KR102534513B1 - Composition showing antioxidant and immunity improving activity containing callophyllis rhynchocarpa extract, and health functional food containing the same - Google Patents

Abstract

The present invention relates to a functional composition containing red beak leaves and a health functional food containing the same. Antioxidant and immune-improving composition according to the present invention is to include a beak red leaf extract.
In the case of using the composition, as a natural material-based material while enhancing antioxidant and immunity-improving activities, it can be taken for a long period of time without side effects and has excellent flavor and palatability.

Description

Composition showing antioxidant and immune-improving activity containing red leaf extract and health functional food containing the same

The present invention relates to a composition for antioxidation and immunity improvement containing a red leaf extract and a health functional food containing the same. More specifically, it is to provide a functional composition showing antioxidant and immune-improving effects and a health functional food containing the same, while containing the fermented red leaf extract or fermented red leaf extract as an active ingredient.

Activities such as hydrogen peroxide (H ₂ O ₂ ), superoxide anion (O ₂ ^- ), and hydroxyl radical (OH), which are inevitable by-products caused by the process of consuming oxygen in the human body Oxygen species (reactive oxygen species; ROS) cause toxicity due to their high reactivity in vivo. An appropriate amount of reactive oxygen species plays an important role in maintaining cellular functions such as electron transport system in mitochondria and activation of white blood cells by acting as intracellular signaling substances in vivo. On the other hand, excessively produced reactive oxygen species cause oxidative stress by breaking the antioxidant balance in the body. The toxicity of reactive oxygen species damages cell components such as lipids, proteins, sugars, and DNA non-selectively and irreversibly, resulting in cancer, brain diseases such as stroke and Alzheimer's disease, heart disease, arteriosclerosis, inflammation, and autoimmune diseases. can cause various diseases and harmonization of

Endogenous antioxidant enzymes such as catalase (CAT), glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) maintain a balance with active oxygen in vivo. Prevents DNA damage, protein modification and lipid peroxidation. Among human body parts, the brain has relatively few endogenous antioxidant enzymes that remove free radicals, and accounts for 20% of the total oxygen consumption, so it is the part where the largest amount of active oxygen is generated in the body. In addition, the brain has a high content of unsaturated fats that are easily oxidized by reactive oxygen species due to its neurotransmission function, so it is very vulnerable to oxidative stress compared to other organs. Excessive reactive oxygen species causes brain damage through the death of nerve cells. . Accordingly, interest in natural materials rich in phenolic compounds, which are reported to protect nerve cells from oxidative stress induced by active oxygen through antioxidant activity and contribute to the prevention of neurodegenerative diseases, is increasing.

Recently, research on the development of various natural components that act as free radical scavengers has been attempted. As a result of these studies, the ingredients that are currently attracting the most attention are phytonutrients, which are plant nutrients, and numerous papers have been published worldwide. Citrus bioflavonoids and A component called pycnogenol extracted from pine bark and grape seeds has been reported. In addition to the above components, there are tocopherol, ascorbic acid and its derivatives, carotenoids, flavonoid compounds, phospholipids, browning reaction products, amino acids, peptides, proteins, spice extracts, and the like.

On the other hand, research on spice extracts has recently been a field of increasing interest since it began in the 1940s, and in particular, rosemary and sage extracts have been approved by the FDA. In addition to the above ingredients, many ingredients such as nutmeg, cloves, and red pepper are known as antiseptic extracts. Although the degree varies depending on the type, the free radical scavenger effect is recognized. In addition, oryzanol and Together, guaiac resin, seshimul, sesamolin, and sesamin in sesame oil are well known, and various herbal medicines and edible plants including balsam extract, rutin, and somok are reported to have this effect.

On the other hand, immunity (Immune) refers to a mechanism for physiologically recognizing and excluding exogenous and endogenous foreign substances in the body of humans and animals, and maintaining homeostasis. Immunity can be largely classified into two types, which are divided into innate immunity, which is present from birth, and acquired immunity, which is obtained through adaptation to life.

Congenital immunity is also called innate immunity. It reacts non-specifically to the antigen and has no special memory function. The innate immune system includes skin and mucous tissues that block the invasion of antigens, acidic gastric acid, and complement present in the blood. Cells include macrophages responsible for phagocytosis, polymorphonuclear leukocytes, and K cells that can kill infected cells. In fact, most infections are defended by this innate immunity.

Acquired immunity is also called acquired immunity. It can remember the first invaded antigen, and when it invades again, it can specifically react and effectively remove the antigen. It plays a role in reinforcing the innate immunity.

Acquired immunity includes T cells and B cells. Cells that differentiate into lymphocytes in the epithelial cells of the thymus gland by the special internal environment and thymic humoral factors are called T cells, and cells that differentiate into lymphocytes in the bone marrow independently of the thymus gland are called B cells.

The role of T cells is to stimulate B cells to produce antibodies and directly attack antigens, such as tumor cells. , to produce and secrete antibodies of IgA and IgE.

'Health' refers to a state in which there is no abnormality in the body, that is, a state in which there is no disease or injury. If your immunity is high, you are less likely to get sick, and even if you get sick, you will recover quickly. It is obvious that maintaining such a healthy state continuously and extending lifespan as one ages. Therefore, immunity is directly related to human health and lifespan, and it can be said that it is essential to manage immunity in order to maintain health and extend lifespan. However, looking at the lifestyles of modern people, many factors that decrease immunity, such as stress, lack of exercise, irregular life rhythm, unbalanced diet, and smoking, account for a large number of factors. There are various pollutions in the environment, and there are many factors that can lower immunity. Therefore, modern people should make efforts to enhance immunity.

Immunity boosters such as zimodulin and broncobaxom can be used to boost immunity. Immunostimulants contain bovine thymus glands or purified bacterial bodies. The mechanism of action in the body of the immune enhancer containing purified bacteria is to train the immune system by injecting weakened bacteria into the body. is to do However, the effect of the immune enhancer is very insignificant, and there are side effects such as a decrease in appetite.

Accordingly, the inventors of the present invention continued research on materials that can exhibit antioxidant activity and immune-improving activity based on various materials based on natural products that can be taken for a long time without side effects, and as a result, the present invention was completed. .

KR 10-2016-0011173 A KR 10-2007-0089288 A

An object of the present invention is to provide a functional composition that can be taken for a long period of time without side effects such as cytotoxicity while using the fermented red leaf extract or fermented red leaf extract as an active ingredient.

An object of the present invention is to provide a functional composition capable of exhibiting antioxidant and immune-improving effects.

An object of the present invention is to provide a red leaf-based functional food or health functional food that is easy to popularize due to its excellent flavor and palatability.

In order to achieve the above object, the composition for improving antioxidant and immunity according to an embodiment of the present invention may include a red beak leaf extract.

In the composition for improving antioxidant and immunity, the extract may be fermented.

In the antioxidant and immune-improving composition, the extract is Lactobacillus casei; Lactobacillus rhamnosus; Bifidobacterium bifidum Bifidobacteria; Bifidobacterium breve Bifidobacteria; Lactobacillus acidophilus; Any one selected from the group consisting of Bacillus subtilis and mixtures thereof may be inoculated and fermented.

In the antioxidant and immunity-improving composition, the extract may be extracted by using any one selected from the group consisting of water, lower alcohols having 1 to 6 carbon atoms, and mixtures thereof as an extraction solvent.

In the antioxidant and immunity-improving composition, the extract may further include any one selected from the group consisting of semogasari extract, rhubarb (Eisenia bicyclis) extract, and mixtures thereof.

Food according to another embodiment of the present invention may include the antioxidant and immunity improving composition.

Health functional food according to another embodiment of the present invention may include the composition for improving the antioxidant and immunity.

Hereinafter, the present invention will be described in more detail.

Antioxidant and immune-improving composition according to an embodiment of the present invention may include a beak red leaf extract.

As used herein, the term 'extract' has a meaning commonly used in the art as a crude extract in the art as described above, but also includes fractions obtained by additional fractionation of the extract in a broad sense. That is, the plant extract includes not only those obtained using the above-described extraction solvent, but also those obtained by additionally applying a purification process thereto. For example, a fraction obtained by passing the extract through an ultrafiltration membrane having a certain molecular weight cut-off value, separation by various chromatography (made for separation according to size, charge, hydrophobicity or affinity), etc. Fractions obtained through various purification methods are also included in the plant extract of the present invention.

The extract used in the present invention may be prepared in a powder state by additional processes such as distillation under reduced pressure and freeze drying or spray drying.

As used herein, the term "immune disease" refers to a disease caused by an excessive response of the body's immune system, and representatively includes allergic disease. However, since the inflammatory response eventually occurs due to the response of the immune system in vivo, inflammatory diseases and immune diseases can be used as terms referring to similar ranges in a broad sense, and are not used as distinct terms in this specification. , I will mix it up.

As used herein, the term "inflammatory disease" is a disease caused by an inflammatory reaction derived from an external substance or an internal substance, and includes neuroinflammatory diseases and arthritis.

In the case of containing the red beak leaf extract, it can produce an effect of enhancing immunity, including antiviral and antibacterial effects, in addition to antioxidant and anti-inflammatory activities.

The composition for improving antibacterial, anti-inflammatory, antiviral and immune functions containing the red beak leaf extract may increase cytokine production of TNF-α and IL-12.

Cytokines are protein immunomodulators secreted from immune cells and bind to specific receptors in the process of autocrine signaling, paracrine signaling, and endocrine signaling to respond to an immune response. get involved in There are various types of cytokines involved in cell proliferation, differentiation, apoptosis, or wound healing, and many of them are particularly involved in immunity and inflammation.

Among these, the most important immune function is performed by a group of cytokines known as interleukin (Interleukin-1, -6, -12), interferon gamma (IFN-γ), and tumor necrosis factor (TNF).

Interleukin (IL) is secreted from leukocytes and is involved in the immune system regulation function, and more than 30 types are currently known. Interleukin is a substance that plays an important role in the body's defense by controlling the immune response by acting on various stages of the immune response, and its types include IL-1, 6, and 12.

Among them, IL-12 is a cytokine composed of p35 and p40 proteins. It plays a very important immunological function by linking innate immunity and acquired immunity, and various effector cells, such as macrophages, involved in the elimination of endogenous antigens. ), induces activation of NK cells and T-cells. NK cells and T-cells activated by stimulation of IL-12 stimulate the production of IFN-γ, and thus have properties as a major factor mediating the initial innate immune response. It is also the most important cellular immune mediator because it induces the immune machinery of T-cells to be mainly Th-1-oriented.

Interferon (IFN) functions to prevent the proliferation of viruses or control cell proliferation, and also plays an important role in the immune system. There are alpha (α), beta (β), gamma (γ), and omega (ω) types of interferons. Interferon-α is mainly secreted by macrophages, and interferon-β is mainly secreted by fibroblasts. Interferon-γ is secreted by Th1 cells and NK cells, and interferon-ω is mainly secreted by trophoblasts or trophoblasts. In particular, interferon-γ (IFN-γ) is produced by T lymphocytes and macrogranulocytes and is a substance that induces the killing of phagocytic microorganisms or antigens. In addition, it has various immunomodulatory effects and activates macrophages.

Tumor necrosis factor (TNF) is a cell signaling protein associated with an inflammatory response and is a cytokine that induces an acute phase response. Tumor necrosis factor is divided into tumor necrosis factor-α (TNF-α) and tumor necrosis factor-β (TNF-β) called lymphotoxin, but generally tumor necrosis factor refers to tumor necrosis factor-α .

Therefore, in the case of the immune function improvement effect by the antibacterial, anti-inflammatory, antiviral and immune function improving composition of the present invention, TNF-α and IL-12 that activate the immune response by using natural products that do not have toxicity and side effect problems in the human body. increases cytokine production.

The composition is to prevent a disease selected from the group consisting of allergy, inflammatory disease and immune disease.

Allergy refers to various symptoms caused by an overreaction of the immune system to a substance that is not a particular problem for most people. These include hay fever, food allergies, atopic dermatitis, asthma and other anaphylaxis, and show symptoms such as congested eyes, itchy skin rashes, runny nose, difficulty breathing and edema.

Inflammation is a kind of in vivo response to damage or infection in a specific tissue, and the main mediator is immune cells. It is divided into acute inflammation and chronic inflammation depending on the damaged or infected part of the tissue. . When the tissue damage or infection is small or temporary, it is resolved by acute inflammation, but when the inflammation is large or chronically infected, acute inflammation is not resolved and progresses to chronic inflammation. In the case of chronic inflammation, it can progress to a chronic inflammatory disease.

Immune disease refers to a disease that becomes a problem when a specific immune response occurs, but is not limited thereto, and preferably includes autoimmune disease, transplant rejection, and graft-versus-host disease. Atopy, rheumatoid arthritis, Hashimoto's thyroiditis, pernicious anemia, Addison's disease, type 1 diabetes, lupus, chronic fatigue syndrome, fibromyalgia, hypothyroidism and hyperthyroidism, scleroderma, Behcet's disease, inflammatory bowel disease, multiple sclerosis, myasthenia gravis, It may be Meniere's syndrome, Guilian-Barre syndrome, Sjogren's syndrome, vitiligo, endometriosis, psoriasis, vitiligo, systemic scleroderma, asthma or ulcerative colitis.

Antioxidant and immune-improving composition of the present invention is to include a beak red leaf extract.

The beak red leaf (Callophyllis rhynchocarpa) corresponds to the algae Callimenia and Chinuari, and the body is a linear flat membrane, densely branched, expanding in a fan shape, 5 to 7 cm in height, and 3 to 3.5 mm in width at the bottom. The second phase gradually diverges into the second phase, such as multi-chasang or small idol. Cystocarp is hemispherical, formed on the surface close to the upper margin, and has two projections. The color is bright red, the quality is membranous, and it does not adhere well to the ground when dry.

It is mainly distributed in Japan, Sakhalin, Sea of Okhotsk, and Ulleungdo, Mokdo, Busan, and Wando in Korea.

In the composition for improving antioxidant and immunity, the extract may be fermented. The fermented extract increases the activity of active ingredients exhibiting antioxidant and immune-improving effects, and can have the effect of neutralizing the strong aroma and fishy taste unique to red beak leaves.

In the antioxidant and immune-improving composition comprising the beak red leaf extract, the extract is Lactobacillus casei; Lactobacillus rhamnosus; Bifidobacterium bifidum Bifidobacteria; Bifidobacterium breve Bifidobacteria; Lactobacillus acidophilus; Any one selected from the group consisting of Bacillus subtilis and mixtures thereof may be inoculated and fermented.

Preferably, the extract may be inoculated with and fermented by Lactobacillus casei and Bifidobacterium bifidum. In the case of fermentation by the microorganism, it is possible to increase the antioxidant activity and improve the immunity improvement effect, while improving the unique flavor of the red leaf.

In the antioxidant and immunity-improving composition, the extract may be extracted by using any one selected from the group consisting of water, lower alcohols having 1 to 6 carbon atoms, and mixtures thereof as an extraction solvent.

The extract is obtained by using a polar solvent such as alcohol having 1 to 10 carbon atoms such as water, ethanol, methanol, etc. or a mixed solvent having a mixing ratio of 1:0.1 to 1:10 of alcohol and water with respect to the pulverized product of the red leaf. It can be eluted, and more specifically, water can be used as an extraction solvent. At this time, the extraction temperature may be 10 ℃ to 100 ℃, preferably the extract extracted at room temperature.

The extraction solvent may be used in an amount of 2 to 50 times, preferably 2 to 20 times, based on the weight of the sample. For extraction, the sample may be left for leaching in the extraction solvent for 1 to 72 hours, specifically 1 to 24 hours. It may also be filtered after extraction.

It may be a product obtained by concentrating the filtered extract under reduced pressure with a vacuum rotary evaporator, but is not limited thereto, and includes all extracts, diluted or concentrated solutions of extracts, dried products obtained by drying extracts, and crude or purified products thereof.

In the antioxidant and immunity-improving composition, the extract may further include any one selected from the group consisting of semogasari extract, rhubarb (Eisenia bicyclis) extract, horse chestnut extract, and mixtures thereof.

Preferably, it may further include 1 to 10 parts by weight of semogasari extract, 1 to 10 parts by weight of rhubarb (Eisenia bicyclis) extract, and 10 to 20 parts by weight of horse chestnut extract based on 100 parts by weight of the red leaf extract. In the case of the above range, the antioxidant activity and immunity improvement effect may be higher.

Preferably, the extract is fermented, and may further include a mixture of rice water and molasses. When the mixture of rice water and molasses is included, it is possible to provide food with high palatability by removing the strong or fishy scent peculiar to the red beak leaf while doubling the antioxidant activity and immune activity.

The mixture of rice water and molasses is a mixture of 1 to 10 parts by weight of molasses based on 100 parts by weight of rice water, and the mixture may be included in 50 to 200 parts by weight based on 100 parts by weight of the red leaf extract. In the case of the above range, a synergistic effect and an effect of improving palatability can be produced.

The rice water can be obtained in the process of washing rice or rice bran.

Food according to another embodiment of the present invention may include the antioxidant and immunity improving composition.

Health functional food according to another embodiment of the present invention may include the composition for improving the antioxidant and immunity.

As a specific example, a processed food capable of enhancing antibacterial, anti-inflammatory, antiviral and immune function improving effects can be prepared using the food composition. Such processed foods include, for example, sweets, tea, beverages, alcoholic beverages, fermented foods, canned foods, processed milk products, processed meat products, noodles, and the like. Sweets include biscuits, pies, cakes, breads, candies, jellies, chewing gum, cereals, and the like. Beverages include drinking water, carbonated beverages, functional ionic beverages, functional ionic beverages, juice (eg, apple, pear, grape, aloe, tangerine, peach, carrot, tomato juice, etc.), sikhye, and the like. Liquor includes rice wine, whiskey, soju, beer, liquor, fruit wine, and the like. Fermented foods include soy sauce, soybean paste, red pepper paste, and the like. Canned food includes canned seafood (e.g., canned tuna, mackerel, saury, canned conch, etc.), canned livestock products (canned beef, pork, chicken, turkey, etc.), canned agricultural products (canned corn, peaches, canned pineapple, etc.). Milk processed foods include cheese, butter, yogurt, and the like. Processed meat products include pork cutlet, beef cutlet, chicken cutlet, and sausage. Includes sweet and sour pork, nuggets, nubani, etc. It includes noodles such as sealed packaged raw noodles. In addition to this, the composition can be used for retort food, soup, and the like.

A food composition according to another embodiment of the present invention is prepared by including the antibacterial, anti-inflammatory, antiviral and immune function improving composition as an active ingredient.

Health functional food according to another embodiment of the present invention may include a composition for antibacterial, anti-inflammatory, antiviral and immune function improvement including the red beak leaf.

The health functional food is not particularly limited as long as it can be ingested to improve antibacterial, anti-inflammatory, antiviral and immune activity.

When using the health functional food composition of the present invention as a food additive, the health functional food composition may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to conventional methods. Active ingredients can be appropriately used depending on their purpose of use (prevention or improvement). In general, when preparing food or beverage, the health functional food composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material. However, in the case of long-term intake for health purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

There is no particular limitation on the type of health functional food. The health functional food composition of the present invention can be made into food, particularly functional food. The functional food of the present invention

It includes components commonly added during food preparation, and includes, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, when prepared as a drink, natural carbohydrates or flavors may be included as additional ingredients in addition to active ingredients. The natural carbohydrates are monosaccharides (eg, glucose, fructose, etc.), disaccharides (eg, maltose, sucrose, etc.), oligosaccharides, polysaccharides (eg, dextrins, cyclodextrins, etc.) or sugar alcohols (eg, maltose, sucrose, etc.) , xylitol, sorbitol, erythritol, etc.) are preferred. As the flavoring agent, natural flavoring agents (eg, thaumatin, stevia extract, etc.) and synthetic flavoring agents (eg, saccharin, aspartame, etc.) may be used.

In addition to the health functional food composition, various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonic acid It may further contain a carbonation agent used in beverages and the like.

The present invention provides a functional composition that can be taken for a long period of time without side effects such as cytotoxicity while using the fermented red leaf extract or fermented red leaf extract as an active ingredient.

The present invention provides a functional composition capable of exhibiting antioxidant and immune-enhancing effects.

The present invention provides a red leaf-based functional food or health functional food that is easy to popularize due to its excellent flavor and palatability.

1 relates to evaluation results of antioxidant activity according to an embodiment of the present invention.
Figure 2 is a graph measuring the concentration of nitric oxide present in the culture medium treated with the composition according to an embodiment of the present invention.
Figure 3 is a graph measuring the concentration of TNF-α present in the culture medium treated with the composition according to an embodiment of the present invention.
Figure 4 is a graph measuring the concentration of interleukin-12 present in the culture medium with the composition according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily implement the present invention. However, the present invention may be embodied in many different forms and is not limited to the embodiments described herein.

[Preparation Example: Preparation of extract]

1. Preparation of red beak leaf extract

The washed and dried red leaves were pulverized, extracted under pressure with hot water, concentrated and filtered to prepare a red leaf extract (P1).

2. Preparation of other extracts

Semogasari extract (R1), rhubarb (Eisenia bicyclis) extract (R2), and horse chestnut extract (R3) were prepared in the same manner as the red beak leaf extract.

3. Preparation of mixed extract and fermented extract

In order to confirm the synergistic effect by mixing, a mixed extract was prepared according to the composition shown in [Table 1] below.

M1 M2 M3 M4 M5 P1 100 100 100 100 100 R1 - 0.1 One 10 20 R2 - 0.1 One 10 20 R3 - One 10 20 30

(unit: parts by weight)

4. Preparation of fermented extract

In order to confirm the synergistic effect by fermentation of the extract, fermentation was performed according to [Table 2] below. Synergistic effect while inoculating S1 (Lactobacillus casei), S2 (Bifidobacterium bifidum bifidus bacteria), and S3 (mixture of S1 and S2) as fermentation (S) strains Confirmed. On the other hand, the mixture (U) of rice water and molasses was mixed with 2 parts by weight of molasses based on 100 parts by weight of rice water.

B1 B2 B3 B4 extract M4 M4 M4 M4 S S1 S2 S3 S3 U - - - U

(unit: parts by weight)

[Experimental Example: Antioxidant and immune enhancing effect]

1. Evaluation of antioxidant activity

In order to evaluate the antioxidant activity of the extract according to each preparation example, a DPPH radical scavenging activity test was conducted. DPPH radical scavenging activity was measured by the method of Yamaguchi et al. 60 μl DPPH solution (60 μM) was added to the 60 μl sample solution, stirred for 10 seconds, and then the mixed solution was transferred to a quartz capillary tube and measured by ESR after 2 minutes.

The results are shown in [Figure 1] below.

Referring to the following [Figure 1], it can be confirmed that the antioxidant activity in the production example according to the present invention, in particular, it can be confirmed that the synergistic effect appears in the combination range of M3 to M4. In addition, it can be confirmed that B3 and B4 by fermentation show an additional synergistic effect. In particular, it can be confirmed that the antioxidant activity of B4 is the most excellent.

2. Measurement of nitric oxide production

In order to confirm the ability of the composition according to the preparation example to have an immune activity, griess reagent [1% (w / v) sulfanilamide in 5% (v / v) phosphoric acid (phosphoric acid) and 0.1% (w / v) naphthyl Ethylenediamine-hydrochloric acid (naphtylethylenediamine-HCl)] was used to measure the production concentration of NO ^2- present in the cell culture medium. Specifically, RAW 264.7 cells were dispensed in a 6-well plate at a concentration of 5X10 ⁵ cell/well, cultured for 24 hours, and then each prepared sample was treated. As a normal control, the sample was treated with 0ug/mL, and a positive control The lipopolysaccharide was treated with 1ug/mL and cultured for 24 hours. After mixing the supernatant of the cell culture medium and the same amount of griess reagent and reacting at room temperature for 15 minutes, the absorbance was measured at 540 nm, and the nitric oxide concentration was obtained and converted from a standard curve prepared using sodium nitrite.

The results are shown in [Figure 2] below.

The production of nitric oxide is known as an important signaling substance that acts as a defense against tumor cells or intracellularly infected microorganisms in the immune system. It was confirmed that the activity appeared, and it was confirmed that less than 40% of nitric oxide was produced compared to the positive control group throughout the examples.

In particular, it was confirmed that a synergistic effect appeared in the range of combinations of M3 and M4, and it was confirmed that an additional synergistic effect was exhibited in the case of B3 and B4 by fermentation. In particular, it can be confirmed that the activity of suppressing the production of nitric oxide of B4 is the most excellent. Excessive production of nitric oxide causes systemic inflammation and has various negative effects on the living body, but production of an appropriate amount of nitric oxide is considered an important factor in innate immunity, so the composition can exhibit excellent immune activity.

3. Measurement of TNF-α and IL-12 production ability

The secretion of TNF-α and IL-12 in the cell culture medium was measured using ELISA. Capture antibodies (anti-mouse TNF-α and IL-12) were each dispensed on a 96-well plate and coated at 4 °C. It was washed with washing buffer (PBS containing 0.05% Tween-20), blocked with phosphate-buffered saline (PBS) containing 10% FBS for 1 hour, and then washed with washing buffer. Each microplate was dispensed with 100 uL of the supernatant of the same cell culture as in which nitric oxide was measured, reacted at room temperature for 2 hours, and washed with washing buffer. Each diluted biotinylated anti-mouse TNF-α and interleukin-6 detection antibody and streptavidin-horseradish peroxidase conjugate solution were added and reacted at room temperature for 1 hour. After washing the reacted plate with washing buffer, tetramethylbenzidine (TMB) substrate solution was added to develop color in a dark room for 30 minutes, and 1 M phosphoric acid was added to stop the color reaction, and absorbance was measured at 450 nm.

The results are shown in [Figure 3] below. Referring to [Figure 3] below, as a result of examining the immune activity for the ability to produce TNF-α, an inflammatory cytokine, in the case of M3 and M4, secretion of TNF-α by macrophage stimulation is increased confirmed to do so. In addition, it can be confirmed that in the case of B1 to B4, the secretion of TNF-α produced by the positive control group (lipopolysaccharide) is promoted more.

In addition, referring to [Figure 4], it was confirmed that the production of IL-12, an immune mediator, increased in a concentration-dependent manner in M3 and M4. In addition, it can be seen that the production of IL-12 is more promoted when B3 and B4 are used as fermented compositions. It can be understood that the above composition can play an important role in the immune response by increasing the production of interleukin-6 and differentiating B lymphocytes.

4. Evaluation of palatability

Ten adults who responded that they had a strong sense of aversion to the strong scent and taste of seaweeds were selected as test subjects, and the extract according to the preparation example was provided as a tea drink, and then the overall acceptability including flavor and taste was evaluated. The overall acceptability was evaluated with an index of 1 to 10, and the higher the index, the better the acceptability.

The results are shown in [Table 3] below.

M1 M2 M3 M4 M5 B1 B2 B3 B4 preference 1.1 1.2 1.1 1.3 1.4 2.3 1.6 2.8 5.9

(Unit: index)

Referring to [Table 3], in the case of the embodiment of the present invention, the palatability was evaluated to be low for consumers who have a reluctance to the strong and fishy scent of red leaves, but in the case of B3 fermented in a certain range, the fishy scent is somewhat neutralized was able to confirm that In particular, in the case of B4 to which a mixture of rice water and molasses was added in the fermented composition, a significant portion of flavor and taste was improved.

Therefore, in the case of using the above range, it can be confirmed that it can be more easily supplied as a functional food or health functional food for improving antioxidant and immune function with high palatability while using red leaves.

Although the preferred embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements of those skilled in the art using the basic concept of the present invention defined in the following claims are also made according to the present invention. falls within the scope of the rights of

Claims (7)

Contains red leaf extract,
Further comprising semogasari extract, rhubarb (Eisenia bicyclis) extract and horse chestnut extract,
1 to 10 parts by weight of semogasari extract, 1 to 10 parts by weight of rhubarb (Eisenia bicyclis) extract, and 10 to 20 parts by weight of horse chestnut extract, based on 100 parts by weight of the red leaf extract.
A composition for antioxidant and immunity improvement. According to claim 1,
The extract is fermented
A composition for antioxidant and immunity improvement. According to claim 1,
The extract is Lactobacillus casei; Lactobacillus rhamnosus; Bifidobacterium bifidum Bifidobacteria; Bifidobacterium breve Bifidobacteria; Lactobacillus acidophilus; Any one selected from the group consisting of Bacillus subtilis and mixtures thereof is inoculated and fermented
A composition for antioxidant and immunity improvement. According to claim 1,
The extract is extracted by using any one selected from the group consisting of water, lower alcohols having 1 to 6 carbon atoms, and mixtures thereof as an extraction solvent.
A composition for antioxidant and immunity improvement. delete A food product comprising a composition according to any one of claims 1 to 4. A health functional food comprising the composition according to any one of claims 1 to 4.

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