KR20100011580A - Composition for enhancing immune system function comprising extract of abalone viscera - Google Patents

Abstract

PURPOSE: A composition containing abalone viscera extract as an active ingredient is provided to enhance Th1 immune response(cellular immune response) and Th2 immune response(humoral immunity reaction). CONSTITUTION: A composition for enhancing immunity as an active ingredient contains Haliotis discus hannai, Holiotis gigantean, Haliotis discus, Haliotis sieboldi, or Haliotis diversicolor super-texta. The abalone viscera extract obtained using water, anhydrous or hydrous low alcohol of 1-4 carbon atoms, mixture solvent of low alcohol and water, acetone, ethyl acetate, chloroform, 1,3-butyleneglycol, hexane, diethylether, or butylacetate. The composition enhances immune response Th1 immune response, Th2 immune response or both of the immune response of the T1 and T2.

Description

Composition for Enhancing Immune System Function Comprising Extract of Abalone Viscera}

The present invention relates to an immunopotentiated composition comprising an abalone viscera extract as an active ingredient.

Overture drinks with "Abalone research on the processing of vitamin B ₁ and vitamin B ₂ Lots of calcium and phosphorus, etc., and the mineral-rich healthy food is a basic research on some component analysis D performed two euros bars, rollover There is a study on the lawsuit of 'but there is still a lack of scientific evidence on the functionality. Dried abalone is known to be effective in dissolving gallstones and improving liver detoxification with taurine, lowering cholesterol, improving heart function, and restoring vision, but developing processed foods and functional food materials using abalone in Korea. Very little research has been done.

Abalone is rich in protein and vitamins, so it is effective in skin care, nourishing tonic and postpartum cooking, and in particular, it is rich in taurine, which is effective in protecting liver, fatigue and myocardial infarction. In terms of taste and taste, it is incomparable with other seafood.

   The efficacy of Korean abalone, which has been recognized for its excellence since ancient times, has been described as “Efficacy of abalone for lightening the body and brightening the eyes” in the Oriental medicine book, Myungbyeong Myungbyeong and Kyuhap. . In the Joseon Dynasty, Seokjeong Jeong Yakjeon introduces abalone in the name of blowfish, saying, `` Red meat is sweet and can be eaten raw or cooked, but the best way is to make pho. The intestines can be eaten, soaked in salted seafood, and boiled. In the spring and summer, there is a poison, and when it comes into contact with it, the flesh boils and the affected area bursts. In general, shellfish are known to be effective in restoring tired nerves, and abalone is known to be effective in relieving dull optic nerve fatigue. The abalone shell is also used in Korean medicine, so it is useful for conjunctivitis and cataracts. It is used to relieve the symptoms of burning or chest swelling, strengthens the liver function, and boils abalone when the body is weak. It is known to be good for urine, jaundice and cystitis.

Abalone has excellent nutritional value, even as feces, which can be used to treat diabetes and high blood pressure. Abalone is rich in protein, rich in the amino acids that make up it, and also rich in minerals and vitamins such as phosphorus, iron and iodine. For this reason, abalone has been treated as a high-grade seafood since ancient times, and has excellent efficacy in skin beauty, nourishing tonic, postpartum cooking, and weak constitution, and has been widely used for edible as well as medicinal purposes.

A lack of iodine and thyroid hormones can lead to obesity due to a slowing of metabolism or the accumulation of fat in the blood vessels. In addition, thyroid hormones are involved in the physical, mental and sexual growth of growing children. Lack of iodine can lead to significant hypothyroidism, resulting in rough skin and thinning hair. Therefore, eating iodine with foods like seaweed is a safe way. Abalone is known to increase the strength of the liver because it has a high content of iodine, pain in the head, ringing, and dry tongue and throat. It is said that orphan feeding abalone when mother's milk does not come out within 7 days of old, has a great effect, because there are many minerals in abalone. In addition, abalone is a high protein ingredient, so high absorption in the body is good food for pregnant women, obesity, liver cirrhosis. It is also used as a medicine for liver function recovery and pulmonary tuberculosis. Abalone is rich in sulfur-containing amino acids such as methionine and cysteine, which is good for restoring and restoring fatigue after illness. As such, the efficacy and function of abalone are known in folk remedies and some ancient texts that have been used for a long time, but there are no data that have proven this fact based on scientific facts. Therefore, it is urgent to prove and reevaluate the efficacy of abalone through more scientific and empirical research.

Throughout this specification, many papers and patent documents are referenced and their citations are indicated. The disclosures of cited papers and patent documents are incorporated herein by reference in their entirety, and the level of the technical field to which the present invention belongs and the contents of the present invention are more clearly described.

The present inventors tried to identify the physiological activity of Abalone viscera extract. As a result, the present inventors have completed the present invention by elucidating that the abalone viscera extract has the effect of enhancing the activity of B cells and T cells involved in the immune response in vivo, and thus improving the immune activity in vivo. .

It is therefore an object of the present invention to provide an immunopotentiated composition.

Other objects and advantages of the present invention will become apparent from the following detailed description, claims and drawings.

According to an aspect of the present invention, it provides an immunopotentiated composition comprising an abalone viscera extract as an active ingredient.

The present inventors tried to identify the physiological activity of Abalone viscera extract. As a result, the inventors have found that the abalone viscera extract has the effect of enhancing the activity of B cells and T cells involved in the immune response in vivo, and thus improving the immune activity in vivo.

The present invention is based on the immune activation of abalone viscera extract. Abalone viscera extract used as an active ingredient in the composition of the present invention can be obtained using a variety of abalone. According to a preferred embodiment of the present invention, the abalone used in the present invention is abalone ( Haliotis) discus hannai), horses overturned (Holiotis gigantea), Blackcurrant Overture (Haliotis discus ), Haliotis sieboldi or Haliotis diversicolor super - texta , more preferably true abalone, horse abalone, black abalone or sibolt abalone, and even more preferably a true abalone or a horse abalone More preferably, even more preferably is abalone, and most preferably is abalone abalone inhabited or cultured in Jindo-gun, Jeollanam-do, Korea.

The composition of the present invention includes the abalone viscera extract used as an active ingredient. The term “extract” as used herein referring to abalone viscera is an abalone viscera workpiece formulated (eg, powdered) for administration to an animal of the abalone viscera itself as well as the extraction result obtained by treating the extract solvent in the abalone viscera. Also has a meaning to include.

If the abalone viscera extract used in the composition of the present invention is obtained by treating the abalone viscera with an extracting solvent, various extracting solvents may be used. Preferably, (a) water, (b) anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms (e.g. methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol and normal-butanol, etc.), (c) Mixed solvent of the lower alcohol and water, (d) acetone, (e) ethyl acetate, (f) chloroform, (g) 1,3-butylene glycol, (h) hexane, (i) diethyl ether, or (j) Butyl acetate may be treated with abalone intestine to obtain an extract.

As used herein, the term "extract" has a meaning commonly used as a crude extract in the art as described above, but broadly includes a fraction additionally fractionating the extract. That is, the abalone viscera extract includes not only those obtained by using the above-described extraction solvent, but also those obtained by additionally applying a purification process thereto. For example, fractions obtained by passing the extract through an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity), etc. The fraction obtained through the purification method is also included in the abalone viscera extract of the present invention.

Abalone viscera extract used in the present invention may be prepared in a powder state by an additional process such as vacuum distillation and freeze drying or spray drying.

Alternatively, abalone viscera extract may be used abalone viscera processing formulated (eg, powdered) to administer the abalone viscera itself to the animal.

The composition of the present invention is administered to a living body to function to enhance immunity. As used herein, the term "immune potentiation" is meant to increase the immune response or activity of the immune system in vivo.

In vivo immune responses can be broadly divided into Th1 and Th2 immune responses. As used herein, the term “Th1 immune response” refers to a response that induces a cellular immune response, and “Th2 immune response” refers to a response that promotes a humoral immune response.

According to a preferred embodiment of the present invention, the composition of the present invention enhances a Th1 immune response or a Th2 immune response. More preferably, the compositions of the present invention enhance both Th1 immune and Th2 immune responses.

The composition of the present invention promotes the expression of various cytokines involved in the Th1 immune response. For example, the compositions of the present invention enhance the expression of interleukin-17 (IL-17), interferon gamma (IFN-γ) or tumor necrosis factor-α (TNF-α). The composition of the present invention promotes the expression of various cytokines involved in the Th2 immune response. For example, the compositions of the present invention enhance the expression of IL-4 or IL-13.

According to a preferred embodiment of the present invention, the composition of the present invention exhibits an excellent effect of promoting the proliferation of B cells, T cells or B cells and T cells.

According to a preferred embodiment of the present invention, the abalone viscera extract increases the production of IgG (Immunoglobulin G) or IgE (Immunoglobulin E) antibodies of B cells. IgG is the most abundant antibody in the human body, and is responsible for removing these toxic substances by inducing phagocytosis when invading bacteria and viruses. IgE is a type of antibody that plays an important role in protecting our bodies, especially from parasitic infections. This IgE is also secreted in large amounts during the infection of the parasites to activate the phagocytosis of immune cells to induce the removal of infected cells and parasites.

The composition of the present invention may be provided in a pharmaceutical composition or a food composition.

When the composition of the present invention is made into a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention may be administered orally or parenterally, and preferably applied by oral administration.

Suitable dosages of the pharmaceutical compositions of the present invention may vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to response of the patient. Can be. Typical dosages of the pharmaceutical compositions of the invention are in the range of 0.001-100 mg / kg on an adult basis.

The pharmaceutical compositions of the present invention may be prepared in unit dose form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporation into a multi-dose container. The formulation may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of extracts, powders, powders, granules, tablets or capsules, and may further comprise dispersants or stabilizers.

When the composition of the present invention is prepared as a food composition, as an active ingredient, as well as abalone viscera extract, it contains components that are commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavors Include the first. Examples of the above carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And sugars such as conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and xylitol, sorbitol, erythritol. As the flavoring agent, natural flavoring agents (tauumatin, stevia extract (for example rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

The features and advantages of the present invention are summarized as follows:

(Iii) The present invention provides an immunopotentiated composition using the abalone viscera extract.

(Ii) The composition of the present invention comprising the abalone viscera extract as an active ingredient greatly improves the immune activity in vivo by enhancing the Th1 immune response (ie, cellular immune response) and Th2 immune response (ie, humoral immune response) do.

(Iii) The present invention provides basic data as medicines and foods of abalone viscera extract having an immune enhancing function.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Example

Experimental material

Abalone  And mouse

The abalone abalone ( Haliotis discus hannai) was used as a sample of cultured abalone produced in Jindo-gun, Jeollanam-do, and the abalone viscera was extracted from the abalone and then lyophilized to convert it into a powder form. Mice for the study used Balb c / J mice (SLC, Japan).

Isolation of T and B Cells

Two groups of mice were orally administered with PBS or abdominal intestine for 20 days, and spleen and intestinal lymph nodes were extracted from each mouse. The lymph nodes thus harvested were made to single cell level using collagenase V (Sigma, USA). For specific separation of T cells and B cells from single cells, T cells are CD4 ⁺ isolation magnetic beads (Milteny Biotech, Germany, Cat No. 130-049-201), and B cells are B220 magnetic beads (Milteny). Biotech, Germany), and columns (milteny Biotech, Germany, Cat No. 130-042-401) were used to acquire lymph node T cells and B cells.

Experiment method

Measurement of Cytokine Expression

To measure the amount of cytokines expressed by T cells and B cells, RNA was isolated from the isolated T and B cells using a Trizol reagent (Molecular Research Center), and 1 µg of the isolated RNA was oligo (dT) primer ( CDNA was synthesized using Promega) and the Improm-II Reverse Transcription System (Promega).

Quantitative quantification of the amount of cytokines expressed by each cell using quantitative real-time PCR using various primers that can specifically amplify specific cytokines using the synthesized cDNA Analyze by. Quantitative real-time PCR was incubated at 95 ° C. for 10 minutes, followed by 40 cycles of 95 ° C. 30 sec, 62 ° C. 30 sec, and 72 ° C. 30 sec. The sequence of the primers used is as follows: IL-2 (forward primer 5′-CCT GAG CAG GAT GGA GAA TTA CA-3 ′ and reverse primer 5′-TCC AGA ACA TGC CGC AGA G-3 ′); IL-4 (forward primer 5 'ACA GGA GAA GGG ACG CCA T-3' and reverse primer 5'-GAA GCC CTA CAG ACG AGC TCA-3 '); IL-10 (forward primer 5'-ATA ACT GCA CCC ACT TCC CA-3 'and reverse primer 5'-TCA TTT CCG ATA AGG CTT GG-3'); IL-17A (forward primer 5'-TTC ATC TGT GTC TCT GAT GCT-3 'and reverse primer 5'-TTG ACC TTC ACA TTC TGG AG-3'), IFN-γ (forward primer 5'-TCA AGT GGC ATA GAT GTG GAA GAA-3 'and reverse primer 5'-TGG CTC TGC AGG ATT TTC ATG-3'), TNF-α (forward primer 5'-CAT CTT CTC AAA ATT CGA GTG ACA A-3 'and reverse primer 5 '-TGG GAG TAG ACA AGG TAC AAC CC-3'), IL-13 (forward primer 5'-GCA ACA TCA CAC AGG ACC AGA-3 'and reverse primer 5'-GTC AGG GAA TCC AGG GCT AC-3' )

IgG ( immunoglobulin  G) and IgE ( immunoglobulin  Determination of the secretion capacity of E)

B cells isolated from each mouse for determination of the amount of antibody secreted by the B cells were collected from lipopolisaccharide (LPS) (Sigma, USA), CD40 antibody (BD Bioscience, USA), IgM antibody (BD Bioscience, USA) and Various stimuli via IL-4 (InterLukin-4; (BD Bioscience, USA) were incubated in the incubator for a total of three days.To determine the amount of antibody present in the harvested cultures, the IgG antibody was subjected to mouse IgG ELISA. (Enzyme-Linked Immunosorbent Assay) kit (BD Bioscience, USA) was used, and the IgE antibody was measured using the mouse IgG ELISA kit (Bethyl Laboratories, Inc., USA) to measure the amount of IgG and IgE in the culture.

Measurement of Cell Differentiation Capacity

To measure the differentiation capacity of T cells and B cells, T cells use CD3 antibodies (BD Pharmigen, USA; Cat No. 553058) and CD28 antibodies (BD Pharmigen, USA; Cat No. 553294), and B cells are lipopolysaccharides. After stimulation with various CD40 antibodies, IgM antibodies, and IL-4, the cells were cultured for 60 hours. The cultured cells were cultured again by adding 0.5 μCi of [H ³ ] -thymidine (NEN Life science, Boston, Mass., USA), which is a radioisotope, and cultured again for 16 hours. ) Values were measured using a scintillator.

Observation of Weight Changes and Intestinal Condition in Mice

The body weight of the mice was measured every day before oral administration to monitor the change in mice after oral administration of PBS or abalone viscera extract for 20 days. At the end of the last administration, the intestines were removed from the mice in each group and the intestine condition was visually confirmed.

Animal Experiment Time Schedule

The 6-week-old mice were divided into two groups, one of which was a control group, PBS, and the experimental group received oral administration of the same amount of abalone viscera extract using a syringe in each group of mice daily for 20 days (FIG. 1). At this time, the weight change of the two groups of mice was monitored before administration. After 20 days of oral administration, T cells and B cells were separated from the spleen and intestinal lymph nodes of each group of mice in the same manner. In order to measure the activity of T cells, experiments were performed to measure the level of T cell proliferation and cytokine expression, and to measure the level of B cell proliferation, cytokine expression, and antibody secretion. It was.

Experiment result

1. Influence of Abalone Visceral Extracts on Mouse Growth

Abalone and abalone viscera are currently used as foods, but foods can also cause abdominal pain and upset stomach in general when excessive doses are taken. Therefore, prior to this study, the abdominal viscera extract (1 mg / mouse) was tested for oral administration in mice. To this end, two groups of mice orally administered PBS and abalone viscera extracts were fasted for 1 hour before oral administration, and the weight of the mice in each group was measured immediately before the administration, thereby monitoring the change in the weight of the mice during oral administration. It was.

Compared to the mouse group administered orally with PBS, the control group, the abalone viscera extract showed a significantly higher weight gain after 6 days of administration (FIG. 2). As shown in FIG. 2, it was further observed that the oral administration of the abalone oral mice increased by about 1 g (5% of body weight) in the control mice on the 20th day of the last oral administration. there was.

In addition, after the last oral administration in order to confirm the presence of more accurate toxicity, the mice of each group was sacrificed, the intestines were extracted from each group to determine whether the intestinal tissue state changes according to oral administration. Large differences were not observed in the intestines of the mice administered PBS and the intestines of the abalone viscera extract (FIG. 3).

The results showed that the 1 mg abalone viscera extract used in this study did not show toxicity during oral administration (1 dose / day) for 20 days in mice. The increase rate could be confirmed. This phenomenon is inferred due to the high nutritional function (high protein food) of abalone viscera extract, as it is known so far, and indirectly confirmed the high pharmacologic function of abalone, which has been oral without scientific proof. .

2. Evaluation of the Effect of Abalone Visceral Extracts on T Cells

A Study on the Change of T Cell Proliferation in Abalone Intestine Extracts

The activity of T cells could be evaluated by the degree of proliferation by which the cells themselves proliferate and the degree of secretion of immune mediators secreted by the cells. Therefore, in this experiment, the change in the degree of proliferation of T cells that may appear upon ingestion of abalone viscera extract was confirmed. In general, immune cells begin to proliferate only when given an external stimulus, such as invading pathogens, and the extent to which they respond to the external stimulus depends on how proliferates the same stimulus. Therefore, in order to give the same stimulation as this external stimulation, in this experiment, the stimulation was performed using CD3 and CD28 antibodies, which can stimulate T cells, and then the change in the degree of T cell proliferation was measured using radioisotopes. saw.

As can be seen in Figure 4, there was little differentiation of the two groups of T cells in the absence of stimulation. However, when stimulated with CD3 antibody, T cells of mice treated with Abalone viscera extract showed about 100% increase in cell proliferation compared to T cells of control group, using stronger CD3 and CD28 antibodies. Even when stimulated, the proliferation of T cells treated with Abalone viscera extract was consistently high by about 20%. This suggests that ingestion of abalone viscera increases T cell proliferation.

Changes in the Expression of Immune Intermediates in T Cells by Ingestion of Abalone Visceral Extracts

The results of splenic position T cells showed the effect of oral administration of abalone viscera extract on the total immunity, and the results of intestinal lymph node position T cells showed the effect on the vasculature (Fig. 5). To evaluate the effect of abalone viscera extract on the whole immune system, splenic CD4 + T cells were isolated from two groups of mice, and CD4 + T cells were stimulated with CD3 and CD28 antibodies for 12 hours before RNA was isolated. Afterwards, the expression patterns of various functions of cytokines were compared with T cells of the control group using quantitative rea-time PCR.

As a result, the expression level of IL-4 (Interleukin-4), IL-17 (Interleukin-17) and TNF-α (Tumor Necrosis Factor-α) in the mouse spleen T cells orally administered abalone viscera extract It was confirmed that the increase significantly compared to (Fig. 5a). IL-4 is an immune mediator that induces a representative humoral immune response (Th1-mediated immune response). In the case of IL-17 and TNF-α, it is an immune mediator that induces a cellular immune response (Th2-mediated immune response). It is a substance. Therefore, the intake of abalone viscera extract can be confirmed to increase both humoral and cellular immunity by T cells.

In addition, the same method was used to extract intestinal lymph nodes from two groups of mice, isolate CD4 + T cells, stimulate CD4 + T cells, and measure changes in cytokine expression using quantitative rea-time PCR. It was. As can be seen in Figure 5b, it was confirmed that the expression of IL-4 and IL-17 significantly increased in the case of intestinal lymph node position T cells similarly to the splenic position T cells, and interferon-gamma (Interferon- gamma: IFN-γ) expression was also significantly increased. It can be concluded that ingestion of abalone viscera extract improves immune function of T cells not only in intestinal immune system but also in whole immune system.

3. Evaluation of the Effect of Abalone Intestine Extract on B Cells

A study on the degree of proliferation of B cells after ingestion of abalone viscera extract

Through the previous experiment, the effect of abalone viscera extract or immune response on T cells was evaluated. In this experiment, the effect on the immune response capacity of B cells was evaluated. Experimental method was made in the same manner as described above, and after stimulating the cells under various conditions after separating the B cells, the extent of B cell proliferation was measured using a radioisotope.

After oral administration of abalone viscera extract, B cells proliferated about 30% higher than B cells in the control group when stimulated with CD40 antibody and IgM antibody and when stimulated with CD40 antibody, IgM antibody and IL-4, respectively. It could be confirmed that the degree. It can be seen that uptake of abalone viscera increases the proliferation of B cells (FIG. 6).

Evaluation of the Effect of Abalone Visceral Extracts on Antibody Production of B Cells

The main task of B cells in the immune system is to produce antibodies, neutralize various toxins, and induce humoral immune responses through reactions with foreign antigens. Therefore, in this experiment, the change of antibody secretion of B cells was measured by ingestion of abalone viscera extract for proper evaluation of B cell function.

Briefly describing the experimental method, in order to induce the production of the antibody from the two groups of B cells separated by various stimulation by the same method described above, the cells were cultured in vitro for 3 days. The culture solution of the cells thus cultured was collected and the amount of antibody contained therein was accurately quantified by ELISA.

IgG is the most abundant antibody in the human body, and is responsible for removing these toxic substances by inducing phagocytosis upon invasion of bacteria and viruses. In experiments that measured the amount of IgG antibodies secreted by B cells after stimulating B cells using various kinds of stimuli, the B cells of mice with abalone viscera extracts were significantly higher than those of the control group. It was confirmed that the level has the ability to secrete IgG antibody (Fig. 7a).

Next, the degree of IgE secreted by B cells was compared. IgE is a type of antibody that plays an important role in protecting our bodies, especially from parasitic infections. This IgE is also secreted in large amounts during the infection of the parasites to activate the phagocytosis of immune cells to induce the removal of infected cells and parasites. As can be seen in Figure 7b it was confirmed that the amount of IgE secreted by the B cells of the mice ingested abalone viscera extract significantly higher than the control. Through this, intake of abalone viscera extract was confirmed to increase the antibody production capacity of B cells to a high level.

Changes in the Expression of Immune Intermediates in B Cells by Ingestion of Abalone Intestine Extracts

The function of B cells can be confirmed not only by the cell proliferation capacity and the antibody production capacity described above, but also by the degree of secretion of cytokines, which are immune regulators. Therefore, in this experiment, we confirmed the changes in cytokine expression of B cells that are changed by ingestion of abalone viscera extract.

As a result, oral administration of abalone viscera extract significantly increased the expression level of TNF-α specifically in splenic B cells (FIG. 8A), and IL-13 (Interleukin-13) in intestinal lymph node B cells. It was confirmed that the expression of increased to a significant level (Fig. 8b). Based on these results, it was confirmed that ingestion of abalone viscera extract increased the expression of specific cytokines in splenic and intestinal lymph node position B cells.

conclusion

In conclusion, continuous ingestion of abalone viscera extract was found to enhance the function of T cells and B cells in the human immune system. The present invention is the first data to scientifically prove the efficacy of abalone viscera extract.

Review

To date, abalone has been studied in terms of its pharmacological function, but there are no studies on its function in immunological aspects.

The present invention carried out an evaluation study on the effect of the intake of abalone viscera extract in immunological aspects in order to prepare a more empirical and scientific basis for the function of abalone viscera extract. To evaluate the toxicity of the abalone viscera extract used in this experiment, mice were orally administered various abalone viscera extracts, and even at the highest concentration of 1 mg / ml, any weight loss was observed during the 20-day oral administration period. No weight gain, as well as a higher level of weight gain compared to the control. In addition, when comparing the state of the intestines at the expense of mice after 20 days of administration, no significant difference was found between the two groups. Through this study, it was confirmed that the abalone viscera extract used in this study was not toxic and had high nutritional function. Again indirectly confirmed.

Evaluation of the Effect of Abalone Visceral Extracts on T-cell Functions In conclusion, we concluded that continuous uptake of abalone viscera extracts increased T-cell function in both the overall and intestinal immune systems. First, the evaluation of the effect of intake of abalone viscera extract on the proliferation of T cells showed that sustained intake increased the proliferation of T cells to a significant level compared to the control group. A second evaluation of the effect of the extract on the level of cytokine expression in T cells revealed that abalone visceral extract significantly improved the humoral immune cytokine IL-4 and cellular immune cytokines in both spleen and lymph node T cells. It was confirmed that the expression levels of the kine IFN-γ, IL-17 and TNF-α can be increased. Through this, it was confirmed that continuous intake of abalone viscera extract improved T cell function of total immune and intestinal immune system.

In addition, the results of the evaluation of the effect on the function of B cells confirmed that the intake of abalone viscera extracts not only T cells but also B cells improved its immune function, and the experiments measuring the proliferation of B cells Compared to the B cells of the control group, the proliferation of the B cells of the experimental group was confirmed to increase to a significant level. As a result, the production capacity of the antibody was also significantly increased compared to the B cells of the control group, it was confirmed that both the IgG antibody and IgE antibody secreted much in response to the stimulation.

Lastly, the expression level of IL-13 and IFN-γ in both spleen and intestinal lymph node position B cells of oral administration of abdominal visceral extracts were investigated through a study on the change in the expression level of cytokine, an immunomodulator. It was confirmed that this increased to a meaningful level.

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

1 is a schematic diagram showing the process of the animal experiment method.

Figure 2 is a graph showing the weight change of mice after oral administration of PBS or abalone viscera extract (40 ㎍ / g) once daily for 20 days. The control group is a group administered with PBS, and the sample is a group administered with abalone viscera extract.

Figure 3 is an image showing the appearance of the intestine collected from mice orally administered PBS or abalone viscera extract (40 ㎍ / g) once daily for 20 days.

4 is a result of stimulating T cells of mice injected with oral PBS or abalone viscera extracts with CD3 antibody and CD28 antibody for 56 hours, and then measuring the proliferation change of T cells with a radioisotope using a scintillator. The control group is a group administered with PBS, and the sample is a group administered with abalone viscera extract.

5 is a result of analyzing the amount of cytokines expressed in T cells located in (a) spleen and (b) intestinal lymph nodes of mice administered orally with PBS or abalone viscera extract, respectively, by real-time PCR. The control group is a group administered with PBS, and the sample is a group administered with abalone viscera extract.

Figure 6 is stimulated with LPS, CD40 antibody, IgM antibody and IL-4 for 3 days in B cells of mice orally administered PBS or abdominal viscera extract, respectively, the growth of T cells using a radioisotope scintillator ( scintillator). The control group is a group administered with PBS, and the sample is a group administered with abalone viscera extract.

Figure 7 is stimulated with LPS, CD40 antibody, IgM antibody and IL-4 in B cells of mice orally administered PBS or abalone viscera extract, respectively, and cultured for 3 days (a) IgG and (b) IgE contained in the culture medium It is the result of measuring the quantity of the antibody of ELISA. The control group is a group administered with PBS, and the sample is a group administered with abalone viscera extract.

FIG. 8 shows the results of real-time PCR analysis of the amount of cytokines expressed in B cells located in (a) spleen and (b) intestinal lymph nodes of mice administered orally with PBS or abalone viscera extract. The control group is a group administered with PBS, and the sample is a group administered with abalone viscera extract.

Claims (8)

Immunostimulating composition comprising an abalone viscera extract as an active ingredient. The method of claim 1, wherein the abalone is Haliotis discus hannai , Holiotis gigantea , Haliotis discus , Haliotis sieboldi or Haliotis diversicolor super-texta Immunopotentiating composition. 3. The immunopotentiated composition according to claim 2, wherein the abalone is abalone. 2. The immunopotentiating composition of claim 1, wherein the composition enhances a Th1 immune response, a Th2 immune response, or both a Th1 immune response and a Th2 immune response. The immunopotentiating composition according to claim 4, wherein the composition promotes proliferation of B cells, T cells or B cells and T cells. According to claim 4, wherein the composition enhances the expression of cytokine IL-17, IFN-γ or TNF-α involved in the Th1 immune response, and the composition of the cytokine IL-4 or IL-13 involved in the Th2 immune response Immunopotentiating composition, characterized in that to enhance expression. 5. The immunopotentiated composition of claim 4, wherein the composition increases production of IgG or IgE antibodies. 2. The immunopotentiating composition according to claim 1, wherein the composition is a food composition.

Images