shows the effect by the macomb fermented material using Bacillus natto, and showed systolic blood pressure. It is a graph which shows the effect by the macomb fermented material using natto bacteria, and showed a blood pressure rise value.

Claims (3)

A composition for suppressing an increase in blood pressure , which comprises a fermented macomb using natto bacteria as an active ingredient . The composition for suppressing an increase in blood pressure according to claim 1, comprising an extract of the fermented macomb as an active ingredient. The composition for suppressing an increase in blood pressure according to claim 1 or 2, which is used in the form of a functional food, food material, pet food, food or drink, or pharmaceutical.

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