KR19990070812A - Functional stress relief drink - Google Patents

Abstract

The present invention relates to a functional stress-relieving beverage, comprising seven kinds of herbal extracts such as ginseng and donkey, named mind tonic, and 12 kinds of food components such as soybean germ and bamboo sprout. The functional stress-relieving beverage, which was prepared by mixing, suppresses neutral lipids and LDL-cholesterol, which are known as a causative agent of adult diseases, and increases the content of HDL-cholesterol that prevents adult diseases. In addition, it inhibits the production of active oxygen such as superoxide radical (0₂-) and hydroxy radical (• 0H), which promotes aging, and increases the activity of catalase in the free radical scavenging enzyme. In addition, noradrenaline (NA) increased by physical and psychological stress, and MHPG-SO (3-methoxy-4-hydroxyphenylethyleneglycolsulfate) and corticosterone, which are increased by physical and psychological stress, were effectively reduced. It was.

Description

Functional stress relief drink

The present invention relates to a functional stress releasing beverage.

Until now, stress had been known for physical, chemical, and mental stress, but did not consider the development of stress-relieving beverages because they were not so seriously regarded as stress.

Recently, as stress is found to be the cause of all illnesses, the need for stress relief is emphasized. However, the need for stress relief drinks was not recognized because it was not possible to solve stress through beverages.

But the question is: 1) How do you measure stress? It is very difficult to study the problem of human beings and people who have complicated social life. 2) How to load stress through animal experiments and 3) How to evaluate the stress relief.

Until now, it has been focused on the fact that under stress, gastrointestinal disorders occur.

Resolving Functional Stress The biggest problem in the development of beverages is how animal testing can induce sociopsychological stress.

A communication box, which can be solved through animal experiments, was developed in 1992 by Prof. Masahiro Ishikawa, Ehime University School of Medicine. The device (64 × 64 × 40 cm) consists of eight rooms with foot-shock stress and eight rooms with no socio-psychological stress, with regular installations of copper wires of diameter 0.5 cm at 1.3 cm intervals. It is.

Put the mouse in the foot-shock stress and the sociopsychological stress, then switch on the electric switch. Only the black part is subjected to 2 mA electric shock for 10 seconds on the four feet (foot-shock stress) and 120 seconds for electricity. Not flowing is repeated for 1 hour. At this time, the white rat was not directly shocked, but the stress of the black rat was considered to be a social psychological stress. And the control just puts an hour in the white part. At this time, the experiment is most effective between 09:00 and 11:00 am.

The present invention relates to a functional stress-relieving beverage prepared by mixing herbal extracts such as ginseng radix, named mind tonic, and food ingredients such as soybean germ and bamboo shoots.

In the verification of the effect of the functional beverage of the present invention, the stress of the human should be premised, but the alternative method of causing the sociopsychological stress that the human will receive through animal experiments has been described above. The animal testing method developed by Masahiro Ishikawa's team was used.

In addition, stress experiments to date have focused mainly on the disorder of the stomach.

However, the present invention systematically evaluates the effects of functional stress-relieving beverages of the present invention with the main evaluation of the physiological changes of sympathetic activity and endocrine system as the autonomic nervous system.

The functional stress-relieving beverage of the present invention is based on 7 kinds of herbal extracts (mind tonic), and 12 kinds of food ingredients such as soybean germ and bamboo shoots are mixed.

That is, it is a herbal medicine named mind tonic, 0.5 to 25 weight of ginseng, wolfberry, Schisandra chinensis, jujube 0.3 to 5.0 weight, donkey 0.5 to 20.0 weight, elder berry 0.6 to 25.0 weight, cola fruit (nuts of kola) 18.0 to 30.0 parts by weight, extracted and concentrated with ethanol, followed by 0.8 to 25.0 weights of propylene glycol, 1.0 to 20.0 weights of glycerol, 2.5 to 35.0 weights of starch syrup, and 1.0 to 20.0 parts of purified water. Used to be added.

0.01-10.0 parts by weight of soybean embryos, bamboo shoots and shiitake mushroom extracts and 0.01-5.0 parts by weight of ginger extract are added to 0.01-10.0 parts by weight of the mind tonic, and asparagine, tyrosine and tryptophan are added again. 0.01 to 5.0 parts by weight of taurine, calcium lactate and sodium benzoate, 0.0001 to 0.05 parts by weight of potassium iodide, 0.01 to 5.0 parts by weight of vitamins C and B ₁ , 0.01 to 5.0 parts by weight of citric acid, and 0.01 to 5.0 parts by weight of sorbitol Add. 0.5 to 20.0 parts by weight of white sugar and 0.5 to 10.0 parts by weight of fructose are added and mixed. 0.5 to 20.0 parts by weight of grape juice is added and mixed. Was prepared.

As a test for the effect of the functional stress-relieving beverage of the present invention, a sociopsychological stress experiment apparatus developed by Masahiro Ishikawa, a professor's team of the Department of Medicine, Ehime University, was used.

After administering the functional beverage of the present invention to mice in a predetermined period of time, stress was applied to the mice, and the stress relief was determined by the content of noradrenaline (NA), which is known to be reduced by physical and psychological stresses. Physiological Effects of Drinks for Functional Stress Relief by Testing the Significance of MHPG-SO ₄ (3-methoxy-4-hydroxyphenylethyleneglyc ol sulfate) and Corticosterone Known to Increase Was evaluated.

The contents of the herbal medicines and food ingredients used in the present invention, and the experimental materials, experimental methods and results are as follows.

1. Experimental Materials

(1) functional stress relief drink

Herbal medicine named mind tonic, extracted with ethanol with 1.8 weight of ginseng, wolfberry, Schisandra chinensis, jujube, 3.6 weight, Angelica 5.4 weight, elder berry 6.0 weight, nuts of kola 18.0 weight Concentrated, and then used as 8.0 parts by weight of propylene glycol (propylene glycol), 9.0 weight of glycerol, 25.0 weight of starch syrup, 100 parts by weight of purified water 17.0 parts by weight.

To 0.1% and 0.5% of this mind tonic, 0.05 parts by weight of soybean embryos, bamboo shoots and shiitake mushroom extracts were added as food ingredients, and 0.03 parts by weight of ginger extract, respectively, followed by asparagine, tyrosine, tryptophan, taurine, and lactic acid. 0.05 parts by weight of calcium and sodium benzoate, 0.03 parts by weight of vitamins C and B ₁ , 0.07 parts by weight of citric acid and 0.5 parts by weight of sorbitol were added, and then 2.5 parts by weight of white sugar and 4.5 parts by weight of fructose were added and mixed. The grape juice was added and mixed to remove the precipitate by centrifugation at 1,000 × g for 10 minutes to prepare a functional stress relief beverage of the present invention by 120ml as a supernatant.

(2) Animal testing for effect test

Experimental animals used in this animal experiment,

1) For the adult disease and anti-aging effect test, a 42-day Sprague-Dawley rat (rats: 120 ± 10g, ♂) was used for 42 days.

2) For the stress-relieving effect experiment, the corresponding drink was administered for 4 weeks using 4 weeks old ICR mice (mouse: 25 ± 3g, ♂).

The next three days induce stress for 1 hour per day in a social and psychological stress box, then collect blood and brain to measure corticosterone in plasma and noradrenaline in brain fraction. NA) and MHPG-SO ₄ (3-methoxy-4-hydroxyphenylethyleneglycol sulfate) were measured to evaluate the stress relief effect.

(3) Reagent

As a reagent used in this experiment, noradrenaline (NA), MHPG-SO ₄ (3-methoxy-4-hydroxyphenylethyleneglycol sulfate), and corticosterone (Sigma) express reagents were used to measure and evaluate the stress relief effect. For other assay reagents, more than 1 grade of reagent was used.

2. Experiment Method

(1) odor and separation of blood

The animals were anesthetized with ether, and blood was collected from the heart, left for 2 hours at low temperature, centrifuged at 700 × g for 10 minutes, and the supernatant was treated with 0.05 ml of blood per 1.0 ml of anticoagulant (sodium heparin). The bottle was placed in a complete blood cell count (green cross) and used for analysis of corticosterone while cryopreservation at -70 ° C.

(2) fraction of brain tissue

Aliquot the brain tissue and place it in buffer solution (1.15% KCl / 10mM phosphate buffer + 5mM EDTA, pH 7.4). Before analyzing the stress relief effect, the contents of noradrenaline (NA) and MHPG-SO ₄ were analyzed according to the following pretreatment, respectively.

(3) measurement of corticosterone

Corticosterone content of plasma isolated from blood was measured according to the method of Sliber (1958). After shaking 1.0 ml of plasma vigorously for 30 seconds in three times the amount of petroleum ether, the solvent layer is discarded after centrifugation. After diluting the purified plasma with 5.0 ml of distilled water, the mixture was extracted by shaking with 15.0 ml of methylene chloride for 30 seconds in a centrifuge tube.

The extract is added to a tube containing 1.0 ml of 0.1N NaOH solution, shaken quickly and centrifuged. After centrifugation, the alkali layer is discarded and 10.0 ml of methylene chloride is placed in a tube containing 2.0 ml of 30N sulfuric acid. After shaking for about 30 seconds, after centrifugation for 1 to 2 minutes, the upper solvent layer was discarded and the lower layer was 2.0 ml and left at room temperature for 30 to 90 minutes, and measured at excitation 470 nm and emission 530 nm.

Corticosterone standard solution was dissolved in 5.0 ml of corticosterone in 5.0 ml of ethanol, diluted with 1,000 ml of distilled water, and then 0.1 to 0.5 ug. Was quantified.

(4) measurement of noradrenaline

The content of noradrenaline (NA) in the brain was measured according to the method of Kohno (1979). Brain tissue is ground in a low temperature room with 4.0 ml of 0.2 N sulfuric acid containing 0.1% sodium metabis sulfate (Na ₂ S ₂ O ₅ ), and then centrifuged at 8000 × g for 5 minutes. The supernatant is taken 2.0 ml each to simultaneously measure NA and MHPG-SO ₄ . Next, 3.0 ml of 0.4N PCA containing 0.1% Na ₂ S ₂ O ₅ and 0.05% EDTA was added. After centrifugation at 8000 × g for 10 minutes, the supernatant was transferred to a cap tube and stored at −45 ° C. until the experiment.

The pyrex column (diameter 6 mm, height 25 cm) for NA measurement is filled with glass wool and filled with alumina activated at pH 7.5-8.0 at a height of exactly 6.0 cm. NA is extracted from the column by 0.05 ml PCN 2.0 ml. After extraction, the solution is converted to a fluorescent compound by 2.0 ml of iodine reagent.

Fluorescence of NA was measured at excitation 380 nm and emission 495 nm. The NA standard was prepared at a concentration of 100 ug / ml and the content of noradrenaline (ng / g brain) was quantified according to the above procedure by a standard calibration curve.

(5) Measurement of MHPG-SO ₄

The content of MHPG-SO ₄ in brain tissue was measured according to the method of Kohno (1979). Brain tissue is ground in a low temperature room with 4.0 ml of 0.2 N sulfuric acid containing 0.1% sodium metabis sulfate (Na ₂ S ₂ O ₅ ) and centrifuged at 8000 × g for 5 minutes. The supernatant is taken 2.0 ml each to simultaneously measure NA and MHPG-SO ₄ . Each sample is adjusted to pH 6.0-6.5 with 0.3 N barium hydroxide solution, and then all samples are made to the same amount with distilled water. After centrifugation at 8000 x g for 10 minutes, the supernatant was transferred to a cap tube and stored at -45 ° C until the experiment.

The pyrex column (diameter 6 mm, height 25 cm) for the measurement of MHPG-SO ₄ is covered with glass wool and filled with DEAE sephadex A-25 at exactly 6.0 cm. MHPG-SO ₄ is extracted from the column as 2.0 ml 0.4N PCA with 0.1% sodium metabissulphate.

Immediately after separation of MHPG-SO ₄ , 0.1 ml of 0.2% cysteine and 0.1 ml of 70% PCA are added, and the reaction mixtures are boiled in a water bath at 90-95 ° C. for 25 minutes. In this case, blank is added without heating. After adding 0.3 ml of ethylenediamine, all samples were placed in a hot bath for 5 minutes, and then cooled.

Fluorescence of MHPG-SO ₄ was measured at excitation 325 nm and emission 465 nm. MHPG-SO ₄ standard solution (100ug / ml) was prepared with 0.5% sodium metabis sulfate (Na ₂ S ₂ O ₅ ) concentration and the content of MHPG-SO ₄ (ng / g brain) by the above procedure according to the standard calibration curve. Was quantified.

3. Evaluation of Experimental Results

(1) Reducing Effect of Corticosterone

Corticosterone, which is present in plasma, is a hormone involved in the metabolism of glucose in the adrenal cortex and is used as an indicator of stress.

In other words, this corti is not only significantly increased under physical stress (foot-shock stress) loads such as electric shocks, but also significantly increases under sociopsychological stress (non-foot-shock stress) loads. Decreasing the amount of cholesterol can be a way to relieve stress.

The mind tonic-added beverage as a stress-relieving beverage of the present invention was administered to an ICR mouse for 4 weeks, followed by an electric shock of 2 mA for 1 hour from 10:00 to 11:00 for 3 days (120 after 10 seconds of shock). After taking a second break), the psychotic stress is reported, and then the effect of the mind tonic-added beverage on the content of corticosterone (corticosterone) is shown in Table 1 below.

Table 1. Effect of beverage of the present invention on the content of corticosterone

Content vs. Content of Corticosterone

(ug / dl plasma) (%)

Physical stress load group 31.59 ± 6.38-

(foot-shock stress)

Psychological stress load group 26.03 ± 0.56 100.0% (-)

(psychological stress)

Stress relief drink group

0.1% -mind tonic dose group 21.06 ± 1.15 80.9% (-19.1%) ^**

(0.1% -mind tonic)

0.5% -mind tonic dose group 15.12 ± 3.21 58.1% (-41.9%) ^***

(0.5% -mind tonic)

Significance test: Psychological stress group ^** p0.01; ^*** p0.001

Psychological stress under the psychological stress of the electrical shock load (foot-shock stress) as a control group (26.03 ± 0.56ug / dl plasma: 100%) compared the administration group of the beverage for stress relief As shown, the 0.1% and 0.5% mind-tonic-added beverages were 21.06 ± 1.15ug / dl plasma and 15.12 ± 3.21ug / dl plasma, respectively, 80.9% and 58.1%, respectively, with corticosterone content of 19.1%, respectively. And 41.9% significant reduction, scientifically proven to effectively relieve stress.

(2) increase effect of noradrenaline

Noradrenaline (NA) is a hormone secreted with adrenaline and is secreted as a neurotransmitter at the sympathetic nerve endings of brain tissue. Among these neurotransmitters, noradrenaline is also used as an indicator of stress.

In other words, noradrenaline is significantly reduced in physical stress (foot-shock stress) loads such as electric shocks, and significantly reduced in sociopsychological stress (non-foot-shock stress) loads. The ability to increase the content of can be a way to effectively relieve stress.

A mind tonic-added beverage, the functional stress-relieving beverage of the present invention, was administered weekly to an ICR mouse, followed by an electric shock of 2 mA for 1 hour from 10:00 to 11:00 for 3 days (after 10 seconds of shock). After 120 seconds of rest) and seeing this, psychological stress is observed, and the effect of the mind tonic-added beverage on the content of noradrenaline is shown in Table 2.

Table 2. Effect of beverage of the present invention on the content of noradrenaline

Content Ratio of Noradrenaline

(ng / g brain) (%)

Physical stress load group 299.4 ± 7.8-

(foot-shock stress)

Psychological stress load group 306.3 ± 21.7 100.0%

(psychological stress)

Stress relief drink group

0.1% Mind Tonic Group 378.1 ± 48.8 123.4% (+ 23.4%) ^***

(0.1% -mind tonic)

0.5% Mind Tonic Group 416.2 ± 8.5 135.9% (+ 35.9%) ^***

(0.5% -mind tonic)

☞ Significance test: compared to psychological stress group ^*** p0.001

Mind tonic, a stress-relieving beverage, using the psychological stress group that received the psychological stress of foot-shock stress as a control group (306.3 ± 21.7 ng / g brain: 100%) When comparing the administration effects of) -added beverages, 0.1% and 0.5% -mind tonic-added beverages were 378.1 ± 48.8ng / g brain and 416.2 ± 8.5ng / g brain, respectively, 123.4% and 135.9%, respectively. It has been scientifically proven that the adrenaline content is significantly increased by 23.4% and 35.9%, respectively, to relieve stress very effectively.

(3) reduction effect of MHPG-SO ₄

MHPG-SO ₄ (3-methoxy-4-hydroxyphenylethyleneglycol sulfate) is a metabolite of noradrenaline (NA), which is secreted as a neurotransmitter at the sympathetic nerve endings of brain tissue, and is used as an indicator of stress like NA. .

This is because MHPG- not only increases significantly during physical stress (foot-shock stress) load such as electric shock but also significantly increases under sociopsychological stress (non-foot-shock stress) load. Reducing the content of SO ₄ may be an effective way to relieve stress.

The mind tonic-added beverage, the functional stress-relieving beverage of the present invention, was administered to an ICR mouse for 4 weeks, followed by an electric shock of 2 mA for 1 hour until 10: 00-11: 00 for 3 days (after 10 seconds of shock). After 120 seconds of rest) and seeing this, psychological stress is observed, and then the effect of MHPG-SO ₄ as a dose effect of the mind tonic-added beverage is shown in Table 3 below.

Table 3. Effect of Drink of the Invention on the Content of MHPG-SO ₄

Classification Content of MHPG-SO ₄

(ng / g brain) (%)

Physical stress load group 226.4 ± 39.7-

(foot-shock stress)

Psychological stress load group 163.2 ± 15.6 100.0%

(psychological stress)

Stress relief drink group

0.1% Mind Tonic Group 119.0 ± 2.7 72.9% (-27.1%) ^***

(0.1% -mind tonic)

0.5% Mind Tonic Group 131.2 ± 7.8 80.4% (-19.6%) ^***

(0.5% -mind tonic)

☞ Significance test: Psychological stress group ^*** p0.001

Mind tonic developed as a stress-relieving beverage using the control group (163.2 ± 15.6 ng / g brain: 100%) as the control group (163.2 ± 15.6ng / g brain: 100%) who reported the impact of foot-shock stress. tonic) -added beverages compared to the administration of 0.1% and 0.5% -mind tonic-added drinks were 119.0 ± 2.7ng / g brain and 131.2 ± 7.8ng / g brain, respectively, 72.9% and 80.4%, respectively. It has been scientifically proven that MHPG-SO ₄ content is significantly reduced by 27.1% and 19.6%, respectively, to effectively relieve stress.

In summary, the stress-relieving beverage of the present invention prepared with 12 kinds of herbal ingredients such as 7 herbal medicines and soybean germ extracts named mind tonic is effective in relieving stress and at the same time carrying out animal experiments. It has been proved that the induction of social and psychological stress of people who have complicated social life is very effective. In addition, the results of the administration of the mind tonic-added stress releasing beverage of the present invention, the content of noradrenaline significantly reduced during physical and psychological stress load effectively increased while corti significantly increased during physical and psychological stress load The content of costosterone (corticosterone) or MHPG-SO ₄ is effectively reduced, and the stress relief effect of the beverage of the present invention is expected to be very large.

Embodiments of the present invention are as follows.

(Example 1)

The biggest problem in developing functional beverages to relieve stress is how animal testing can induce sociopsychological stress.

As a device that can be solved through animal experiments, a sociopsychological stress communication box developed by Masahiro Ishikawa, a professor of medicine at Ehime University, Japan, was used.

The functional stress-relieving beverage of the present invention comprises 12 kinds of foods such as soybean germ and bamboo shoots in 0.5 parts by weight of 7 kinds of mind tonics such as ginseng (ginseng radix) and angelicae radix. 0.03 to 0.05 parts by weight of the component is added and mixed, and then 0.05 parts by weight of calcium lactate and sodium benzoate, 0.03 parts by weight of vitamin C and B ₁ , 0.07 parts by weight of citric acid, 0.0002 parts by weight of potassium iodide, and sorbitol 0.5 Add parts by weight. 2.5 parts by weight of white sugar and 4.5 parts by weight of fructose were added and mixed, and 5.0% grape juice was added and mixed, followed by centrifugation at 1,000 × g for 10 minutes to remove the precipitate, so that the whole was 120 ml as a supernatant solution for stress relief. A beverage was prepared.

(Effect test)

As an animal experiment for the effect test of the beverage of the embodiment of the present invention, the stress-relieving effect test for 4 weeks using the 4 weeks old ICR mice (mouse: 25 ± 3g, ♂), then 3 During the 1 hour period from 10:00 to 11:00 during the day, the psychosomatic stress communication box was continuously subjected to 2 mA electric shock (120 seconds rest after a 10 second shock), followed by psychological stress, and then the mind tonic -The stress relief effect of the added beverage was measured.

Corticosterone content (ug / dl plasma) of plasma isolated from blood was measured at excitation 470nm and emission 530nm after pretreatment according to the method of Sliber et al., And the concentration of noradrenaline (NA) in brain cells The content (ng / g brain) was pretreated according to Kohno's method, and the fluorescent material was measured at excitation 380 nm and emission 495 nm. The content of MHPG-SO ₄ (ng / g brain) was determined according to Kohno's method. Fluorescence was measured at excitation 325 nm and emission 465 nm.

As described above, 0.1% of the functional stress-relieving beverage of the present invention was analyzed by using a psychosocial stress load box (communication box) on the content of corticosterone in plasma as a stress index component. And 0.5% -mind tonic-added beverages were 21.06 ± 1.15ug / dl plasma and 15.12 ± 3.21ug / dl plasma, respectively, 80.9% and 58.1% compared to the control group with 19.1% and 41.9% respectively. Significantly decreased.

In addition, the analysis of the effect on the content of noradrenaline in the brain cells, 0.1% and 0.5% -mind tonic-added beverages were 378.1 ± 48.8ng / g brain and 416.2 ± 8.5ng / g brain, respectively. 123.4% and 135.9%, respectively, significantly increased the noradrenaline content by 23.4% and 35.9%, respectively, and the developmental stress-relieving beverages affected the content of MHPG-SO ₄ as a metabolite of noradrenaline. As a result of comparing the effects, the 0.1% and 0.5% -mind tonic-added beverages were 119.0 ± 2.7ng / g brain and 131.2 ± 7.8ng / g brain, respectively, 72.9% and 80.4%, respectively, MHPG-SO ₄ Were significantly decreased by 27.1% and 19.6%, respectively.

In conclusion, the functional stress releasing beverage of the present invention has been scientifically proven to have an effect of releasing stress.

Claims (1)

Ginseng 0.5-25 weight, wolfberry, schizandra, jujube 0.3-5.0 weight, Angelica 0.5-20.0 weight, elder berry 0.6-25.0 weight, nuts of kola 18.0-30.0 weight Next, 0.01-10.0 parts by weight of the herbal extract prepared from 0.8 to 25.0 weights of propylene glycol, 1.0 to 20.0 weights of glycerol, 2.5 to 35.0 weights of starch syrup, and 1.0 to 20.0 parts by weight of purified water. Add 0.01 to 5.0 parts by weight of soybean embryos, bamboo shoots, and shiitake mushroom extracts, and 0.01 to 5.0 parts by weight of ginger extract, and add asparagine, tyrosine, tryptophan, taurine, calcium lactate, and sodium benzoate, respectively. the 0.01 to 5.0 parts by weight, from 0.0001 to 0.05 parts by weight of potassium iodide, vitamin C and B ₁ are added 0.01 to 5.0 ssikeul parts by weight of citric acid 0.01 to 5.0 parts by weight, and 0.01 to 5.0 parts by weight of sorbitol. 0.5 to 20.0 parts by weight of white sugar and 0.5 to 10.0 parts by weight of fructose are added and mixed, and 0.5 to 20.0 parts by weight of grape juice is added and mixed to produce a functional stress-relieving beverage.