KR20180047602A - FOOD COMPOSITION CONTAINING HYDROLYSATE DERIVED FROM Hippocampus abdominalis AND EXTRACT OF RED GINSENG FOR RELIEVING FATIGUE - Google Patents

Abstract

The present invention provides a health functional food composition for relieving fatigue and a health food for relieving fatigue containing a mixture of Hippocampus abdominalis hydrolysate and red ginseng extract, wherein Hippocampus abdominalis dried sample is treated with Alcalase protein hydrolyzing enzyme to obtain a hydrolysate, and the hydrolysate and red ginseng extract are mixed at a weight ratio of 1:1.5-4, and a concentration of the mixture is 200-500 μg/ml. The composition has an effect of reducing and relieving fatigue accumulation.

Description

TECHNICAL FIELD [0001] The present invention relates to a health functional food composition for improving fatigue, which contains a mixture of a hydrolyzate of hippocampus and a red ginseng extract. BACKGROUND OF THE INVENTION 1. Field of the Invention [0002]

The present invention relates to a health functional food composition containing a mixture of a hippocampus hydrolyzate and a red ginseng extract prepared using a hydrolyzate, more particularly, to a horseradish hydrolyzate prepared by using an Alcalase hydrolyzing enzyme, The present invention relates to a health functional food composition for improving fatigue containing a mixture of hippocampal hydrolyzate and red ginseng extract having an effect of improving fatigue by mixing hippocampal hydrolyzate and red ginseng extract in accordance with the domestic situation which should be used at 50% or less in food.

The hippocampus is a tibia fish belonging to the spindle group, and among the 52 genera 215 species, 32 species of hippocampus are classified. In Korea, there are five species of hippocampus, hippocampus, coral hippocampus, and coral hippocampus.

In Europe and North America, horticulture has been used as a medicine in Southeast Asia, including China, because it is said to be effective for asthma, liver dysfunction, sexual dysfunction and general pain, while ornamental use and drying to commercialize it as a souvenir or antique.

The major constituents of the hippocampus are known as steroids, minerals and amino acid peptide fatty acids, and they focus on the efficacy of the hippocampus as drugs. Conventional studies have shown that anti-cancer, anti-aging, antithrombotic, antiarthritic, antioxidant, erectile dysfunction, And the effect was revealed.

Hippocampus abdominalis belongs to Syngnathifromes and Syngnathidae. It lives in temperate and tropical waters. It has the largest size among hippocampus and has beautiful body color and body shape, It is also popular in the coronary market.

Hippocampus farming has already been tried in New Zealand, Australia and China, and in recent years, full breeding of the Big Veli hippocampus designated as edible has been successful in Jeju. However, the research on physiological activity of the Bigbellie hippocampus has been lacking compared with that of the Hippocampus kuda Bleeker, Hippocampus guttu- latus and Short-snouted seahorse (Hippocampus hippocampus) species,

The inventors of the present invention have found that the production of Bigvalley hippocampal hydrolysates using two digestive enzymes (trypsin, pepsin) and five food hydrolytic enzymes (neutrase, protamex, alcalase, koji-zyme and flavourzyme) It is known to cause inflammation and damage intracellular tissues by using fractions of respective molecular weights derived from hydrolyzate and causes diseases such as aging and cancer, diabetes, stroke, arteriosclerosis, cardiovascular disease, anemia, gastric ulcer and dementia Studies on the antioxidative activity of the hippocampus by confirming the scavenging ability against the free radicals have been conducted, but research on the anti-fatigue efficacy of the hydrolyzate of the hippocampus of the present invention has not been conducted.

Fatigue can be divided into physical fatigue and mental fatigue. Symptoms can vary from helpless to drowsiness, muscle aches, and body aches accompanied by chills. Repeated fatigue can cause chronic fatigue syndrome. Modern people in a busy life are becoming tired easily due to lack of activity, mental stress, lazy habits and obesity, and interest and desire to maintain health without disease are getting higher and higher.

In general, fatigue causes a rapid increase in oxygen demand for proper energy supply of cells during exercise. If the metabolism of the cells does not satisfy the oxygen demand, the cells become temporarily supplied with energy by oxygen-free metabolism, Causing fatigue. Thus, many people prefer to take health-functional foods to supplement their nutritional support for fatigue, health promotion and disease prevention.

In order to satisfy the condition that the use of hippocampus for food is limited to less than 50%, the present invention relates to a method for determining the compounding ratio of the best hippocampal hydrolyzate having a synergistic effect by combining with red ginseng which has already been recognized as having an anti- Thus, it is intended to provide a composition for improving fatigue including hydrolysates of Bigbell horse seaweed and extracts of red ginseng.

Korean Patent Laid-Open No. 10-2009-0094072 relates to a peptide for treating or preventing arthritis having an amino acid sequence LEDPFDKDDWDNWK isolated from hippocampal protein-derived enzyme hydrolyzate, and a pharmaceutical and health functional food composition containing the peptide as an active ingredient , The hippocampal hydrolyzate derived peptide according to the present invention induces differentiation of osteogenic MG-63 and cartilage-absorbing SW-1353 cells and inhibits NO production and inhibits p38 kinase / NF-κB and SW-1353 The present invention provides a harmful peptide of hippocampal hydrolyzate for treating arthritis, which can effectively reduce MAP Ks / NF-κB in cells and can be used as a therapeutically effective agent capable of controlling the differentiation and inflammatory activity of arthritis . In Korean Patent Laid-Open Publication No. 10-2014-0042148, seahorse-derived biofunctional peptides are capable of protecting brain cells from neurotoxic substances and oxidative stress, inhibiting the expression of inflammatory mediators and expressing anti-inflammatory cytokines To prevent neurodegenerative diseases including degenerative neurodegenerative diseases including an active ingredient of a bioactive peptide capable of inhibiting the expression of cathepsin B and cathepsin D, which are involved in the production of neurotoxin A? 42, Or therapeutic compositions. Korean Patent No. 10-1596728 discloses a food composition and a pharmaceutical composition for preventing and ameliorating menopausal symptoms including a zinc / red ginseng mixture, an octacosanol mixture, and a vitamin mixture. The present invention also relates to a zinc / red ginseng mixture, Kits for the prevention and amelioration of male menopausal symptoms comprising a mixture of octacosanol and vitamin and a kit for the prevention and treatment of male menopausal symptoms comprising zinc / red ginseng, octacosanol and vitamins. Korean Patent Laid-Open Publication No. 10-2003-0041370 relates to a herbal composition for treating and preventing sexual dysfunction, including medicinal herb medicine, antler, echinacea, phlegm, lysozyme, ginseng and hippocampus, wherein the herbal composition is selected from the group consisting of corn oil, A herbal composition for the treatment of sexual dysfunction which can further comprise at least one ingredient selected from the group consisting of liquorice, Angelica keiskei, Bacillus subtilis, Jahragah, coriander, and seaweed as auxiliary ingredients. However, the prior art does not disclose the structure related to the health functional food composition for improving fatigue, which contains a mixture of the hippocampus hydrolyzate and the red ginseng extract of the present invention.

According to the present invention, hyaluronic acid hydrolyzate prepared by using Alcalase hydrolysis enzyme is mixed with red ginseng extract, which is known for its fatigue improvement effect, to be 50% or less And a mixture of the hippocampal hydrolyzate and the red ginseng extract, which has the maximum fatigue improving effect while satisfying the content of the hyaluronic acid hydrolyzate and the red ginseng extract.

As a means for solving the above problem, the present invention provides a health functional food composition for improving fatigue containing a mixture of hippocampal hydrolyzate and red ginseng extract. The hippocampus is Hippocampus abdominalis , the hippocampus hydrolyzate is obtained by treating the hippocampal dry sample with Alcalase protein hydrolyzing enzyme, the hippocampus hydrolyzate and the red ginseng extract are mixed at a weight ratio of 1: 1.5-4, and the concentration of the mixture is 200 to 500 ㎍ / ml .

Also provided is a health food for improving fatigue containing a mixture of hippocampus hydrolyzate and red ginseng extract containing the composition.

As described in the following examples, a health functional food composition for fatigue improvement containing a mixture of hippocampal hydrolyzate and red ginseng extract was treated with a mouse-derived myoblast C2C12 cell line to confirm cell proliferation efficacy, When the hydrolyzate and red ginseng extract were combined at a weight ratio of 1: 1.5-4, it was confirmed that the activity of improving fatigue was higher than that of red ginseng 100% extract.

In the in vivo model, glycogen and ATP content and lactate content were measured by using Zebrafish. When the hippocampus hydrolyzate and red ginseng extract were combined at a weight ratio of 1: 1.5-4, the content of glycogen was high and the fatigue inducing substance, L- Lactate content was remarkably low and ATP content was high.

The results of the above Examples show that the composition of the present invention containing the mixture of hippocampus hydrolyzate and red ginseng extract at a weight ratio of 1: 1.5-4 and containing a concentration of 200 to 500 / / ml has the effect of improving fatigue .

FIG. 1 shows the cytotoxicity evaluation results of the concentration ratio of Alcalase hydrolyzate and red ginseng extract in horses.
Fig. 2 shows the cell proliferation efficacy results by the concentration ratio of Alcalase hydrolyzate and red ginseng extract of hippocampus.
FIG. 3 shows fatigue-improving efficacy results by the concentration ratio of Alcalase hydrolyzate and red ginseng extract.
Figure 4 shows a running water bath for measuring fatigue improving activity in an in vivo model.
Figure 5 shows the results of a swimming analysis using the EthoVision system of Zebrafish, an in vivo model.
Figure 6 shows the glycogen content in the in vivo model zebrafish muscle.
Fig. 7 shows the content of L-lactate in the in vivo model zebrafish muscle.
Figure 8 shows the ATP content in the zebrafish muscle, an in vivo model.

Hereinafter, the present invention will be described in detail with reference to specific examples.

One. Big Valley  hippocampus Hydrolyzate and  Preparation of a composition mixed with red ginseng extract

1.1 Big Valley  hippocampus Hydrolyzate  Produce

The experimental fishes used in the examples of the present invention were the Big-belly hippocampus ( Hippocampus abdominalis ) were lyophilized, pulverized and stored at -70 ° C.

Alcalase hydrolysis enzyme was used to adjust the substrate to enzyme ratio to 100: 1. Hydrolysis was performed for 24 hours. The enzyme was inactivated at 100 ° C for 10 minutes to stop the enzyme reaction, and the pH was adjusted to 7.0. After centrifugation for 20 minutes, the supernatant, except for the residue, was filtered through filter paper, lyophilized, and powdered.

In addition, the red ginseng extract used in the present invention was purchased from a commercial concentrated red ginseng concentrate. The hippocampus hydrolyzate and red ginseng extract were mixed at a weight ratio of 1: 1.5-4.

1.2 Cytotoxicity assay

The mouse-derived myoblast C2C12 cell line was cultured in DMEM (Dulbeccos'modified Eagle'medium) medium containing 10% bovine serum and L-glutamine at 37 ° C in a 5% CO ₂ incubator.

The cells were divided into 48 well plates at 5 × 10 ⁴ cells / ml, and after 16 hours, they were changed to DMEM containing 10% horse serum. The samples were treated with the hydrolyzate of Alcalase hydrolyzate and red ginseng extract (20% to 80% based on the mixing ratio of red ginseng) and the concentration (50, 100, 200, 250 and 500 / / ml) For example, the red ginseng extract content of 20% means that the hydrolysis extract of hippocampus consists of 80%.

The MTT assay was performed to measure the cytotoxicity and the principle was that the soluble yellow MTT [3- (4,5-dimethylthiaxol-2-yl) -2, 5-diphenyl tetrazolium bromide] is reduced to measure purple colored water-insoluble formazan.

After 24 hours, add 50 μl of MTT solution (2 mg / ml) and allow to react for 2 hours. The supernatant was completely removed and dimethylsulfoxide (DMSO) was added to completely dissolve the precipitate and the absorbance was measured at 540 nm.

FIG. 1 shows the cytotoxicity evaluation results of the concentration ratio of Alcalase hydrolyzate and red ginseng extract in horses. As a result of the experiment, it was confirmed that there was no cytotoxicity at the treatment rate of the hyaluronic acid Alcalase hydrolyzate and red ginseng extract.

2. Seahorse Alcalase Hydrolyzate and  Improvement of fatigue of composition composed of red ginseng extract

2.1 Cell proliferation efficacy

The mouse-derived myoblast C2C12 cell line was cultured in DMEM (Dulbeccos'modified Eagle'medium) medium containing 10% bovine serum and L-glutamine at 37 ° C in a 5% CO ₂ incubator.

The cells were divided into 48 well plates at 5 × 10 ⁴ cells / ml, and after 16 hours, they were changed to DMEM containing 10% horse serum. 300, 400, 500 ㎍ / ㎖) for each concentration of horseradish Alcalase and red ginseng extract (0%, 20%, 40%, 60%, 80% ml and cultured for a total of 4 days.

During the 4-day incubation, the medium was replaced once every two days and the samples were again treated. MTT assay was used to measure cell proliferation after 4 days of culture.

Fig. 2 shows the cell proliferation efficacy results by the concentration ratio of Alcalase hydrolyzate and red ginseng extract of hippocampus. As a result, the cell proliferation efficiency was highest at 200 and 250 ㎍ / ml when the ratio of the hyaline Alcalase hydrolyzate and the red ginseng extract was 0% and 20%, respectively. As a result, the hippocampal Alcalase hydrolyzate itself exhibited cell proliferation It seems to be effective.

However, as described above, in order to use hippocampus as a food in domestic, it is necessary to use less than 50% of hippocampus extract. Therefore, the ratio of hydrolyzate of hippocampal Alcalase hydrolyzate and red ginseng extract is 60% (60% of red ginseng extract, 40%), 80% (red ginseng extract 80%, hippocampal hydrolysis 20%). When the blending ratio was 60% and 80%, the cell proliferation efficiency was increased depending on the concentration.

The cell proliferation efficiency of mouse derived myoblasts is indirectly shown by the fact that when the proliferation of muscle cells is increased, the amount of muscle mass is increased, so that the mixture ratio of Alcalase hydrolyzate and red ginseng extract of 60% and 80% .

2.2 Fatigue improving activity

The mouse-derived myoblast C2C12 cell line was cultured in DMEM (Dulbeccos'modified Eagle'medium) medium containing 10% bovine serum and L-glutamine at 37 ° C in a 5% CO ₂ incubator. The cells were divided into 48 well plates at 5 × 10 ⁴ cells / ml, and after 16 hours, they were changed to DMEM containing 10% horse serum.

(200, 250, 300, 400, 500 ㎍ / ml) of the hydrolysates of Alcalase hydrolyzate and red ginseng extract (0%, 20%, 40%, 60%, 80% And cultured for a total of 4 days.

During the 4-day incubation, the medium was replaced once every two days and the samples were again treated. After 4 days of incubation, the media and samples were changed. After 1 hour, the cells were treated with 100 μM H ₂ O ₂ as a fatigue inducing substance and MTT assay was performed to measure fatigue improving activity 24 hours later.

FIG. 3 shows fatigue-improving efficacy results by the concentration ratio of Alcalase hydrolyzate and red ginseng extract. As a result of the above experiment, it was found that the fatigue improving effect was better than that of 100% of red ginseng when the ratio of hyaluronic acid hydrolyzate and red ginseng extract was 60% and 80%, respectively. And a significant difference in activity was observed between the two groups. These results suggest that the combination of hyaline Alcalase hydrolyzate and red ginseng extract has a synergistic effect on fatigue improving activity.

2.3 In vivo  Evaluation of fatigue improvement activity in model

Zebrafish was used as an in vivo model, and zebrafish was vertebrae of similar genetic homology to humans. Figure 4 shows a running water bath for measuring fatigue improving activity in an in vivo model. Fifty animals of Zebrafish were divided into groups according to the mixing ratio of Alcalase hydrolyzate and red ginseng extract (Control, 0%, 60%, 80%, 100% (500 / / ml).

Control group is a group that does not feed Alcalase hydrolyzate and red ginseng extract. The flow rate of the aquarium is 0.3 to 0.4 m ² / sec. When divided into zones 1 and 2, every two days of exercise is monitored for 30 minutes using the EthoVision® system to determine which zone the zebrafish stays in and travels more. And the muscle tissue of zebrafish was obtained on the eighth day after the start of the experiment. Analysis of glycogen and ATP content and lactate content in muscle were analyzed.

Figure 5 shows the results of a swimming analysis using the EthoVision system of Zebrafish, an in vivo model. As a result of the experiment, the result of (A) shows the number of zebrafish movements in Zone 1 and Zone 2, and the movement of water flow from Zone 1 to Zone 2 can be seen in the running water tank. And thus it can be confirmed that fatigue improving effect is obtained.

The frequency of staying in zone 1 increased and the frequency of staying in zone 2 decreased from 0% and 100% when the ratio of Alcalase hydrolyzate and red ginseng was 60% and 80%, respectively. The result of (B) shows the same pattern as (A) as a result of analyzing the time of zebrafish staying in each zone.

Figure 6 shows the glycogen content in the in vivo model zebrafish muscle. Analysis of glycogen content in the muscles of zebrafish revealed that glycogen content was higher than that of 0% and 100% when the ratio of hydrolysates of Alcalase hydrolyzate and red ginseng was 60% and 80%, respectively.

These results suggest that the higher the glycogen content, the more energy is present in the muscle. Therefore, it is considered that fatigue is less when the ratio of Alcalase hydrolyzate and red ginseng extract is 60% and 80%.

Fig. 7 shows the content of L-lactate in the in vivo model zebrafish muscle. As a result of the above experiment, the content of L-lactate present in the muscle of zebrafish was analyzed. L-lactate is one of the most important factors in the measurement of fatigue-improving activity as it induces fatigue when intracellular production and accumulation occurs.

L-lactate content was significantly lower than that of 0% and 100% when the ratio of Alcalase hydrolyzate and red ginseng extract was 80%.

Figure 8 shows the ATP content in the zebrafish muscle, an in vivo model. As a result of the above experiment, the result of analyzing the ATP content in the muscle in zebrafish. In general, when the cells are metabolized smoothly, the production of lactate is increased and the production of ATP is increased.

These results show that when the compounding ratio of Alcalase hydrolyzate and red ginseng extract is 80%, the ATP content is higher than 0% and 100%, and the fatigue accumulation is not observed.

By providing the functional food containing the Big Bell hippocampus and the red ginseng extract, which are known to be effective in improving the fatigue, the functional food product development industry as well as the aquaculture industry can be developed and the effect of the income increase can be industrially applicable .

Claims (4)

Hippocampus abdominalis ) health-functional food composition for improving fatigue containing a mixture of hydrolyzate and red ginseng extract
[Claim 2] The method according to claim 1, wherein the hippocampus and red ginseng extract are mixed at a weight ratio of 1: 1.5-4 and the hippocampus hydrolysis extract is less than 50 wt% Health functional food composition for improving fatigue
3. The fatigue-improving health functional food composition according to claim 2, wherein the concentration of the mixture is 200 to 500 占 퐂 / ml, wherein the composition contains a mixture of hydrolysates of hippocampus and red ginseng extract
A fatigue-improving health food containing a mixture of a hippocampal hydrolyzate and a red ginseng extract comprising the composition of any one of claims 1 to 3

Images