WO2018182151A1 - Functional food composition for prevention and alleviation of dementia - Google Patents

Functional food composition for prevention and alleviation of dementia Download PDF

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Publication number
WO2018182151A1
WO2018182151A1 PCT/KR2018/000503 KR2018000503W WO2018182151A1 WO 2018182151 A1 WO2018182151 A1 WO 2018182151A1 KR 2018000503 W KR2018000503 W KR 2018000503W WO 2018182151 A1 WO2018182151 A1 WO 2018182151A1
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root
hot water
fruit
food composition
honey
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PCT/KR2018/000503
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French (fr)
Korean (ko)
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강기원
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강기원
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/38—Other non-alcoholic beverages
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/52—Adding ingredients
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L21/00—Marmalades, jams, jellies or the like; Products from apiculture; Preparation or treatment thereof
    • A23L21/20—Products from apiculture, e.g. royal jelly or pollen; Substitutes therefor
    • A23L21/25—Honey; Honey substitutes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P10/00—Shaping or working of foodstuffs characterised by the products
    • A23P10/20—Agglomerating; Granulating; Tabletting
    • A23P10/28—Tabletting; Making food bars by compression of a dry powdered mixture
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/322—Foods, ingredients or supplements having a functional effect on health having an effect on the health of the nervous system or on mental function
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2250/00—Food ingredients
    • A23V2250/20—Natural extracts
    • A23V2250/21—Plant extracts
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2250/00—Food ingredients
    • A23V2250/20—Natural extracts
    • A23V2250/21—Plant extracts
    • A23V2250/2124—Ginseng
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2300/00—Processes
    • A23V2300/14—Extraction
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2300/00—Processes
    • A23V2300/50—Concentrating, enriching or enhancing in functional factors

Definitions

  • the present invention is to prevent and improve immunity enhancement or dementia, containing hot water extracts of eight kinds of raw plants yellow root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and baekshao root as an active ingredient
  • the present invention relates to a food composition, and more particularly to a functional food composition for preventing and improving dementia such as Alzheimer's using hot water extracts of the eight herbal plants.
  • This dementia is a kind of syndrome in which people who have been living normally have impaired brain function due to various causes, and the cognitive function is deteriorated and overall decreases as compared to the past. Pathology is remembered by damage and loss of nerve cells. This is a brain disease in which severe disorders such as concentration, concentration, speech and cognition are progressively progressed over a long period of time, ultimately leading to death.
  • Alzheimer's disease Vascular dementia, Parkinson's disease, Lewy body dementia, Huntington's disease, Creutzfeldt-Jakob disease Jacob disease, Pick's disease and the like are known.
  • AD Alzheimer's disease
  • a ⁇ deposition in Alzheimer's disease is detrimental to neurons and stromal cells in the brain, resulting in brain inflammation and neuronal death, which are characteristic features of Alzheimer's disease.
  • Senile plaque and neurofibrillary tangles are pathological features in the brains of patients who die from AD.
  • the senile plaque is formed by accumulation of proteins and dead cells, etc. outside the cell, and its main component is a peptide called amyloid-beta (A ⁇ ).
  • a ⁇ amyloid precursor protein
  • APP amyloid precursor protein
  • BSCE amyloid precursor protein
  • nicotine receptor inhibitor ABT-418 muscarinic receptor inhibitor zanomeline, acetylcholine precursor lecithin, acetyl-L-carnitine, metal chelator desperioxamine, rioquinol, beta-sheet breaker iA ⁇ 5, iA ⁇ 11, Antioxidants Vitamin E, Ginkgo Biloba extract, Melatonin, Idebenone, sAPP release nicotine, acetylcholine, carbacol, beta secretase or gamma secretase inhibitors OM99-1, OM99-2, OM99-3, Z-VLL -CHO, NSAIDs, an anti-inflammatory drug, are being developed, but the effects are still effective enough to alleviate or slow down the pathological symptoms, and they are insignificant or difficult to apply due to their toxicity. Development is urgent.
  • Rehmannia glutinosa is the root of Rehmannia glutinosa Libosch, a plant of the Scrophulariaceae, and its main ingredients are Catalpol, Verbascose mannitol, glucose, and Maninotriose. (Maninotriose) and Vitamin A (Vitamin A).
  • Saengjihwang is a component that is full of flavor, sweet, nonpoisonous, releasing all heat, breaking blood, moaning, boosting jeong and blood, and used to control the blood.
  • Red ginseng is red ginseng dried by rooting Panax ginseng C.A. Meyer.
  • the main components of red ginseng are glycosides, panacen, polyacetylene compounds, nitrogen-containing compounds, flavonoids, vitamins (group B), trace elements, enzymes, antioxidants, organic acids and amino acids, like white ginseng.
  • Red ginseng has a sedative and arousing effect on the central nervous system, and is known to act on the circulatory system to prevent hypertension or atherosclerosis. In addition, it lowers blood sugar levels, protects the liver, and acts on the endocrine system to indirectly affect sexual and reproductive effects. It has anti-inflammatory and anti-tumor effects, protects against radiation and protects the skin. There is an action.
  • functionalities were recognized for improving blood flow through fatigue, enhancement of immunity, and inhibition of platelet aggregation.
  • Baekbokyeong (Poria cocos Wolf), also known as white sol pungryok, is a parasitic mushroom in the roots of pine trees. It is a medicinal herb that strengthens the rain, protects the middle element, removes moisture, helps sap metabolism, and stabilizes the mind. It grows and grows where pine trees are cut from all over Japan. It grows large like a hump from the roots of pine trees that have been cut off for many years, and is found by stabbing a pine tree stump around it. It is a mass about 30 ⁇ 50cm in diameter, rough on the outside like pine bark, and white on the inside. It is parasitic at the root of red pine, tastes sweet and boring, has a flat nature, and acts as a diuretic, but not strong.
  • White wheat (Apis cerana) is a medicinal herb that has the effect of adding qi, nourishing the middle, nourishing the lungs, softening the dry, softening the intestines, relieving pain, and detoxifying insecticides.
  • Wolfberry (Lycium chinense) is a fruit of the wolfberry family belonging to the branch family.
  • the wolfberry is a round or oval berry, and when dried, it is crumpled and contains many seeds.
  • Wolfberry contains 3.39 mg% of carotene, 0.23 mg% of vitamin B1, 0.33 mg% of vitamin B2, 1.7 mg% of nicotin acid, and 3 mg% of vitamin C. It contains a large amount of various inorganic components such as ⁇ -sitosterol and linoleic acid.
  • tonic bosin and squeezing nominal effect it has the effect of tonic bosin and squeezing nominal effect, and it is effective in protecting liver by suppressing and decreasing intestinal absorption of low-weight cholesterol, and it has excellent prevention and treatment effect on adult diseases such as hypertension, angina pectoris, arteriosclerosis and diabetes. It is also known to be excellent in eye protection.
  • Cornus officinalis is a dried fruit of Cornus officinalis Sieb. Et.Zucc, a deciduous arborescent belonging to the Cornaceae family.
  • the flesh is separated from the flesh and seeds.
  • the pulp contains glycosides such as cornin, morroniside, loganin, tannin, saponin, and organic acids such as wine, malic acid, tartaric acid, etc.
  • glycosides such as cornin, morroniside, loganin, tannin, saponin, and organic acids such as wine, malic acid, tartaric acid, etc.
  • seeds contain palmitic acid, oleic acid, linoleic acid, and the like.
  • cornine is known to have an excitatory action of parasympathetic nerves. From ancient times, the herbal medicine was medicinal.
  • Schisandra chinensis is a ball-shaped, about 1cm in diameter, dark red. The inside contains 1 to 2 red juice and reddish brown seeds. It has five flavors: sweet, sour, bitter, salty, and spicy, with strong acidity.
  • Kinds include Schisandra chinensis (North Schisandra chinensis), South Schisandra chinensis, Black Schisandra chinensis. It contains ingredients such as xanthrin, gomisin, citric acid, malic acid, and citric acid, which strengthens the heart, lowers blood pressure, and boosts immunity. Strong lung function, cough, expectorant action to help treat coughs and thirst. Soak dried fruit in cold water, add honey and sugar to reddish water, drink it as a drink, or make a flower or starch. Boil chestnuts, jujube, and samsam together to make tea or make alcohol.
  • Baek Shou (Cynanchum wilfordii) is used for nourishment, tonic, blood and ripening. It contains lecithin and corticosteroids. It reduces cholesterol and prevents atherosclerosis, promotes peristalsis to make pain, stabilizes nervous breakdown and treats premature ejaculation. Dongbobom is to remove inflammation, remove sputum and phlegm, and to treat boils, hemorrhoids, body dryness with chronic fatigue, postpartum disease, and lobster.
  • the prior art related to the present invention is known in the Republic of Korea Patent Publication No. 10-2017-0003395 is known in the pharmaceutical composition for the prevention and treatment of vascular dementia or Alzheimer's disease containing pepper extract as an active ingredient and Republic of Korea Patent Publication No. 10-2012-0099960 It is known to the composition for the prevention and treatment of dementia which uses flavonol derived from a plant as an active ingredient.
  • it is possible to prevent and improve immunity or dementia by using hot water extracts of 8 kinds of herbal plants of raw turmeric root, red ginseng root, baekbokryeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and baekshao root. None known.
  • an object of the present invention is to prevent and improve immunity or dementia by using hot water extracts of 8 kinds of herbal plants of raw turmeric root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and Baekshao root.
  • hot water extracts of 8 kinds of herbal plants of raw turmeric root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and Baekshao root To provide a functional food composition.
  • the above object of the present invention is based on the total weight of the food composition, 20% by weight of raw yellow sulfur root, 20% by weight of red ginseng root, 10% by weight of baekbokyeong root, 10% by weight of white wheat (honey), 10% by weight of Goji berries, 10% by weight of cornus fruit, Schisandra chinensis Washing with hot water 20 times (w / w) of purified water in a mixture containing 10% by weight of fruit and 10% by weight of white roots after washing with water, and then extracting hot water with a shaker for 3 hours; Filtering the hot water extract of the herb plant obtained in the above step with a double ply gauze and then concentrating under reduced pressure with a rotary evaporator; Concentrating the decompressed hot water extract obtained in the above step through the animal experiment and the cell test to test the efficacy, using the hot water extract obtained as the test material to perform immuno-enhanced and dementia prevention and improvement experiments and the results Achieved by evaluation.
  • the present invention is a weight ratio of 2: 2: 1: 1: 1: 1: 1 raw saengjihwang root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and eight kinds of baekshao root It has the effect of providing the hot water extract of and has an excellent effect of providing a functional food composition for preventing and improving immunity or dementia.
  • 1 is a graph showing the weight change according to the intraperitoneal administration of hot water extracts of raw Gohwang-root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and Baek-Shao roots in accordance with the present invention. to be.
  • FIG. 2 is a graph showing the DPPH radical scavenging ability of the hot water extracts of raw jihwang root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and Baekshui root according to the present invention.
  • Figure 3 is a graph showing the NO release amount when administration of raw water extracts of raw jihwang root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and Baekshao root according to the present invention to RAW 264.7 cells to be.
  • Figure 4 is B- when the hot-water extracts of the green goji root, red ginseng root, baekbokyeong root, white wheat (honey), Goji berries, cornus fruit, Schisandra chinensis and Baekshui root according to the present invention to C57BL / 6 mouse splenocytes. It is a graph showing lymphocyte activity.
  • 5 is a graph showing the macrophage activity when the hot-water extracts of Gojihwang root, red ginseng root, Baekbokryeong root, white wheat (honey), Goji berries, cornus fruit, Schisandra chinensis and Baeksiao root were intraperitoneally administered to 5xFAD mice according to the present invention. to be.
  • FIG. 6 shows the amount of SIRT1 expression when the hot-water extracts of the Gojihwang root, red ginseng root, Baekbokyeong root, white wheat (honey), Goji berries, cornus fruit, Schisandra chinensis and Baeksiao root were intraperitoneally administered to 5xFAD mice according to the present invention. It is a graph.
  • Figure 8 shows the expression level of sAPP ⁇ when administered the HEK293 cells of the hot-water extracts of fresh green grass roots, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and Baekshao root according to the present invention. It is a graph.
  • Figure 9 shows the expression level of sAPP ⁇ when administered the HEK293 cells with the hot-water extracts of the Gojihwang root, red ginseng root, Baekbokryeong root, white wheat (honey), Goji berries, cornus fruit, Schisandra chinensis and Baekshao root according to the present invention. It is a graph.
  • the present invention provides a food composition for preventing and improving dementia.
  • the food composition is extracted from a mixed herbaceous plant consisting of raw jihwang root, red ginseng root, baekbokyeong root, white wheat (honey), goji berries, cornus fruit, Schisandra chinensis and Baekshou root.
  • Saengjihwang Root 200g, Red Ginseng Root 200g, Baekbokyeong Root 100g, White Wheat (Honey) 100g, Wolfberry Fruit 100g, Cornus Fruit 100g, Schizandra Fruit 100g and 100g White Sewage Root were weighed and blended (Table 1) After hydrolyzing 20 times (w / w) of weight, hot water was extracted for 3 hours using a gas bath. The hot water extract was cooled, filtered through a double gauze, and concentrated under reduced pressure with a rotary evaporator to obtain 2 kg of a concentrate of the hot water extract.
  • Hot water extract raw material blending ratio of 8 herb plants of the present invention (wt%) designation content Skeletal Root 20 Red ginseng root 20 Baekbokyeong Root 10 White wheat (honey) 10 Wolfberry fruit 10 Cornus Fruit 10 Schisandra berries 10 Bai Shao Root 10 Total amount 100
  • Experimental animals purchased 7-week-old male 5xFAD mice and C57BL / 6 to adjust to a new breeding environment (temperature 22 ⁇ 3 °C, humidity 50 ⁇ 10%) for one week, and water and feed were freely fed.
  • Example 2 In order to confirm the toxicity of the eight hot water extracts prepared according to Example 1, the experimental animals of Example 2 were intraperitoneally administered at a concentration of 1, 10, 100 mg / mL, and the weight of the experimental animals was measured for 1 week. Change was confirmed. The control group was intraperitoneally administered saline solution.
  • the toxicity of the hot water extracts of the eight herbal plants obtained in Example 1 was measured.
  • RAW 264.7 cells were aliquoted into a 24 well plate to be 5 x 10 4 cells / well and incubated for 24 hours.
  • the cultured cells were cultured by exchanging hot water extracts of the eight herbal plants of the present invention with cell culture solutions containing 1, 10 and 100 mg / mL concentrations. After culturing the cells for 48 hours, MTT assay was performed to dissolve live cells in formazan in isopropanol, and the absorbance was measured at a wavelength of 570 nm. As a control, physiological saline was used.
  • DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity was measured.
  • 1 mL of 200 uM DPPH and 1 mL of hot water extracts of the eight herbal plants of the present invention obtained in Example 1 were mixed and reacted for 30 minutes at room temperature and in the dark, and then absorbance was measured at 517 nm.
  • Ascrobic acid was used as a positive control and LPS was stimulated as a negative control.
  • DPPH radical scavenging activity (%) was calculated by the method of the following equation (1).
  • B-lymphocyte stimulatory activity against the hot water extracts of the eight kinds of herbal plants of the present invention raw sulfur root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, schisandra chinensis and baekshou roots obtained in Example 1
  • Splenocytes were cultured in RPMI1640 medium to which 10% FBS was added, transferred to a 48well plate, and then stimulated with LPS at a concentration of 25 ug / mL, and 10 mg / mL of the hot water extracts of the eight herb plants obtained in Example 1 were obtained. Treatment with concentration was incubated for 2 days. The control group was incubated with saline.
  • the raw water extracts of the eight kinds of herbal plants of the present invention raw sulfur root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis fruit and Rootshao root obtained in Example 1 at a concentration of 100 mg / kg
  • Intraperitoneally administered to 5xFAD mice of Example 2 the control group was administered with saline under the same conditions. Mice were sacrificed 2 days after intraperitoneal administration to obtain macrophage.
  • the obtained macrophages were redispersed in RPMI 1640 medium at a concentration of 1 ⁇ 10 8 / mL, the dispersions were dispensed in 200 uL, and then cultured in a 5% CO 2 incubator at 37 ° C. for 24 hours, and 0.1% Triton X-100 was added.
  • the amount of lysosomal phosphatase dissolved and secreted was measured by absorbance at 405 nm with an ELISA reader to measure immune activity by enhancing macrophage activity.
  • Example 1 In order to confirm the effect of preventing and improving dementia of the eight herbal plants, including the present invention raw jihwang root, red ginseng root, baekbokyeong root, white wheat (honey), goji berries, cornus fruit, Schisandra chinensis fruit and Schisandra chinensis roots obtained in Example 1
  • the extract was intraperitoneally administered to the 5xFAD mouse of Example 2 at a concentration of 100 mg / kg to confirm the expression level of the SIRT1 gene.
  • the amount of protein was determined by BCA quantification of the brain tissue extracted in the above step, and the expression level of SIRT1 was confirmed by Western blot using 50% of protein using 10% SDS-PAGE gel.
  • Physiological saline was administered as a negative control and resveratrol 10 uM concentration as a positive control.
  • Example 1 Determination of the present invention obtained in Example 1 to determine the effect of inhibiting the production of A ⁇ , which causes dementia, and the present invention obtained in Example 1, the root of the present invention, yellow ginseng root, red ginseng root, Baekbokyeong root, white wheat (honey), goji fruit, cornus fruit,
  • the hot water extracts of the eight herb plants of Schisandra chinensis and Baekshui root were intraperitoneally administered to the 5xFAD mice of Example 2 at a concentration of 100 mg / kg, and the saline was administered to the control group under the same conditions. Mice were sacrificed 2 days after intraperitoneal administration to obtain blood. Only the supernatant (serum) in the obtained blood was isolated and the amount of A ⁇ was quantified using the A ⁇ quantification Elisa kit (Thermo fisher).
  • HEK293 cells were plated with DMEM medium containing 10% FBS and 1% streptomycin added to the plate at 37 °C and 5%.
  • the cells were recovered in 0.9 mL DMEM medium, and then the present invention of the present invention obtained in Example 1 of the root, red ginseng root, Baekbokryeong root, white wheat (honey) and wolfberry
  • Eight kinds of herbal plant hot water extracts of the fruit, cornus fruit, Schisandra chinensis and Baek Shou roots were added to the suspension of 0.1 mL of 10 mg / mL and cultured for 24 hours.
  • the control group was used by adding 0.1mL of DMEM medium instead of the eight herbal plant hot water extracts.
  • the culture cultured in the above step was performed by Western blot using a 10% SDS-PAGE gel to confirm the expression level of sAPP ⁇ and sAPP ⁇ .
  • the composition of the present invention containing the hot water extracts of the eight kinds of herbal plants of the present invention raw sulfur root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis fruit and baekshao root as an active ingredient Edible carriers, excipients or diluents which may be included in lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpi Preferred are lollidon, cellulose, polyvinylpyrrolidone, cellulose, water, sugar syrup, methylcellulose, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate and mineral oil.
  • it may further include any one of the components selected from commercially available lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives.
  • the following is a health food containing hot water extracts of the eight kinds of herbal plants of the present invention raw jihwang root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, schisandra chinensis and baekshou roots prepared according to the present invention Describe the specific composition of the composition formulation.
  • Health supplement containing the concentrate of the hot-water extracts of the eight herbal plants of the present invention Goji sulfur root, red ginseng root, baekbokyeong root, white wheat (honey), Goji fruit, cornus fruit, Schisandra chinensis and Baekshao root obtained in Example 1 Food was prepared according to the conventional health food production method with the ingredient content of Table 3. This product can be formulated into formulations such as pills and capsules.
  • the beverage containing the concentrate of the hot-water extracts of the eight kinds of herbal plants of the present invention raw jihwang root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and Baekshao root obtained in Example 1
  • To the content of Table 4 was prepared according to the conventional beverage production method. This product was formulated in the formulation of a pack.
  • the hot water extracts of the eight kinds of herbal plants of the present invention green grass roots, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis fruit and Schisandra chinensis root are used for prevention and improvement of dementia as well as enhancing immune activity. Since it is an excellent effect, it is a very useful invention in the functional food industry.

Abstract

The present invention relates to a hot water extract of eight herb plants, which are fresh Rehmanniae Radix roots, white poria roots, white honey (honey), Lycium barbarum fruits, Cornus officinalis fruits, Schisandra chinensis fruits, and white Pleuropterus multiflorus roots. More specifically, the present invention has an excellent effect of being usable as a functional health supplement food composition, containing a concentrate of a hot water extract of eight herb plants, which are effective in the enhancement of immune activity or in the prevention and alleviation of dementia.

Description

치매 예방 및 개선 기능성 식품 조성물Dementia Prevention and Improvement Functional Food Composition
본 발명은 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리 8종의 허브식물의 열수추출물을 유효성분으로 함유하는 면역증강 또는 치매 예방 및 개선 기능성 식품 조성물에 관한 것으로, 더욱 상세하게는 상기 8종의 허브식물의 열수추출물을 이용하여 알츠하이머와 같은 치매 예방 및 개선 기능성 식품 조성물에 관한 것이다.The present invention is to prevent and improve immunity enhancement or dementia, containing hot water extracts of eight kinds of raw plants yellow root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and baekshao root as an active ingredient The present invention relates to a food composition, and more particularly to a functional food composition for preventing and improving dementia such as Alzheimer's using hot water extracts of the eight herbal plants.
최근 산업화와 환경의 변화 속에서 현대인들은 다양한 원인에 의한 면역력 저하로 인해 각종 질병에 노출되어 있으며, 이러한 질병 치료를 위해 막대한 비용을 소모하고 있다. 특히, 최근 오염된 공기 및 신생활 변화 등의 이유로 알러지성 질환은 여러 알러지 유발인자에 의해 인체 면역체계를 과도하게 항진시켜 대식세포와 같은 면역세포에서 TNF-α(tumor necrosis factor-α), IL-1(interleukin-1), IL-6(interleukin-6), 프로스타글란딘(prostagladin), 류코트리엔(luecotriene), MMP-9(matrix metalloproteinase-9) 및 산화질소(nitric oxide, NO)와 같은 염증유발 물질을 과도하게 유도하여 이는 염증반응으로 진행되어 여러가지 질병 및 면역 과민반응 증상 등을 일으키는 것으로 알려져 있다.In the recent industrialization and changes in the environment, modern people are exposed to various diseases due to a decrease in immunity due to various causes, and are spending enormous costs for treating such diseases. In particular, allergic diseases due to the recent contaminated air and new life changes excessively stimulate the human immune system by various allergens, causing TNF-α (tumor necrosis factor-α) and IL in immune cells such as macrophages. Inflammatory substances such as interleukin-1, IL-6, interstaglandin, prostagladin, luecotriene, matrix metalloproteinase-9, and nitric oxide (NO) It is known to cause excessively induced inflammatory reactions, causing various diseases and immune hypersensitivity symptoms.
또, 이러한 면역체계의 붕괴로 인해 신경세포의 손상 및 손실로 뇌기능이 손상되면서 치매가 발병하고 있으며 그 치매의 발병시기도 점차 빨라지고 있다.In addition, the deterioration of the immune system due to the damage and loss of nerve cells damage the brain function, dementia is onset and the onset of dementia is getting faster.
이러한 치매(dementia)는 정상적으로 생활해오던 사람이 다양한 원인에 인해 뇌기능이 손상되면서 이전에 비해 인지 기능이 지속적이고 전반적으로 저하되는 일종의 증후군으로, 병리학적으로는 신경세포의 손상 및 손실에 의해 기억, 집중, 언어 및 인지 등에 심한 장애가 오랜 시간 동안 점진적으로 진행되어 궁극적으로는 사망에 이르게 하는 뇌 질환이다. 이러한 치매의 원인 질환들로는 알츠하이머병(Alzheimer's disease), 혈관성 치매(Vascular dementia), 파킨슨씨 병(Parkinson's disease), 루이체 치매(Lewy body dementia), 무도병(Huntington's disease), 크루츠펠트-야콥병(Creutzfeldt-Jacob disease), 픽스씨 병(Pick's disease) 등이 알려져 있다.This dementia is a kind of syndrome in which people who have been living normally have impaired brain function due to various causes, and the cognitive function is deteriorated and overall decreases as compared to the past. Pathology is remembered by damage and loss of nerve cells. This is a brain disease in which severe disorders such as concentration, concentration, speech and cognition are progressively progressed over a long period of time, ultimately leading to death. Alzheimer's disease, Vascular dementia, Parkinson's disease, Lewy body dementia, Huntington's disease, Creutzfeldt-Jakob disease Jacob disease, Pick's disease and the like are known.
그 중 알츠하이머 질환(AD)은 가장 흔한 퇴행성 뇌질환으로 점진적인 신경세포의 퇴화로 인해 인지능력 상실을 가져온다. 뇌 조직내 아밀로이드 플라그 및 아밀로이드로서 존재하는, 원섬유 형태의 아밀로이드-β 단백질 (Aβ)로 명명되는 39-43개 아미노산 펩티드의 축적을 특징으로 한다. 알츠하이머 병에서 Aβ 침착은 뇌의 신경세포 및 간질세포에게 유해하고, 그 결과 알츠하이머 병의 특징적인 특성인 뇌 염증 및 신경세포 사멸을 일으킨다. Alzheimer's disease (AD) is the most common degenerative brain disease resulting in cognitive loss due to progressive neuronal degeneration. It is characterized by the accumulation of 39-43 amino acid peptides called amyloid-β protein (Aβ) in fibrillar form, present as amyloid plaque and amyloid in brain tissue. Aβ deposition in Alzheimer's disease is detrimental to neurons and stromal cells in the brain, resulting in brain inflammation and neuronal death, which are characteristic features of Alzheimer's disease.
AD로 사망한 환자의 두뇌에서는 노인성 플라그(senile plaque)와 신경원섬유의 엉킴(neurofibrillary tangles)이 병리학적 특성으로 나타난다. 이 중 노인성 플라그는 세포 외부에 단백질과 죽은 세포 등이 축적되어 형성되는 것으로, 주 구성 성분은 아밀로이드-베타(Amyloid-β(Aβ)라는 펩티드이다.Senile plaque and neurofibrillary tangles are pathological features in the brains of patients who die from AD. Among them, the senile plaque is formed by accumulation of proteins and dead cells, etc. outside the cell, and its main component is a peptide called amyloid-beta (Aβ).
AD 환자의 주요 특징인 인지 작용의 점진적 상실은 비정상적으로 축적된 Aβ에 의해 유발되는 것으로 보인다. Aβ는 아밀로이드 전구 단백질(amyloid precursor protein: 이하 "APP"라 칭함)로부터 단백질 분해(proteolysis) 과정을 통해 생성된다. 전구물질인 APP는 β-세크라타제(BSCE) 및 γ-세크라타제에 의해 분해되어 Aβ가 생성된다. 그러나 α-세크라타제에 의해 APP가 절단될 경우 Aβ의 생성을 억제하고 신경세포 보호 효과를 나타내는 APPα의 생성을 유도하므로 α-세크라타제의 활성을 증가시키는 것이 알츠하이머병의 치료 또는 개선의 방법이다. 또한, 체내에서 SIRT1이 활성화되면 다양한 전사조절인자들을 탈아세틸화시킴으로써 α-세크라타제를 활성화시킨다.The gradual loss of cognitive function, a major feature of AD patients, appears to be caused by abnormally accumulated Aβ. Aβ is produced through proteolysis from amyloid precursor protein (hereinafter referred to as "APP"). The precursor APP is degraded by β-secretase (BSCE) and γ-secretase to produce Aβ. However, when APP is cleaved by α-secretase, it inhibits the production of Aβ and induces the production of APPα, a neuroprotective effect. Therefore, increasing the activity of α-secretase is a method of treating or improving Alzheimer's disease. to be. In addition, activation of SIRT1 in the body activates α-secretase by deacetylating various transcriptional regulators.
한편 니코틴 수용체 저해제인 ABT-418, 무스카린 수용체 저해제인 자노멜린, 아세틸콜린 전구체인 레시틴, 아세틸-L-카르니틴, 금속 킬레이터인 데스페리옥사민, 리오퀴놀, 베타-쉬트 브레이커인 iAβ5, iAβ11, 항산화제인 비타민 E, 은행잎 추출물, 멜라토닌, 이데베논, sAPP 방출제인 니코틴, 아세틸콜린, 카바콜, 베타 세크라타제 또는 감마 세크라타제 저해제인 OM99-1, OM99-2, OM99-3, Z-VLL-CHO, 항염증제인 NSAIDs계 약물 등이 개발되고 있으나, 아직 효과가 약간의 병리적 증상을 완화 또는 진행 정도를 늦추는 정도로 생겨 미미하거나 자체의 독성때문에 실제 적용이 어려운 물질이 대부분이어서 안정하며 효과적인 AD 치료제의 개발이 시급하다.On the other hand, nicotine receptor inhibitor ABT-418, muscarinic receptor inhibitor zanomeline, acetylcholine precursor lecithin, acetyl-L-carnitine, metal chelator desperioxamine, rioquinol, beta-sheet breaker iAβ5, iAβ11, Antioxidants Vitamin E, Ginkgo Biloba extract, Melatonin, Idebenone, sAPP release nicotine, acetylcholine, carbacol, beta secretase or gamma secretase inhibitors OM99-1, OM99-2, OM99-3, Z-VLL -CHO, NSAIDs, an anti-inflammatory drug, are being developed, but the effects are still effective enough to alleviate or slow down the pathological symptoms, and they are insignificant or difficult to apply due to their toxicity. Development is urgent.
생지황(Rehmannia glutinosa)은 현삼과(Scrophulariaceae) 식물인 지황 (Rehmannia glutinosa Libosch.)의 근경으로, 주성분으로는 카탈폴(Catalpol), 베르바스코스 만니톨(Verbascose mannitol), 글루코스(Glucose), 마니노트리오스(Maninotriose), 비타민 A(Vitamin A)를 포함하고 있다. 생지황은 성분이 차고 맛이 달며 독이 없고 모든 열을 풀어주고 피를 깨뜨리며, 신음을 보하고 정과 혈을 북돋아 주며, 혈증을 다스리는데 쓰인다.Rehmannia glutinosa is the root of Rehmannia glutinosa Libosch, a plant of the Scrophulariaceae, and its main ingredients are Catalpol, Verbascose mannitol, glucose, and Maninotriose. (Maninotriose) and Vitamin A (Vitamin A). Saengjihwang is a component that is full of flavor, sweet, nonpoisonous, releasing all heat, breaking blood, moaning, boosting jeong and blood, and used to control the blood.
홍삼은 수삼(Panax ginseng C.A. Meyer)의 뿌리를 쪄서 말린 붉은 인삼을 말한다. 홍삼의 주요 성분으로는 백삼과 같이 배당체(glycosides), 인삼향성분(panacen), 폴리아세틸렌계 화합물, 함질소성분, 플라보노이드, 비타민(B군), 미량원소, 효소, 항산화 물질과 유기산 및 아미노산 등이 있다. 홍삼은 중추신경에 대해서 진정작용과 흥분작용이 있으며, 순환계에 작용하여 고혈압이나 동맥경화의 예방효과가 있다고 알려져 있다. 그외에도 혈당치를 저하시켜 주고, 간을 보호하며, 내분비계에 작용하여 성행동이나 생식효과에 간접적으로 유효하게 작용하며, 항염 및 항종양작용이 있고, 방사선에 대한 방어효과와 피부를 보호하며 부드럽게 하는 작용이 있다. 또한 피로회복, 면역력 증진, 혈소판 응집 억제를 통한 혈액 흐름 개선에 관한 기능성이 인정되었다.Red ginseng is red ginseng dried by rooting Panax ginseng C.A. Meyer. The main components of red ginseng are glycosides, panacen, polyacetylene compounds, nitrogen-containing compounds, flavonoids, vitamins (group B), trace elements, enzymes, antioxidants, organic acids and amino acids, like white ginseng. There is this. Red ginseng has a sedative and arousing effect on the central nervous system, and is known to act on the circulatory system to prevent hypertension or atherosclerosis. In addition, it lowers blood sugar levels, protects the liver, and acts on the endocrine system to indirectly affect sexual and reproductive effects. It has anti-inflammatory and anti-tumor effects, protects against radiation and protects the skin. There is an action. In addition, functionalities were recognized for improving blood flow through fatigue, enhancement of immunity, and inhibition of platelet aggregation.
백복령(Poria cocos Wolf)은 흰솔풍령이라고도 불리며 소나무 뿌리에 기생하는 버섯으로써, 비를 튼튼하게 하고 중초를 보해주며 습을 빼주고 수액대사를 돕고 정신을 안정시켜주는 효능이 있는 약재이다. 전국 각지의 소나무를 베어낸 곳에 자라며 재배도 한다. 베어 낸 지 여러 해 지난 소나무 뿌리에 기생하여 혹처럼 크게 자라는데, 소나무 그루터기 주변을 쇠꼬챙이로 찔러서 찾아낸다. 지름 30~50㎝ 쯤의 덩어리로 겉은 소나무 껍질처럼 거칠고 속은 희다. 적송의 뿌리에 기생하며 맛은 달고 심심하며 성질이 평하여 보하는 작용이 있고 이뇨작용을 하지만 강하지 않다. 옛날부터 강장제로 사용되었다. 폐경·비경·심경·신경·방광경에 작용하여 비장을 보하고 가래를 삭이며 정신을 안정시킨다. 약리실험 결과에 의하면 이뇨작용과 혈당량을 낮추는 작용, 진정작용 등을 하는 것으로 밝혀졌으며 면역 부활작용을 하는 것으로도 알려져 있다. 비장이 허하여 몸이 붓는 경우와 담음병에 사용한다. 또, 게우는 데나 복수·설사·건망증·수면장애 등에도 처방한다. 위·간·이자·신장 등의 질병에도 탁월한 효과가 있는 것으로 알려져 있으며, 질병을 앓고 난 후 허약한 사람이나 만성 위장병 환자 등의 치료를 위한 약재로 이용되고 남녀의 임독성 질병은 물론 모든 성병과 부인병에도 효과가 있는 것으로 알려져 있다. Baekbokyeong (Poria cocos Wolf), also known as white sol pungryok, is a parasitic mushroom in the roots of pine trees. It is a medicinal herb that strengthens the rain, protects the middle element, removes moisture, helps sap metabolism, and stabilizes the mind. It grows and grows where pine trees are cut from all over Japan. It grows large like a hump from the roots of pine trees that have been cut off for many years, and is found by stabbing a pine tree stump around it. It is a mass about 30 ~ 50㎝ in diameter, rough on the outside like pine bark, and white on the inside. It is parasitic at the root of red pine, tastes sweet and boring, has a flat nature, and acts as a diuretic, but not strong. It has been used as a tonic since ancient times. It acts on menopause, parenteral, heart, nerve and bladder to protect the spleen, sputum and stabilize the mind. The results of pharmacological experiments have been shown to act as a diuretic, lowering blood sugar, sedation, and also known as an immune resurrection. If the spleen is swelling and the body is swollen and it is used for Damping disease. It is also prescribed for craving, ascites, diarrhea, forgetfulness and sleep disorders. It is known to have an excellent effect on diseases such as stomach, liver, interest, and kidneys.It is used as a medicine for the treatment of fragile people and patients with chronic gastrointestinal disorders after suffering from diseases. It is also known to be effective for women's diseases.
백밀(Apis cerana)은 기를 더해주고 중초를 보해주고 폐를 자양하며 마른 것을 부드럽게 해주고 장을 부드럽게 하며 통증을 멎게하고 해독 살충하는 효능이 있는 약재이다. White wheat (Apis cerana) is a medicinal herb that has the effect of adding qi, nourishing the middle, nourishing the lungs, softening the dry, softening the intestines, relieving pain, and detoxifying insecticides.
구기자(Lycium chinense)는 가지 과에 속하는 구기자나무의 열매이다. 구기자는 원형 또는 타원형의 장과이고 건조시켰을 때에 겉이 쭈글쭈글하고 속에는 많은 씨가 들어있다. 구기자의 성분은 Carotene 3.39㎎％, vitamin B1 0.23㎎％, vitamin B2 0.33㎎％, nicotin acid 1.7㎎％, vitamin C 3㎎％을 함유하고, β-sitosterol, linoleic acid 등 각종 무기성분 등을 다량 함유하고 있어 강장보신 및 강압명목의 효과를 지니고 있고, 저비중 콜레스테롤의 장내흡수를 억제 강하하여 간장보호에 효과적이며, 고혈압, 협심증, 동맥경화 및 당뇨병 등의 성인병에 탁월한 예방 및 치료 효과를 지니고 있는 동시에, 시력보호 효과도 뛰어난 것으로 알려져 있다.Wolfberry (Lycium chinense) is a fruit of the wolfberry family belonging to the branch family. The wolfberry is a round or oval berry, and when dried, it is crumpled and contains many seeds. Wolfberry contains 3.39 mg% of carotene, 0.23 mg% of vitamin B1, 0.33 mg% of vitamin B2, 1.7 mg% of nicotin acid, and 3 mg% of vitamin C. It contains a large amount of various inorganic components such as β-sitosterol and linoleic acid. It has the effect of tonic bosin and squeezing nominal effect, and it is effective in protecting liver by suppressing and decreasing intestinal absorption of low-weight cholesterol, and it has excellent prevention and treatment effect on adult diseases such as hypertension, angina pectoris, arteriosclerosis and diabetes. It is also known to be excellent in eye protection.
산수유(Cornus officinalis)는 층층나무과(Cornaceae)에 속한 낙엽소교목인 산수유나무 (Cornus officinalis Sieb. et.Zucc)의 성숙한 열매를 건조한 것으로, 과육과 씨앗을 분리하여 육질은 술과 차 및 한약의 재료로 사용하며 과육에는 코르닌(cornin), 모로니사이드(Morroniside), 로가닌(Loganin), 타닌(tannin), 사포닌(Saponin) 등의 배당체와 포도주산, 사과산, 주석산 등의 유기산이 함유되어 있고, 그밖에 비타민 A와 다량의 당도 포함되어 있다. 종자에는 팔미틴산, 올레인산, 리놀산 등이 함유되어 있다. 성분 중 코르닌은 부교감신경의 흥분작용이 있는 것으로 알려져 있다. 예로부터 한방에서는 과육을 약용하였다. "동의보감" "향약집성방" 등에 의하면 강음, 신정과 신기보강, 수렴 등의 효능이 있다고 한다. 두통, 이명, 해수병, 해열, 월경과다 등에 약재로 쓰이며 식은땀, 야뇨증 등의 민간요법에도 사용된다. 차나 술로도 장복하며, 지한, 보음 등의 효과가 있다. Cornus officinalis is a dried fruit of Cornus officinalis Sieb. Et.Zucc, a deciduous arborescent belonging to the Cornaceae family. The flesh is separated from the flesh and seeds. The pulp contains glycosides such as cornin, morroniside, loganin, tannin, saponin, and organic acids such as wine, malic acid, tartaric acid, etc. In addition, it contains vitamin A and a large amount of sugar. Seeds contain palmitic acid, oleic acid, linoleic acid, and the like. Among the components, cornine is known to have an excitatory action of parasympathetic nerves. From ancient times, the herbal medicine was medicinal. According to the "consensus relief" and "hyangjeokbangbang" such as strong sound, Shinjung and reinforcement, it has the effect of convergence. It is used as a medicine for headache, tinnitus, seawater disease, fever and excessive menstruation. It is also used for folk remedies such as cold sweat and nocturnal enuresis. Tea and liquor is also blessed, the cold, the effect of the sound.
오미자(Schisandra chinensis)는 공 모양으로 지름 약 1cm이고 짙은 붉은 빛깔이다. 속에는 붉은 즙과 불그스레한 갈색 종자가 1∼2개 들어 있다. 단맛, 신맛, 쓴맛, 짠맛, 매운맛의 5가지 맛이 나며 그 중에서도 신맛이 강하다. 종류에는 오미자(북오미자), 남오미자, 흑오미자 등이 있다. 시잔드린, 고미신, 시트럴, 사과산, 시트르산 등의 성분이 들어 있어 심장을 강하게 하고 혈압을 내리며 면역력을 높여 주어 강장제로 쓴다. 폐 기능을 강하게 하고 진해, 거담 작용이 있어서 기침이나 갈증 등을 치료하는 데 도움이 된다. 말린 열매를 찬물에 담가 붉게 우러난 물에 꿀, 설탕을 넣어 음료로 마시거나 화채나 녹말편을 만들어 먹는다. 밤, 대추, 미삼을 함께 넣고 끓여 차를 만들거나 술을 담그기도 한다.Schisandra chinensis is a ball-shaped, about 1cm in diameter, dark red. The inside contains 1 to 2 red juice and reddish brown seeds. It has five flavors: sweet, sour, bitter, salty, and spicy, with strong acidity. Kinds include Schisandra chinensis (North Schisandra chinensis), South Schisandra chinensis, Black Schisandra chinensis. It contains ingredients such as xanthrin, gomisin, citric acid, malic acid, and citric acid, which strengthens the heart, lowers blood pressure, and boosts immunity. Strong lung function, cough, expectorant action to help treat coughs and thirst. Soak dried fruit in cold water, add honey and sugar to reddish water, drink it as a drink, or make a flower or starch. Boil chestnuts, jujube, and samsam together to make tea or make alcohol.
백하수오(Cynanchum wilfordii)는 자양, 강장, 보혈, 익정의 용도로 사용되며, 레시틴성분과 부신피질호르몬 형태의 물질이 함유되어 있다. 콜레스테롤을 감소시켜 동맥경화증을 예방하고 연동운동을 촉진시켜 통변시키며, 신경쇠약을 안정시키고 조루증을 치료한다. 동의보감에는 염증을 삭히고 가래와 담을 없애며 종기, 치질, 만성피로로 몸이 마르는 것, 산후병, 대하 등을 치료한다.Baek Shou (Cynanchum wilfordii) is used for nourishment, tonic, blood and ripening. It contains lecithin and corticosteroids. It reduces cholesterol and prevents atherosclerosis, promotes peristalsis to make pain, stabilizes nervous breakdown and treats premature ejaculation. Dongbobom is to remove inflammation, remove sputum and phlegm, and to treat boils, hemorrhoids, body dryness with chronic fatigue, postpartum disease, and lobster.
본 발명과 관련된 선행기술로는 대한민국 공개특허 10-2017-0003395호에 고추 추출물을 유효성분으로 함유하는 혈관성 치매 또는 알츠하이머의 예방 및 치료용 약학적 조성물에 공지되어 있고 대한민국 공개특허 10-2012-0099960호에 식물에서 유래한 플라보놀을 유효성분으로 하는 치매 예방 및 치료용 조성물에 관한 것이 공지되어있다. 그러나 현재까지 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리 8종의 허브식물의 열수추출물을 이용하여 면역증강 또는 치매 예방 및 개선 식품 조성물에 대하여는 공지된 바 없다.The prior art related to the present invention is known in the Republic of Korea Patent Publication No. 10-2017-0003395 is known in the pharmaceutical composition for the prevention and treatment of vascular dementia or Alzheimer's disease containing pepper extract as an active ingredient and Republic of Korea Patent Publication No. 10-2012-0099960 It is known to the composition for the prevention and treatment of dementia which uses flavonol derived from a plant as an active ingredient. However, up to now, it is possible to prevent and improve immunity or dementia by using hot water extracts of 8 kinds of herbal plants of raw turmeric root, red ginseng root, baekbokryeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and baekshao root. None known.
따라서, 본 발명의 목적은 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리 8종의 허브식물의 열수추출물을 이용하여 면역증강 또는 치매 예방 및 개선 기능성 식품 조성물을 제공하는 데 있다.Accordingly, an object of the present invention is to prevent and improve immunity or dementia by using hot water extracts of 8 kinds of herbal plants of raw turmeric root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and Baekshao root. To provide a functional food composition.
본 발명의 상기 목적은 식품 조성물 총 중량에 대하여 생지황 뿌리 20중량%, 홍삼근 20중량%, 백복령 뿌리 10중량%, 백밀(꿀) 10중량%, 구기자 열매 10중량%, 산수유 열매 10중량%, 오미자 열매 10중량% 및 백하수오 뿌리 10중량%를 수세건조 후 세밀한 것을 배합한 혼합물에 상기 혼합물 총 중량의 20배(w/w)의 정제수를 가수하여 약탕기로 3시간 동안 열수추출하는 단계와; 상기 단계에서 얻은 허브식물의 열수추출물을 2겹 거즈로 여과한 다음 회전식 증발기로 감압농축하는 단계와; 상기 단계에서 얻은 감압농축한 열수추출물을 동물실험 및 세포실험을 통해 효능을 시험하는 단계로 이루어지고 상기에서 얻은 열수추출물을 공시재료로 사용하여 면역증강과 치매 예방 및 개선 실험을 수행하고 그 결과를 평가함으로써 달성하였다.The above object of the present invention is based on the total weight of the food composition, 20% by weight of raw yellow sulfur root, 20% by weight of red ginseng root, 10% by weight of baekbokyeong root, 10% by weight of white wheat (honey), 10% by weight of Goji berries, 10% by weight of cornus fruit, Schisandra chinensis Washing with hot water 20 times (w / w) of purified water in a mixture containing 10% by weight of fruit and 10% by weight of white roots after washing with water, and then extracting hot water with a shaker for 3 hours; Filtering the hot water extract of the herb plant obtained in the above step with a double ply gauze and then concentrating under reduced pressure with a rotary evaporator; Concentrating the decompressed hot water extract obtained in the above step through the animal experiment and the cell test to test the efficacy, using the hot water extract obtained as the test material to perform immuno-enhanced and dementia prevention and improvement experiments and the results Achieved by evaluation.
본 발명은 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리 8종의 허브식물을 중량비 2:2:1:1:1:1:1:1의 열수추출물을 제공하는 효과가 있고 면역증강 또는 치매 예방 및 개선 기능성 식품 조성물을 제공하는 뛰어난 효과가 있다.The present invention is a weight ratio of 2: 2: 1: 1: 1: 1: 1: 1 raw saengjihwang root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and eight kinds of baekshao root It has the effect of providing the hot water extract of and has an excellent effect of providing a functional food composition for preventing and improving immunity or dementia.
도 1은 본 발명에 따른 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리 8종의 열수추출물을 실험동물에 복강투여에 따른 체중 변화를 나타낸 그래프이다.1 is a graph showing the weight change according to the intraperitoneal administration of hot water extracts of raw Gohwang-root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and Baek-Shao roots in accordance with the present invention. to be.
도 2는 본 발명에 따른 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리 8종의 열수추출물의 DPPH 라디칼 소거능을 나타낸 그래프이다.2 is a graph showing the DPPH radical scavenging ability of the hot water extracts of raw jihwang root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and Baekshui root according to the present invention.
도 3은 본 발명에 따른 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리 8종의 열수추출물을 RAW 264.7 세포에 투여했을 때 NO 방출량을 나타낸 그래프이다.Figure 3 is a graph showing the NO release amount when administration of raw water extracts of raw jihwang root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and Baekshao root according to the present invention to RAW 264.7 cells to be.
도 4는 본 발명에 따른 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리 8종의 열수추출물을 C57BL/6 마우스 비장세포에 투여했을 때 B-림프구 활성을 나타낸 그래프이다.Figure 4 is B- when the hot-water extracts of the green goji root, red ginseng root, baekbokyeong root, white wheat (honey), Goji berries, cornus fruit, Schisandra chinensis and Baekshui root according to the present invention to C57BL / 6 mouse splenocytes. It is a graph showing lymphocyte activity.
도 5는 본 발명에 따른 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리 8종의 열수추출물을 5xFAD 마우스에 복강투여했을 때 마크로파지 활성을 나타낸 그래프이다.5 is a graph showing the macrophage activity when the hot-water extracts of Gojihwang root, red ginseng root, Baekbokryeong root, white wheat (honey), Goji berries, cornus fruit, Schisandra chinensis and Baeksiao root were intraperitoneally administered to 5xFAD mice according to the present invention. to be.
도 6은 본 발명에 따른 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리 8종의 열수추출물을 5xFAD 마우스에 복강투여했을 때 SIRT1 발현량을 나타낸 그래프이다.FIG. 6 shows the amount of SIRT1 expression when the hot-water extracts of the Gojihwang root, red ginseng root, Baekbokyeong root, white wheat (honey), Goji berries, cornus fruit, Schisandra chinensis and Baeksiao root were intraperitoneally administered to 5xFAD mice according to the present invention. It is a graph.
도 7은 본 발명에 따른 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리 8종의 열수추출물을 5xFAD 마우스에 복강투여했을 때 Amyloid beta의 발현량을 나타낸 그래프이다.7 is the expression level of Amyloid beta when the hot-water extracts of raw Goji Rhizome root, red ginseng root, Baekbokryeong root, white wheat (honey), Goji berries, cornus fruit, Schisandra chinensis and Baeksiao root were intraperitoneally administered to 5xFAD mice according to the present invention. This is a graph.
도 8은 본 발명에 따른 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리 8종의 열수추출물을 HEK293 세포에 투여했을 때 sAPPα의 발현량을 나타낸 그래프이다.Figure 8 shows the expression level of sAPPα when administered the HEK293 cells of the hot-water extracts of fresh green grass roots, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and Baekshao root according to the present invention. It is a graph.
도 9는 본 발명에 따른 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리 8종의 열수추출물을 HEK293 세포에 투여했을 때 sAPPβ의 발현량을 나타낸 그래프이다.Figure 9 shows the expression level of sAPPβ when administered the HEK293 cells with the hot-water extracts of the Gojihwang root, red ginseng root, Baekbokryeong root, white wheat (honey), Goji berries, cornus fruit, Schisandra chinensis and Baekshao root according to the present invention. It is a graph.
본 발명은 치매 예방 및 개선용 식품 조성물을 제공한다.The present invention provides a food composition for preventing and improving dementia.
본 발명에 따르면 상기 식품 조성물은 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리로 이루어진 혼합 허브식물에서 수추출한다.According to the present invention, the food composition is extracted from a mixed herbaceous plant consisting of raw jihwang root, red ginseng root, baekbokyeong root, white wheat (honey), goji berries, cornus fruit, Schisandra chinensis and Baekshou root.
본 발명은 하기의 구체적인 실시예 및 실험예에 의해 보다 구체적으로 설명되나, 하기 실시예 및 실험예는 본 발명을 예시하기 위한 것으로 본 발명의 권리범위가 이에 한정하는 것으로 의도되지는 않는다.The present invention is described in more detail by the following specific examples and experimental examples, but the following examples and experimental examples are intended to illustrate the present invention and are not intended to limit the scope of the present invention.
실시예1. 본 발명 8종의 허브식물의 열수추출물의 제조Example 1 Preparation of hot water extracts of eight herbal plants of the present invention
생지황 뿌리 200g, 홍삼근 200g, 백복령 뿌리 100g, 백밀(꿀) 100g, 구기자 열매 100g, 산수유 열매 100g, 오미자 열매 100g 및 백하수오 뿌리 100g을 칭량하여 세밀하고 배합한 다음(표1) 증류수를 허브식물 총 중량의 20배(w/w)를 가수한 후 가스 약탕기로 3시간 동안 열수추출하였다. 상기 열수추출물을 냉각한 후 2겹 거즈로 여과한 다음 회전식 증발기로 감압농축하여 열수추출물의 농축액 2kg을 수득하였다.Saengjihwang Root 200g, Red Ginseng Root 200g, Baekbokyeong Root 100g, White Wheat (Honey) 100g, Wolfberry Fruit 100g, Cornus Fruit 100g, Schizandra Fruit 100g and 100g White Sewage Root were weighed and blended (Table 1) After hydrolyzing 20 times (w / w) of weight, hot water was extracted for 3 hours using a gas bath. The hot water extract was cooled, filtered through a double gauze, and concentrated under reduced pressure with a rotary evaporator to obtain 2 kg of a concentrate of the hot water extract.
본 발명 8종의 허브식물의 열수추출물 원료성분 배합비(중량%)Hot water extract raw material blending ratio of 8 herb plants of the present invention (wt%)
명칭designation 함량content
생지황 뿌리 Skeletal Root 2020
홍삼근 Red ginseng root 2020
백복령 뿌리 Baekbokyeong Root 1010
백밀(꿀)White wheat (honey) 1010
구기자 열매 Wolfberry fruit 1010
산수유 열매 Cornus Fruit 1010
오미자 열매 Schisandra berries 1010
백하수오 뿌리 Bai Shao Root 1010
Total amount Total amount 100100
실시예2. 실험동물 준비Example 2. Laboratory Animal Preparation
실험동물은 7주령의 수컷 5xFAD 마우스와 C57BL/6를 구입하여 1주일간 새로운 사육환경(온도 22±3℃, 습도 50±10%)에 적응시키고 물과 사료는 자유롭게 섭식하도록 하였다. Experimental animals purchased 7-week-old male 5xFAD mice and C57BL / 6 to adjust to a new breeding environment (temperature 22 ± 3 ℃, humidity 50 ± 10%) for one week, and water and feed were freely fed.
실험예1. 본 발명 8종의 허브식물의 열수추출물 투여시 체중변화Experimental Example 1. Body weight change during administration of hot water extracts of eight herbal plants of the present invention
상기 실시예1에 따라 제조된 본 발명 8종의 열수추출물의 독성을 확인하기 위해 상기 실시예2의 실험동물에 1, 10, 100mg/mL 농도로 복강투여하였고 1주일간 실험동물의 체중을 측정하여 변화를 확인하였다. 대조군은 생리식염수를 복강투여하였다.In order to confirm the toxicity of the eight hot water extracts prepared according to Example 1, the experimental animals of Example 2 were intraperitoneally administered at a concentration of 1, 10, 100 mg / mL, and the weight of the experimental animals was measured for 1 week. Change was confirmed. The control group was intraperitoneally administered saline solution.
실험 결과, 도1에서 알 수 있듯이 본 발명 8종의 허브식물의 열수추출물 1~100mg/mL 농도에서 실험동물의 급격한 체중변화가 나타나지 않아 8종의 허브식물의 열수추출물 투여가 건강에 유해성이 없음을 확인하였다.As a result, as shown in Figure 1 there is no rapid weight change of the experimental animal at the concentration of 1 ~ 100mg / mL of the hot water extracts of the eight herbal plants of the present invention do not have the harmful effects on the health of the hot water extracts of the eight herbal plants It was confirmed.
실험예2. 본 발명 8종의 허브식물의 열수추출물의 독성Experimental Example 2. Toxicity of Hot Water Extracts of 8 Herb Plants
본 발명 상기 실시예1에서 얻은 8종의 허브식물의 열수추출물의 독성을 측정하였다. 이를 위해 RAW 264.7 세포가 5 x 104cells/well이 되도록 24 well plate에 분주하고 24시간 배양하였다. 배양한 세포에 본 발명 8종의 허브식물의 열수추출물을 1, 10 및 100mg/mL 농도로 함유한 세포배양액으로 교환하여 세포를 배양하였다. 세포를 48시간 배양한 후 MTT assay를 실시하여 살아있는 세포수를 formazan을 isopropanol에 용해시킨 다음 570nm의 파장에서 흡광도를 측정하였다.대조군으로는 생리식염수를 사용하였다.The toxicity of the hot water extracts of the eight herbal plants obtained in Example 1 was measured. To this end, RAW 264.7 cells were aliquoted into a 24 well plate to be 5 x 10 4 cells / well and incubated for 24 hours. The cultured cells were cultured by exchanging hot water extracts of the eight herbal plants of the present invention with cell culture solutions containing 1, 10 and 100 mg / mL concentrations. After culturing the cells for 48 hours, MTT assay was performed to dissolve live cells in formazan in isopropanol, and the absorbance was measured at a wavelength of 570 nm. As a control, physiological saline was used.
실험 결과, 하기 표2에서 알 수 있듯이 본 발명 8종의 허브식물의 열수추출물은 독성이 없는 무독성(non-toxic)임을 확인하였다As a result, as shown in Table 2, it was confirmed that the hot water extracts of the eight plant species of the present invention are non-toxic (non-toxic).
SalineSaline 8종의 허브식물의 열수추출물(ug/mL)Hot Water Extract of 8 Herb Plants (ug / mL)
1One 1010 100100
1One 1.1961.196 0.9870.987 0.9650.965 0.9440.944
22 1.1611.161 1.0611.061 0.9870.987 0.9510.951
33 1.1481.148 0.9710.971 0.9610.961 0.9910.991
MeanMean 1.1681.168 1.0061.006 0.9710.971 0.9620.962
% of control% of control 100.0100.0 86.186.1 83.183.1 82.382.3
실험예3. 항산화활성 측정Experimental Example 3. Antioxidant Activity Measurement
상기 실시예1에서 수득한 본 발명 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리의 8종의 허브식물의 열수추출물의 항산화능을 측정하기 위해 DPPH(1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity를 측정하였다. 이를 위해 200uM DPPH 1mL와 상기 실시예1에서 수득한 본 발명 8종의 허브식물의 열수추출물 1mL를 혼합하여 상온, 암소에서 30분 동안 반응시킨 후 517nm에서 흡광도를 측정하였다. 양성대조군으로는 ascrobic acid를 사용하였고 음성대조군은 LPS로 자극을 주었다. DPPH radical 소거능(%)은 하기 수학식1의 방법으로 계산하였다.In order to measure the antioxidant activity of the hot water extracts of the eight kinds of herbal plants of the present invention Goji sulfur root, red ginseng root, baekbokyeong root, white wheat (honey), Goji fruit, cornus fruit, Schisandra chinensis and Baekshao root obtained in Example 1 DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity was measured. To this end, 1 mL of 200 uM DPPH and 1 mL of hot water extracts of the eight herbal plants of the present invention obtained in Example 1 were mixed and reacted for 30 minutes at room temperature and in the dark, and then absorbance was measured at 517 nm. Ascrobic acid was used as a positive control and LPS was stimulated as a negative control. DPPH radical scavenging activity (%) was calculated by the method of the following equation (1).
실험 결과, 도2에서 알 수 있듯이 음성 대조군에 비해 본 발명 8종의 허브식물의 열수추출물의 DPPH radical 소거능이 증가되어 항산화 기능성 물질이 함유되었음을 확인하였다.As a result, as shown in Figure 2, compared to the negative control, the DPPH radical scavenging ability of the hot water extracts of the eight species of the present invention was confirmed to contain an antioxidant functional material.
Figure PCTKR2018000503-appb-M000001
Figure PCTKR2018000503-appb-M000001
실험예4. NO 방출량을 통한 면역활성 측정Experimental Example 4. Immune activity measurement through NO release
상기 실시예1에서 수득한 본 발명 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리의 8종의 허브식물의 열수추출물을 이용하여 NO 방출량을 측정하여 면역활성을 확인하였다. 이를 위해 RAW 264.7 세포를 LPS 100ng/mL 농도로 자극을 준 후 본 발명 8종의 허브식물의 열수추출물을 10mg/mL 농도로 처리하여 NO 방출량을 측정하였다. 대조군으로는 LPS 무처리군을 이용하였다.Measurement of NO release using hot water extracts of the eight herbs of the present invention raw jihwang root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and Baek sewha root obtained in Example 1 Immune activity was confirmed. To this end, after stimulating RAW 264.7 cells at a concentration of 100ng / mL LPS, the amount of NO release was measured by treating the hot water extracts of the eight herbal plants of the present invention at a concentration of 10mg / mL. LPS untreated group was used as a control.
실험 결과, 도3에서 알 수 있듯이 10mg/mL 농도에서 8종의 허브식물의 열수추출물의 NO 방출량이 74.3%로 다량의 NO를 방출시켜 면역 활성이 현저히 증진됨을 확인하였다.As a result, as shown in Figure 3 it was confirmed that the release of a large amount of NO of the hot water extracts of the eight herbal plants at a concentration of 10mg / mL to 74.3% to release a large amount of NO significantly enhanced immune activity.
실험예5. B-림프구 자극활성 측정Experimental Example 5. B-lymphocyte stimulation activity measurement
상기 실시예1에서 수득한 본 발명 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리의 8종의 허브식물의 열수추출물에 대한 B-림프구 자극활성을 확인하기 위해 상기 실시예2에 기재된 C57BL/6 마우스를 비장적출실험에 사용하였다. 비장세포는 10% FBS가 첨가된 RPMI1640 배지에서 배양하여 48well plate에 옮긴 후 LPS를 25ug/mL 농도로 자극을 준 후 상기 실시예1에서 얻은 본 발명 8종의 허브식물의 열수추출물을 10mg/mL 농도로 처리하여 2일동안 배양하였다. 대조군은 생리식염수를 처리하여 배양하였다.B-lymphocyte stimulatory activity against the hot water extracts of the eight kinds of herbal plants of the present invention raw sulfur root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, schisandra chinensis and baekshou roots obtained in Example 1 To confirm the C57BL / 6 mice described in Example 2 was used in the spleen extraction experiment. Splenocytes were cultured in RPMI1640 medium to which 10% FBS was added, transferred to a 48well plate, and then stimulated with LPS at a concentration of 25 ug / mL, and 10 mg / mL of the hot water extracts of the eight herb plants obtained in Example 1 were obtained. Treatment with concentration was incubated for 2 days. The control group was incubated with saline.
실험결과, 도4에서 알 수 있듯이 대조군에 비해 본 발명 8종의 허브식물의 열수추출물에서 B-림프구가 활성화되어 면역활성이 현저히 증강되는 것을 확인하였다.As a result, as shown in Figure 4 it was confirmed that the B-lymphocytes are activated in the hot water extracts of the eight herbal plants of the present invention compared to the control group is significantly enhanced immune activity.
실험예6. 마크로파지 활성을 통한 면역활성 측정Experimental Example 6. Determination of immune activity through macrophage activity
상기 실시예1에서 수득한 본 발명 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리의 8종의 허브식물의 열수추출물을 100mg/kg 농도로 상기 실시예2의 5xFAD 마우스에 복강투여하고 대조군에는 생리식염수를 동일조건으로 투여하였다. 마우스에 복강 투여 2일 후 희생시켜 마크로파지(macrophage)를 수득하였다. 수득한 마크로파지를 1x108 개/mL 농도로 RPMI 1640 배지에 재분산시키고 분산액을 200uL씩 분주한 다음 37℃에서 24시간 동안 5% CO2 배양기에서 배양시켰고 0.1% Triton X-100을 첨가하여 세포막을 용해시켜 분비되는 리소좀의 포스파타아제의 양을 ELISA reader기로 405nm에서 흡광도를 측정하여 마크로파지 활성 증강을 통해 면역활성을 측정하였다. The raw water extracts of the eight kinds of herbal plants of the present invention raw sulfur root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis fruit and Rootshao root obtained in Example 1 at a concentration of 100 mg / kg Intraperitoneally administered to 5xFAD mice of Example 2, the control group was administered with saline under the same conditions. Mice were sacrificed 2 days after intraperitoneal administration to obtain macrophage. The obtained macrophages were redispersed in RPMI 1640 medium at a concentration of 1 × 10 8 / mL, the dispersions were dispensed in 200 uL, and then cultured in a 5% CO 2 incubator at 37 ° C. for 24 hours, and 0.1% Triton X-100 was added. The amount of lysosomal phosphatase dissolved and secreted was measured by absorbance at 405 nm with an ELISA reader to measure immune activity by enhancing macrophage activity.
실험 결과, 도5에서 알 수 있듯이 양성 대조군에 비해 본 발명 8종의 허브식물의 열수추출물을 투여한 실험군에서 마크로파지의 활성이 증가하는 것이 확인되어 면역활성 증강효과가 입증되었다.As a result, as shown in Figure 5 it was confirmed that the activity of the macrophage increased in the experimental group administered the hot water extract of the eight herbs of the present invention compared to the positive control, it was confirmed that the immune activity enhancing effect.
실험예7. SIRT1 수준 증가 측정Experimental Example 7. SIRT1 level increase measurement
치매 예방 및 개선 효과를 확인하기 위해 상기 실시예1에서 수득한 본 발명 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리 등 8종의 허브식물의 추출물을 100mg/kg 농도로 상기 실시예2의 5xFAD 마우스에 복강투여하여 SIRT1 유전자의 발현량을 확인하였다. 마우스에 복강 투여 2일 후 희생시켜 뇌조직을 적출하여 액화질소로 급속 냉각시켰고 실험시 사용하였다. 상기단계에서 적출한 뇌조직을 BCA 정량법으로 단백질량을 정량하였고 50ug의 단백질을 10% SDS-PAGE 젤을 이용하여 웨스턴블롯을 시행하여 SIRT1의 발현량을 확인하였다. 음성 대조군으로는 생리식염수를 양성 대조군으로는 resveratrol 10uM 농도로 투여하였다.In order to confirm the effect of preventing and improving dementia of the eight herbal plants, including the present invention raw jihwang root, red ginseng root, baekbokyeong root, white wheat (honey), goji berries, cornus fruit, Schisandra chinensis fruit and Schisandra chinensis roots obtained in Example 1 The extract was intraperitoneally administered to the 5xFAD mouse of Example 2 at a concentration of 100 mg / kg to confirm the expression level of the SIRT1 gene. Two days after the intraperitoneal administration to the mice, the brain tissue was sacrificed and rapidly cooled with liquid nitrogen, which was used for the experiment. The amount of protein was determined by BCA quantification of the brain tissue extracted in the above step, and the expression level of SIRT1 was confirmed by Western blot using 50% of protein using 10% SDS-PAGE gel. Physiological saline was administered as a negative control and resveratrol 10 uM concentration as a positive control.
실험 결과, 도6에서 알 수 있듯이 본 발명 8종의 허브식물의 열수추출물이 양성 대조군에 비해 SIRT1 발현량을 증가시키는 것을 확인하였다. 이로써 본 발명 8종의 허브식물의 열수추출물의 치매 예방 및 개선효과를 확인하였다.As a result, as shown in Figure 6 it was confirmed that the hot water extract of the eight herbal plants of the present invention increased the amount of SIRT1 expression compared to the positive control. This confirmed the dementia prevention and improvement effect of the hot water extract of the eight herbs of the present invention.
실험예8. Aβ 억제효과 측정Experimental Example 8. Measurement of Aβ Inhibitory Effect
치매의 원인이 되는 Aβ의 생성억제 효과를 측정하여 치매 예방 및 개선효과를 확인하기 위해 상기 실시예1에서 수득한 본 발명 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리의 8종의 허브식물의 열수추출물을 100mg/kg 농도로 상기 실시예2의 5xFAD 마우스에 복강투여하고 대조군에는 생리식염수를 동일조건으로 투여하였다. 마우스에 복강 투여 2일 후 희생시켜 혈액을 수득하였다. 수득한 혈액 중 상층액(serum)만을 분리해 Aβ 정량 Elisa kit(Thermo fisher)를 사용하여 Aβ량을 정량하였다.Determination of the present invention obtained in Example 1 to determine the effect of inhibiting the production of Aβ, which causes dementia, and the present invention obtained in Example 1, the root of the present invention, yellow ginseng root, red ginseng root, Baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, The hot water extracts of the eight herb plants of Schisandra chinensis and Baekshui root were intraperitoneally administered to the 5xFAD mice of Example 2 at a concentration of 100 mg / kg, and the saline was administered to the control group under the same conditions. Mice were sacrificed 2 days after intraperitoneal administration to obtain blood. Only the supernatant (serum) in the obtained blood was isolated and the amount of Aβ was quantified using the Aβ quantification Elisa kit (Thermo fisher).
실험 결과, 도7에서 알 수 있듯이 생리식염수를 투여한 대조군에 비해 본 발명 8종의 허브식물의 열수추출물을 투여한 실험군에서 Aβ의 생성이 억제되는 것을 확인하였다. 이로써 본 발명 8종의 허브식물의 열수추출물에 의해 치매의 원인이 되는 Aβ의 생성이 현저히 억제하는 것을 확인하였다.As a result, as shown in Figure 7, it was confirmed that the production of Aβ is suppressed in the experimental group administered the hot water extracts of the eight herbs of the present invention compared to the control group administered with saline. As a result, it was confirmed that the hot water extract of the eight herb plants of the present invention significantly suppressed the production of Aβ, which causes dementia.
실험예9. sAPPα 및 sAPPβ 발현량 측정Experimental Example 9. Measurement of sAPPα and sAPPβ Expression
sAPPα 및 sAPPβ의 발현량을 측정하여 치매 예방 및 개선효과를 확인하기 위해 HEK293 세포를 plate에 10% FBS 및 1% streptomycin이 첨가된 DMEM배지를 이용하여 세포가 80~90% 자라도록 37℃, 5% CO2 배양기에서 배양하였고 상기 단계에서 얻은 배양액에서 배지를 제거 후 세포를 0.9mL DMEM 배지로 회수한 후 상기 실시예1에서 수득한 본 발명 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리의 8종의 허브식물 열수추출물을 10mg/mL 농도로 현탁한 것 0.1mL 첨가하여 24시간 배양하였다. 대조군은 8종의 허브식물 열수추출물 대신 DMEM 배지 0.1mL을 첨가하여 이용하였다. 상기 단계에서 배양한 배양물을 10% SDS-PAGE 젤을 이용하여 웨스턴블롯을 시행하여 sAPPα 및 sAPPβ의 발현량을 확인하였다. To measure the expression of sAPPα and sAPPβ and to determine the effects of dementia prevention and improvement, HEK293 cells were plated with DMEM medium containing 10% FBS and 1% streptomycin added to the plate at 37 ℃ and 5%. After culturing in a% CO 2 incubator and removing the medium from the culture medium obtained in the above step, the cells were recovered in 0.9 mL DMEM medium, and then the present invention of the present invention obtained in Example 1 of the root, red ginseng root, Baekbokryeong root, white wheat (honey) and wolfberry Eight kinds of herbal plant hot water extracts of the fruit, cornus fruit, Schisandra chinensis and Baek Shou roots were added to the suspension of 0.1 mL of 10 mg / mL and cultured for 24 hours. The control group was used by adding 0.1mL of DMEM medium instead of the eight herbal plant hot water extracts. The culture cultured in the above step was performed by Western blot using a 10% SDS-PAGE gel to confirm the expression level of sAPPα and sAPPβ.
실험 결과, 도8 및 도9에서 알 수 있듯이 대조군에 비해 본 발명 8종의 허브식물의 열수추출물을 첨가한 실험군에서 sAPPα의 발현량은 증가하였고 sAPPβ의 발현량은 억제되는 것을 확인하였다. 이로써 본 발명 8종의 허브식물의 열수추출물에 의해 SIRT1 유전자의 발현이 증가하여 α-세크라타제 발현이 증가하였고 이로써 sAPPα 발현량은 증가, sAPPβ의 발현량은 감소하는 것이 확인되어 치매의 원인이 되는 Aβ의 생성을 현저히 억제하는 것이 입증되었다.As a result, as shown in Figures 8 and 9 it was confirmed that the expression level of sAPPα was increased and the expression level of sAPPβ was increased in the experimental group to which the hot water extracts of the eight herb plants of the present invention were added as compared to the control group. As a result, the expression of SIRT1 gene was increased by the hydrothermal extracts of the eight herbal plants of the present invention, thereby increasing the expression of α-secretase, thereby increasing the expression of sAPPα and decreasing the expression of sAPPβ. Has been shown to significantly inhibit the production of Aβ.
또, 본 발명에 따르면 본 발명 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리의 8종의 허브식물의 열수추출물을 유효성분으로 함유하는 식품 조성물에 포함할 수 있는 식용가능한 담체, 부형제 또는 희석제로 락토즈, 덱스트로즈, 슈크로즈, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산칼슘, 알지네이트, 젤라틴, 규산칼슘, 미세결정성 셀룰로즈, 폴리비닐피롤리돈, 셀룰로즈, 폴리비닐피롤리돈, 셀룰로즈, 물, 설탕시럽, 메틸셀룰로즈, 메틸 하이드록시 벤조에이트, 프로필 하이드록시 벤조에이트, 활석, 스테아트산 마그네슘 및 미네랄 오일이 바람직하였다.In addition, according to the present invention, the composition of the present invention containing the hot water extracts of the eight kinds of herbal plants of the present invention raw sulfur root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis fruit and baekshao root as an active ingredient Edible carriers, excipients or diluents which may be included in lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpi Preferred are lollidon, cellulose, polyvinylpyrrolidone, cellulose, water, sugar syrup, methylcellulose, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate and mineral oil.
또, 상기 성분들 외에도 상업적으로 이용가능한 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 중에서 선택하여 그 어느 하나의 성분을 추가로 더 포함할 수 있었다.In addition to the above components, it may further include any one of the components selected from commercially available lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives.
이하에는 본 발명 방법에 따라 제조된 본 발명 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리의 8종의 허브식물의 열수추출물을 함유하는 건강식품 조성물 제제의 구체적인 조성을 설명한다.The following is a health food containing hot water extracts of the eight kinds of herbal plants of the present invention raw jihwang root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, schisandra chinensis and baekshou roots prepared according to the present invention Describe the specific composition of the composition formulation.
제조예1. 건강 보조 식품의 제조Preparation Example 1. Manufacture of dietary supplements
상기 실시예1에서 수득한 본 발명 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리의 8종의 허브식물의 열수추출물의 농축액을 함유하는 건강 보조 식품은 하기 표3의 성분함량으로 통상의 건강식품 제조방법에 따라 제조하였다. 본 제품은 환제, 캡슐제 등 제형으로 제품화 할 수 있다.Health supplement containing the concentrate of the hot-water extracts of the eight herbal plants of the present invention Goji sulfur root, red ginseng root, baekbokyeong root, white wheat (honey), Goji fruit, cornus fruit, Schisandra chinensis and Baekshao root obtained in Example 1 Food was prepared according to the conventional health food production method with the ingredient content of Table 3. This product can be formulated into formulations such as pills and capsules.
식품 성분 함량Food ingredient content
재료명Material name 함량(중량%)Content (% by weight)
8종의 허브식물의 열수추출물 농축액Hot Water Extract Concentrate of 8 Herb Plants 75.3775.37
비타민A아세테이트Vitamin A Acetate 0.010.01
비타민EVitamin E 0.080.08
비타민B1Vitamin B1 0.010.01
비타민B2Vitamin B2 0.010.01
비타민B6Vitamin B6 0.040.04
비타민B12Vitamin B12 0.000.00
비타민CVitamin C 0.750.75
비오틴Biotin 0.000.00
니코틴산아미드Nicotinic acid amide 0.130.13
엽산 Folic acid 0.000.00
판토텐산 칼슘Calcium Pantothenate 0.040.04
황산제1철Ferrous sulfate 0.130.13
산화아연Zinc oxide 0.060.06
탄산마그네슘Magnesium carbonate 1.911.91
제1인산칼륨Potassium phosphate monobasic 1.131.13
제2인산칼슘Dicalcium Phosphate 4.154.15
구연산칼륨Potassium citrate 6.786.78
탄산칼슘Calcium carbonate 7.547.54
염화마그네슘Magnesium chloride 1.871.87
합계Sum 100.00100.00
제조예2. 발효 음료 제조Preparation Example 2. Fermented beverage manufacturers
상기 실시예1에서 수득한 본 발명 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리의 8종의 허브식물의 열수추출물의 농축액이 함유된 음료를 하기 표4의 함량으로 통상의 음료 제조방법에 따라 제조하였다. 본 제품은 팩의 제형으로 제품화하였다.The beverage containing the concentrate of the hot-water extracts of the eight kinds of herbal plants of the present invention raw jihwang root, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis and Baekshao root obtained in Example 1 To the content of Table 4 was prepared according to the conventional beverage production method. This product was formulated in the formulation of a pack.
음료성분 함량Drink Ingredient Content
재료명Material name 함량(중량%)Content (% by weight)
8종의 허브식물의 열수추출물 농축액Hot Water Extract Concentrate of 8 Herb Plants 10.010.0
액상과당(wt%)Liquid fructose (wt%) 12.012.0
덱스트린(wt%)Dextrin (wt%) 0.50.5
구연산(wt%)Citric Acid (wt%) 0.130.13
젖산칼슘(wt%)Calcium lactate (wt%) 1.61.6
비타민C(wt%)Vitamin C (wt%) 0.030.03
구연산삼나트륨(wt%)Trisodium citrate (wt%) 0.020.02
젤탄검(wt%)Geltan gum (wt%) 0.030.03
정제수(wt%)Purified water (wt%) 잔량Remaining amount
합계(wt%)Total (wt%) 100.0100.0
이상 설명한 바와 같이 본 발명 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리 8종의 허브식물의 열수추출물은 면역활성 증진은 물론 치매 예방 및 개선에 뛰어난 효과가 있으므로 기능성 식품 산업상 매우 유용한 발명인 것이다.As described above, the hot water extracts of the eight kinds of herbal plants of the present invention green grass roots, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis fruit and Schisandra chinensis root are used for prevention and improvement of dementia as well as enhancing immune activity. Since it is an excellent effect, it is a very useful invention in the functional food industry.

Claims (6)

  1. 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리를 중량비 2:2:1:1:1:1:1:1로 정량하여 세밀하는 단계와; 상기 단계에서 얻은 분말을 혼합한 다음 20배(w/w)의 정제수를 가수하여 가스 약탕기로 열수추출하는 단계와; 상기 단계에서 얻은 추출물을 2겹 거즈로 여과한 다음 회전식 증발기로 감압농축하는 단계로 이루어진 8종의 허브식물의 열수추출물의 제조방법.Quantifying and fine-graining the roots of raw sulfur, red ginseng root, baekbokyeong root, white wheat (honey), goji fruit, cornus fruit, Schisandra chinensis fruit and Baek Shou root in a weight ratio of 2: 2: 1: 1: 1: 1: 1: 1: 1; Mixing the powder obtained in the above step, and then extracting hot water with a gas bath using 20 times (w / w) of purified water; Filtration of the extract obtained in the above step with a two-ply gauze, and the method of producing a hot water extract of the eight kinds of herbal plants consisting of the step of concentrating under reduced pressure with a rotary evaporator.
  2. 제 1항의 방법에 따라 제조된 생지황 뿌리, 홍삼근, 백복령 뿌리, 백밀(꿀), 구기자 열매, 산수유 열매, 오미자 열매 및 백하수오 뿌리 8종의 허브식물의 열수추출물.Hot-water extracts of 8 kinds of raw plant roots, red ginseng root, baekbokyeong root, white wheat (honey), Goji fruit, cornus fruit, Schisandra chinensis and Baekshao root prepared according to the method of claim 1.
  3. 제 2항의 열수추출물이 함유된 면역활성 증진 기능성 식품 조성물.Immune activity enhancing functional food composition containing the hot water extract of claim 2.
  4. 제 2항의 열수추출물이 함유된 치매 예방 및 개선 기능성 식품 조성물.Dementia prevention and improvement functional food composition containing the hot water extract of claim 2.
  5. 제 3항 또는 제 4항에 있어서, 상기 식품조성물이 음료 또는 환제 중에서 선택되는 어느 하나의 제형인 식품 조성물.The food composition according to claim 3 or 4, wherein the food composition is any one of a formulation selected from beverages or pills.
  6. 제 5항에 있어서, 상기 식품 조성물은 담체, 부형제 또는 희석제로 락토즈, 덱스트로즈, 슈크로즈, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산칼슘, 알지네이트, 젤라틴, 규산칼슘, 미세결정성 셀룰로즈, 폴리비닐피롤리돈, 셀룰로즈, 폴리비닐피롤리돈, 셀룰로즈, 물, 설탕시럽, 메틸셀룰로즈, 메틸 하이드록시 벤조에이트, 프로필 하이드록시 벤조에이트, 활석, 스테아트산 마그네슘 및 미네랄 오일 중에서 선택되는 어느 하나의 첨가제가 더 포함되는 것이 특징인 식품 조성물.The method of claim 5, wherein the food composition is a carrier, excipient or diluent, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, Any one selected from polyvinylpyrrolidone, cellulose, polyvinylpyrrolidone, cellulose, water, sugar syrup, methylcellulose, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate and mineral oil Food composition characterized in that it further comprises an additive.
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