KR102632034B1 - Composition for preventing, ameliorating or treating arthritis and joint pain comprising mixed extract of Alpiniae Oxyphyllae, Paeonia lactiflora and Glycyrrhiza uralensis as effective component - Google Patents

Abstract

The present invention relates to a composition for preventing, ameliorating or treating arthritis and joint pain comprising a mixed extract of Alpiniae oxyphylla, Paeonia and Glycyrrhiza uralensis as an effective component. The mixed extract of Alpiniae oxyphylla, Paeonia and Glycyrrhiza uralensis of the present invention has an excellent effect of inhibiting the expression of cartilage matrix lyase (MMP-13), exhibits an analgesic effect in a pain-induced animal model, increases a weight load rate, reduces the expression level of inflammatory factors (TNF-α, IL-6, IL-1β, 5-LOX, COX-2, PGE2) and cartilage degeneration-inducing factors (MMP-2, MMP-9), and increases the expression level of cartilage formation indicators (aggrecan (ACAN), collagen type ii (COL2A1)). Therefore, the composition of the present invention can be usefully used for preventing, alleviating or treating arthritis and joint pain.

Description

Composition for preventing, ameliorating or treating arthritis and joint pain comprising mixed extract of Alpiniae Oxyphyllae, Paeonia lactiflora and Glycyrrhiza uralensis as active ingredients containing mixed extract of Alpiniae Oxyphyllae, Paeonia lactiflora and Glycyrrhiza uralensis as active ingredients effective component}

The present invention relates to a composition for preventing, improving or treating arthritis and joint pain containing mixed extracts of ripe perilla, peony and licorice as active ingredients.

As we enter an aging society, inflammatory diseases, including arthritis, are becoming a social issue regardless of gender. Inflammatory diseases caused by this inflammatory reaction include gastritis, colitis, arthritis, nephritis, hepatitis, arteriosclerosis, cancer, and degenerative diseases. Among them, effective drugs or treatments for preventing and treating arthritis have not been developed to date. Arthritis refers to inflammatory changes occurring within the joint due to various causes such as aging, mechanical damage, and immune abnormalities.

Among the above-mentioned arthritis, osteoarthritis is a type of arthritis, also called degenerative arthritis, and refers to arthritis that occurs due to degenerative changes in the cartilage and surrounding bone in synovial joints. In other words, osteoarthritis is a disease characterized by gradual loss of joint cartilage, hypertrophy of bones located below the cartilage, bone formation at the joint edges, and non-specific synovial inflammation. Osteoarthritis is a disease that occurs when cartilage is damaged due to aging or excessive physical pressure (eg, obesity, trauma, etc.). Therefore, osteoarthritis causes severe pain and movement disorders in joints that receive a lot of weight, such as knee joints and hip joints, and if left untreated for a long period of time, it can even lead to deformation of the joints.

In addition, gouty arthritis is an inflammation that occurs when uric acid is deposited in the spaces and tissues within joints. As the concentration of uric acid (a product left over from the human body metabolizing a substance called purine ingested through food) increases in the blood, uric acid salts increase. (Uric acid exists in the form of urate in blood, body fluids, and joint fluid) It is a disease in which crystals are deposited in the cartilage, tendons, and surrounding tissues of joints. This phenomenon causes inflammation of the joints, causing recurrent attacks accompanied by extreme pain.

Meanwhile, Alpiniae Oxyphyllae is a plant belonging to the ginger family and mainly grows in Guangdong Province and Hainan Province, China. In particular, the ripe fruit is called Alpiniae Oxyphyllae Fructus, and has a spherical or oval shape with slightly pointed ends, 1 to 2 cm in length and 7 to 10 mm in diameter.

Peony ( Paeonia lactiflora ) is a perennial herbaceous plant belonging to the Ranunculaceae family. Its large and attractive flowers are why it is also called hamburger flower, and its roots are mainly used as a medicine. It is known to have an analgesic effect, improve digestive disorders, and calm muscle spasms in the arms and legs caused by anemia.

Licorice ( Glycyrrhiza uralensis ), also known as Ural licorice or curved licorice, treats heat and morale in the internal organs of the body and has diuretic and anti-inflammatory effects. It acts as an antitussive expectorant, relaxes muscles, improves blood circulation, strengthens muscles, strengthens bones, detoxifies, and improves skin. It is known to have such effects.

Meanwhile, Korean Patent No. 2048565 discloses 'Composition for preventing or treating osteoarthritis containing mixed extracts of Samchae and turmeric', and Korean Patent No. 1023487 discloses 'Mixed herbal extracts of Schisandra chinensis, Goldenrod and seaweed bark are effective.' Although a 'composition for preventing or treating arthritis containing the present invention as an active ingredient' has been disclosed, a composition for preventing, improving or treating arthritis and joint pain containing the mixed extract of Rikiji, peony and licorice of the present invention as an active ingredient has not been disclosed.

The present invention was developed in response to the above-mentioned needs. The present invention relates to a composition for preventing, improving, or treating arthritis and joint pain containing mixed extracts of raw root, peony, and licorice as active ingredients. The mixed extract of peony and licorice has an excellent effect of inhibiting the expression of cartilage matrix degrading enzyme (MMP-13), has an analgesic effect in pain-induced animal models, increases weight bearing rate, and reduces inflammation-causing factors (TNF-α). , IL-6, IL-1β, 5-LOX, COX-2, PGE2) and cartilage degeneration-inducing factors (MMP-2, MMP-9), and chondrogenesis indicators (aggrecan (ACAN), collagen) The present invention was completed by confirming that it has the effect of increasing the expression level of type II (COL2A1)).

In order to achieve the above object, the present invention provides a health functional food composition for preventing or improving arthritis and joint pain containing mixed extracts of ripe, peony, and licorice as active ingredients.

Additionally, the present invention provides a pharmaceutical composition for the prevention or treatment of arthritis and joint pain containing mixed extracts of ripe, peony, and licorice as active ingredients.

In addition, the present invention provides a feed additive for preventing or improving arthritis and joint pain containing mixed extracts of ripe perilla, peony, and licorice as active ingredients.

In addition, the present invention provides a veterinary composition for the prevention or treatment of arthritis and joint pain containing mixed extracts of ripe, peony, and licorice as active ingredients.

The present invention relates to a composition for preventing, improving or treating arthritis and joint pain containing a mixed extract of Rhodiola root, peony and licorice as an active ingredient. The mixed extract of Rhodiola root, peony and licorice of the present invention is a cartilage matrix degrading enzyme (MMP- 13), has an excellent effect of inhibiting the expression of Effect of reducing the expression level of COX-2, PGE2) and cartilage degeneration inducers (MMP-2, MMP-9) and increasing the expression level of cartilage formation indicators (aggrecan (ACAN), collagen type II (COL2A1)) There is.

Figure 1 shows the results of confirming the change in the content of MMP-13 in human chondrocytes (SW1353) according to treatment with the mixed extract of Rikiji, peony, and licorice of the present invention. #### indicates that the MMP-13 content of the IL-1β treated group was significantly increased compared to the untreated control group (Con), p<0.0001. **** indicates a significant decrease in MMP-13 content in the group treated with the mixed extract of ripe perilla, peony and licorice, the group treated with the mixed extract of peony and licorice, or the group treated with the ripe extract compared to the IL-1β treated group, p<0.0001. . $$$ indicates that the MMP-13 content of the group treated with the mixed extract of ripe peony, peony, and licorice was significantly decreased compared to the group treated with the mixed extract of peony and licorice or the group treated with the raw extract, p<0.001.
Figure 2 shows the results of confirming the analgesic effect on pain caused by administration of mixed extracts of Rikiji, peony, and licorice in an animal model. * indicates that after inducing pain, the number of body twists for 10 minutes was significantly reduced in the experimental group administered the mixed extract of raw rhododendron, peony, and licorice compared to the control group (con), which was not treated with the mixed extract of rhizome, peony, and licorice. p<0.05, and # indicates that after inducing pain, the number of body twists for 10 minutes was significantly reduced in the experimental group administered the mixed extract of ripe orchid, peony, and licorice compared to the experimental group administered the extract of ripe orchid or mixed extract of peony and licorice. That is, p<0.05.
Figure 3 shows the results showing the hind limb weight bearing ratio (%) of SD rats. Con is the negative control group, MIA (monosodium iodoacetate) is the osteoarthritis-inducing group, GLM (green lipped mussel) is the positive control group and is the MIA+GLM administration group, and Rikji + Peony + Licorice are MIA and 75, 150 or 300 mg/kg. This group was administered concurrently with mixed extracts of Rikji, peony, and licorice. #### indicates that the weight-bearing ratio of the group with osteoarthritis induced by MIA was significantly reduced compared to the negative control group, p<0.0001, *, **, ***, **** are MIA+GLM. Alternatively, the weight-bearing ratio of the MIA+Rikji, peony, and licorice mixed extract group was statistically significantly increased compared to the osteoarthritis-induced group by MIA treatment. * means p<0.05, ** means p<0.01, ** * means p<0.001, and **** means p<0.0001.
Figure 4 shows the results of confirming the expression of inflammatory factors (TNF-α, IL-6, IL-1β, 5-LOX, COX-2, PGE2) by mixed extracts of Rikji, Peony, and Licorice root in an osteoarthritis animal model. Con is the negative control group, MIA (monosodium iodoacetate) is the osteoarthritis-inducing group, GLM (green lipped mussel) is the positive control group and is the MIA+GLM administration group, and Rikji + Peony + Licorice are MIA and 75, 150 or 300 mg/kg. This group was administered concurrently with mixed extracts of Rikji, peony, and licorice. ###, #### indicate that the expression of inflammatory factors significantly increased during MIA treatment compared to Con, the negative control group, ### is p<0.001, and #### is p<0.0001. *, **, ***, **** indicate a statistically significant decrease in the expression level of inflammatory factors in the MIA+GLM or MIA+Rikji, peony, and licorice mixed extract group compared to the MIA group. p<0.05, ** is p<0.01, *** is P<0.001, and **** is P<0.0001.
Figure 5 shows the results of confirming the expression of cartilage degeneration-inducing factors (MMP-2, MMP-9) by mixed extracts of ripe perilla, peony, and licorice in an osteoarthritis animal model. Con is the negative control group, MIA (monosodium iodoacetate) is the osteoarthritis-inducing group, GLM (green lipped mussel) is the positive control group and is the MIA+GLM administration group, and Rikji + Peony + Licorice are MIA and 75, 150 or 300 mg/kg. This group was administered concurrently with mixed extracts of Rikji, peony, and licorice. ### indicates that the expression level of cartilage degeneration-inducing factors significantly increased during MIA treatment compared to Con, the negative control group, p<0.001. *, ** indicate a statistically significant decrease in the expression level of cartilage degeneration-causing factors in the MIA+GLM or MIA+Rikji, peony, and licorice mixed extract group compared to the MIA group, * means p<0.05, and ** means p<0.01.
Figure 6 shows the results of confirming the expression of cartilage formation indicators (aggrecan (ACAN), collagen type II (COL2A1)) by mixed extracts of ripe perilla, peony, and licorice in an osteoarthritis animal model. Con is the negative control group, MIA (monosodium iodoacetate) is the osteoarthritis-inducing group, GLM (green lipped mussel) is the positive control group and is the MIA+GLM administration group, and Rikji + Peony + Licorice are MIA and 75, 150 or 300 mg/kg. This group was administered concurrently with mixed extracts of Rikji, peony, and licorice. # indicates that the expression level of cartilage formation index significantly decreased during MIA treatment compared to Con, the negative control group, p<0.05. *, **, *** indicate a statistically significant increase in the expression level of cartilage formation indicators in the MIA+GLM or MIA+Rikji, peony and licorice mixed extract group compared to the MIA group, * means p<0.05; ** is p<0.01, *** is p<0.001.

In order to achieve the object of the present invention, the present invention provides a health functional food composition for preventing or improving arthritis and joint pain containing mixed extracts of ripe perilla, peony and licorice as active ingredients.

The arthritis is preferably osteoarthritis, and the joint pain may be joint pain caused by osteoarthritis, but is not limited thereto.

The mixed extract of ripe perilla, peony and licorice is preferably a mixture of ripe perilla, peony and licorice in a weight ratio of 6 to 8:1 to 2:1 to 2, more preferably 7:1.5: It is mixed at a weight ratio of 1.5, but is not limited thereto.

Any part of the ripe perilla, peony or licorice can be used, preferably the fruit of the ripe perch, the root of peony or the root of licorice, but is not limited thereto.

The extraction solvent for the mixed extract of ripe apricot, peony, and licorice is preferably water, a C ₁ to C ₄ lower alcohol, or a mixture thereof, and more preferably ethanol, but is not limited thereto.

The mixed extract of ripe, peony, and licorice is characterized by suppressing the expression of inflammatory cytokines.

The composition is preferably manufactured in a dosage form selected from powder, granule, pill, tablet, capsule, candy, syrup and beverage, but is not limited thereto.

When using the health functional food composition of the present invention as a food additive, the composition can be added as is or used together with other foods or food ingredients, and can be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment). Generally, when producing a food or beverage, the composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw materials. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

There are no special restrictions on the types of foods above. Examples of foods to which the health functional food composition can be added include meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, and tea. , drinks, alcoholic beverages, and vitamin complexes, etc., and includes all health functional foods in the conventional sense.

Additionally, the health functional food composition of the present invention can be manufactured into food, especially functional food. The functional food of the present invention includes ingredients commonly added during food production, and includes, for example, proteins, carbohydrates, fats, nutrients and seasonings. For example, when manufactured as a drink, natural carbohydrates or flavoring agents may be included as additional ingredients in addition to the active ingredient. The natural carbohydrates include monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose, etc.), oligosaccharides, polysaccharides (e.g., dextrins, cyclodextrins, etc.), or sugar alcohols (e.g., For example, xylitol, sorbitol, erythritol, etc.) is preferable. The flavoring agent may be a natural flavoring agent (e.g., thaumatin, stevia extract, etc.) or a synthetic flavoring agent (e.g., saccharin, aspartame, etc.).

In addition to the above health functional food composition, various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonic acid. It may further contain carbonating agents used in beverages. The ratio of the ingredients added is not very important, but is generally selected in the range of 0.01 to 0.1 parts by weight based on 100 parts by weight of the health functional food composition of the present invention.

Additionally, the present invention provides a pharmaceutical composition for the prevention or treatment of arthritis and joint pain containing mixed extracts of ripe, peony, and licorice as active ingredients.

The pharmaceutical composition according to the present invention can be formulated and used in the form of oral dosage forms such as capsules, powders, granules, tablets, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions according to conventional methods. there is.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier, excipient, or diluent in addition to the above active ingredients.

Carriers, excipients and diluents that may be included in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate. , cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. in the pharmaceutical composition. It is prepared by mixing. Additionally, in addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, oral solutions, emulsions, syrups, etc. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. . Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, wethepsol, macrogol, Tween 61, cacao, laurin, glycerogelatin, etc. can be used.

The appropriate dosage of the pharmaceutical composition of the present invention may be prescribed in various ways depending on factors such as formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity. You can.

The pharmaceutical composition of the present invention can be administered orally or parenterally, and in the case of parenteral administration, it can be administered topically on the skin, intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc.

In addition, the present invention provides a feed additive for preventing or improving arthritis and joint pain containing mixed extracts of ripe perilla, peony, and licorice as active ingredients.

The feed additive of the present invention corresponds to supplementary feed under the Feed Management Act. In the present invention, the term 'feed' may mean any natural or artificial diet, meal, etc., or a component of the meal, for or suitable for eating, ingestion, and digestion by animals. The type of feed is not particularly limited, and feed commonly used in the art can be used. Non-limiting examples of the feed include plant feeds such as grains, roots and fruits, food processing by-products, algae, fiber, pharmaceutical by-products, oils and fats, starches, cucurbits or grain by-products; Examples include animal feeds such as proteins, inorganic substances, fats and oils, minerals, oils and fats, single-cell proteins, zooplanktons or foods. These may be used alone or in combination of two or more types.

In addition, the present invention provides a veterinary composition for the prevention or treatment of arthritis and joint pain containing mixed extracts of ripe, peony, and licorice as active ingredients.

The veterinary composition of the present invention may further include appropriate excipients and diluents according to conventional methods. Excipients and diluents that may be included in the veterinary composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate, cetanol, stearyl alcohol, liquid paraffin, sorbitan monostearate. , polysorbate 60, methylparaben, propylparaben, and mineral oil. The veterinary composition according to the present invention may further include fillers, anti-aggregants, lubricants, wetting agents, spices, emulsifiers, preservatives, etc. The veterinary composition according to the present invention provides rapid and sustained release of the active ingredient after administration to an animal. or may be formulated using methods well known in the art to provide sustained release, the dosage form being powders, granules, tablets, capsules, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, It may be in the form of a suppository, sterile injectable solution, or sterile topical medication. The effective amount of the veterinary composition according to the present invention can be appropriately selected depending on the individual animal. Severity of the disease or condition, sensitivity to the active ingredient of the present invention depending on the individual's age, weight, health condition or gender, administration route, administration period, factors including other compositions mixed or used simultaneously with the composition, and other physiological or It can be determined based on factors well known in the veterinary field.

Hereinafter, the present invention will be described in more detail using examples. These examples are only for illustrating the present invention in more detail, and it is obvious to those skilled in the art that the scope of the present invention is not limited thereto.

Example 1. Preparation of mixed extract of ripe apricot, peony and licorice

15 liters of 50% (v/v) ethanol was added to 1 kg of ripe fruit, and 15 liters of 30% (v/v) ethanol was added to each peony root or licorice root, and extracted at 85°C for 3 hours. The filtrate was concentrated and dried under reduced pressure at 45°C to obtain ripe extract, peony extract, and licorice extract, respectively. Afterwards, the ripe root: peony: licorice extracts were mixed at a weight ratio of 7:1.5:1.5 to prepare a mixed extract of ripe root, peony, and licorice.

Example 2. Confirmation of the effect of reducing the level of MMP-13 expression mediated by IL-1β

When MMP (matrix metalloproteinase) is synthesized following the secretion of inflammatory cytokines, the cartilage matrix molecules (extracellular matrix, ECM) that make up cartilage tissue are decomposed.

An ELISA kit (R&D Systems) was used to determine whether mixed extracts of ripe, peony, and licorice extract affect the expression of MMP-13 in human chondrocytes (SW1353). Human chondrocytes (SW1353) were cultured using DMEM/F12 (Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12) culture medium supplemented with 10% FBS, then pretreated with mixed extracts of ripe apricots, peony, and licorice and incubated for 2 hours. After culturing for a period of time, the cells were treated with IL-1β (10 μg/ml) for 24 hours. A peony and licorice mixture prepared by mixing 30㎍/㎖ peony extract and 30㎍/㎖ licorice extract and 140㎍/㎖ ripe extract were used as comparison groups.

As a result, the expression level of MMP-13 increased in the culture supernatant treated with IL-1β for 24 hours, but the expression level of MMP-13 due to IL-1β was significantly decreased in cells pretreated with mixed extracts of ripe perilla, peony, and licorice. . In particular, the inhibitory effect on MMP-13 expression of the mixed extract of ripe perilla, peony and licorice was confirmed to be significantly better than that of the mixed extract of peony and licorice or the extract of ripe perilla alone (Figure 1).

Example 3. Evaluation of analgesic efficacy in an animal model of acetic acid-induced pain

After acclimatizing 7-week-old ICR mice for a week, mixed extracts of ripe, peony, and licorice were administered, and 1 hour later, 0.75% (v/v) acetic acid (acetic acid) was intraperitoneally administered at a dose of 10 ml/kg to the mouse abdominal cavity. For intraperitoneal acetic acid administration, the subjects were fasted for more than 18 hours. Five minutes after acetic acid was administered, the number of times the mouse twisted its body was observed for 10 minutes. A peony and licorice mixture prepared by mixing 30㎍/㎖ peony extract and 30㎍/㎖ licorice extract and 140㎍/㎖ ripe extract were used as comparison groups.

As a result, intraperitoneal administration of acetic acid caused damage to capillaries and caused pain, increasing the number of times the mouse twisted its body, and when mixed extracts of ripe lychee, peony, and licorice were administered, the number of times the mouse twisted its body increased. decreased (Figure 2).

In addition, the pain inhibition rate of the group administered mixed extracts of ripe perilla, peony, and licorice can be calculated using Colby's equation in Equation 1 below (Reference: S.R., Colby, Weeds 15 (1967), 20-22) . As a result of calculation using the method of Equation 1 below, it was confirmed that the pain suppressing effect of the mixed extract of ripe, peony, and licorice had a synergistic effect compared to the mixed extract of peony, peony, and licorice, or the extract of ripe, alone (Table 1).

[Equation 1]

Pain suppression effect increase rate (%) = A+B-(AXB/100)

A = observed efficacy of active ingredient A at the same dose used in the mixture

B=observed efficacy of active ingredient B at the same dose used in the mixture

If the actual effect exceeds the calculated value, the effect of the mixture is superadditive, i.e. a synergistic effect exists. In this case, the actual observed effect should be greater than the value of the expected effect (E) calculated using the above equation.

Results of synergistic effect analysis of mixed extracts of ripe, peony, and licorice extracts division ripe
extract peony and licorice
mixed extract Ripe apricots, peonies and licorice
mixed extract Dosage (mg/kg) Ripe 140 Peony 30 + Licorice 30 Ripe 140 + Peony 30 + Licorice 30 Pain suppression rate (%) 14.60 22.63 40.88 Pain suppression effect increase rate (%) - - 33.9

Example 4. Confirmation of the effect of mixed extracts of ripe, peony, and licorice on weight bearing in an animal model of osteoarthritis induced by MIA (monosodium iodoacetate).

To determine the effect of mixed extracts of ripe perilla, peony and licorice on weight bearing in an animal model of MIA-induced osteoarthritis, MIA (60 mg/ml in 0.9% saline, an osteoarthritis-causing substance) was placed in the joint space of the right hind limb of 7-week-old SD rats. After inducing osteoarthritis by administering 50㎕ (diluted to a concentration of The load factor was measured. Liprinol green lipped mussel oil (GLM, 100 mg/kg) was used as a positive control.

Hind limb weight bearing was measured using a paw weight tester (Incapacitance tester, Linton instrument Co., UK, Ser No. 01/45/25). The rat with osteoarthritis induced in the tester's holder stood on its normal foot to which MIA had not been administered due to pain, so the weight of both feet was unbalanced, and the weight of the foot to which MIA had been administered was measured to be relatively lighter compared to the weight of the normal foot. When measuring the weight of the feet, the weight (g) of both feet was measured while the SD rat's abdomen did not touch the sensor of the device, and using the measured weight of the feet, the weight bearing ratio (%) was calculated using the method of Equation 2 below. . The weight bearing is the force of supporting and pressing with the foot. Normally, the weight of both feet is balanced and the weight bearing rate of one foot is 50%. However, as the pain becomes worse due to osteoarthritis, the weight bearing rate (%) of the hind limb caused by osteoarthritis decreases.

[Equation 2]

Weight bearing ratio (%) = [Weight of hind limbs causing osteoarthritis / (Weight of both hind limbs)] × 100

As a result, it was confirmed that compared to the normal SD rat group, the weight bearing ratio (%) of the group in which osteoarthritis was induced by administering MIA decreased by about half compared to the control group over time. In contrast, the weight bearing rate (%), which had been reduced, increased in the group administered MIA+GLM or MIA+ripe, peony, and licorice mixed extract (Figure 3).

Example 5. Confirmation of the effect of mixed extracts of Rikji, Peony, and Licorice root on inflammatory factors and cartilage degeneration factors in an animal model of osteoarthritis induced by MIA (monosodium iodoacetate).

Inflammatory factors (TNF-α, IL-6, IL-1β, 5-LOX, COX-2, PGE2) and cartilage degeneration factors (MMP-2, MMP-9) in the blood and joints of animals with osteoarthritis. The expression level was confirmed using ELISA technique.

As a result, when osteoarthritis was induced by MIA, it was confirmed that the expression level of inflammatory factors and cartilage degeneration-inducing factors increased, while the expression level of the increased factors was confirmed to decrease when mixed extracts of ripe root, peony, and licorice were treated in parallel. (Figures 4 and 5). Through this, it was found that mixed extracts of ripe perilla, peony, and licorice can suppress inflammation and joint cartilage damage caused by osteoarthritis.

Example 6. Confirmation of the effect of mixed extracts of Rikji, Peony, and Licorice root on cartilage formation indices in an animal model of osteoarthritis induced by MIA (monosodium iodoacetate).

The expression levels of cartilage formation indicators (aggrecan (ACAN), collagen type II (COL2A1)) in the joints of animals with osteoarthritis induced by administration of mixed extracts of Rhodiola chinensis, peony, and licorice were measured using the primers shown in Table 2 below. Confirmed.

Primers used to measure the expression level of cartilage formation indicator genes gene name direction Sequence (5'->3') sequence number Aggrecan forward 5′-GAAGTGGCGTCCAAACCAA-3′ One reverse 5′-CGTTCCATTCACCCCTCTCA-3′ 2 collagen type II forward 5′-GCAACAGCAGGGTTCACGTACA-3′ 3 reverse 5′-TCGGTACTCGATGATGGTCTTG-3′ 4 GAPDH forward 5′-GGGTGTGAACCACGAGAAAT-3′ 5 reverse 5′-ACTGTGGTCATGAGCCCTTC-3′ 6

As a result, it was confirmed that the expression of cartilage formation indicators (aggrecan (ACAN), collagen type II (COL2A1)) was decreased in samples in which osteoarthritis was induced by MIA, whereas cartilage formation indicators were confirmed when combined extracts of ripe perilla, peony, and licorice were combined. It was confirmed that the expression level increased (Figure 6). Through this, it was found that the mixed extract of Rikji, peony, and licorice had a cartilage-protecting effect in an osteoarthritis animal model.

Claims (11)

A health functional food composition for the prevention or improvement of arthritis and joint pain containing as an active ingredient a mixture of ripe, peony and licorice extracts mixed in a weight ratio of 6 to 8:1 to 2:1 to 2. The health functional food composition for preventing or improving arthritis and joint pain according to claim 1, wherein the arthritis is osteoarthritis, and the joint pain is joint pain caused by osteoarthritis. delete The health functional food composition for preventing or improving arthritis and joint pain according to claim 1, wherein the extraction solvent of the extract is water, a lower alcohol of C ₁ to C ₄ , or a mixture thereof. The health functional food composition for preventing or improving arthritis and joint pain according to claim 1, wherein the active ingredient inhibits the expression of inflammatory cytokines. The health functional food composition for preventing or improving arthritis and joint pain according to claim 1, wherein the composition is manufactured in any one formulation selected from powder, granule, pill, tablet, capsule, candy, syrup, and beverage. . A pharmaceutical composition for the prevention or treatment of arthritis and joint pain, containing as an active ingredient a mixture of ripe, peony and licorice extracts mixed in a weight ratio of 6 to 8:1 to 2:1 to 2. The pharmaceutical composition for preventing or treating arthritis and joint pain according to claim 7, wherein the composition further comprises a pharmaceutically acceptable carrier, excipient, or diluent in addition to the active ingredient. The pharmaceutical composition for preventing or treating arthritis and joint pain according to claim 7, wherein the composition is prepared in any one formulation selected from capsules, powders, granules, tablets, suspensions, emulsions, syrups, and aerosols. A feed additive for the prevention or improvement of arthritis and joint pain containing as an active ingredient a mixture of ripe, peony and licorice extracts mixed in a weight ratio of 6 to 8:1 to 2:1 to 2. A veterinary composition for the prevention or treatment of arthritis and joint pain, containing as an active ingredient a mixture of ripe, peony and licorice extracts mixed in a weight ratio of 6 to 8:1 to 2:1 to 2.