JP2014181214A - Dopamine production promoter - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a novel dopamine production promoter.SOLUTION: A dopamine production promoter comprises miso extracts as an active ingredient.

Description

  The present invention relates to a dopamine production promoter.

  Miso, the representative of Japanese food culture, was originally a luxury and valuable food that was valued by temples and aristocratic classes, and was used as a side dish and medicine rather than being cooked as miso soup. Since the samurai's dietary habit of “one soup one dish” was established during the Kamakura period and it became popular to eat in the form of miso soup, it is now an indispensable item for the Japanese diet.

  The main ingredient of miso is soybean, which is rich in nutrients by mixing koji and salt and fermenting it, soy protein is easy to digest, and amino acids that are the source of umami are released in large quantities. In addition, many nutrients such as saponins, isoflavones, enzymes, vitamins, potassium, and fiber are included, and effects on health maintenance and prevention of lifestyle-related diseases are expected. On the other hand, there is a disagreement that the health effects of miso have been underestimated due to concerns about the salt contained. Miso contains many nutrients in addition to salt, and is expected to suppress salt absorption in the gastrointestinal tract, dilate blood vessels, protect endothelial cells, and protect blood vessels and organs beyond the effects of salt. It is considered high. However, the health benefits of miso have often been inferred from the constituent nutrients of miso so far, and long-term ingestion of miso exerts a protective effect on vascular disorders that are actually seen in hypertension, and contributes to brain, heart and kidney function. It is not always clear whether it is useful or not.

  Under such circumstances, it has been reported by the present inventor that long-term miso intake attenuates salt-sensitive hypertension in male Dahl rats and exhibits an organ protecting action (Non-patent Document 1).

Uehara Y et al., Nutrition 2012; 28: 924-931.

  An object of the present invention is to provide a novel dopamine production promoter.

In order to solve the above-described problems, the present invention provides a dopamine production promoter containing a miso extract as an active ingredient.
A preferred embodiment of the dopamine production promoter of the present invention is a dopamine production promoter for promoting dopamine production in the kidney.

  According to the present invention, a dopamine production promoter containing a miso extract as an active ingredient is provided as a novel dopamine production promoter.

FIG. 1 is a graph showing cumulative salt intake (g) in each experimental group (control group, 0.5% DS group, MISO group). FIG. 2 is a diagram showing changes in blood pressure (mmHg) in each experimental group (control group, 0.5% DS group, MISO group). FIG. 3A is a diagram showing urinary adrenaline excretion (μg / day) in each experimental group (control group, 0.5% DS group, MISO group). FIG. 3B is a graph showing urinary noradrenaline excretion (μg / day) in each experimental group (control group, 0.5% DS group, MISO group). FIG. 3C is a diagram showing urinary dopamine excretion (μg / day) in each experimental group (control group, 0.5% DS group, MISO group). FIG. 4 is a diagram showing creatinine clearance (mL / min) in each experimental group (control group, 0.5% DS group, MISO group). FIG. 5 is a diagram showing free water reabsorption (mL / min) in each experimental group (control group, 0.5% DS group, MISO group). FIG. 6 is a diagram showing FENa (%) in each experimental group (control group, 0.5% DS group, MISO group).

Hereinafter, the present invention will be described in detail.
The dopamine production promoter of the present invention contains a miso extract as an active ingredient. Here, the “miso extract” includes any form of a miso extract, a diluted solution of the miso extract, a concentrated solution or a dried product, or a purified product thereof.

  The miso used as an extraction raw material when obtaining the miso extract is not particularly limited as long as it is an edible miso. As miso, for example, rice miso (soybeans (excluding defatted soybeans; the same applies hereinafter) cooked in rice and cultured with koji mold (hereinafter referred to as “rice koji”) is added. Salt mixed), barley miso (soybean soup, barley or bare wheat steamed and koji mold cultured (hereinafter referred to as “wheat cake”) mixed with salt), Bean miso (soybean boiled and cultured with koji mold (hereinafter referred to as “bean koji”) mixed with salt), mixed miso (rice miso, wheat miso or bean miso mixed, rice koji and wheat Rice miso, wheat miso, and beans miso, etc., etc. (classified based on raw materials), sweet miso, dry miso (classified based on taste), red Miso, light-colored miso, white miso (classified based on color) Of these, US miso are preferred, more preferably dry miso of rice miso. Miso can be used as an extraction raw material alone or in combination of two or more.

The miso used as an extraction raw material when obtaining the miso extract may be either a homemade miso brewed according to a conventional method or a commercially available miso.
A general method for brewing miso is a koji-making process in which various koji such as rice koji, wheat koji, and soybean cake are obtained by adding koji fungus to rice or wheat and other cereals or beans cooked and cultivated, and soybean Steamed rice (If necessary, you may add rice or wheat steamed grains or beans), various rice cakes obtained in the koji making process, salt, water, other materials in the desired ratio And a fermentation process for obtaining a brewed product by fermenting and aging the koji mixed material.

  Examples of cereals used for brewing miso include rice, wheat (for example, barley, bare wheat, hard wheat, rye, wheat, etc.), corn, buckwheat, millet, millet, and the like. Examples of beans include soybeans, mung beans, peas, black beans, red beans, broad beans, and the like. There are no particular limitations on the varieties and production areas of cereals and beans.

  The koji mold added and cultured to obtain various koji is not particularly limited as long as it is a koji mold generally used for brewing miso. Examples of Aspergillus include Aspergillus oryzae, Aspergillus sojae, Aspergillus tamari and the like. The koji mold may be added to steamed cereals and / or beans in the form of seed pods. The temperature at which the koji mold is added and cultured depends on the optimum growth temperature of the koji mold used, but is usually 20 to 45 ° C., preferably 25 to 40 ° C.

  The salt used for brewing miso is not particularly limited as long as it is an edible salt, and examples thereof include salt, special grade salt, rock salt, common salt, and white salt.

  Examples of other materials used for brewing miso include salt-tolerant yeast and salt-tolerant lactic acid bacteria, and these may be further added and mixed in the preparation step to prepare a koji-mixed material. . Examples of the salt-tolerant yeast include Zygosaccharomyces rouxii, Candida versatilis, Candida etchelsi, and the like, Examples thereof include Tetragenococcus halophilus and Pediococcus acidilactici.

  Various materials (cereals, beans, salt, other materials, etc.) used for brewing miso can be blended in proportions appropriately adjusted according to the type of miso to be brewed. For example, the blending ratio of koji is usually 50% to 20%, preferably 50%. The compounding ratio of the salt is usually 5% by weight or more, preferably 8 to 13% by weight. The amount of water contained in the soot mixed material is usually adjusted to a range of 35 to 55% by weight in consideration of the amount of water contained in all materials.

  A brewed product, that is, miso is brewed by fermenting and aging the koji mixed material obtained in the preparation process in the fermentation and aging process. The fermentation / ripening step is usually performed at 10 to 80 ° C, preferably 25 to 35 ° C or 45 to 60 ° C. You may stir suitably as needed in the middle of a fermentation / ripening process. The period of the fermentation / ripening step depends on the ripening temperature, but is usually about 1 month to 1 year at 25 to 35 ° C. and usually about 1 to 7 days at 45 to 55 ° C.

  The solvent used as the extraction solvent when obtaining the miso extract is preferably a polar solvent. Examples of the polar solvent include water, a polar organic solvent, and a mixed solution thereof, and water is preferable. Examples of water include tap water, deionized water, and distilled water. Examples of polar organic solvents include monohydric alcohols, polyhydric alcohols, and ketones. Examples of the monohydric alcohol include lower fats having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, isopropyl alcohol, butyl alcohol, isobutyl alcohol, sec-butyl alcohol, tert-butyl alcohol, pentyl alcohol, and isopentyl alcohol. Examples of the polyhydric alcohol include C2-C5 polyhydric alcohols such as ethylene glycol, propylene glycol, 1,3 butylene glycol, and glycerin. Examples of the ketone include acetone. , Lower aliphatic ketones such as methyl ethyl ketone and diethyl ketone. These solvents are usually used as extraction solvents at room temperature or at temperatures below the boiling point of the solvent. When using the liquid mixture of 2 or more types of solvents as an extraction solvent, the mixing ratio can be adjusted suitably.

  The extraction method for obtaining the miso extract is not particularly limited as long as the miso component soluble in the polar solvent can be eluted, and a normal extraction method used when separating the component from the food may be used. it can. For example, the extraction raw material is put into a treatment tank filled with an extraction solvent, and after elution of soluble components with occasional stirring as necessary, solids are removed by standing, filtration or centrifugation to obtain an extract. be able to. The amount of extraction solvent is usually 5 to 30 times (weight ratio), preferably 10 to 25 times (weight ratio) of the extraction raw material. When water is used as the extraction solvent, the extraction temperature is usually 4 to 40 ° C, preferably 15 to 35 ° C. Centrifugation conditions are usually 5000-15000 g, preferably 8000-12000 g, and usually 5-15 minutes, preferably 8-12 minutes.

  When the solvent is distilled off from the obtained extract, a paste-like concentrate is obtained, and when this concentrate is further dried, a solid extract is obtained. The miso extract may be purified by a method such as activated carbon treatment, adsorption resin treatment, ion exchange resin, liquid-liquid counter-current distribution, etc., as long as the physiological activity is not lowered.

  Miso extract has a dopamine production promoting effect. Although the details of the active ingredient have not been clarified at present, it is clear from Examples described later that the miso extract has a dopamine production promoting action.

  Miso extract can be used as it is as a dopamine production promoter, but it can be used as a tablet, capsule, powder, granule, syrup, troche using pharmaceutically acceptable carriers and other auxiliaries. It can also be used by formulating it into a desired dosage form such as an agent, a drink, an injection, a drip, a suppository. Examples of the auxiliary agent used for the formulation include excipients, binders, disintegrants, lubricants, stabilizers, extenders, diluents, flavoring and flavoring agents, and the like.

  The administration route of the dopamine production promoter of the present invention is not particularly limited, but is preferably oral administration. The dose and frequency of administration of the dopamine production promoter of the present invention can be appropriately adjusted according to the administration subject, administration route and the like. The miso extract, which is an active ingredient of the dopamine production promoter of the present invention, is non-toxic to the human body, so there is no particular limitation on the amount of intake thereof. In terms of the amount of the product, it can be administered about 1 to 3 times a day so that the daily dose is 8 to 24 g / day.

  The dopamine production promoter of the present invention can promote dopamine production in vivo (particularly the kidney). Here, “promoting dopamine production” means promoting dopamine production with a statistically significant difference compared to the control. The dopamine production promoter of the present invention preferably promotes the dopamine production amount to 120% or more, more preferably 1000% or more, more preferably 1200% or more, when the control dopamine production amount is 100%. It is even more preferable to promote it. Examples of the method for measuring the dopamine production amount used when comparing the dopamine production amount include HPLC method, microdialysis method, polarography method and the like.

  The dopamine production promoter of the present invention can enhance the physiological activity of dopamine by promoting dopamine production in vivo. The physiological activity of dopamine includes, for example, water / natriuretic action, vasodilatory action, etc. By enhancing the physiological activity of dopamine, for example, blood pressure decrease in hypertension, prevention or treatment of arteriosclerosis, brain damage, cardiac It is possible to prevent disorders and kidney disorders. For example, the dopamine production promoter of the present invention can cause water / Na diuresis by promoting dopamine production in the kidney, and can lower blood pressure through an increase in free water clearance and a decrease in the amount of fluid. . In addition, it is thought that the direct vasodilatory effect of dopamine is also concerned in the fall of blood pressure.

1. Experimental Method (1) Equipment In this experiment, the following equipment was used.
・ Homogenizer Kinematica CH-6010 (manufactured by POLYTRON)
・ High-pressure steam sterilizer HV HV-25 (manufactured by HIRAYAMA)
・ Stirred Ultrafiltration Cells-8050 (MILLIPORE)
・ Sphygmomanometer MANOMETER-TACHOMETER KN-210-1 (manufactured by NATSUME)
・ Small high-speed cooling centrifuge MX-30 (made by TOMY)

(2) Oral administration experiment 20 6-week-old female Dahl salt-sensitive rats (purchased from Sankyo Lab Service Co., Ltd.)
(A) Tap water administration group (hereinafter referred to as “control group”),
(B) 0.5% saline administration group (hereinafter referred to as “0.5% DS group”),
(C) 4% miso aqueous solution (corresponding to 0.5% saline) administration group (hereinafter referred to as “MISO group”)
The animals were reared for 12 weeks under constant temperature and humidity (23 ± 2 ° C.) and automatic lighting (8:00 am to 8:00 pm).

Rats of any experimental group were allowed to freely feed the solid feed CE-7 (Japan Clare Co., Ltd.) for long-term breeding.
The 0.5% saline was prepared by dissolving sodium chloride (Wako Pure Chemical Industries, Ltd.) in water and freely ingested by the 0.5% DS group.

  A 4% miso aqueous solution was prepared by dissolving 40 g of miso with 1000 mL of tap water at about 25 ° C., pulverizing with a homogenizer, and applying to an autoclave, and the MISO group freely ingested. As the miso, commercially available dry rice miso (boiled rice paste: 5 steps, salt content: 12.5%, raw materials: rice (domestic), salt (domestic), soybean (produced in North America)) was used.

The water in the control group and the saline in the 0.5% DS group were changed once every two days, and the aqueous miso solution in the MISO group was changed daily.
The intake of tap water, saline or miso aqueous solution (hereinafter referred to as “drinking water”) was measured every 2 days, and blood pressure and body weight were measured every 2 weeks. At the 12th week, Dahl rats were placed in a metabolic cage, and after a one-day habituation period, the amount of drinking water, food intake and urine volume for 24 hours were measured.

(3) Measuring method <blood pressure>
After warming the rat, blood pressure was measured by tail-cuff method using MANOMETER-TACHOMETER KN-210-1 (manufactured by NATSUME). The blood pressure measurement value was measured three times, and the average value was adopted.

<Urine catecholamine excretion>
Urinary adrenaline excretion, urinary noradrenaline excretion and urinary dopamine excretion were measured by HPLC method (Ushio et al .: Clinical Laboratory Instruments and Reagents 11-4-635-641 1988, Seiichi Oishi et al .: Japanese Clinical 53-Increase- 507-510 1995) was entrusted to SRL.

<Creatinine Clearance (Ccr)>
At 12 weeks, Dahl rats were placed in a metabolic cage, and after a period of one day of habituation, urine excreted for 24 hours was collected, and blood was collected from Dahl rats under pentobarbital anesthesia. From the blood and urine creatinine values, creatinine clearance was calculated using the following formula.
Ccr = Ucr × V / Scr
[Where Ccr is creatinine clearance (mL / min), Scr is serum creatinine concentration (mg / dL), Ucr is urine creatinine concentration (mg / dL), and V is urine volume (mL / min) . ]
The serum method and urine creatine concentration were outsourced to SRL, Inc. for enzymatic measurement.

<Free water reabsorption>
The free water reabsorption amount was calculated from the following equation.
TCH ₂ O = (Uosm−V) / Posm−V
[Wherein, TCH ₂ O is the amount of free water reabsorption (mL / min), Uosm is urine osmotic pressure (mOsm / KgH ₂ O), Posm is plasma osmotic pressure (mOsm / KgH ₂ O), V is urine for 1 minute Represents volume (mL / min). ]
About urine osmotic pressure and plasma osmotic pressure, the measurement by the freezing point depression method was entrusted to SRL.

<FENa (Fractional Excretion of Sodium)>
FENa (urinary sodium excretion rate) was calculated using the following formula.
FENa (%) = (U _Na × S _cr ) / (U _cr × S _Na ) × 100
[In the formula, U _Na is urinary Na concentration (mEQ / L), S _Na is serum Na concentration (mEQ / L), S _cr is serum creatinine concentration (mg / dL), U _cr is urine creatinine concentration (Mg / dL).
The urinary Na concentration and serum Na concentration were measured by the electrode method in the Kyoritsu Women's University Clinical Nutrition Laboratory. For serum and urine creatinine concentrations, enzymic method measurement was outsourced to SRL.

(4) Statistical analysis Data were expressed as mean ± standard deviation (SD). Tested by analysis of variance, Post-hoc (LSD) using STATISTICA. Two-sided test determined p <0.05 as significant.

2. Results (1) Oral administration experiment <Total salt intake>
The cumulative salt intake (g) in each experimental group (control group, 0.5% DS group, MISO group) is shown in FIG. The salt intake was calculated from the amount of water consumed. As shown in FIG. 1, the salt intake increased in a time-dependent manner in the MISO group and the 0.5% DS group, and the MISO group showed a significantly higher value than the 0.5% DS group.

<Blood pressure>
FIG. 2 shows changes in blood pressure (mmHg) in each experimental group (control group, 0.5% DS group, MISO group). As shown in FIG. 2, the time-dependent blood pressure increase was observed in the 0.5% DS group as compared with the control group. On the other hand, the increase in blood pressure was significantly suppressed in the MISO group compared to the 0.5% DS group (179 ± 2.966 mmHg), and decreased to 170 ± 3.068 mmHg at the end of the experiment (p <0.05).

<Urine catecholamine excretion>
Urinary adrenaline excretion (μg / day), urinary noradrenaline excretion (μg / day) and urinary dopamine excretion (μg / day) in each experimental group (control group, 0.5% DS group, MISO group) Are shown in FIGS. 3A, 3B and 3C, respectively.
As shown in FIGS. 3A and 3B, urinary excretion of adrenaline and noradrenaline was significantly higher in the MISO group than in the 0.5% DS group (0.231 ± 0.082 μg / day vs. 0.016 ± 0.009 μg / day, p <0.05; 1.288 ± 0.378 vs 0.157 ± 0.144 μg / day, p <0.05).
As shown in FIG. 3C, urinary excretion of dopamine was significantly higher in the MISO group than in the 0.5% DS group (11.686 ± 5.285 vs 1.325 ± 0.996 μg). / Day, p <0.05).

<Creatinine Clearance (CCr)>
FIG. 4 shows the creatinine clearance (mL / min) in each experimental group (control group, 0.5% DS group, MISO group). As shown in FIG. 4, creatinine clearance was significantly higher at 0.5% DS compared to the control group (0.865 ± 0.208 vs 1.412 ± 0.426 mL / min, p < In addition, the MISO group showed a significantly higher value compared to 0.5% DS (1.798 ± 0.365 mL / min, p <0.05).

<Free water reabsorption>
FIG. 5 shows the free water reabsorption (mL / min) in each experimental group (control group, 0.5% DS group, MISO group). As shown in FIG. 5, free water reabsorption in the kidneys was significantly higher in the 0.5% DS group compared to the control group (0.017 ± 0.008 vs. 0.035 ± 0.005 mL). / Min, p <0.05), the MISO group showed a significantly lower value than the 0.5% DS group (0.021 ± 0.008 vs. 0.035 ± 0.005 mL / min, p <0.05).

<FENa>
FIG. 6 shows FENa (%) in each experimental group (control group, 0.5% DS group, MISO group). FENa represents the proportion of sodium excreted in the urine out of the sodium that has emerged from the glomeruli into the Bowman's sac. As shown in FIG. 6, FENa was significantly higher in the 0.5% DS group compared to the control group (0.257 ± 0.117% vs 1.074 ± 0.403%, p <0 .05), the MISO group showed significantly higher values than the 0.5% DS group (2.517 ± 0.444%, p <0.05).

3. Discussion In the oral administration experiment, since the miso aqueous solution was freely ingested, more miso aqueous solution was ingested compared to the saline (0.5% w / v), resulting in 0.5% DS. The salt intake in the MISO group was greater than that in the group. However, the blood pressure was lower in the MISO group than in the 0.5% DS group. This was the same result as the previous study on male Dahl salt-sensitive rats. Using the relational expression (Y = 157.82 + 0.42091 × X) between the salt intake (X) and the blood pressure value (Y), the blood pressure expected value was calculated from the salt intake of the MISO group, which was 176 mmHg. The actual measurement value of blood pressure in the MISO group was 170 mmHg, which was lower than the predicted value. Further, when the salt intake was calculated from the measured value of blood pressure using the same mathematical formula, it was predicted to be 29 g, but the actual salt intake of the MISO group was 44 g. From this, it is considered that the influence of 34% of the ingested amount of salt was cut in the MISO group.

  When the relationship between the change in body fluid volume and the activity of the sympathetic nervous system was examined, in the oral administration experimental group, the urinary Na excretion rate was significantly higher than that in the 0.5% DS group, and the urinary dopamine excretion amount was also high. Significantly higher. Considering that renal dopamine has the effect of promoting water / Na excretion, the miso component causes water / Na diuresis by promoting dopamine production in the kidney, resulting in increased free water clearance and fluid volume It is thought that the blood pressure was lowered through the decrease in blood pressure. It is considered that the direct vasodilatory effect of dopamine is also involved in lowering blood pressure.

  On the other hand, urinary excretion of adrenaline and noradrenaline was significantly higher in the MISO group than in the 0.5% DS group. This was thought to be because adrenaline and noradrenaline were increased in reactivity by lowering blood pressure. On the contrary, it was suggested that the miso component has no effect of suppressing such baroreceptor reflex. In addition, as can be seen from the creatinine clearance, the MISO group maintains glomerular filtration rate compared to the 0.5% DS group, indicating that it acts suppressively on hypertensive nephropathy. It was done. About such an organ protective effect, it was the same result as the previous study using a male Dahl rat.

  From the above, miso ingredients promote kidney dopamine production, increase blood flow, increase glomerular filtration, cause water / Na diuresis, thereby increasing free water clearance and reducing body fluid volume It was suggested that the blood pressure rise was suppressed.

Claims (2)

  A dopamine production promoter containing a miso extract as an active ingredient.   The dopamine production promoter of Claim 1 for promoting dopamine production in the kidney.

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