KR20190078911A - Food composition for improvement of aging and immunity with the hot water extract of Cirsium setidens Nakai and Ligularia fischeri - Google Patents

Abstract

The present invention relates to a food composition for alleviating aging or for enhancing immune function, which comprises a hot water extract of Cirsium setidens and Ligularia fischeri. The composition of the present invention exhibits excellent aging alleviation efficacy and immune function enhancing efficacy.

Description

TECHNICAL FIELD The present invention relates to a food composition for enhancing aging or improving immune function, which comprises a hot water extract of Gonad and a hot water extract of Ganoderma lucidum.

The present invention relates to a food composition for improving the aging function or improving the immune function, and more particularly, to a food composition for improving the aging function or improving the immune function, which comprises a ghodnia hot-water extract and a hot water extract.

Gondeure consider Thistle (Cirsium genus Chris Leeum (Cirsium) setidens Nakai. It contains polyphenols, dietary fibers, minerals and vitamins, and is known to exhibit various physiological activities including anticancer activity.

The gondol is picked up every month in May and is used for food. The soft young leaves and soft stems that spring up in the spring are used as a bit of herb or soup, dried and dried. Fried, stuffed, roasted, and dipped in the shell of the stem is used to remove the cooked, has a unique flavor, and it was also used as a car to feel good.

On the other hand, Asteraceae is a kind of stigma characterized by broad leaves of Compositae, and its scientific name is Ligularia fischeri ), and is native to Japan, China, and eastern Siberia. In Korea, it grows mainly in the deep mountainous highlands of the country, and artificial cultivation is mainly performed in Gangwon province. It is widely used in the form of green herbs, herbs and spices, or green juice.

It has a relatively high nutrient content compared to other vegetables, because it contains various vitamins and minerals. It has been known for some time that cough, sputum, leg pain, back pain, headache, whooping cough, and asthma are effective, and blood circulation is active. In the private sector, it was used for jaundice, hypertension, liver disease, and infectious dermatoses and dandruff. In addition, proliferation inhibitory effects of cancer extract on cancer cells have been reported.

Korean Patent Registration No. 10-1470888 (Registration Date 2014.12.08) discloses a liver protection and liver function improving food containing fermented sea mustard which is cultured by inoculation with Lactobacillus planta (Lactobacillus plantarum) Compositions are described. Korean Patent Laid-Open Publication No. 10-2017-0115168 (published on Oct. 17, 2017) discloses a drying method for drying a ghred having excellent antioxidant ability, and describes a drying condition in which the ghdra palatability and quality are excellent.

The present invention aims to develop and provide a food composition capable of improving aging and improving immunity by using ghdra or shiitake.

The present invention is characterized in that it comprises all of the gondola hot-water extract and the hot-water extract of gingko extract, wherein the gingle extract is dried and then dried, and the gingko extract is dried after being roasted A food composition for improving aging is provided.

In the food composition for improving aging according to the present invention, the gondola hot-water extract or the hot-water extract of gingko may be preferably extracted at a temperature of 80 to 100 ° C for about 2 to 4 hours.

The present invention is characterized in that it comprises all of the gondola hot-water extract and the hot-water extract of gingko extract, wherein the gingle extract is dried and then dried, and the gingko extract is dried after being roasted There is provided a food composition for improving immune function.

In the food composition for improving immune function of the present invention, the gonadal hot-water extract or the ganoderma hot-water extract may be preferably extracted at a temperature of 80 to 100 ° C for about 2 to 4 hours.

In the food composition for improving immune function of the present invention, the gonad extract and the god extract may preferably exhibit an immune function improving effect by proliferating spleen cells. At this time, the splenocyte may more preferably be an enhanced secretion of IL6, which is a cytokine.

The food composition of the present invention comprising the hot water extract of ghred and the hot water extract of the gherkin exhibits an excellent aging improving effect and an immune function improving effect.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a process flow diagram of a dry gonad extract and a dried gonad extract.
Fig. 2 shows the cytotoxicity evaluation results of the gherkin extract and the dried gherkin extract.
Fig. 3 shows the results of evaluation of the NO production ability of the dry gonad extract and the dry gonad extract.
Fig. 4 shows the cytokine production ability of macrophylla extracts of Ganoderma lucidum extract and Ganoderma lucidum extract.
Fig. 5 shows the results of measuring the splenocyte proliferative activity of the dried gonad extract and the dried gonad extract.
Fig. 6 shows the results of measuring the effect of the gherkin extract and the dried gherkin extract on cytokine secretion.

The present invention is characterized in that it comprises all of the gondola hot-water extract and the hot-water extract of gingko extract, wherein the gingle extract is dried and then dried, and the gingko extract is dried after being roasted A food composition for improving aging is provided.

In addition, the present invention is characterized in that it comprises all of the gondola hot-water extract and the gyoza hot-water extract, wherein the gondre extract is dried after pounding the gondola, and the gassy extract is dried To provide a food composition for improving immune function.

In the present invention, it was intended to develop a food having an aging improving effect or an immune function improving effect by using an oxidizing agent. In the present invention, experiments have been carried out with various wild plants, and it has been confirmed that they have excellent antioxidative and immunological improving ability against ganoderma and flavorings, compared to other wild plants. In the present invention, the antioxidant activity of Gonadal hot-water extract and Ganoderma lucidum extract was confirmed. Since the oxidative mechanism is one of the main mechanisms of aging, the Gonadal hot-water extract and the ganoderma hot-water extract of the present invention, It was evaluated as having.

The present invention is characterized by including both a gondola hot water extract and a hot water extract of Ganoderma lucidum. It is possible to exert excellent antioxidant ability (aging improving ability) and immunity improving ability by including both of them, .

On the other hand, the present invention is characterized in that, instead of obtaining a hot-water extract from raw ghreaves or raw gruel, the gruel or raw gruel is dried and then dried to obtain a hot-water extract. The wild-caught extracts or gingiva extracts were obtained by using dried leaves, which were called dried fruits and dried fruits. The extracts were found to have superior antioxidative activity and improved immunity.

In the present invention, water is added to a gondola or sea urchin in the form of an achene, and hot water extraction is performed. The hot water extraction is not necessarily limited to specific conditions, but is preferably extracted at a temperature of 80 to 100 캜 for about 2 to 4 hours It is good. If necessary, it may be extracted by applying ultrasonic waves or the like.

Meanwhile, the present invention includes gondola hot water extract and ginger hot water extract. Preferably, gondola hot water extract and ginger hot water extract comprise 0.1 to 50 wt% of the food composition of the present invention. When the amount is less than 0.1% by weight, the effect is insufficient. When the amount is more than 50% by weight, the increase in the effect on the usage amount is insignificant, which is uneconomical.

On the other hand, the food composition of the present invention is preferably used in foods, cereals, caffeinated beverages, general drinks, chocolate, breads, And supplementary foods. However, the present invention is not limited thereto.

Hereinafter, the present invention will be described in more detail with reference to the following Examples or Experimental Examples. However, the scope of the present invention is not limited to the following embodiments and experimental examples, and includes modifications of equivalent technical ideas.

[ Example  One: Gunnedra  And From Gun Heat number  Preparation of extract]

First, the gondre and the shrimp were pitted, and then the dried gondre and the gundam were prepared. After soaking, dried herbs are sometimes referred to in the art as so-called 'silkworms'.

In this Example, the extracts were prepared from the above-prepared dry ganoderma sp. The preparation of the extract was carried out by weighing, extraction, filtration and freeze-drying as shown in Fig. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a process flow diagram of a dry gonad extract and a dried gonad extract. Specifically, after drying, the dried gondola or goryeo sample was pulverized, and about 50 g of each sample was weighed and used as an extraction raw material. A reflux extractor (DH.WEB01008 (WiseBath)) was used and water was extracted using a solvent. The extraction was carried out in the same manner as in Table 1, except that the solvent was added and the temperature and the time were matched with each other (Table 1).

Extraction condition Name of sample Extraction sample amount
(g) Extraction solvent
(mL) Extraction temperature and time
(° C, h) Gunnedra 50.0250 500 90 ° C, 3h Dry 50.1113

After extraction, the extract was centrifuged at 5,000 rpm for 30 minutes using a large-capacity centrifuge (2236HR, Gyrozen). The extract samples were loaded into a freeze dryer (Ilshin Bio-Base Co., Ltd.) and lyophilized at 35 ° C for 3 days at an internal pressure of 0.5 Torr. The yield of each extract was calculated as the weight of lyophilisate relative to the sample before extraction (Table 2).

Extraction yield Name of sample Extract (g) yield(%) Gunnedra 3.70 7.40 Dry 5.00 9.98

[ Experimental Example  One: Gunnedra  The extract and Dry  Evaluation of antioxidant efficacy of extracts]

 (One) DPPH  Evaluation of radical scavenging activity

The antioxidant efficacy of the extracts of Gunnigan and Ganoderma lucidum were evaluated by the known DPPH radical scavenging activity (Table 3).

DPPH radical scavenging activity Name of sample EC ₅₀ (μg / ml) ascorbic acid 8.41 + - 0.10 Gunnedra 237.48 ± 1.41 Dry 176.97 ± 1.43

As a result, the ascorbic acid EC ₅₀ value of the antioxidant was 8.41 ㎍ / ㎖. The EC ₅₀ value of the hot - water extract was 176.97 ㎍ / ㎖ for dry and 237.48 ㎍ / / Ml.

Inhibition of MMP-1 expression by antioxidative and UVA-mediated antioxidant activity, J. Korean Society of Cosmetic Scientists, 2006, 32 (3): 129-134) The DPPH radical scavenging ability of the dried ginseng extract, which is not the silkworm, was 58.3% at 1 mg / ml (EC ₅₀ value is the concentration at the point where the absorbance is 50% Compared with the extracts used in the present invention, the extracts of the present invention exhibited antioxidant activity of about 5 times higher than those of the extracts of the present invention. Activity. ≪ / RTI >

 (2) ABTS  Evaluation of radical scavenging activity

The antioxidant efficacy of Gunnigan's extract and dried green onion extracts was evaluated by the ABTS radical scavenging activity test (Table 4).

ABTS radical scavenging activity Name of sample EC ₅₀ (μg / ml) ascorbic acid 18.76 ± 0.17 Gunnedra 331.33 + - 7.30 Dry 110.78 + - 4.54

The EC ₅₀ value of ascorbic acid (control) was 18.76 ㎍ / ㎖, which was 110.78 ㎍ / ㎖ dry and 331.33 ㎍ / ㎖ dry.

Ganoderma sp. And Ganoderma lucidum were excellent antioxidant activity, considering that Ganoderma sp. And Ganoderma sp. Were not single compounds. In particular, it was confirmed that the dried pine needle had a DPPH radical scavenging activity and an ABTS radical scavenging activity, which are superior to those of Gunned gore.

 (3) Total Phenolic compound  Content evaluation

Total phenolic compounds of Gunnigan 's extract and Gunnorrhoid extract were determined by the total phenolic compounds assay. A standard curve using Gallic acid was calculated and the total phenolic compound content was calculated (Table 5).

Determination of total phenolic compounds Name of sample Content of extract
(Gallic acid, mg / 100g) Gunnedra 564.84 + - 9.98 Dry 1317.96 + 26.15

 The results showed that gum arabic was 1,317.96 ㎍ / 100㎎ and gonadra was 564.84 ㎍ / 100㎎.

 (4) Evaluation of total flavonoid compound content

Total flavonoid compounds of Gunnigan 's extract and Gunnogi extract were measured by total flavonoid assay. Using Quercetin as a standard reagent, a standard curve was drawn and the total phenolic compound content was calculated (Table 6).

Total flavonoid compound measurement Name of sample Content of extract
(Quercetin, μg / 100g) Gunnedra 111.1 + - 20.3 Dry 1532.2 ± 18.3

As a result of the experiment, it was 1532.2 ㎍ / 100g dry matter and 111.1 ㎍ / 100g dry matter gonad.

[ Experimental Example  2: evaluation of immunity-enhancing effect of the extract]

 (1) Evaluation of macrophage cytotoxicity

To evaluate the toxicity of each extract to cells, the ability of mitochondria to reduce Tetrazolium salts (WST-1), a water soluble substrate of yellow by dehydrogenase action, to formazan, The survival rate was measured by the WST-1 cell proliferation assay. RAW264.7 cells were seeded at a density of 1 × 10 ⁴ / well in a 96-well plate. After 24 hours, the extracts were treated for each concentration and cultured for 24 hours at 37 ° C. in 5% CO ₂ . Then, WST-1 reagent was added thereto, reacted at 37 ° C. for 1 hour, and the absorbance was measured at 450 nm through an ELISA reader and expressed as a percentage (FIG. 2). Fig. 2 shows the cytotoxicity evaluation results of the gherkin extract and the dried gherkin extract.

As a result of cytotoxicity, cell viability was 80% or higher at a concentration of 200 ug / ml or less. Therefore, it was evaluated as a material with high safety.

 (2) macrophage NO Generation  Evaluation (immunity enhancement)

For macrophage culture, DMEM medium containing 10% FBS and 1% penicillin-streptomycin was used and the cells were cultured in a CO ₂ incubator (37 ° C, 5% CO ₂ ). Raw 264.7 cell lines were stimulated by treatment with LPS (lipopolysaccharide) for the production of NO. Raw 264.7 cells were plated at 1 × 10 ⁶ cells / well in a 96-well plate and cultured for 16 hours. Then, the sample and 10 μl of LPS were added and cultured for 24 hours.

50 μl of Griess reagent (1% sulfanilamide, 0.1% N-1-naphthylethyenediamine and 5% phosphoric acid) was added to 50 μl of the supernatant of the cultured mixture and reacted at room temperature for 5 minutes. Absorbance was measured at 550 nm. The concentration of NO was determined by preparing a standard curve using sodium nitrate (NaNO ₂ ).

As shown in FIG. 3, it was confirmed that the concentration of NO (nitrite) was increased in a concentration-dependent manner from the lowest concentration of 25 μg / ml to the concentration of 500 μg / ml, I could. From these results, it was confirmed that the dry gonad extract and the dry gonad extract of the present invention had immunological activity. Fig. 3 shows the results of evaluation of the NO production ability of the dry gonad extract and the dry gonad extract.

 (3) intracellular cytokine ( Cytokine ) Generation  evaluation

(CBA) Mouse Th1 / Th2 cells were cultured in the presence of 100%, 200%, and 500 μg / ml of gonad extracts and 20% (IL-10, IL-17A, TNFα, IFN-γ, IL-6, IL-4 and IL4) secreted in the medium with the Th17 Cytokine Kit.

CBA kit was measured using Flow Cytometry Analyzer (BD Accuri C6) and analyzed with FCAP Array v3.0 software (BD). Data were analyzed by two-way ANOVA using GraphPad PRISM 6 software (* p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001, . Fig. 4 shows the cytokine production ability of macrophylla extracts of Ganoderma lucidum extract and Ganoderma lucidum extract.

As shown in FIG. 4, when the extract of Gunnigan gland extract or dried ganoderma extract was treated with macrophages, the amount of cytokines other than IL-17A was significantly increased compared with the control group. Particularly, IL-10 production was significantly increased in the cytokine IL-10 depending on the treatment concentration in the ganoderma extract and the ganoderma extract. When the ganoderma extract was treated at a concentration of 500 μg / ml, Which is about 2,000 times higher than the untreated group. TNF-α increased to a concentration of 4,000 pg / ml, which was about 10 times higher than that of the untreated group. INF-γ (Interferon-γ) showed a significant increase at a concentration of 200 μg / ㎖ of the dried persimmon extract. IL-6 showed a significant increase according to the treatment concentration in Ganoderma extract and Ganoderma lucidum extract. IL-4 showed a significant increase from 200 μg / ㎖ concentration in dried persimmon extract. IL-2 showed a significant increase from 500 μg / ㎖ concentration in dried persimmon extract.

As a result of evaluation of the cytokine production ability in macrophages as above, it was judged that both dry and dry gonad extract had immunity enhancing effect.

 (4) Gunnedra  The extract and Dry  The extract Splenocyte  Evaluation of efficacy on proliferation

The spleen is the main lymphatic organ that collects the antigen from the blood, and after the maturation of the T and B cells and the stimulation from the antigen, it divides and differentiates. The spleen cell proliferation is very important for the activation of the immune system. In order to examine the effect of the extracts on the proliferative capacity of mouse spleen cells in the present invention, the proliferative activity was observed by WST1 assay after treatment at various concentrations.

To isolate mouse spleen cells, BALB / c mice were sacrificed by cervical dislocation. The spleens were aseptically harvested, washed with RPMI 1640 medium, and homogenized with a Tissue grinder to separate the cells. The separated cell suspension was passed through a 40 μm mesh sieve, centrifuged at 3,000 rpm for 5 minutes, and red blood cell lysis buffer was added for 5 minutes to remove red blood cells. The cells were centrifuged twice with RPMI 1640 medium, diluted with RPMI 1640 medium containing 10% FBS to a concentration of 5 × 10 ⁶ cells / ml, and dispensed in a volume of 100 μl into a 96-well plate. Were used for the measurement of proliferative activity. 1 μg / ml Concanavalin A (ConA) was used as a positive control for cell proliferation. After each sample treatment, the cells were cultured for 48 hours and the proliferative capacity was measured using the WST1 assay.

As a result of the experiment, it was not more than the group treated with ConA, but the extracts of GuanGDREE and Ganoderma lucidum showed a growth rate of over 150% at 500 ug / mL compared with the control group. Fig. 5 shows the results of measuring the splenocyte proliferative activity of the dried gonad extract and the dried gonad extract.

 (5) Splenocyte  culture Supernatant  Cytokine measurement

IL2 and IL6 play an important role in the immune response by carrying out intracellular signal transduction in the early inflammatory response and are thus actively used as an indicator of immune enhancement in studies examining the immune function of many foods. Therefore, IL6 and IL2 were measured in order to determine whether the extracts of Ganoderma extract and Ganoderma sp. In addition, IgG and IgA were measured by ELISA assay kit.

The mouse spleen cells were separated in the same manner as described above, and the separated splenocytes were dispensed into a 24-well plate at 2 × 10 ⁶ cells / well. The prepared cells were treated with 1 μM methotrexate (MTX) for 24 hours, and each extract was added at a concentration of 500 μg / ml. After culturing for 24 hours, the cytokine secreted in the culture solution was measured. The amounts of IgG, IgA, IL2, and IL6 were measured using an ELISA cytokine kit (Koma Biotech).

As shown in FIG. 6, no difference was observed in IL2, IgG, and IgA, but it was confirmed that IL-6 had an effect of enhancing IL-6 secretion. Fig. 6 shows the results of measuring the effect of the gherkin extract and the dried gherkin extract on cytokine secretion.

Claims (6)

Gondola hot water extract, and ginger hot water extract,
The gonad extract may contain,
And after drying the raw ghred, it is dried.
The above-
Wherein the raw food is dried and then dried.
The method according to claim 1,
The gondola hot-water extract or hot-
Wherein the composition is extracted at a temperature of 80 to 100 DEG C for about 2 to 4 hours.
Gondola hot water extract, and ginger hot water extract,
The gonad extract may contain,
And after drying the raw ghred, it is dried.
The above-
Wherein the composition is dried and then dried.
The method of claim 3,
The gondola hot-water extract or hot-
Wherein the composition is extracted at a temperature of 80 to 100 占 폚 for about 2 to 4 hours.
The method of claim 3,
The gondre extract and the ginger extract have the following characteristics:
And exhibiting an immune function improving effect by proliferating spleen cells.
The method of claim 3,
The splenocyte can be,
Wherein secretion of IL6 as a cytokine is enhanced.

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