KR102098998B1 - Medicinal herbs composition for enhancing immune activity - Google Patents

Abstract

The present invention relates to a medicinal herb composition for enhancing immune activity, including roe deer mushroom, zinnia mushroom, red ginseng, Bokryeong, Cheonmun-dong, McMun-dong, Gojija, Chosukjam, raw red yeast, and bee-mil, and the herbal composition for enhancing immune activity of the present invention is inflammatory In addition to reducing the cytokine (cytokine), TNF-α (tumor necrosis factor α), IL-2 and IL-6, and immunoglobulin E (Immunoglobulin E, IgE), as well as reducing white blood cells (White Blood) Cell, WBC), neutrophils (neutrophils), lymphocytes (lymphocytes), and monocytes (monocytes) to increase the effect.

Description

Medicinal herbs composition for enhancing immune activity

The present invention relates to an herbal composition for enhancing immune activity, and more specifically, to a medicinal herb composition for enhancing immune activity, including roe deer mushroom, red oyster mushroom, red ginseng, boknyeong, astronomical dong, maekmun-dong, goji berry, chosukjam, raw green beans and beeswax. It is about.

As interest in health increases, interest in foods, preparations, and treatment methods for improving immunity is also increasing.

Gyeongokgo (瓊 玉 膏) is one of the herbal medicine prescriptions, and consists of ginseng, baekbokryong, saengjihwang, and bongmil. It is said that Gyeongokgo fills the jeong and replenishes the water, blackens the hair, makes the teeth come out, and removes the disease by making the Manshin (구 神) prosperous. Iksuyoungjingo (것이 永 眞 膏) is the addition of cheonmun-dong, macmun-dong, and goji berry to Gyeongok High School. Iksuyoungjingo is a preparation that enhances the effect of stimulating and nurturing genitals by adding materials that can be bore in Gyeongokgo (Kim MD.The literature study on the efficacy and manufacturing process of Gyeongoggo. J Oriental Medical Classics. 2011; 24 (2): 51-64). Gyeongokgo and Iksuyoungjingo are representative high-form preparations that complement the deficiency.

In order to enhance the effectiveness of gyeongokgo or iksuyoungjingo, techniques for adding functional raw materials or other medicines have been developed. Korean Registered Patent No. 10-1198211 (2012.11.07.) Discloses a method of manufacturing gyeongokgo using sea cucumber, which boils the fleshy part of sea cucumber in which the gut is separated in a mixture of Chihwang extract, red ginseng powder and Baekboknyeong powder. After drying and pulverization to provide a jadeite made by mixing the sea cucumber powder obtained. Korean Registered Patent No. 10-1016195 (2011.02.22.) Discloses Cheonma gyeongokgo, which crushes ginseng and Baekbokryeong, produces raw saengwanghwang juice, and infiltrates the ginseng powder and Baegbokgryeong powder into the saengjihwang juice and Cheongmagyi and refrigerates It provides a cheonma gyeongokgo prepared by adding the cheonma gyo during the process of preparing saccharified solution with glutinous rice by storing, immersing, cooling and tangling.

Thus, the inventors of the present invention, as an additional raw material to the composition constituting Iksuyoungjingo (red ginseng, Baekbokryeong, raw sake juice, beekmil, cheonmun-dong, maekmun-dong, gojija), the herbal composition containing roe deer mushroom, zinnia mushroom, and zinnia sleep is immune active It has been confirmed that it promotes and has led to the present invention.

Korean Registered Patent No. 10-1198211 (2012.11.07.) Korean Registered Patent No. 10-1016195 (2011.02.22.)

An object of the present invention is to provide an herbal composition for enhancing immune activity.

Another object of the present invention is to provide a method for preparing an herbal medicine for enhancing immune activity.

In order to achieve the above object, the present invention provides a herbal medicine composition for enhancing immune activity, including roe deer mushroom, matsutake mushroom, red ginseng, bokryeong, cheonmun-dong, maekmun-dong, goji berry, chosukjam, saengjihwang, and beekmil.

In one embodiment of the present invention, the bokryeong may be baekbokryeong, bokryeongpi or a mixture thereof.

In one embodiment of the present invention, the raw geohwang may be raw geohwang juice, geohwang foil or a mixture thereof.

In one embodiment of the present invention, the composition for medicinal herbs for enhancing immune activity is 1 to 7% by weight of yellow beetle mushroom, 0.5 to 5% by weight of zinnia mushroom, 2 to 10% by weight of red ginseng, 5 to 15% of bokkryeong Weight%, 0.5 to 5% by weight, munmundong 0.5 to 5% by weight, 0.5 to 5% by weight goji wolfberry, 0.5 to 5% by weight choknapjam, 30 to 53% by weight of raw green beans and 25 to 40% by weight sealing .

In one embodiment of the present invention, the herbal composition for enhancing the immune activity may be fermented with a Leuconostoc mesenteroides subsp. Dextranicum strain.

In one embodiment of the invention, the fermentation may be carried out at 33 to 38 ℃ for 3 to 7 days.

In one embodiment of the present invention, the herbal composition for enhancing immune activity may be included in cosmetics, health supplements, or pharmaceutical compositions.

In one embodiment of the present invention, the herbal composition for enhancing immune activity may be prepared in a soft extract, pill, tablet or high dosage form.

In order to achieve another object of the present invention, the present invention provides a method of manufacturing a high-medicinal herb for enhancing immune activity by using the Chinese herbal medicine method for enhancing the immuno-activity.

The herbal composition for enhancing immune activity of the present invention reduces inflammatory cytokines, TNF-α (tumor necrosis factor α), IL-2 and IL-6, and immunoglobulin E (Immunoglobulin E, IgE), an immune antibody. In addition to having a reducing effect, white blood cells (WBC), neutrophils (neutrophils), lymphocytes (lymphocytes), and monocytes (monocytes) increase the effect.

In addition, the herbal composition for enhancing the immune activity of the present invention is excellent in acid resistance and sufficiency and has antibacterial activity against harmful bacteria such as E. coli Leuconostoc mesenteroides subsp. Dextranicum strain By being fermented using, Leukonostock mesenteroides subspheris dextranicum strain survives through the stomach and digestive tract to the intestine, thereby suppressing harmful microorganisms in the intestine and preventing pathogenic microbial infections, which is beneficial for gut health and probiotics ( probiotics).

The herbal composition for enhancing immune activity of the present invention may be used in cosmetics, health supplements, or pharmaceutical compositions.

The herbal composition for enhancing immune activity of the present invention can not only reduce the cost but also prevent the environmental pollution by reducing the generation of waste by utilizing the donation of blossom mushrooms that were previously disposed of, Bokryeongpi or Chihwang.

1 is a phylogenetic analysis according to 16S rRNA sequencing of Leuconostoc mesenteroides subsp. Dextranicum strains finally selected in the present invention.
Figure 2 is a graph showing the effect of the immune enhancing composition of the present invention on the TNF-α content in the serum of rats.
Figure 3 is a graph showing the effect of the immune-enhancing composition of the present invention on the serum IL-2 content of rats.
Figure 4 is a graph showing the effect of the immune-enhancing composition of the present invention on the serum IL-6 content of rats.
5 is a graph showing the effect of the immune enhancing composition of the present invention on the IgE content in the serum of rats.
6 is a graph showing the effect of the immune enhancing composition of the present invention on the spleen weight of rats.
7 is a graph showing the effect of the immune enhancing composition of the present invention on leukocytes in rat blood.
8 is a graph showing the effect of the immune enhancing composition of the present invention on red blood cells in rat blood.
9 is a graph showing the effect of the immune enhancing composition of the present invention on the weight gain of rats.
10 is a graph showing the effect of the immune enhancing composition of the present invention on the serum aminotransferase content of rats.

Herbal medicine composition for enhancing the immune activity of the present invention is characterized in that it contains roe deer mushroom, matsutake mushroom, red ginseng, Bokryeong, Cheonmun-dong, McMun-dong, goji berry, Chosukjam, saengjihwang, and beeswak.

In the present invention, the powder form of roe, mushroom, red ginseng, Bokryeong, astronomical dong, McMun-dong, goji berry, and Chomnapjam are preferably used.

In the present invention, the composition for medicinal herbs for enhancing immunity activity is 1 to 7% by weight of yellow beetroot mushroom, 0.5 to 5% by weight of zinnia mushroom, 2 to 10% by weight of red ginseng, 5 to 15% by weight of bokryeong, 5 to 15% by weight, cheonmundong It is preferable to include 0.5 to 5% by weight, 0.5 to 5% by weight of McMundong, 0.5 to 5% by weight of goji berry, 0.5 to 5% by weight of Chomnapam, 30 to 53% by weight of raw sulcus, and 25 to 40% by weight of sealed wheat.

In the present invention, the blossom mushrooms may be used alone or as a mixture of the fruit body or the stem of the blossom mushroom.

The base of the blossom mushroom is obtained through a process of separating the fruit from the harvested blossom mushroom. When the mushrooms are harvested by the sawdust cultivation method, it is preferable that the base separated from the fruiting body has undergone a pre-treatment process for removing sawdust with a dry grinder.

In the present invention, the Bokryeong may be used in Baekbokyeong, Bokryeongpi or a mixture thereof. When Baeknyeongnyeong and Boknyeongpi are used in combination, it is preferable to contain 3 to 13% by weight of Baengnyeongnyeong and 0.5 to 5% by weight of Boknyeongpi based on the total weight of the composition.

In the present invention, the raw geohwang may be used in combination with raw geohwang juice, geohwang foil or these. It is squeezed raw saenghwang and the raw saenghwang juice is squeezed out and left as a residue is jihwang foil. When used in a mixture of raw aquatic sulfur juice and citrine foil, it is preferable to include 25 to 40% by weight raw aquatic citrate juice and 10 to 25% by weight citrate foil based on the total weight of the composition.

In the present invention, the composition for medicinal herbs for enhancing the immune activity is 1 to 7% by weight of yellow beetroot mushrooms, 0.5 to 5% by weight of zinnia mushrooms, 2 to 10% by weight of red ginseng, 3 to 13% by weight of Baekbokryeong, Bokryeongpi 0.5 to 5% by weight, 0.5 to 5% by weight in Astronomical Building, 0.5 to 5% by weight in McMundong, 0.5 to 5% by weight wolfberry, 0.5 to 5% by weight, Chomyeonjam 25 to 40% by weight, 10 to 25% by weight of yellow citrine juice And 25-40% by weight of sealing.

On the other hand, the herbal composition for enhancing the immune activity of the present invention may be fermented into Leuconostoc mesenteroides subsp. Dextranicum strain.

In the present invention, the fermentation is preferably performed for 3 to 7 days at 33 to 38 ° C, and more preferably for 4 to 6 days at 36 to 37 ° C.

The herbal composition for enhancing the immune activity of the present invention may be prepared in a soft extract, pill, tablet or high dosage form.

In particular, the herbal composition for enhancing immune activity of the present invention may be prepared in a high dosage form using a double boiling method.

Bokryeong is a mushroom of the genus genus Mushrooms of the genus Dermatophyte, which is parasitic on the roots of pine trees. The surface of the mushroom shade is reddish brown, light brown, and dark brown. The flesh is white and gradually turns pink. The white one is called Baekbokryeong, and the red one is called Jeokbokryeong. As the pharmacological action of Bokryeong, diuretic action, bactericidal action, relaxation of the intestinal tract, ulcer prevention effect, hypoglycemic action, increased cardiac contractility, immune-enhancing action, and anti-tumor action are known.

Bokryeongpi is about 20% of the total weight of Bokryeong. Normally, it has been reported that the bokryongpi can be suppressed in allergies by inducing conditioned T cells in vivo.

Roe deer mushroom is a white, white, thick mushroom that grows on the trunks of broad-leaved trees, such as oak and oak trees, from summer to autumn. Most mushrooms have up to two types of active polysaccharides, but roe deer mushrooms contain five types of active polysaccharides, which exert excellent anti-cancer effects. In addition, research results have shown that roe deer mushroom is good for digestive diseases by strengthening blood vessels surrounding the stomach wall, and is also effective in preventing dementia and improving diabetes. In addition, it has the effects of improving chronic enteritis, increasing immune function, and suppressing dementia, and has been spotlighted as a healthy food due to its low calorie and abundant dietary fiber.

Blossom mushrooms have a high beta-glucan content of at least 40% of the dry weight. Beta-glucan is a polysaccharide, a substance that exists in cell walls, mushrooms, and grains. It does not directly attack cancer cells, but activates the immune function of normal cells in humans with a non-specific immune response, suppresses the proliferation and recurrence of cancer cells, and activates macrophages. It enters the body with cancer cells and promotes the secretion of various cytokines, thereby activating the immune function of immune cells T cells and B cells. Beta glucan, a major component of the blossom mushroom, has a limitation in effective intake or hydration in a general way, so it is preferable to perform a particle size of 200 to 440 mesh when crushing the blossom mushroom in the present invention.

Cho-Zam-Jam is a perennial plant belonging to the family of honeybees of the dicotyledonous plant, contains a component called phenylethanoid, which activates brain function, and is rich in choline that can prevent dementia. Chorus sleep also helps to prevent edema and stroke, and improves liver cirrhosis and arteriosclerosis and suppresses the formation of fatty liver by smoothing blood circulation. In addition, it has the effect of helping to stop bleeding and boils, and is also effective for headaches, sore throat, and bronchitis, including colds.

Hereinafter, the herbal composition for enhancing the immune activity of the present invention through the following examples will be described in more detail.

<Example 1> Selection of microorganisms suitable for fermentation of herbal composition

1-1. Strain separation from natural products

A total of 17 fermented foods (6 kinds of cabbage kimchi, 3 kinds of fresh kimchi, 3 kinds of Dongchimi, bachelor kimchi, radish kimchi, cucumber pickle, plum liquid, yogurt) were used as samples. In order to separate microorganisms from the fermented food, each sample is diluted in decimal and smeared on Lactobacilli MRS agar (Difco Co., USA) and incubated at 37 ° C for 2 days to randomly separate and separate lactic acid bacteria. Silver was passaged and stored at 4 ° C for use.

1-2. Separation of lactic acid bacteria suitable for fermentation starters

Among the strains isolated from MRS agar, lactic acid bacteria are separated using MRS plate medium containing pH6, which is a pH indicator of 0.006% BCP (Bromocresol Purple, Sigma, USA), and the purple medium of BCP is yellow. Lactobacillus was isolated repeatedly until a pure strain was obtained by confirming the colonies changing to.

1-3. Acid resistance analysis

In order to analyze the acid resistance, the isolates were inoculated 10% in 0.05 M sodium phosphate buffer solution adjusted to pH 2.0 and 4.0 with 0.05 M sodium phosphate buffer solution pH 7.0 and HCl, and then cultured with shaking at 37 ° C for 2 hours, followed by 600 Survivability (%) was calculated by measuring absorbance at nm.

The pH of gastric juice is pH 0.78 to 0.90 for fasting, but when food is consumed, the pH of the stomach rises to 2.0 to 3.0 and the time for food to pass through the stomach is 2 to 4 hours. In order to show the effect as a probiotic, it must survive below pH 3 and reach the small intestine through the stomach. Strains having a survival rate of 70% or more at pH 2 were selected.

1-4. Bile resistance analysis

In order to analyze the bile resistance, the isolates were inoculated 10% in 0.3% (w / v) oxgall added with Lactobacilli MRS broth and 0.45 μm membrane filter, and incubated at 37 ° C. for 8 hours, followed by incubation at 37 ° C. for 600 hours. The absorbance was measured at and the survival rate (%) was calculated.

1-5. Antibacterial activity against E. coli

The strain used for screening for antibacterial activity was obtained by inoculating the culture medium of 2-3 mL of cultured on a plate medium and inoculating it into a 10 mL nutrient broth bacterial growth liquid medium, incubating at 37 ° C. for 24 hours, and then activating 0.2 mL of the test solution. Then, it was coated with a sterilized glass rod to spread evenly on the medium for the base layer, and then measured by agar plate diffusion method using a paper disc of Piddok.

1-6. Identification of selected strains

When the results of acid resistance, bile resistance and antibacterial activity were synthesized, the most efficient 5-1 strain was finally selected and the finally selected strain was analyzed by 16S rRNA partiall sequencing. 1 shows a phylogenetic analysis diagram according to 16S rRNA sequencing of the finally selected strain in the present invention. As a result of microbial identification, it was confirmed that the final selection strain 5-1 showed 99% similarity to the Leuconostoc mesenteroides subsp. Dextranicum strain (Korea Microbiological Conservation Center). In FIG. 1, the scale bar represents a 0.002% nucleotide difference.

<Example 2> Preparation of herbal composition for enhancing immune activity

2-1. Ingredient preparation

Sparassis crispa was purchased from (Young) Baek-Asan Blossom Mushroom (Hwasun, Korea), and Hericium erinaceus was purchased from Eulim Bio (Pyeongteck, Korea). Ginseng (Red Ginseng radix), Poria cocos (Poria cocos), cheonmundong (Asparagi radix), maekmundong (Liriopis tuber), Goji (Lycii fructus), the cornerstone sleep herbal medicines distribution center (Stachys sieboldii Miq), Honey (Honey) (Hwasun, Korea) and Omni Herb (Daegu, Korea). Rehmanniae radix was purchased from Saengjihwang Farming Association (Geumsan, Korea). All materials were made in Korea.

Roe deer mushroom, zinnia mushroom, red ginseng, Bokryeong, Cheonmun-dong, McMun-dong, goji berry, Choseokjam, and raw sulfur were washed with ultrasonic cleaner. The bokryeong was peeled and divided into bokryeongpi (15% of the bokryeong raw material) and white bokryeong (85% of the bokryeong raw material). Among the above-mentioned materials, solid materials such as red beetroot mushroom, matsutake mushroom, red ginseng, Baekbok-ryeong, Bokryeong-pi, cheonmun-dong, maekmun-dong, gojija, and choseokjam were respectively pulverized and finely powdered. The raw turmeric was compressed by crushing and then separated into a raw turmeric juice (60% juice rate) and a crimson foil.

2-2. Preparation of herbal composition for enhancing immune activity

The ingredients prepared above were mixed in the amounts shown in Table 1 to prepare an herbal composition for enhancing immune activity.

Prepared material amount (g) Red ginseng 900 Sealing 6000 Astronomical building 150 McMundong 150 Wolfberry 300 Blossom mushroom 200 Roe deer mushroom 700 Sleep 300 Baekbongnyeong 1530 Saengjihwang Juice 5760 Bokryeongpi 270 Jihwang Park 3840 system 20,100
<Example 3> Preparation of herbal composition for enhancing immune activity

delete

The ingredients prepared in Example 2 were mixed in the amounts shown in Table 1 to prepare an herbal composition for enhancing immune activity.

<Example 4> Preparation of herbal composition for enhancing immune activity

4-1. Strain preparation

A starter strain was prepared by inoculating MRS medium with the Leuconostock mesenteroides subspheris destranicum strain finally selected in Example 1 in MRS medium for 48 hours.

4-2. Preparation of herbal composition for enhancing immune activity

The materials prepared above were mixed in the amounts shown in Table 1. A 2% (v / v) inoculation of the pre-cultured Leuconostock mesenteroides subspheris dextranicum strain was fermented at 37 ° C. for 110 hours to prepare an herbal composition for enhancing immune activity.

<Example 5> Preparation of herbal composition for enhancing immune activity

A herbal medicine composition for enhancing immune activity was prepared in the same manner as in Example 4, except that the mixture was mixed in the amounts shown in Table 1.

<Example 6> Immunity-promoting high production

The herbal composition for enhancing immune activity prepared in Example 2 was kneaded well and placed in a pot. The pot was placed in a water-filled water bath. The primary bath was carried out while maintaining at 98-102 ° C for 3 days using a temperature measuring device (KP3001, Ziraffe, China), and the water filled in the bath was supplemented so that the pottery could be kept over 80%. After the 1st bath, the 1st aging process was carried out while leaving at room temperature for 1 day. Subsequently, the secondary bath was performed under the same conditions as the primary bath for 1 day, and then the secondary aging process was performed under the same conditions as the primary aging for 1 day to produce an immune-enhanced gob (IYG-gami). .

<Example 7> Immunity-promoting high production

Immune-enhancing high (膏) was prepared in the same manner as in Example 6, except that the herbal composition for enhancing immune activity prepared in Example 3 was used (M-IYG-gami).

<Example 8> Immunity-promoting high production

Immune-promoting high (膏) was prepared in the same manner as in Example 6, except that the herbal medicine composition for enhancing immune activity prepared in Example 4 was used (IYG-gami-F).

<Example 9> Preparation of high immunity

Immune-enhancing high (膏) was prepared in the same manner as in Example 6, except that the herbal medicine composition for enhancing immune activity prepared in Example 5 was used (M-IYG-gami-F).

<Experimental Example> Immunity enhancement effect through animal experiments

1-1. Experimental method

1) Animal

Sprague Dawley-based rats weighing about 170-180 g were fed solid feed (Pellet, GMO, Korea) and water in a breeding facility (room temperature 24-26 ° C, humidity 40-60%) in a constant temperature and humidity environment. Adapted to the laboratory environment for more than a week while supplying enough, it was used for experiments, and water and solid feed were freely consumed during the experiment.

2) Induced immune depression

For immunosuppression, methotrexate (MTX) (Sigma, USA) powder was defined as a concentration of 2 mg / Kg. Oral administration was performed once a day for 4 days, once every 4 days from the 8th to the 11th of the 20th of the entire experimental period. Was implemented.

3) Group separation (n = 5)

The experimental group was a normal group that did not induce immunosuppression (Normal), an untreated control group (Control) after inducing MTX immunosuppression, and an induction of MTX immunosuppression Example 6 Sample group administered experimental group A (IYG-gami), induced MTX immunosuppression After Example 8 Sample administration group B (IYG-gami-F), after administration of MTX immunosuppression Example 7 Sample administration group C (M-IYG-gami), after administration of MTX immunosuppression Example 9 sample administration group D (M-IYG-gami-F).

4) Sample administration

Each sample was prepared at a concentration of 200 mg / Kg and then administered with drinking water for the entire experimental period (20 days).

5) Weight and spleen weight measurement

Body weights were measured on the 1st, 5th, 10th, 15th, and 20th days of the experiment using an electronic balance (Cass, China). Spleen weight was measured by separating the spleen after the experiment.

6) Blood and serological tests

About 100 μl of blood obtained by blood collection was put into EDTA-bottle, and immediately injected into Multi species Hematology Analyser (950, Hemavet, USA) to measure Leukocytes, Erythrocytes, WBC, RBC, HGB, and HCT. The rest of the blood was separated for 20 minutes at 3,500 rpm in a high-speed centrifuge (Centricon T-42K, Italy), and serum was analyzed by Dri-chem 4000i (Fujifilm corp. Japan).

7) ELISA measurement

① TNF-a

TNF-α was measured using Rat TNF-α (Invitrogen, USA). To the microplate coated with Rat TNF-α, 100 μl of Rat TNF-α standard, serum, and control were added, and after tapping with a plate cover, mixed for 1 minute and left at room temperature for 2 hours. After washing 4 times with 400 μl of Washing solution, 100 μl of Rat TNF-α Biontin Conjugate was added, the plate cover was covered, and left at room temperature for 1 hour. After washing 4 times with 400 μl of Washing solution, 100 μl of Streptavidin-HRP Working solution was added, the plate cover was covered, and left at room temperature for 30 minutes. After washing 4 times with 400 μl of Washing solution, 100 μl of Stabilized Chromogen was added, the plate cover was covered, and left at room temperature (dark) for 30 minutes. After adding 100 μl of Stop solution to the plate to stop the color reaction, OD (Optical Density) was measured at 450 nm with a microplate reader (EZ Read 400, Biochrom, US). A standard curve was created to assay the amount of TNF-α in the sample.

② Interleukin-2 (IL-2)

IL-2 was measured using Rat IL-2 (R & D Systems, USA). 50 ul of Assy Diluent RD1-21 was added to each well of the microplate), 50 ul of rat IL-2 standard, control, serum was added, and after tapping with a plate cover, mixed for 1 minute and left at room temperature for 2 hours. After washing 5 times with 400 ul of Washing solution, 100 ul of Rat IL-2 conjugate was added and the plate cover was covered and left at room temperature for 2 hours. After washing 5 times with 400 ul of Washing solution, 100 ul of Substrate solution was added, the plate cover was covered, and left at room temperature (dark condition) for 30 minutes. After adding 100 ul of Stop solution to the plate to stop the color reaction, OD (Optical density) was measured at 450 nm with a microplate reader (EZ Read 400, Biochrom, US).

③ Interleukin-6 (IL-6)

IL-6 was measured using Rat IL-6 (R & D Systems, USA). 50 ul of Assy Diluent RD1-21 was added to each well of the microplate, and 50 ul of rat IL-10 standard, control, serum was added, and after tapping with a plate cover, mixed for 1 minute and left at room temperature for 2 hours. After washing 5 times with 400 ul of Washing solution, 100 ul of Rat IL-10 conjugate was added, the plate cover was covered, and left at room temperature for 2 hours. After washing 5 times with 400 ul of Washing solution, 100 ul of Substrate solution was added, the plate cover was covered, and left at room temperature (dark condition) for 30 minutes. After adding 100 ul of Stop solution to the plate to stop the color reaction, OD (Optical density) was measured at 450 nm with a microplate reader (EZ Read 400, Biochrom, US).

④ Immunoglobulin E (Ig E)

Ig E was measured using a Rat Ig E Elisa Kit (Abcam, UK). After adding 100 μl of Ig E standard, control, and serum to each well of the microplate, tapping with a plate cover, mixing for 1 minute, and allowed to stand at room temperature for 1 hour. After washing 3 times with 1X Wash Buffer, 100 µl of 1X Enzyme-Antibody conjugate was added, the plate cover was covered, and left at room temperature (dark) for 1 hour. After washing 3 times with 1X Wash Buffer, 100 µl of TMB Substrate solution was added, the plate cover was covered, and left at room temperature (dark condition) for 10 minutes. After adding 100 μl of Stop solution to the plate to stop the color reaction, OD (Optical Density) was measured at 450 nm with a microplate reader (EZ Read 400, Biochrom, US).

1-2. Statistics processing

All measured values were expressed as mean and standard error using the Excel statistic program (Microsoft, USA). Statistical analysis between each experimental group was non-parametric using SPSS for Windows (SPSS, USA). Mann-Whitney U test was performed. Each experimental group was tested for significance at α = 0.05 level (P <0.05) and α = 0.01 level (P <0.01) compared to the control group.

1-3. Experiment result

1) Effect on TNF-α

The TNF-α content showed a significant increase in the non-treated control group after induction of immunosuppression compared to the normal group that did not induce immunosuppression. All of the experimental groups A to D in which the herbal composition for enhancing the immune activity of the present invention was administered were significantly reduced compared to the control group (Table 2 and FIG. 2).

2. Effect on Interleukin-2 (IL-2)

In IL-2, the control group showed a significant increase compared to the normal group, and all of the experimental groups showed a significant decrease compared to the control group (Table 3 and FIG. 3).

3. Effect on Interleukin-6 (IL-6)

In IL-6, the control group showed a significant increase compared to the normal group, and all of the experimental groups showed a significant decrease compared to the control group (Table 4 and FIG. 4).

4. Effect on Immunoglobulin E (IgE)

In IgE, the control group showed a significant decrease compared to the normal group, and the experimental groups B, C, and D showed a significant decrease compared to the control group (Table 5 and FIG. 5).

5. Effect on spleen weight

As a result of observing the effect on the spleen weight change according to the herbal composition of the present invention and its fermentation, the normal group was 0.83 ± 0.03 g, the control was 0.71 ± 0.03 g, and the control group showed a significant decrease compared to the normal group, and IYG -gami group 0.78 ± 0.06 g, IYG-gami-F group 0.78 ± 0.05 g, M-IYG-gami group 0.78 ± 0.03 g, M-IYG-gami-F group 0.81 ± 0.02 g compared to the control group The IYG-gami-F group showed a significant increase (Table 6 and Figure 6).

6. Effect on changes in blood Leukocytes content

As a result of observing the effect on changes in blood Leukocytes according to the herbal composition of the present invention and its fermentation, the IYG-gami-F group showed a significant increase in the case of WBC and Lymphocytes compared to the control group (Table 7 and FIG. 7).

7. Effect on blood Erythrocytes content change

As a result of observing the effect on the change of blood Erythrocytes according to the herbal composition of the present invention and its fermentation, in the case of MCV, the experimental group showed a significant decrease, and in the case of MCH, the IYG-gami group showed a significant decrease. (Table 8 and Figure 8).

8. Effect on Change of Weight Gain

As a result of observing the effect on the weight gain change according to the herbal composition of the present invention and its fermentation, the control group showed a significant decrease on the 15th and 20th days compared to the normal group, and the IYG-gami-F group was 15 compared to the control group. On the first day, the 20th day showed a significant increase, and the M-IYG-ami-F group and the M-IYG-gami-F group showed a significant increase on the 15th day (Table 9 and FIG. 9).

9. Effect on serum aminotransferase content change

Iksuyoungjin Gogami Complex (Red Ginseng, Baekbokryeong, Raw Jihwang Juice, Bongmil, Cheonmun-dong, Macmun-dong, Gojija, Flowering Mushroom, Roe Mushroom Mushroom, Choseokjam), Iksu-young Jingogami Complex, and these according to fermentation As a result of observing changes in serum aminotransferase content of the complex of the immune function, aspartate aminotransferase (AST), 75.6 ± 3.4 U / l in the normal group, 86.6 ± 6.6 U / l in the control group, and IYG-gami group 83.8 ± 3.2 U / l, 83.2 ± 4.4 U / l for IYG-gami-F group, 88.6 ± 9.7 U / l for M-IYG-gami group, 83.8 ± 6.3 U / l for M-IYG-gami-F group In the alanine aminotransferase (ALT), 43.6 ± 3.5 U / l in the normal group, 47.7 ± 4.4 U / l in the control group, and 42.6 ± 3.2 U in the IYG-gami group. / l, 40.8 ± 2.7 U / l in the IYG-gami-F group, 46.2 ± 7.0 U / l in the M-IYG-gami group, and 45.2 ± 1.8 U / l in the M-IYG-gami-F group. None of the experimental groups showed significant changes ( Table 10 and Figure 10).

Although specific examples of the present invention have been presented above, the present invention is not limited to the above, and various modifications are possible within the technical scope of the present invention, and such modifications will belong to the claims of the present invention described below.

Claims (3)

It consists of a herbal medicine composition for enhancing immune activity, including roe deer mushroom, zinnia mushroom, red ginseng, Baekbokryeong, Bokryeongpi, Cheonmun-dong, McMun-dong, Gojija, Chosukjam, saengjihwang juice, Jihwang Park, and beeswax.
The composition for medicinal herbs for enhancing immunity is 1 to 7% by weight of yellow beetroot mushroom, 0.5 to 5% by weight of zinnia mushroom, 2 to 10% by weight of red ginseng, 3 to 13% by weight of Baengboknyeong, 0.5 to 5% by weight of Bokryeongpi , 0.5 to 5% by weight, Astronomical dong 0.5 to 5% by weight, 0.5 to 5% by weight wolfberry, 0.5 to 5% by weight Chomyeonjam, 25 to 40% by weight raw saengjuk juice, 10 to 25% by weight sulphate foil and 25 to 40 sealed seals Weight percent,
The herbal composition for enhancing the immune activity is fermented into Leuconostoc mesenteroides subsp. Dextranicum strain,
The fermentation is a herbal medicine composition for enhancing immune activity, characterized in that is carried out for 4 to 6 days at 36 to 37 ℃. delete delete

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