KR20210058752A - Health functional food for ameliorating stress comprising extract of Salvia plebeia - Google Patents

Abstract

The present invention relates to a health functional food for alleviating stress containing a Salvia plebeia extract. The Salvia plebeia extract suppress weight loss and immune function degradation due to stress, and has an effect of regulating the secretion of IL-6, TNF-α, MMP-2, MMP-3, MMP-8, MMP-13, and lipocalin-2 increased due to stress, thereby being able to be usefully used as the health functional food for alleviating stress.

Description

{Health functional food for ameliorating stress comprising extract of Salvia plebeia}

The present invention relates to a health functional food for improving stress, and more specifically, to a health functional food for improving stress including an extract of baeam chazugi.

In the modern society, an increasing number of people complain of anxiety disorders and mental disorders caused by various stresses. In general, our living body produces several reactions against stress, which leads to changes in the endocrine system and autonomic nervous system. In particular, persistent or chronic stress is accompanied by psychological changes such as depression, anxiety, and anger, and the pathogenesis of various diseases such as mental disorders, digestive disorders such as central nervous system, immune system disorders, nervous gastric ulcers, and cardiovascular disorders such as angina. It is closely related.

Chronic psychological stress consequently causes depression.In Western society, drugs related to depression are among the drugs with the highest prescription rate among prescription drugs, and the World Health Organization (WHO) reports that by 2020, depression in the U.S. and Western Europe. It is estimated to be the disease with the highest prevalence. In Korea, as urbanization accelerates, the gap between the rich and the poor and feelings of alienation are deepening, and the incidence of depression is rapidly increasing. Currently, Korea is one of the countries with the highest suicide rate among OECD countries.

Specifically, depression (or severe depressive disorder) is a serious mental disorder that accompanies physical and cognitive dysfunction along with subjective emotions such as sadness and lethargy. The most important symptom of depression is the loss of interest and enjoyment for universal stimuli. When depression occurs, daily life becomes difficult, resulting in huge social and economic losses, and death as 60 to 70% of the causes of suicide are depression. It is also a disease that leads to. Therefore, although there is a need for and development of antidepressants, some antidepressants have severe side effects or may not meet expectations.

It usually takes about 2 weeks to improve symptoms such as improvement of depression by administration of antidepressants, and there are many patients who do not respond well to antidepressants, or recurrence of depression, and are unable to take drugs due to severe side effects. Specifically, tricyclic antidepressants such as imipramine and desipramine have severe side effects such as hypotension and heart dysfunction, and are the most widely used selective drugs such as fluoxetine or sertraline. Serotonin absorption inhibitors cause nausea, gastrointestinal bleeding or sexual dysfunction. Therefore, the development of new drugs that are more effective and have fewer side effects than existing antidepressants has been continuously attempted, but the development costs are enormous, so pharmaceutical companies cannot easily develop them.

Therefore, it is necessary to develop a health functional food that can prevent the development of stress stimulation into mental disorders without the problems that appear in the antidepressants. Natural products contain various unknown ingredients and have the advantage of being relatively safe. , It has a very high potential for stress improvement. In this regard, referring to the specific prior art, grapefruit oil; And there is a patent (KR 10-2010-0106062 A) on a fragrance extract containing bergamot oil, which is said to have stress relief and soothing effects, and can be used in various fields such as beauty and cosmetics. However, the stress referred to in the above patent provides an instant refreshing sensation and does not substantially alleviate mental stress.

On the other hand, Salvia plebeia R.Br. is a plant belonging to the Lamiaceae family (Labiatae), and is expressed as a common sage in English. It grows wild in all areas of Korea, and is an annual or biennial upright vegetation that is distributed in the wet waters and ditches in the lowlands. It has the tinnitus of Seolgyeoncho, Gwadongcheong, or Bambaechu, and in the private sector, it is also called as leprosy cabbage and gombo cabbage. It has been reported that the main components of Baeam Chazugi include flavonoids and phenolic substances, essential oils, saponins, cardiac glycosides, and unsaturated sterols.

From ancient times, Baeam Chazgi has been known to be effective in clearing blood, coughing, asthma, inflammation, high blood pressure, bronchial disease, women's diseases, infertility, diabetes, and poor circulation. However, there is no prior art for a health functional food for improving stress, including baeam chazugi extract. Thus, the present inventors were studying to find an extract for improving stress from natural substances, and confirmed that the Baeam Chazugi extract has the effect of inhibiting weight loss due to stress and controlling the secretion of cytokines, MMPs, and lipocalins. By doing this, the present invention was completed.

An object of the present invention is to provide a health functional food for improving stress.

Another object of the present invention is to provide a pharmaceutical composition for improving stress.

To achieve the above object,

One aspect of the present invention provides a health functional food for improving stress containing a baeam chazugi extract as an active ingredient.

Another aspect of the present invention provides a pharmaceutical composition for improving stress containing Baiam chazugi extract as an active ingredient.

Baiam chazugi extract of the present invention inhibits weight loss and immune function decline due to stress, and increases IL-6, TNF-α, MMP-2, MMP-3, MMP-8, MMP-13, due to stress. Since the effect of controlling the secretion of lipocalin-2 is excellent, it can be usefully used as a health functional food for stress improvement.

1 is a graph measuring the weight of rats of each group prepared in an experimental animal model after breeding for 12 weeks.
FIG. 2 is a graph showing the weight of the spleen extracted by sacrificing the rats of each group prepared in the experimental animal model after 12 weeks of breeding.
3 is a graph showing serum IL-6 levels in blood by analyzing serum obtained from rats of each group prepared in an experimental animal model after 12 weeks of breeding.
FIG. 4 is a graph showing serum TNF-α levels in blood by analyzing serum obtained from rats of each group prepared in an experimental animal model after breeding for 12 weeks.
FIG. 5 is a graph showing serum MMP-2 levels in blood by analyzing serum obtained from rats of each group prepared in an experimental animal model after 12 weeks of breeding.
FIG. 6 is a graph showing serum MMP-3 levels in blood by analyzing serum obtained from rats of each group prepared in an experimental animal model after breeding for 12 weeks.
FIG. 7 is a graph in which serum MMP-8 levels in blood were measured by analyzing serum obtained from rats of each group prepared in an experimental animal model after breeding for 12 weeks.
FIG. 8 is a graph showing serum MMP-13 levels in blood by analyzing serum obtained from rats of each group prepared in an experimental animal model after breeding for 12 weeks.
FIG. 9 is a graph showing serum lipocalin-2 levels in blood by analyzing serum obtained from rats of each group prepared in an experimental animal model after breeding for 12 weeks.

Hereinafter, the present invention will be described in detail.

One aspect of the present invention provides a health functional food for improving stress, including a baeam chazugi extract.

The baiam chazugi extract may be prepared in the form of a liquid or powder.

The baeam chazugi extract may be prepared by a manufacturing method comprising the step of preparing an extract by adding an extraction solvent to baeam chazugi.

The baiam chazugi extract may be prepared by including one or more of filtration, concentration under reduced pressure, and drying, if necessary.

The Baeam Chazugi extract may be prepared in the form of a powder by adding an extraction solvent to Baeam Chazugi to prepare an extract, followed by filtration, concentration under reduced pressure, and lyophilization.

In the above manufacturing method, the baeam chazugi may be used without limitation, such as cultivated or commercially available ones.

In addition, the extraction solvent may be water, _{a lower alcohol of C 1} to C ₃ , or a mixed solvent thereof, and specifically, the _{lower alcohol of C 1} to C ₃ may be ethanol. Further, the extraction solvent may be ethanol or an aqueous ethanol solution, and the aqueous ethanol solution is 20 to 90% (v/v), 30 to 80% (v/v), 20 to 80% (v/v), 30 to 70% (v/v) or 40 to 60% (v/v).

Here, the extraction method may be immersion, shaking extraction, Soxhlet extraction, or reflux extraction. At this time, the extraction temperature is preferably 5 to 50 °C, more preferably 20 to 30 °C, but is not limited thereto. At this time, the extraction time is 10 to 96 hours; 15 to 72 hours; 20 to 48 hours; Alternatively, it may be 23 to 25 hours, but is not limited thereto. The extraction may be repeated 1 to 5 times. In addition, the extraction solvent is preferably extracted by adding 2 to 20 times the weight of the baiam chazugi used for extraction, more preferably 5 to 15 times to extract, and 8 to 12 times to extract. Although particularly preferred, it is not limited thereto. In a range outside the conditions of the extraction time or the number of extractions, extraction may not be sufficiently performed, so that the content of the active ingredient in the Baiam Chazgi extract may be insignificant. Alternatively, the extraction yield may no longer increase, and thus the efficiency of the extraction operation may decrease.

On the other hand, the filtration may use a gravity filter, a vacuum filter, a pressurized filter, a compressed filter, a centrifugal filter, or a membrane separation filter, but is not limited thereto, and the vacuum concentration may be performed using a rotary vacuum concentrator, a vacuum vacuum concentrator, or a vacuum rotary evaporator. However, the drying is not limited thereto, and the drying may be reduced pressure drying, vacuum drying, boiling drying, spray drying, or freeze drying, and specifically, may be freeze drying, but is not limited thereto.

In addition, the extract of the present invention includes not only the extract by the above-described extraction solvent, but also an extract obtained through another conventional extraction method, or an extract that has undergone purification and fermentation processes. It includes not only the extract by the decompression by carbon dioxide, the extraction by the supercritical extraction method by high temperature, the extraction by the extraction method using ultrasonic waves, but also the fractions such as the fraction using the difference in solvent polarity for the crude extract. Separation using an ultrafiltration membrane having a certain molecular weight cut-off value, separation by various chromatography, or extracts obtained by fermentation products using natural or various microorganisms are also included in the extract of the present invention.

The baiam chazugi extract can suppress weight loss due to stress.

In a specific embodiment of the present invention, the weight of the rat decreases due to the pain stress caused by performing a knee incision, but it was found that the stress was suppressed and weight loss was suppressed when the extract of Baeam Chazgi was administered to the stress-induced rat (Experimental See Example 1 and Fig. 1).

The baiam chazugi extract can suppress the decrease in immune function due to stress. In a specific embodiment of the present invention, the weight of the spleen, which is the immune tissue of the rat, is reduced due to the pain stress caused by performing a knee incision, but the stress is suppressed and the reduction of the spleen weight is suppressed by feeding the extract of Bacillus Chazugi to the stress-induced rat. It was found (see Experimental Example 2 and Fig. 2).

The baiam chazugi extract may reduce cytokines increased due to stress. Here, the cytokine may be one or more selected from the group consisting of IL-6 and TNF-α.

The baiam chazugi extract may reduce matrix metallopeptidase (MMP) increased due to stress. Here, the MMP may be at least one selected from the group consisting of MMP-2, MMP-3, MMP-8, and MMP-13.

The baiam chazugi extract may reduce lipocalin increased due to stress. Here, the lipocalin may be lipocalin-2.

In a specific embodiment of the present invention, levels of IL-6, TNF-α, MMP-2, MMP-3, MMP-8, MMP-13 and lipocalin-2 in the blood of rats due to pain stress induced by performing a knee incision However, feeding the extract of Baeam Chazugi to stress-inducing rats suppressed stress and decreased the levels of IL-6, TNF-α, MMP-2, MMP-3, MMP-8, MMP-13 and lipocalin-2 in the blood. (See Experimental Examples 3 to 5 and FIGS. 3 to 9).

From this, the Baiam Chazugi extract of the present invention has the effect of inhibiting weight loss and immune tissue weight reduction due to stress induction, and blood IL-6, TNF-α, MMP-2, MMP-3, MMP- increased by stress induction. 8, Since it has the effect of reducing MMP-13 and lipocalin-2, it can be seen that it can be usefully used as a health functional food for stress improvement.

"Health functional food" as used herein refers to a food manufactured using nutrients that are liable to be deficient in daily meals or raw materials or ingredients having useful functions for the human body, and refers to foods that help maintain the health of the human body, but is not limited thereto. It is not used in a sense that includes all health foods in the usual sense.

The form and type of the health functional food is not particularly limited. Specifically, the health functional food may be in the form of tablets, capsules, powders, granules, liquids, and pills. The health functional food may contain various flavoring agents, sweetening agents, or natural carbohydrates as additional ingredients. The sweetener may be a natural or synthetic sweetener, and examples of the natural sweetener include taumatin and stevia extract. Meanwhile, examples of synthetic sweeteners include saccharin and aspartame. In addition, the natural carbohydrates may be monosaccharides, disaccharides, polysaccharides, oligosaccharides and sugar alcohols.

In addition to the above-described additional ingredients, the health functional food of the present invention includes nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pexane and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, and preservatives. , Glycerin, alcohol, and the like may be further included. These ingredients can be used independently or in combination. The ratio of the additive may be selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the extract of the present invention.

The baiam chazugi extract of the present invention may be added to food as it is or used with other foods or food ingredients. At this time, the content of the added active ingredient may be determined according to the purpose. In general, the content of the health functional food may be 0.01 to 90 parts by weight of the total food weight. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of health control, the amount may be below the above range, and if there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

Another aspect of the present invention provides a pharmaceutical composition for improving stress containing Baiam chazugi extract as an active ingredient.

The description of the pear-am chazugi extract is the same as the description of the pear-am chazugi extract in the health functional food.

The pharmaceutical composition for improving stress containing the extract of Baeam Chazugi of the present invention as an active ingredient contains 0.1 to 99.9% by weight of the extract of the present invention as an active ingredient based on the total weight of the composition for administration, and is pharmaceutically acceptable. Possible carriers, excipients or diluents may be included. The pharmaceutically acceptable carrier is not particularly limited as long as it is suitable for in vivo delivery of the composition, and is, for example, selected from the group consisting of saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol and ethanol. Any one of these components or one or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as needed. In addition, a diluent, a dispersant, a surfactant, a binder, and a lubricant may be additionally added to form an injectable formulation such as an aqueous solution, a suspension, an emulsion, a powder, a tablet, a capsule, a pill, a granule, or an injection solution. Furthermore, it can be preferably formulated according to each disease or component using an appropriate method in the art.

In addition, the pharmaceutical composition for improving stress containing the extract of Bacillus Chazugi of the present invention as an active ingredient can be administered orally or parenterally, and when administered parenterally, subcutaneous injection, intravenous injection, intramuscular injection, or intraperitoneal injection injection method It is preferable to choose. In order to formulate a formulation for parenteral administration, the composition is mixed with the composition in water to prepare a suspension, and it is formulated in a unit dosage form of an ampoule or vial.

The pharmaceutical composition for improving stress containing the extract of Baeam Chazugi of the present invention as an active ingredient is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is the type of patient's disease, severity, drug activity, Sensitivity to drugs, time of administration, route of administration and rate of excretion, duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field. The composition may be administered as an individual therapeutic agent or administered in combination with other therapeutic agents, may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. It is important to administer an amount capable of obtaining the maximum effect in a minimum amount without side effects in consideration of all the above factors, and this can be easily determined by a person skilled in the art.

Hereinafter, the present invention will be described in detail by the following examples and experimental examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the contents of the present invention are not limited by the following Examples and Experimental Examples.

<Example 1> Preparation of Baeam Chazugi Extract

Salvia plebeia R.Br., grown in Jangseong, Jeollanam-do, was washed with water to wash off soil and dirt. To the washed embryonic teaser, 10 times the weight of alcohol (50% (v/v)) was added, and the primary extraction was performed at room temperature for 24 hours. The first extract was recovered, 10 times the weight of the remaining residue was added to alcohol (50% (v/v)), and the second extraction was performed at room temperature for 24 hours. The first extract and the second extract were mixed, and concentrated under reduced pressure. The concentrate was lyophilized and powdered to obtain a Baiam chazugi extract, which was used in the following experiment.

<Experiment preparation>

1. Preparation of experimental animal model

In order to confirm the stress improvement effect of Baeam Chazugi extract, a stress-induced rat model was prepared. The normal rat model was Sprague-Dawley (SD) male white rat, and the stress-induced rat model had pain stress by performing an anterior cruciate ligament transaction on the rat. During the breeding period of normal rats and stress-induced rats, general solid feed (central laboratory animal, Seoul, Korea) and water were freely ingested, and the temperature of the breeding environment was 23±2℃ and the humidity was 50±10%. The contrast period was adjusted in units of 12 hours.

The Baeam Chazugi extract prepared in Example 1 was dissolved in a solvent (Phosphate Buffer Saline, PBS), and 150 mg of Baeam Chazugi extract per 1 kg of the weight of the stress-inducing rat was fed to Baeam Chazugi Group 1, and Baeam Chazugi 2 The group was fed 300 mg of Baeam Chazgi extract per 1 kg of the weight of stress-induced rats, and the control group was fed only the same amount of PBS to the stressed rats. That is, the experiment was conducted by dividing it into a normal group, a control group, a group of 1 group of Baeam Chazugi and 2 groups of Baeam Chazgi.

This was bred for a total of 12 weeks, and after the breeding was completed, the body weight was measured _{, anesthetized with CO 2} , and blood was collected. The collected blood was placed in a centrifugal test tube and centrifuged for 15 minutes at 2000 rpm and 4°C to obtain serum. The collected rats were _{anesthetized with CO 2} and euthanized, and the spleen and thymus were excised. The weight was measured using.

<Experimental Example 1> Analysis of the effect of inhibiting weight loss by inducing stress

In order to confirm whether the extract according to the present invention has a stress suppressing effect, an experiment was performed to analyze the effect of suppressing weight loss caused by stress induction by the following method, and the results are shown in FIG. 1.

For the normal group, the control group, the group 1 group and the group 2 of the embryonic tea prepared as in the experimental animal model, the body weight was measured every week for 12 weeks, and the body weight at the 12th week at the end of the experiment period is shown in FIG. 1 Shown in. The weight of the rat was measured and recorded in a state where the numerical value was stabilized on a scale capable of measuring to one decimal place.

Compared to the normal group, the control group lost 8.4% in weight. In addition, the body weight of the group 1 was higher than that of the control group, and the group 2 had a body weight higher than that of the control group (see Fig. 1).

The above results indicate that the weight of the rat is reduced due to the pain stress caused by performing a knee incision, but the stress is suppressed and the weight loss is suppressed when the extract of Baeam Chazgi is administered to the stress-induced rat. From this, it can be seen that the baiam chazugi extract of the present invention has an effect of suppressing weight loss due to stress induction, and thus can be usefully used as a health functional food for improving stress.

<Experimental Example 2> Analysis of the inhibitory effect on reducing immune tissue weight by inducing stress

In order to confirm whether the extract of Baeam Chazugi according to the present invention has a stress suppressing effect, an experiment was performed to analyze the effect of suppressing immune tissue weight reduction caused by stress induction by the following method, and the results are shown in FIG. 2. When stress is induced, the function of the spleen, which is an immune tissue, decreases and an abnormality occurs in immunity, so the following experiment was performed.

According to the preparation of the experimental animal model, the normal group, the control group, the Baeam Chazgi group 1 and Baeam Chazgi group 2 were reared, euthanized after 12 weeks of breeding was completed, and the spleen, an immune organ, was extracted, and the weight was weighed with a chemical balance. It was measured.

Compared to the normal group, the control group decreased the spleen weight by 13.5%. And, compared to the control group, the spleen weight was 17% higher in the group 1, and the second group had a 7% higher spleen weight compared to the control group (see FIG. 2).

The above results indicate that the spleen weight of the rat is reduced due to the pain stress caused by performing a knee incision, but the stress is suppressed and the reduction of the spleen weight is suppressed by feeding the extract of Baeam Chazugi to the stress-induced rat. From this, it can be seen that the extract of Baeam Chazugi of the present invention has an inhibitory effect on reducing the weight of immune tissues caused by stress induction, and thus can be usefully used as a health functional food for improving stress.

<Experimental Example 3> Analysis of the modulatory effect of IL-6 and TNF-α in blood increased by stress induction

In order to check whether the Baiam chazugi extract according to the present invention has a stress-inhibiting effect, regulation of IL-6 (Interleukin-6) and TNF-α (Tumor necrosis factor-α) in blood, which are increased by stress induction by the following method An experiment to analyze the effect was performed, and the results are shown in FIGS. 3 and 4. When stress is induced, cytokines IL-6 and TNF-α are secreted in the body, so they are well known as representative blood stress indicators.

In accordance with the preparation of the experimental animal model, the normal group, the control group, the Baeam Chazgi group 1 and Baeam Chazgi group 2 were reared, blood was collected after 12 weeks of breeding was completed, and the collected blood was centrifuged to obtain serum. , This was taken as a sample.

Quantitative analysis was performed using a mouse enzyme-linked immnunosorbent assay (ELISA) kit. The sample was placed on a plate coated with a monoclonal antibody, and then reacted at room temperature for 150 minutes in a 400 rpm rotary incubator, and then washed three or more times with the provided washing buffer. Then, polyclonal antibody was added to attach IL-6 (abcam, ab119548) to the plate, reacted at room temperature for 1 hour at 400 rpm, washed three or more times with the provided washing buffer, and then added the conjugate solution of IL-6. It was reacted for 30 minutes at room temperature at 400 rpm. The reacted plate was washed three or more times with a washing buffer and color was developed at room temperature with a substrate solution, and then a stop solution was added to stop the color reaction, and the absorbance was measured at 450 nm. In addition, an experiment for TNF-α (abcam, ab46070) was performed in the same way.

Compared to the normal group, the control group had a 25% increase in blood IL-6 (pg/mL) levels. In addition, the level of IL-6 (pg/mL) in the blood was decreased by 15% in the group 1 of the embryonic challenge, compared to the control group, and the level of IL-6 (pg/mL) in the blood of the group 2 decreased by 10% compared to the control group. (See Fig. 3).

In addition, compared to the normal group, the control group had a 31.1% increase in blood TNF-α (pg/mL) levels. In addition, the level of TNF-α (pg/mL) in the blood was decreased by 1% in the group 1 of the embryonic challenge, and the level of TNF-α (pg/mL) in the blood of the group 2 decreased by 23% compared to the control group. (See Fig. 4).

The above results show that the levels of IL-6 and TNF-α in the blood of rats are increased due to the pain stress caused by performing a knee incision, but the stress is suppressed and the blood IL-6 and TNF-α levels are suppressed when the extract of Bacillus Chazgi is administered to the stress-induced rats. It indicates a decrease in the -α level. From this, it can be seen that the baiam chazugi extract of the present invention has a modulating effect of IL-6 and TNF-α in blood, which is increased by inducing stress, and thus can be usefully used as a health functional food for improving stress.

<Experimental Example 4> Analysis of the modulating effect of MMP in blood increased by inducing stress

In order to confirm whether the Baiam chazugi extract according to the present invention has a stress-inhibiting effect, an experiment was conducted to analyze the modulating effect of matrix metallopeptidase (MMP) in blood, which is increased by inducing stress in the following method, and the results are shown in FIG. 5 It is shown in to 8. Since it is well known that the MMP level in the body increases when stress is induced, experiments for MMP-2, MMP-3, MMP-8, and MMP-13 in blood were performed as follows.

According to the preparation of the experimental animal model, a normal group, a control group, a group of 1 group of Baeam Chazugi and 2 group of Baeam Chazugi were bred, blood was collected after 12 weeks of breeding, and the collected blood was centrifuged to obtain serum. .

Dispense 100 μL of rat serum into each well in a 96 well plate of the MMP-8 ELISA kit (abcam, ab213910), react at 37°C for 120 minutes, and dispense 250 μL of wash buffer into each well. Leave to be washed for 1 minute to 2 minutes, and repeat the process of wiping off on a sterile tissue 5 or more times. Apply 100 μL of biotinylated antibody to each well, react at 37°C for 60 minutes, dispense 250 μL of wash buffer, leave to wash for 1 to 2 minutes, repeat the process of wiping off on sterile tissues 5 or more times. After that, it is left to dry for at least 5 minutes.

100 μL of Streptavidin peroxidase was dispensed into each well and reacted at 37° C. for 30 minutes, then 250 μL of streptavidin peroxidase was dispensed into each well and washed for 1 minute to 2 minutes, and shaken off with a sterile tissue. 100 μL of 3,3',5,5'-tetramethylbenzidine substrate was dispensed into each well, left at 37°C for 20 minutes, and 100 μL of stopping solution was dispensed. After confirming the stopping reaction changing from blue to yellow, the absorbance (optical density) of each well was measured at 450 nm in a microwell plate reader (Molecular Devices, Sunnyvale, CA, USA) within 30 minutes, and each concentration group for the control group The percentage of absorbance was calculated and evaluated as the relative activity of MMP-8. In addition, the quantitative experiments of MMP-2, MMP-3, and MMP-13 were performed in the same manner as this except for using the ELISA kit corresponding to each MMP.

Compared to the normal group, the control group had a 3.4% increase in blood MMP-2 (ng/mL) levels. In addition, the level of MMP-2 (ng/mL) in the blood was decreased by 4% in the group 1, compared to the control group, and in the group 2, the level of MMP-2 (ng/mL) in the blood was decreased by 9% compared to the control group (Fig. 5). Reference).

In addition, compared to the normal group, the control group had a 64.5% increase in blood MMP-3 (ng/mL) levels. In addition, the blood MMP-3 (ng/mL) level was reduced by 40% in the group 1, compared to the control group, and in the group 2, the level of MMP-3 (ng/mL) in the blood was decreased by 47% compared to the control group (Fig. 6). Reference).

In addition, compared to the normal group, the control group had a 39.1% increase in blood MMP-8 (ng/mL) levels. And, compared with the control group, the level of MMP-8 (ng/mL) in the blood MMP-8 (ng/mL) was decreased by 18% in the group 1, and the level of MMP-8 (ng/mL) in the blood was decreased by 7% in the group 2 (Fig. 7). Reference).

Finally, compared to the normal group, the control group had a 40.6% increase in blood MMP-13 (ng/mL) levels. In addition, the level of MMP-13 (ng/mL) in the blood was decreased by 24% in the group 1 of the embryonic challenge, and the level of MMP-13 (ng/mL) in the group 2 decreased by 23% compared to the control group (Fig. 8). Reference).

The above results show that the levels of MMP-2, MMP-3, MMP-8, and MMP-13 in the blood of the rats increased due to pain stress caused by performing a knee incision. It was suppressed, indicating a decrease in the levels of MMP-2, MMP-3, MMP-8 and MMP-13 in the blood. From this, it can be seen that the baiam chazugi extract of the present invention has a modulating effect of increasing blood MMP due to stress induction, and thus can be usefully used as a health functional food for improving stress.

<Experimental Example 5> Analysis of the modulating effect of lipocalin-2 in blood increased by stress induction

In order to confirm whether the extract according to the present invention has a stress suppressing effect, an experiment was performed to analyze the modulating effect of lipocalin-2 in blood, which is increased by inducing stress by the following method, and the results are shown in FIG. 9. I got it. Since it is well known that lipocalin-2 is upregulated in the body by stress, the following experiment was performed.

According to the preparation of the experimental animal model, a normal group, a control group, a group of 1 group of Baeam Chazugi and 2 group of Baeam Chazugi were bred, blood was collected after 12 weeks of breeding, and the collected blood was centrifuged to obtain serum. . Thereafter, a quantitative experiment was performed in the same manner as in Experimental Example 4, except that the lipocalin-2 ELISA kit was used.

Compared to the normal group, the control group had a 2.6% increase in the level of lipocalin-2 (ng/mL) in the blood. In addition, the level of lipocalin-2 (ng/mL) in blood was decreased by 3% in the group 1 of the embryonic challenge, and the level of lipocalin-2 (ng/mL) in the blood of the group 2 decreased by 7% compared to the control group (Fig. 9). Reference).

The above results indicate that the level of lipocalin-2 in the blood of the rat is increased due to the pain stress caused by performing a knee incision, but the level of lipocalin-2 in the blood is reduced by suppressing the stress when feeding the extract of Bacillus Chazugi to the stress-induced rat. . From this, it can be seen that the baiam chazugi extract of the present invention has a modulating effect of lipocalin-2 in blood, which is increased by inducing stress, and thus can be usefully used as a health functional food for stress improvement.

Claims (12)

Health functional food for improving the lowering of immunity due to stress, containing pear-am chazugi extract as an active ingredient.
The method of claim 1,
The baeam chazugi extract is a health functional food for improving immunity reduction due to stress, characterized in that the extract is extracted with _{water, C 1} to C _{3 lower alcohol or a mixed solvent thereof.}
The method of claim 2,
The baeam chazugi extract is a health functional food for improving immunity reduction due to stress, characterized in that extracted with ethanol or an aqueous ethanol solution.
The method of claim 1,
The baeam chazugi extract is a health functional food for improving immunity reduction due to stress, characterized in that it suppresses weight loss due to stress.
The method of claim 1,
The baeam chazugi extract is a health functional food for improving immunity decrease due to stress, characterized in that it suppresses the decrease in immune function due to stress.
The method of claim 1,
The baeam chazugi extract is a health functional food for improving immunity decrease due to stress, characterized in that it reduces cytokines increased due to stress.
The method of claim 6,
The cytokine is a health functional food for improving immunity reduction due to stress, characterized in that at least one selected from the group consisting of IL-6 and TNF-α.
The method of claim 1,
The baeam chazugi extract is a health functional food for improving immunity reduction due to stress, characterized in that it reduces the MMP (Matrix metallopeptidase) increased due to stress.
The method of claim 8,
The MMP is a health functional food for improving immunity reduction due to stress, characterized in that at least one selected from the group consisting of MMP-2, MMP-3, MMP-8 and MMP-13.
The method of claim 1,
The baeam chazugi extract is a health functional food for improving immunity reduction due to stress, characterized in that reducing the lipocalin increased due to stress.
The method of claim 10,
The lipocalin is a health functional food for improving immunity reduction due to stress, characterized in that lipocalin-2.
A pharmaceutical composition for improving the lowering of immunity due to stress, comprising the extract of Baeam Chazugi as an active ingredient.

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