KR20220100842A - Health functional food for ameliorating stress comprising extract of black rice aleurone - Google Patents

Abstract

The present invention relates to a health functional food for relieving stress comprising black rice aleurone extract, wherein the black rice aleurone extract suppresses weight loss and decline in immune function caused by stress, and has an excellent effect of adjusting a secretion of IL-6, TNF-α, MMP-2, MMP-3, MMP-7, MMP-13, and lipocalin-2 for which increase due to stress, thereby being usefully used as the health functional food for relieving stress.

Description

Health functional food for ameliorating stress comprising extract of black rice aleurone

The present invention relates to a health functional food for improving stress, and more specifically, to a health functional food for improving stress comprising an extract of black rice aloe.

In modern society, people complaining of anxiety disorders and mental disorders due to various stresses are increasing. In general, our living body causes various reactions against stress, which causes changes in the endocrine system and the autonomic nervous system. In particular, continuous or chronic stress accompanies psychological changes such as depression, anxiety, and anger, and the pathogenesis of various diseases such as mental disorders, central nervous system, immune system disorders, digestive disorders such as neurogenic gastric ulcer, and cardiovascular disorders such as angina pectoris. are closely related.

Chronic psychological stress results in 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) predicts that by 2020, depression will be It is estimated to be the most prevalent disease. In Korea, as urbanization accelerates, the inequality between the rich and the poor is deepening, and the incidence of depression is rapidly increasing. Currently, Korea has one of the highest suicide rates among OECD countries.

Specifically, depression (or severe depressive disorder) is a serious mental disorder accompanied by physical and cognitive dysfunction along with subjective emotions such as sadness and helplessness. The most core symptom of depression is loss of interest and pleasure in universal stimuli. When depressed, daily life becomes difficult, resulting in enormous social and economic losses, and death as 60 to 70% of suicide causes 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 have less than expected effects.

It usually takes about 2 weeks to improve symptoms such as improvement of depression by administering antidepressants. Specifically, tricyclic antidepressants such as imipramine and desipramine show severe side effects such as low blood pressure and cardiac dysfunction, and are the most widely used selective agents such as fluoxetine or sertraline. Serotonin uptake inhibitors cause nausea, gastrointestinal bleeding, or sexual dysfunction. Therefore, new drugs that are more effective than existing antidepressants and have fewer side effects are continuously being tried, but the development cost is huge, so pharmaceutical companies cannot easily develop them.

Therefore, it is necessary to develop a health functional food that can prevent stress stimulation from developing into a mental illness without the problems that appear in the antidepressant. Natural products contain various unknown ingredients and have the advantage of being relatively safe. , has a very high potential in improving stress. Referring to the specific prior art in this regard, grapefruit oil; And there is a patent (KR 10-2010-0106062 A) relating to a fragrance extract containing bergamot oil, which has a stress relief and soothing effect, and is said to be useful in various fields such as beauty and cosmetics. However, the stress in the above patent does not actually relieve mental stress as it provides an instant refreshing feeling.

On the other hand, black rice ( Oryza sativa , black rice) is a kind of raw rice, and China is the main production area. As a type of colored rice, it has a unique color and flavor. Anthocyanin, a water-soluble pigment known to have antioxidant, anticancer and anti-ulcer effects, gives it a black color. Black rice contains 4 times more anthocyanin than black soybean. It contains 5 times more than regular rice. Therefore, black rice is known to effectively neutralize free radicals in the body that cause aging and various diseases, as well as prevent heart disease, stroke, adult disease, and cancer.

Aleurone is also a term referring to the aleurone layer, which is a cell layer containing a large amount of aleurone granules. In addition to the storage of nutrients, the aloe layer secretes enzymes such as amylase and supplies them to the embryo, and contains enzymes related to the breakdown of protein granules. The aloe layer can be seen in the outermost layer of endosperm of seeds of Crestedaceae and Coconut plants, and has a high content of protein, fat, and vitamins.

Since there was no prior art for a health functional food for stress improvement containing black rice aloe extract, the present inventors were studying to find an extract for stress improvement from natural substances, black rice aloe extract reduced weight due to stress. The present invention was completed by confirming that there is an effect of inhibiting and regulating the secretion of cytokines, MMP, and lipocalin.

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.

In order to achieve the above purpose,

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

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

Black rice aloe extract of the present invention suppresses weight loss and immune function decline due to stress, and increases IL-6, TNF-α, MMP-2, MMP-3, MMP-7, MMP-13, lipocalin due to stress. As it has an excellent effect of regulating the secretion of -2, it can be usefully used as a health functional food for improving stress.

1 is a graph measuring the weight of rats in each group prepared in an experimental animal model after 12 weeks of breeding.
Figure 2 is a graph measuring the weight of the spleen that was removed after sacrificing the rats of each group prepared in the experimental animal model after 12 weeks of breeding.
Figure 3 is a graph of measuring the weight of the thymus obtained by sacrificing the rats of each group prepared in the experimental animal model after 12 weeks of breeding.
Figure 4 is a graph measuring the level of IL-6 in the blood by analyzing the serum obtained from the rats of each group prepared in the experimental animal model after 12 weeks of breeding.
5 is a graph measuring the level of TNF-α in the blood by analyzing the serum obtained from the rats of each group prepared in the experimental animal model after 12 weeks of breeding.
6 is a graph measuring the level of MMP-2 in the blood by analyzing the serum obtained from the rats of each group prepared in the experimental animal model after breeding for 12 weeks.
7 is a graph measuring the level of MMP-3 in the blood by analyzing the serum obtained from the rats of each group prepared in the experimental animal model after 12 weeks of breeding.
8 is a graph measuring the level of MMP-7 in the blood by analyzing the serum obtained from the rats of each group prepared in the experimental animal model after 12 weeks of breeding.
9 is a graph measuring the level of MMP-13 in the blood by analyzing the serum obtained from the rats of each group prepared in the experimental animal model after 12 weeks of breeding.
Figure 10 is a graph measuring the level of lipocalin-2 in the blood by analyzing the serum obtained from the rats of each group prepared in the experimental animal model after 12 weeks of breeding.

Hereinafter, the present invention will be described in detail.

One aspect of the present invention provides a health functional food for improving stress comprising an extract of black rice aloe.

The black rice aloe extract may be prepared in the form of liquid or powder.

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

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

The black rice aloe extract may be prepared in the form of a powder by adding an extraction solvent to the black rice aloe extract to prepare an extract, filtration, concentration under reduced pressure, and freeze-drying.

In the manufacturing method, black rice can be used without limitation, such as cultivated or commercially available ones.

In addition, the extraction solvent may be water, a C ₁ to C ₃ lower alcohol or a mixed solvent thereof, and specifically, the C ₁ to C ₃ lower alcohol 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 10 to 70 ℃, more preferably 30 to 50 ℃, but is not limited thereto. At this time, the extraction time is 10 to 96 hours; 20 to 82 hours; 30 to 70 hours; or 40 to 56 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 black rice flour used for extraction, more preferably adding 5 to 15 times for extraction, and adding 8 to 12 times for extraction is particularly preferable. Preferably, it is not limited thereto. In a range outside the conditions of the extraction time or the number of times of extraction, the extraction is not sufficiently performed, so that the content of the active ingredient in the black rice aloe extract may be insignificant. Alternatively, the extraction yield may no longer increase and the efficiency of the extraction operation may decrease.

On the other hand, the filtration may use a gravity filter, a vacuum filter, a pressure filter, a compression filter, a centrifugal filter, or a membrane separation filtration, but is not limited thereto, and the reduced pressure concentration may use a rotary reduced pressure concentrator, a vacuum reduced pressure 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 using the above-described extraction solvent, but also extracts obtained through other conventional extraction methods or extracts subjected to purification and fermentation processes. It includes not only extracts by decompression by carbon dioxide, extraction by supercritical extraction by high temperature, extraction by extraction using ultrasonic waves, but also fractions such as fractions using difference in solvent polarity for crude extracts. Separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography methods, or extracts obtained by fermentation products using natural conditions or various microorganisms are also included in the extracts of the present invention.

The black rice aloe extract can suppress weight loss due to stress.

In a specific embodiment of the present invention, the weight of the rat decreased due to the pain stress induced by performing the knee incision, but it was found that the stress was suppressed and the weight loss was suppressed when the black rice aloe extract was fed to the stress-induced rat (Experimental Example) 1 and Figure 1).

The black rice aloe extract can suppress the decrease in immune function due to stress. In a specific embodiment of the present invention, the weight of the spleen and thymus, which are immune tissues of the rat, is reduced due to the pain stress induced by performing knee incision. was inhibited (see Experimental Example 2, FIGS. 2 and 3).

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

The black rice aloe extract can reduce the increased matrix metallopeptidase (MMP) due to stress. Here, MMP may be one or more selected from the group consisting of MMP-2, MMP-3, MMP-7, and MMP-13.

The black rice aloe extract can reduce the increased lipocalin due to stress. Here, lipocalin may be lipocalin-2.

In a specific embodiment of the present invention, the levels of IL-6, TNF-α, MMP-2, MMP-3, MMP-7, MMP-13 and lipocalin-2 in the rat blood due to pain stress induced by knee incision. However, feeding black rice aloe extract to stress-induced rats suppressed stress and decreased the levels of IL-6, TNF-α, MMP-2, MMP-3, MMP-7, MMP-13 and lipocalin-2 in the blood. was found (see Experimental Examples 3 to 5 and FIGS. 4 to 10).

From this, the black rice aloe extract of the present invention has the effect of inhibiting weight loss and immune tissue weight reduction due to stress, and increasing the serum IL-6, TNF-α, MMP-2, MMP-3, MMP-7 by stress. , MMP-13 and lipocalin-2, since it has the effect of reducing it, it can be seen that it can be usefully used as a health functional food for improving stress.

As used herein, the term "health functional food" refers to food manufactured using raw materials or ingredients that are easily deficient in daily meals or have a useful function in the human body, and refers to food that helps maintain human health, but is not limited thereto. and is used in the meaning of including all health foods in the ordinary sense.

The form and type of 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 include 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 thaumatin, stevia extract, and the like. Meanwhile, examples of synthetic sweeteners include saccharin, aspartame, and the like. In addition, the natural carbohydrates may be monosaccharides, disaccharides, polysaccharides, oligosaccharides and sugar alcohols.

The health functional food of the present invention, in addition to the above-described additional ingredients, nutrients, vitamins, electrolytes, flavoring agents, colorants, pextan and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives , glycerin, alcohol, and the like may be further included. These components may be used independently or in combination. The proportion 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.

Black rice aloe extract of the present invention may be added to food as it is or used together with other foods or food ingredients. In this case, the content of the added active ingredient may be determined according to the purpose. In general, the content in 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 health and hygiene or health control, the amount may be less than 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 black rice aloe extract as an active ingredient.

The description of the black rice aloe extract is the same as the description of the black rice aloe extract in the health functional food.

The pharmaceutical composition for improving stress containing the black rice aloe extract 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. carriers, excipients or diluents. The pharmaceutically acceptable carrier is not particularly limited as long as it is suitable for in vivo delivery of the composition, and for example, is selected from the group consisting of saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol and ethanol. Any one or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostats may be added as needed. In addition, diluents, dispersants, surfactants, binders and lubricants may be additionally added to form injectable formulations such as aqueous solutions, suspensions, emulsions, powders, tablets, capsules, pills, granules, or injection solutions. 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 black rice aloe extract 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 is used. It is preferable to choose In order to formulate a formulation for parenteral administration, it is mixed with the composition in water to prepare a suspension, and it is formulated as a unit dosage form in an ampoule or vial.

The pharmaceutical composition for improving stress containing the black rice aloe extract 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, severity, activity of the drug, the type of patient's disease, Sensitivity to the drug, administration time, administration route and excretion rate, treatment period, factors including concurrent drugs and other factors well known in the medical field may be determined. The composition may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered single or multiple. In consideration of all of the above factors, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, which can be easily determined by those 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 only illustrate the present invention, and the content of the present invention is not limited by the following Examples and Experimental Examples.

<Example 1> Preparation of black rice aloe extract

The aleurone layer of black rice (Jindo black rice farming corporation, Jindo) ( Oryza sativa , black rice) was pulverized and extracted with 50% (v/v) alcohol 10 times its weight at 40° C. for 48 hours. . After filtering the extract with a filter paper, it was concentrated, lyophilized, and powdered, which was used in the following experiments.

<Experiment Preparation>

1. Preparation of experimental animal models

In order to confirm the stress improvement effect of black rice aloe extract, a stress-induced rat model was prepared. The normal group rat model used a Sprague-Dawley (SD) species male white rat, and the stress-induced rat model has pain stress by performing an anterior cruciate ligament transaction in the rat. During the breeding period of normal rats and stress-induced rats, the diet was a general solid feed (Central Experimental Animals, Seoul, Korea) and water, and the temperature of the breeding environment was 23±2℃ and the humidity was 50±10%. The light-dark cycle was adjusted in 12-hour increments.

The black rice aloe extract prepared in Example 1 was dissolved in a solvent (Phosphate Buffer Saline, PBS), and 150 mg of black rice aloe extract per 1 kg of stress-induced rat body weight was fed to group 1 of black rice aloe, and stress-induced to group 2 of black rice aloe. 300 mg of black rice aloe extract was fed per 1 kg of rat body weight, and the control group was fed only the same amount of PBS to stressed rats. That is, the experiment was carried out by dividing it into a normal group, a control group, black rice flour 1 group, and black rice flour 2 group.

They were bred for a total of 12 weeks, and after breeding was completed, the body weight was measured, anesthetized with CO ₂ , and blood was collected. The blood collected was placed in _a centrifugation tube and centrifuged at 2000 rpm, 4°C for 15 minutes to obtain serum. was used to measure its weight.

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

In order to confirm whether the black rice aloe extract according to the present invention has a stress suppressive effect, an experiment was performed to analyze the stress-induced weight loss inhibitory effect by the following method, and the results are shown in FIG. 1 .

For the normal group, control group, black rice flour 1 group and black rice flour 2 group prepared as in the experimental animal model, the body weight was measured at a fixed time every week for 12 weeks, and the weight at the 12th week at the end of the experiment period is shown in FIG. It was. The weight of the rat was measured and recorded in a state where the value was stabilized on a scale that can measure up to one decimal place.

Compared to the normal group, the control group lost 8.4% in body weight. And the black rice flour group 1 had a body weight of 0.4% higher than that of the control group, and the black rice flour group 2 group had a body weight that was 7% 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 induced by performing the knee incision, but when the black rice aloe extract is fed to the stress-induced rat, the stress is suppressed and the weight loss is suppressed. From this, it can be seen that the black rice aloe extract of the present invention has an effect of inhibiting weight loss due to stress, and thus can be usefully used as a health functional food for improving stress.

<Experimental Example 2> Analysis of the effect of suppressing the decrease in immune tissue weight due to stress induction

In order to confirm whether the black rice aloe extract according to the present invention has a stress inhibitory effect, an experiment was performed to analyze the effect of inhibiting the decrease in immune tissue weight caused by stress in the following manner, and the results are shown in FIGS. 2 and 3 . . When stress is induced, the function of the spleen or thymus, which is an immune tissue, is lowered, and immunity is abnormal. Therefore, the following experiment was performed.

According to the preparation of the experimental animal model, a normal group, a control group, black rice flour 1 group and black rice flour 2 group were reared, and after 12 weeks of breeding, they were euthanized. measured.

Compared with the normal group, the spleen weight of the control group was decreased by 13%. And the black rice flour group 1 had a spleen weight 10% higher than that of the control group, and the black rice flour group 2 group had a weight 12% higher than the control group (see FIG. 2).

In addition, the thymus weight of the control group was reduced by 23% compared to the normal group. And the black rice flour group 1 had a thymus weight that was 2% higher than that of the control group, and the black rice flour group 2 had a thymus weight that was 19% higher than that of the control group (see FIG. 3).

The above results indicate that the weight of the spleen and thymus of the rat is reduced due to the pain stress induced by performing the knee incision, but when the black rice aloe extract is fed to the stress-induced rat, the stress is suppressed, indicating that the weight loss of the spleen and thymus is suppressed. . From this, it can be seen that the black rice aloe extract of the present invention can be usefully used as a health functional food for improving stress because it has an inhibitory effect on the reduction of immune tissue weight caused by stress.

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

In order to check whether the black rice aloe extract according to the present invention has a stress-suppressing effect, the modulating effect of IL-6 (Interleukin-6) and TNF-α (Tumor necrosis factor-α) in the blood increased by stress induction by the following method An experiment was performed to analyze the , and the results are shown in FIGS. 4 and 5 . 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.

According to the preparation of the experimental animal model, a normal group, a control group, black rice flour 1 group and black rice flour 2 group were bred, blood was collected after 12 weeks of breeding, and the collected blood was centrifuged to obtain serum, which It was used as a sample.

Quantitative analysis was performed using a mouse enzyme-linked immnunosorbent assay (ELISA) kit. Samples were placed on the monoclonal antibody-coated plate, incubated at room temperature for 150 minutes in a 400 rpm rotary incubator, and washed three or more times with the provided washing buffer. Then, the polyclonal antibody was added to attach IL-6 (abcam, ab119548) to the plate, reacted at room temperature for 1 hour at 400 rpm, washed 3 times or more with the provided washing buffer, and then the IL-6 conjugate solution was added The reaction was carried out at 400 rpm at room temperature for 30 minutes. The reacted plate was washed three or more times with washing buffer, and the substrate solution was used to develop color at room temperature, 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 manner as above.

Compared to the normal group, the control group had a 25% increase in the level of IL-6 (pg/mL) in the blood. In addition, the level of IL-6 (pg/mL) in the blood was decreased by 8% in the black rice flour group 1 compared to the control group, and the level of IL-6 (pg/mL) in the blood in the black rice flour group 2 decreased by 11% compared to the control group (Fig. see 4).

In addition, compared to the normal group, the control group increased the level of TNF-α (pg/mL) in the blood by 31.1%. And the black rice flour group 1 had a 7% decrease in blood TNF-α (pg/mL) level compared to the control group, and the 2 group black rice flour reduced blood TNF-α (pg/mL) level by 24% compared to the control group (Fig. 5).

The above results show that the level of IL-6 and TNF-α in the blood increases due to the pain stress induced by knee incision, but when the black rice aloe extract is fed to the stress-induced rat, the stress is suppressed, so that the IL-6 and TNF- It indicates that the α level is reduced. From this, it can be seen that the black rice aloe extract of the present invention has a regulating effect of IL-6 and TNF-α in the blood increased by stress induction, and thus can be usefully used as a health functional food for improving stress.

<Experimental Example 4> Analysis of the regulatory effect of MMP in the blood increased by stress induction

In order to confirm whether the black rice aloe extract according to the present invention has a stress-suppressing effect, an experiment was performed to analyze the regulatory effect of MMP (Matrix metallopeptidase) in the blood increased by stress induction by the following method, and the results are shown in FIGS. 9 is shown. Since it is well known that the level of MMP in the body increases when stress is induced, experiments on MMP-2, MMP-3, MMP-7 and MMP-13 in the blood were performed as follows.

According to the preparation of the experimental animal model, a normal group, a control group, black rice flour 1 group and black rice flour 2 group were reared, and after 12 weeks of breeding was completed, blood was collected, and the collected blood was centrifuged to obtain serum.

100 μL of rat serum was dispensed into each well of a 96-well plate of the MMP-7 ELISA kit (abcam, ab213910), and after reacting at 37° C. for 120 minutes, 250 μL of wash buffer was dispensed into each well. Leave to wash for 1 to 2 minutes, and repeat the process of wiping off with a sterile tissue 5 times or more. 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 minute to 2 minutes, and repeat the shaking process with a sterile tissue at least 5 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 to wash for 1 to 2 minutes, and then wiped off with a sterile tissue. 100 μL of 3,3',5,5'-tetramethylbenzidine substrate was dispensed into each well, and after 20 minutes at 37°C, 100 μL of stopping solution was dispensed. After confirming the stopping reaction that changes 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-7. In addition, quantitative experiments of MMP-2, MMP-3, and MMP-13 were performed in the same manner as in the above, except that the ELISA kit corresponding to each MMP was used.

Compared to the normal group, the control group increased the level of MMP-2 (ng/mL) in the blood by 3.4%. In addition, the level of MMP-2 (ng/mL) in the blood was decreased by 8% in the black rice flour group 1 compared to the control group, and the level of MMP-2 (ng/mL) in the blood in the group black wheat flour 2 decreased by 6% (see FIG. 6). .

In addition, the control group increased the level of MMP-3 (ng/mL) in the blood by 42% compared to the normal group. In addition, the level of MMP-3 (ng/mL) in the blood was decreased by 31% in the black rice aloe group 1 compared to the control group, and the level of MMP-3 (ng/mL) in the blood in the group 2 was decreased by 20% (see FIG. 7). .

And, compared to the normal group, the control group increased the level of MMP-7 (ng/mL) in the blood by 7.8%. And, compared with the control group, the black rice aloe group 1 had a 20% decrease in blood MMP-7 (ng/mL), and the 2 group black rice flour reduced the blood MMP-7 (ng/mL) level by 18% (see FIG. 8). .

Finally, compared to the normal group, the control group increased the level of MMP-13 (ng/mL) in the blood by 40.6%. In addition, the level of MMP-13 (ng/mL) in the blood was decreased by 22% in the black rice flour group 1 compared to the control group, and the level of MMP-13 (ng/mL) in the blood in the group 2 was decreased by 26% (see FIG. 9). .

The above results show that the levels of MMP-2, MMP-3, MMP-7 and MMP-13 in the blood of rats increase due to pain stress induced by knee incision, but the stress is suppressed by feeding black rice aloe extract to stress-induced rats. and decreased blood levels of MMP-2, MMP-3, MMP-7 and MMP-13. From this, it can be seen that the black rice aloe extract of the present invention can be usefully used as a health functional food for improving stress, since it has a regulating effect of MMP in the blood increased by stress induction.

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

In order to confirm whether the black rice aloe extract according to the present invention has a stress-suppressing effect, an experiment was performed to analyze the regulatory effect of lipocalin-2 in the blood increased by stress induction by the following method, and the results are shown in FIG. 10 . . 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, black rice flour 1 group and black rice flour 2 group were reared, and after 12 weeks of breeding was completed, blood was collected, 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. And, compared to the control group, the black rice aloe group 1 had a 7% decrease in blood lipocalin-2 (ng/mL) level, and the 2 group black rice flour reduced the blood lipocalin-2 (ng/mL) level by 10% (see FIG. 10). .

The above results indicate that the level of lipocalin-2 in the blood of rats increases due to the pain stress induced by performing knee incision, but when the black rice aloe extract is fed to the stress-induced rats, the stress is suppressed and the level of lipocalin-2 in the blood is reduced. From this, it can be seen that the black rice aloe extract of the present invention has a regulating effect of lipocalin-2 in the blood increased by stress induction, and thus can be usefully used as a health functional food for improving stress.

Claims (9)

A health functional food for improving stress comprising, as an active ingredient, an extract of black rice aloe extracted with ethanol or 40 to 60% (v/v) ethanol aqueous solution.
The method of claim 1,
The black rice aloe extract is a health functional food for improving stress, characterized in that it suppresses weight loss due to stress.
The method of claim 1,
The black rice aloe extract is a health functional food for improving stress, characterized in that it reduces cytokines increased due to stress.
4. The method of claim 3,
The cytokine is a health functional food for improving stress, characterized in that at least one selected from the group consisting of IL-6 and TNF-α.
The method of claim 1,
The black rice aloe extract is a health functional food for improving stress, characterized in that it reduces the increased matrix metallopeptidase (MMP) due to stress.
6. The method of claim 5,
The MMP is a health functional food for improving stress, characterized in that at least one selected from the group consisting of MMP-2, MMP-3, MMP-7 and MMP-13.
The method of claim 1,
The black rice aloe extract is a health functional food for improving stress, characterized in that it reduces lipocalin increased due to stress.
8. The method of claim 7,
The lipocalin is a health functional food for improving stress, characterized in that it is lipocalin-2.
A pharmaceutical composition for improving stress comprising the black rice aloe extract of claim 1 as an active ingredient.

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