KR20230040903A - Composition for preventing, ameliorating or treating cartilage regeneration, arthritis and pain coused by osteoarthritis comprising Astilbe rubra extract as effective component - Google Patents

Abstract

The present invention relates to a composition containing an extract of Astilbe rubra as an active ingredient for preventing, improving, or treating cartilage regeneration, arthritis, and pain caused by arthritis. The Astilbe rubra extract of the present invention reduces the content of prostaglandin E2 (PGE2), which is an inflammatory mediator, has an excellent effect of inhibiting MMPs, which is cartilage matrix decomposing enzymes, reduces GAG content, and has an analgesic effect in pain-induced animal models. The extract not only relieves edema, but also increases weight-bearing rate, and has the effect of reducing the expression level of inflammatory factors (IL-6, TNF-α, and LTB4) and cartilage formation indicators (aggrecan (ACAN), collagen type II (COL2A1)). Therefore, the composition of the present invention can be usefully used for preventing, improving, or treating arthritis.

Description

Composition for preventing, ameliorating or treating cartilage regeneration, arthritis and pain coused by osteoarthritis comprising Astilbe rubra extract as effective component }

The present invention relates to a composition for preventing, ameliorating or treating pain caused by cartilage regeneration, arthritis and arthritis, containing an extract of deer burdock as an active ingredient.

As we enter an aging society, inflammatory diseases including arthritis are socially emerging regardless of gender. Inflammatory diseases caused by this inflammatory response include gastritis, colitis, arthritis, nephritis, hepatitis, arteriosclerosis, cancer, or degenerative diseases. Among them, effective drugs or treatments for the prevention and treatment of arthritis have not yet been developed. Arthritis refers to an inflammatory change in a joint due to various causes such as aging, mechanical damage, and immune abnormality.

Among the above-mentioned arthritis, osteoarthritis (osteoarthritis) is a kind of arthritis, also called degenerative arthritis, refers to arthritis caused by degenerative changes in cartilage and surrounding bones in synovial joints. That is, osteoarthritis is a disease characterized by gradual loss of articular cartilage, hypertrophy of bones located below the cartilage, bone formation at the joint edge, and non-specific synovial inflammation. Osteoarthritis is a disease caused by damage to cartilage due to aging or excessive physical pressure (eg, obesity, trauma, etc.). Therefore, osteoarthritis shows severe pain and movement disorders in joints that receive a lot of weight, that is, knee (knee) joints, hip (hip) joints, etc., and even causes deformation of the joints when left unattended for a long period of time.

In addition, gouty arthritis is an inflammation caused by the deposition of uric acid in the space and tissues within the joint. As the concentration of uric acid (a product left after the body metabolizes a substance called purine consumed through food) increases, the uric acid (Uric acid exists in the form of uric acid in blood, body fluids, and joint fluid.) It is a disease in which crystals are deposited in joint cartilage, tendons, and surrounding tissues. This phenomenon causes inflammation of the joint, resulting in recurrent attacks with excruciating pain.

On the other hand, roe deer grass ( Astilbe rubra ) is a perennial plant that grows relatively commonly in the mountains of the country, and is also called roe deer urine, red riding horse, king deer urine, or large deer urine. Worldwide, it is distributed in China, Tsushima Island in Japan, northeastern Russia, and India. The stem is upright, 50-70cm high. Root leaves come out 2 times 3 times or rarely 3 times 3 times, and the petiole is long. The small leaf attached to the end is long ovate or ovate oval. Stem leaves are alternate. Flowers hang on the panicle that develops on the top of the flower stem in May-July, and it is pink, but it varies greatly. There are many glandular hairs in the inflorescence, but there are more of them on the branch of the peduncle. The lower branches of the inflorescence do not droop downward. There are few peduncles. There are 5 calyxes, ovate, and petals have rounded ends. There are 10 stamens and 2 styles. The fruit is a capsule, and the end is split into 2 parts. It is used for food and medicine.

As prior art related to arthritis, Korean Patent No. 2231892 discloses a composition for the prevention or treatment of oral diseases containing a deer extract, and Korean Patent No. 0523463 discloses a deer extract or astilbic acid isolated therefrom. An anti-inflammatory or anti-allergic composition containing a peltoboyquinolic acid derivative has been disclosed, but a composition for cartilage regeneration, arthritis, and prevention, improvement, or treatment of pain caused by arthritis, containing the extract of the present invention as an active ingredient, has been disclosed. no bar

The present invention was derived from the above needs, and the present invention provides a composition for cartilage regeneration, prevention, improvement or treatment of arthritis and pain caused by arthritis, containing deer roe deer extract as an active ingredient, and Grass ( Astilbe rubra ) extract reduces the content of prostaglandin E2 (PGE2), an inflammatory mediator, has an excellent inhibitory effect on MMPs, a cartilage matrix decomposition enzyme, reduces the GAG content, and has an analgesic effect in animal models where pain is induced It not only relieves edema, but also increases the weight bearing rate, and the expression level of inflammatory factors (IL-6, TNF-α, and LTB4) and chondrogenic markers (aggrecan (ACAN), collagen type II (COL2A1)) By confirming that there is an effect of reducing, the present invention was completed.

In order to achieve the above object, the present invention provides a health functional food composition for preventing or improving pain caused by cartilage regeneration, arthritis or arthritis, containing an extract of Astilbe rubra as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for preventing or treating pain caused by cartilage regeneration, arthritis or arthritis, containing an extract of blueberry as an active ingredient.

In addition, the present invention provides a feed additive for the prevention or improvement of cartilage regeneration, arthritis or pain caused by arthritis, containing the extract of ermine greens as an active ingredient.

In addition, the present invention provides a veterinary composition for cartilage regeneration, prevention or treatment of arthritis or pain caused by arthritis, containing an extract of blueberry as an active ingredient.

The present invention relates to a composition for cartilage regeneration, arthritis, and prevention, improvement or treatment of pain caused by arthritis, containing an extract of Astilbe rubra as an active ingredient. (PGE2) content, has excellent effect of inhibiting cartilage matrix degradation enzymes, MMPs, reduces GAG content, exhibits analgesic effect in pain-induced animal models, relieves edema, and increases weight-bearing rate It is effective in increasing the expression of inflammation-inducing factors (IL-6, TNF-α and LTB4) and chondrogenic indicators (aggrecan (ACAN), collagen type II (COL2A1)).

Figure 1 is the result of confirming the change in PGE2 content according to the treatment of the extract of the present invention. ## indicates that the PGE2 content of the IL-1β treated group significantly increased compared to the control group (Con) not treated with anything, p<0.01. ** means that the PGE2 content of the deciduous extract-treated group was significantly decreased compared to the IL-1β-treated group, p<0.01.
Figure 2 is the result of confirming the change in the content of MMP-1 and MMP-13 according to the treatment of the extract of the present invention. #### indicates that the MMP-1 and MMP-13 contents of the IL-1β treated group were significantly increased compared to the control group (Con) not treated with anything, p<0.0001. *, **** means that the contents of MMP-1 and MMP-13 in the group treated with deer elegans extract were significantly decreased compared to the group treated with IL-1β, * indicates p<0.05, and **** indicates p<0.0001 am.
Figure 3 is the result of confirming the change in the GAG content according to the treatment of the deer green extract of the present invention. # indicates that the GAG content of the IL-1β treated group was significantly increased compared to the control group (Con) not treated with anything, p<0.05. ** indicates that the GAG content of the deciduous extract treatment group of the present invention was significantly decreased compared to the IL-1β treatment group, p<0.01.
Figure 4 is the result of confirming the analgesic effect on pain according to the administration of the present invention in the animal model. Indo is the positive control indomethacin. *, *** is the number of times the body was twisted for 10 minutes by the positive control group (Indo) compared to the control group (con) not treated with the deciduous extract of the present invention or the experimental group administered with the deciduous extract of the present invention after inducing pain. It means that is significantly decreased, * is p <0.05, *** is p <0.001.
Figure 5 is the result of confirming the ear edema alleviation effect in an animal model inducing inflammation by arachidonic acid (AA) according to the administration of the oxtail extract of the present invention in an animal model. ### indicates that ear edema significantly increased in the arachidonic acid-treated group (AA) compared to the normal group (Nor) not treated with anything, p<0.001. * indicates that ear edema was significantly reduced in the arachidonic acid-treated group (AA) compared to the hepatica hepatica extract treatment group of the present invention, p<0.05.
Figure 6 is a result showing the hind limb weight bearing rate (%) of SD rats. Con is a negative control group, MIA (monosodium iodoacetate) is an osteoarthritis-inducing group, Joins is a positive control group, MIA + JOINS (Joins) administration group, and Nornun grass is a group in which MIA and 100 mg/kg of Nornol grass extract have been concurrently administered. am. #### indicates that the weight bearing ratio of the group induced by osteoarthritis by MIA was significantly decreased compared to the negative control group, p<0.0001, *, **, **** indicate MIA + Joins or MIA + deer extract. The weight bearing ratio of the concurrent administration group was statistically significantly increased compared to the osteoarthritis-induced group by MIA treatment. * means p<0.05, ** means p<0.01, and **** means p<0.0001.
7 shows the results of confirming the expression of inflammatory inducing factors (LTB4, TNF-α and IL-6) and cartilage degeneration inducing factors (MMP-2 and MMP-9) by the deciduous extract in an osteoarthritis animal model. Con is a negative control group, MIA (monosodium iodoacetate) is an osteoarthritis-inducing group, Joins is a positive control group, MIA + JOINS (Joins) administration group, and Nornun grass is a group in which MIA and 100 mg/kg of Nornol grass extract have been concurrently administered. am. #, ##, ### indicates that the expression of inflammatory factors and cartilage degeneration factors increased significantly during MIA treatment compared to the negative control group Con, # is p<0.05, ## is p<0.01, and , ### is p<0.001. *, ** indicate the statistical expression levels of inflammation-inducing factors (LTB4, TNF-α, and IL-6) and cartilage degeneration-inducing factors (MMP-2 and MMP-9) in the MIA+Joins or MIA+Yellow erinaceus extract concurrently administered group compared to the MIA group. It means that it decreased significantly, * is p <0.05, ** is p <0.01.
Figure 8 shows the results of confirming the activity of cartilage formation indexes (aggrecan (ACAN), collagen type II (COL2A1)) by the deer extract in an osteoarthritis animal model. Con is a negative control group, MIA (monosodium iodoacetate) is an osteoarthritis-inducing group, Joins is a positive control group, MIA + JOINS (Joins) administration group, and Nornun grass is a group in which MIA and 100 mg/kg of Nornol grass extract have been concurrently administered. am. #, ### means that the expression level of cartilage formation indexes in the MIA group was significantly decreased compared to Con, # is p<0.05, and ### is p<0.001. * indicates a statistically significant increase in the expression level of cartilage formation indicators in the MIA + Joins or MIA + deer extract parallel administration group compared to the MIA group, p <0.05.

The present invention relates to a health functional food composition for preventing or improving pain caused by cartilage regeneration, arthritis or arthritis, containing an extract of Astilbe rubra as an active ingredient.

The hepatica extract may use all of the whole plant, preferably one or more parts selected from among flowers, leaves, stems and roots, but is not limited thereto. The arthritis is preferably osteoarthritis, but is not limited thereto. The active ingredient is characterized by inhibiting inflammatory cytokines.

The nori extract may be prepared by a method comprising the following steps, but is not limited thereto:

1) extracting by adding an extraction solvent to deer grass;

2) filtering the extract of step 1); and

3) preparing an extract by concentrating the filtered extract of step 2) under reduced pressure and drying it.

In step 1), the extraction solvent is preferably water, C ₁ ~ C ₄ lower alcohol or a mixture thereof, more preferably 70% (v / v) ethanol extract, but is not limited thereto. In the preparation method, all conventional methods known in the art, such as filtration, hot water extraction, immersion extraction, reflux cooling extraction and ultrasonic extraction, can be used for extraction of hepatica. The vacuum concentration in step 3) is preferably performed using a vacuum vacuum concentrator or a vacuum rotary evaporator, but is not limited thereto. In addition, drying is preferably reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, but is not limited thereto. The composition is preferably prepared in any one formulation selected from powder, granule, pill, tablet, capsule, candy, syrup and beverage, but is not limited thereto. When the health functional food composition of the present invention is used as a food additive, the hepatica extract may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to conventional methods. The mixing amount of the active ingredient can be suitably determined according to its purpose of use (prevention, health or therapeutic treatment). In general, when preparing 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 material. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range. There is no particular limitation on the type of food. Examples of foods to which the extract or its fraction can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea , drinks, alcoholic beverages, and vitamin complexes, and includes all health functional foods in the conventional sense. When the composition of the present invention is used as a health drink, it may contain various flavoring agents or natural carbohydrates as additional components, like conventional drinks. The aforementioned natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclotenstrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 g of the composition of the present invention. The composition of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal agents, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonic acid It may contain a carbonation agent used in beverages and the like. In addition, the composition of the present invention may contain fruit flesh for preparing natural fruit juice, fruit juice beverages and vegetable beverages. These components may be used independently or in combination. The ratio of these additives is not very important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

In addition, the present invention relates to a pharmaceutical composition for preventing or treating pain caused by cartilage regeneration, arthritis or arthritis, containing an extract of blueberry as an active ingredient.

The arthritis is preferably osteoarthritis, but is not limited thereto. The composition of the present invention may be prepared in any one formulation selected from capsules, powders, granules, tablets, suspensions, emulsions, syrups and aerosols, but is not limited thereto.

The composition of the present invention may further include a pharmaceutically acceptable carrier, excipient, or diluent in addition to the above active ingredient, and may be in various oral or parenteral formulations. 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 capsules, powders, granules, tablets, pills, and the like, and these solid preparations include at least one excipient in one or more compounds, for example, starch, calcium carbonate, sucrose or lactose ( lactose) and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, emulsions, syrups, aerosols, etc., and various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included in addition to water and liquid paraffin, which are commonly used simple diluents. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried formulations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending solvents. As a base for the suppository, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogelatin, and the like may be used. In the case of parenteral administration, it is preferable to select an external skin or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine intrathecal or intracerebrovascular injection method. The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the level of the effective amount depends on the type, severity, and activity of the drug of the patient's disease. , sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered single or multiple times. Considering all of the above factors, it is important to administer an amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art. The dosage of the composition of the present invention varies in its range depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate and severity of the disease, and the daily dosage is 0.01 to 2,000 mg/kg as a standard, preferably 30 to 500 mg/kg, and more preferably 50 to 300 mg/kg, and may be administered 1 to 6 times a day. The composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers.

In addition, the present invention relates to a feed additive for cartilage regeneration, arthritis, or prevention or improvement of pain caused by arthritis, which contains an extract of blueberry as an active ingredient.

The feed additive of the present invention corresponds to supplementary feed under the Feed Management Act. In the present invention, the term 'feed' may refer to any natural or artificial diet, one meal, etc., or a component of the one meal meal, suitable for or suitable for consumption by animals. The type of feed is not particularly limited, and feeds commonly used in the art may be used. Non-limiting examples of the feed include vegetable feeds such as grains, root fruits, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, meal or grain by-products; Animal feed such as proteins, inorganic materials, oils, mineral oils, oils, single cell proteins, zooplankton, or food may be mentioned. These may be used alone or in combination of two or more.

In addition, the present invention relates to a veterinary composition for cartilage regeneration, prevention or treatment of arthritis or pain caused by arthritis, containing an extract of blueberry as an active ingredient. 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, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, 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-agglomerating agents, lubricants, wetting agents, spices, emulsifiers, preservatives, and the like. Or it may be formulated using a method well known in the art to provide delayed release, and the dosage form may be a powder, granule, tablet, capsule, suspension, emulsion, solution, syrup, aerosol, soft or hard gelatin capsule, It may be in the form of a suppository, a sterile injection solution, or a sterile external preparation. An effective amount of the veterinary composition according to the present invention can be appropriately selected according to the individual animal. The severity of the disease or condition, the sensitivity to the active ingredient of the present invention according to the age, weight, health condition or sex of the subject, the route of administration, the period of administration, factors including other compositions that are combined or used simultaneously with the above composition, and other physiological or It can be determined according to factors well known in the veterinary field.

Hereinafter, the present invention will be described in more detail using examples. These examples are only for explaining 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 Hepatica extract

For 1 kg of deer grass, 15 L of 70% (v/v) ethanol was added, extraction was performed at 85° C. for 3 hours, and the filtrate was concentrated under reduced pressure and dried at 45° C. to obtain an extract of deer green.

Example 2. Evaluation of anti-inflammatory efficacy in chondrocytes

Inhibition of MMPs (MMP-1, MMP-13), which are cartilage matrix degrading enzymes activated by changes in the content of prostaglandin E2 (PGE2), an inflammatory mediator, and increase in inflammatory cytokines, according to extract treatment in human chondrocytes (SW1353) and cartilage A change in the content of GAG (Glycosaminoglycan), a proteoglycan degradation product, was confirmed.

[Cell culture]

After culturing human chondrocytes (SW1353) using a culture medium in which 10% FBS was added to DMEM/F12 (Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12), 25 to 100 μg/ml of deciduous coriander extract was treated. and incubated for 2 hours, and treated with IL-1β (10 μg/ml) for 24 hours.

(1) Confirmation of inhibitory effect on PGE2 production induced by IL-1β

In order to evaluate the anti-inflammatory effect of the extract of wintergreen, the effect of inhibiting the production of PGE2, an inflammatory mediator, was measured using an ELISA kit (R&D Systems).

As a result, PGE2 production was significantly increased by IL-1β treatment in chondrocytes, and PGE2 production was significantly decreased in cells pretreated with 100 μg/ml of crocodile extract (FIG. 1).

(2) Confirmation of IL-1β-mediated reduction of MMP-1 and MMP-13 expression levels

The activation and synthesis of MMPs following the secretion of inflammatory cytokines degrades cartilage matrix molecules (ECM) constituting cartilage tissue. An ELISA kit (R&D Systems) was used to examine the effect of the extract of blueberry extract on the expression of MMPs. It was confirmed using .

As a result, the expression levels of MMP-1 and MMP-13 were increased in the culture supernatant treated with IL-1β for 24 hours. The expression level of was significantly decreased in a concentration-dependent manner (FIG. 2).

(3) Confirmation of GAG (Glycosaminoglycan) content change

GAG is a component constituting cartilage tissue and is a material produced by decomposition of proteoglycan. In order to confirm whether the decomposition of proteoglycan was affected by the decomposition of the blueberry extract, the change in GAG content was confirmed. Specifically, after treatment with IL-1β for 24 hours, the culture supernatant was reacted with blyscan dye to measure absorbance at 656 nm to measure the amount of GAG.

As a result, it was confirmed that the GAG content, which was increased due to the promotion of cartilage degradation due to IL-1β, was reduced by the treatment of the deer extract (FIG. 3).

Example 3. Evaluation of analgesic efficacy in pain-induced animal models by acetic acid induction

After 7-week-old ICR mice were acclimatized for one week, they were orally administered with deciduous extract (150mg/kg, 300mg/kg) and indomethacin (5mg/kg) as a positive control for 4 days, respectively. 1 hour after administration on the 4th day, 0.75% (v/v) acetic acid (acetic acid) was intraperitoneally administered to the mouse at a dose of 10 ml/kg, and fasted for more than 18 hours for intraperitoneal administration of acetic acid. Five minutes after administration of acetic acid, the number of times the mouse twisted its body was observed for 10 minutes.

As a result, intraperitoneal administration of acetic acid caused capillary damage and pain, resulting in an increase in the number of twists of the mouse, and when the extract of erinaceus extract was administered, the number of twists in the mouse was significantly reduced. (Fig. 4).

Example 4. Efficacy evaluation of ear edema relief in inflammation-induced animal models by arachidonic acid

After 7-week-old ICR mice were acclimatized for one week, they were orally administered with deciduous extract (150mg/kg, 300mg/kg) and indomethacin (5mg/kg) as a positive control group for 4 days. 1 hour after administration on the 4th day, 10 μl of 2% arachidonic acid dissolved in acetone was applied to the inner and outer surfaces of both ears, respectively. After 2 hours, they were euthanized, and ear tissue weight was measured using a 5 mm dermal biopsy punch.

As a result, it was confirmed that the ear edema was significantly reduced by the administration of the crocodile extract in the ear edema animal model induced by arachidonic acid (FIG. 5).

Example 5. MIA (monosodium iodoacetate) induced Osteoarthritis Animal Model Confirmation of Effects of Hepatica extract on Weight Bearing

In order to confirm the effect of the hepatica extract prepared in Example 1 on weight bearing in an MIA-induced osteoarthritis animal model, the osteoarthritis-inducing substance MIA (60 mg with 0.9% saline) was placed in the joint cavity of the right hind leg of 7-week-old SD rats. After osteoarthritis was induced by administering 50 μl (diluted to a concentration of mg/ml), 100 mg/kg of a deciduous extract was orally administered once a day for a total of 21 days, and the weight bearing rate was measured at 7-day intervals. As a positive control group, JOINS tablets (20 mg/kg) were used.

Hindlimb weight bearing was measured using a foot weight tester (Incapacitance tester, Linton instrument Co., UK, Ser No. 01/45/25). In the holder of the tester, the osteoarthritis-induced rat stood on its normal foot without MIA administration due to pain, so the weight of both feet lost balance, and the weight of the foot administered with MIA was relatively light compared to the weight of the normal foot. When measuring the weight of the foot, the weight (g) of both feet was measured in a state where the stomach of the SD rat did not touch the sensor of the device, and the weight bearing rate (%) was calculated using the method of Equation 1 below using the measured weight of the foot . The weight bearing is a pressing force supported by the feet. In normal cases, the weight of both feet is balanced and the weight bearing ratio of one foot is 50%.

[Equation 1]

Weight-bearing ratio (%) = [weight of osteoarthritis-induced hind limb / (weight of both hind limbs)] × 100

As a result, as shown in FIG. 6, it was confirmed that the weight bearing rate (%) of the osteoarthritis induced group by administration of MIA was reduced by about half compared to the control group as compared to the normal SD rat group. In contrast, the weight bearing rate (%), which had been reduced, increased in the MIA + Joins or MIA + deer extract administration groups.

Example 6. MIA (monosodium iodoacetate) induced Osteoarthritis Animal Model Confirmation of the Effects of Hepatica extract on Inflammatory Factors and Cartilage Degeneration Inducing Factors

The expression levels of inflammatory factors (IL-6, TNF-α and LTB4) and cartilage degeneration factors (MMP-2 and MMP-9) in blood and joints of animals induced with osteoarthritis were confirmed by ELISA technique.

As a result, as shown in FIG. 7 , when osteoarthritis was induced by MIA, it was confirmed that the expression levels of inflammation inducing factors and cartilage degeneration inducing factors were increased, while the expression levels of the increased factors were decreased when the crocodile extract was treated. Through this, it was found that the extract of oxtail extract inhibits inflammation and articular cartilage damage caused by osteoarthritis.

Example 7. MIA (monosodium iodoacetate) induced Osteoarthritis Animal Model Confirmation of Effects of Hepatica Extract on Cartilage Formation Index

In the joints of animals induced with osteoarthritis, changes in the expression levels of cartilage formation indexes (aggrecan (ACAN), collagen type II (COL2A1)) according to the administration of the extract of erinaceus were confirmed by qRT-PCR technique.

Primer sequences of chondrogenic marker 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′-GCAACAGCAGGTTCACGTACA-3′ 3 reverse 5′-TCGGTACTCGATGATGGTCTTG-3′ 4 GAPDH forward 5′-GGGTGTGAACCACGAGAAAT-3′ 5 reverse 5′-ACTGTGGTCATGAGCCCTTC-3′ 6

As a result, as shown in FIG. 8, it was confirmed that the expression of cartilage formation indicators (aggrecan (ACAN), collagen type II (COL2A1)) was reduced in the sample in which osteoarthritis was induced by MIA, whereas cartilage was treated in parallel with the extract The expression level of the formation index increased. Through this, the chondroprotective effect of the extract of erinaceus was confirmed in an osteoarthritis animal model.

Claims (11)

A health functional food composition for preventing or improving pain caused by cartilage regeneration, arthritis or arthritis, containing an extract of roe deer ( Astilbe rubra ) as an active ingredient. The health functional food composition for preventing or improving pain caused by cartilage regeneration, arthritis or arthritis according to claim 1, wherein the hepatica extract is obtained by extracting one or more parts selected from flowers, leaves, stems and roots. According to claim 1, wherein the arthritis is characterized in that osteoarthritis cartilage regeneration, arthritis or a health functional food composition for preventing or improving pain caused by arthritis. The functional food composition for preventing or improving arthritis according to claim 1, wherein the active ingredient inhibits inflammatory cytokines. The health functional food for preventing or improving pain caused by cartilage regeneration, arthritis, or arthritis according to claim 1, wherein the extraction solvent of the hepatica extract is water, lower alcohol of C ₁ to C ₄ or a mixture thereof. composition. The method of claim 1, wherein the composition is prepared in any one formulation selected from powders, granules, pills, tablets, capsules, candies, syrups and beverages, preventing or improving arthritis or pain caused by arthritis. For health functional food composition. A pharmaceutical composition for preventing or treating pain caused by cartilage regeneration, arthritis or arthritis, containing an extract of ermine greens as an active ingredient. The pharmaceutical composition for preventing or treating pain caused by cartilage regeneration, arthritis or arthritis according to claim 7, wherein the composition further comprises a pharmaceutically acceptable carrier, excipient or diluent in addition to the active ingredient. The method of claim 7, wherein the composition is prepared in any one formulation selected from capsules, powders, granules, tablets, suspensions, emulsions, syrups and aerosols. Therapeutic pharmaceutical composition. A feed additive for cartilage regeneration, arthritis, or preventing or improving pain caused by arthritis, containing an extract of blueberry as an active ingredient. A veterinary composition for cartilage regeneration, prevention or treatment of arthritis or pain caused by arthritis, containing an extract of ermine greens as an active ingredient.

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