KR20090095740A - The healthy and funtional foods for the improvements and prevention of the arthritis symptoms with anti-inflammatory and anti-nociceptive - Google Patents

Abstract

A healthy and functional food for preventing and improving arthritis is provided to show excellent synergistic effects in anti-inflammatory and anti-pain actions using the extracts of Lithospermum erythrorhizon, Chelidonium majus and Cinnamomum cassia. A healthy and functional food for preventing and improving arthritis contains the extracts of Lithospermum erythrorhizon, Chelidonium majus and Cinnamomum cassia. A ratio of Lithospermum erythrorhizon, Chelidonium majus and Cinnamomum cassia is 1~5 : 0.5~2 : 0.5~2, more preferably 1 : 0.5 : 0.5. The extracts are extracted in a low alcoholic solvent such as water, methanol and ethanol. A method for preparing the extracts comprises the following steps of: washing Lithospermum erythrorhizon, Chelidonium majus and Cinnamomum cassia and slicing or chopping each of them; and extracting in 80~100°C of 85% ethanol for 1-3 days. The healthy and functional food can be powder, tablets, granules, pills, hard capsules, soft capsules or liquid. The healthy and functional food represents a raw juice or a drink.

Description

The healthy and funtional foods for the improvements and prevention of the arthritis symptoms with anti-inflammatory and anti-nociceptive}

The present invention relates to a dietary supplement for the prevention and improvement of arthritis having anti-inflammatory analgesic and anti-inflammatory effects from extracts containing essentially the herbaceous, white bran and gyeji.

An inflammatory response is a mechanism for repairing and repairing the damaged area when an invasion that causes any organic change, such as physical action, chemicals, or bacterial infection, is applied to the living body or tissue. Vascular active substances such as serotonin, bradykinin, prostaglandins, hydroxyl-eicosatetraenoic acid (HETE), and leukotriene Increased vascular permeability causes inflammation. Appropriate inflammation, which occurs early in the injury, is a normal response to quick recovery of the damaged area.

However, long-lasting inflammatory reactions can make recovery more difficult by repairing damaged areas and damaging surrounding cells, which can worsen the defenses of surrounding tissues. In addition, prolonged inflammatory reactions can lead to persistent damage to surrounding tissues, resulting in some cancers.

Therefore, tissues with inflammatory reactions should be treated with appropriate anti-inflammatory drugs in order for the recovery program to work well. However, steroidal drugs used to treat inflammatory diseases have many side effects and limit their use. The development of anti-inflammatory agents with excellent effects and low side effects is required (Miller MJ et al., Mediators of inflammation, 4, pp 387-396. 1995: Appleton L. et al., Adv Pharmacol., 35, pp 27). -28, 1996).

On the other hand, arthritis is a disease name that collectively refers to inflammatory changes caused by any cause in the joints. There are many types of degenerative or osteoarthritis, rheumatoid arthritis, etc., but it is called arthritis that causes no loss of the bones of the joint regardless of the cause. Of the types of arthritis, degenerative arthritis is the most common type of arthritis, which may have already begun because of many joints or wounds in youth, but symptoms are mainly seen in older people. It usually occurs in the knee, hip, and spinal joints that carry weight, but can also occur in all joints due to trauma, fractures, and excessive movement.

Rheumatoid arthritis is a long-term disease that affects parts of the human body, including the joints. Rheumatoid arthritis involves swollen joints, invading the surrounding tissues, and secretes certain chemicals that attack and destroy the joint surface. It occurs primarily in the joints of the hands and feet, but sometimes occurs in the hips, knees, and elbows. Even if the joint is not used, swelling, pain, and stiffness may occur. Rheumatoid arthritis occurs in all age groups, but over 70% of all patients are over 30 years old. It may occur simultaneously in many joints.

Treatment is mainly surgical, pharmacotherapy and physiotherapy, but with the exception of some adjuvant therapies are at risk of many side effects. Drugs such as aspirin, ibuprofen, and naproxen, which are used as drug therapy, effectively reduce pain and inflammation, but oral medications may not be safe if the patient has ulcers, asthma, kidney disease, or liver disease. In addition, continuous injection into the same joint area may cause damage to the joint or unexpected gastrointestinal disorders, gastritis and gastric ulcer side effects.

Recently, in order to improve the above side effects, some medicines have been developed with the main ingredient of herbal medicines, but some products are commercially available. There has been a demand for coping with this strong and relatively low side effect dietary supplement and therapeutic agent.

Various biochemical phenomena are involved as a cause of inflammation in living bodies. Macrophage is a multi-functional cell that plays an important role in the inflammatory response by producing various cytokines and NOs under oxidative stress. In particular, iNOS expressed by stimulation such as lipopolysaccharide (LPS), cytokine, TNF-α in macrophages produces a large amount of NO for a long time. This oxidative stress is known to promote NFκB activity, a transcription factor of the inflammatory response. NFκB, composed of p50 and p65 heterodimers, is known to promote gene expression that activates neutrophils such as iNOS and COX-2 after activating and moving to the nucleus. It is known to inhibit (Baeuerle P. et al., Annu. Rev. Immunol., 12 , pp 141-179, 1994; Allenm R. et al, Free Radic. Biol. Med., 28 , pp463-499, 2000) .

Recent studies show that NO is a free radical produced from arginine by three major NOS isoforms: neuronal NOS (nNOS), endothelial NOS (eNOS), and inducible NOS (iNOS). nNOS and eNOS are regulated by Ca ²⁺ / calmodulin, but iNOS is regulated at the level of transcription by inflammatory stimuli such as interleukin, interferon, LPS. NO produced in small amounts by nNOS or eNOS is responsible for normal physiological functions such as vasodilation, neurotransmission, and cellular destruction of pathogens, but NO produced by iNOS in macrophages reacts with superoxide It forms a peroxynitrite, which acts as a powerful oxidant, damaging cells and activating NFκB in macrophages activated by inflammatory stimuli, which is known to be involved in chronic diseases such as inflammatory reactions, cancer, and atherosclerosis. (Lawrence T. et al., Nat Med., 7 , pp1291-1297, 2001; Riehemann K. et al., FEBS Lett., 442 , pp89-94, 1999; Kang JL. Et al., Mol. Cell. Biochem., 215 , pp 1-9, 2000; El-Mahmoudy A. et al., International Immunopharmacology, 2 , pp1603-1611, 2002).

NO is known to be produced by iNOS in various pathophysiological processes, including inflammation and cancer. NOS is an important enzyme that regulates inflammation, vasodilation, neurotransmission, tumor cells and homeostasis of the body. It produces NO from L-arginine, and during inflammatory conditions macrophages produce large amounts of NO by iNOS. It is known to cause various chronic diseases (Kim KM. Et al., Free Radic. Biol. Med., 30 (7) , pp747-756, 2001; Stamler J S. et al., Science, 258 , pp1898-1902 , 1992). Expression of iNOS is induced by NFκB activity, which is an important mechanism by which LPS or cytokine is overproduced in macrophages. NO production in macrophages is directly related to iNOS expression, and LPS is a Gram-negative cell wall material that stimulates immune cells and synergistically increases NO production.

NFκB is a transcription factor involved in various stages such as cytokine response, inflammation, and cell growth regulation. Recent studies strongly suggest that inducible NFκB is involved in the development of atherosclerosis. In many cells, NFκB is a redox-sensitive transcription factor that is involved in various signal transduction from the cytoplasm to the nucleus and is found as a trimer of p50, p65 and IκB subunits in the cytoplasm. When IκB is degraded by oxidative stress, p50 / p65 heterodimers are known to migrate into the nucleus, resulting in continuous DNA binding (Gius D. et al., Toxicol. Lett., 106 , pp93-106, 1999).

Licorice is a perennial herbaceous genus of the dental clinic and corresponds to the roots of lithospermum erythrorhizon. In oriental medicine, it is effective when the repulsion caused by blood fever is dark purple. It has a quick effect on urine bleeding, reduces the incidence of the prevention and treatment of measles, and invades or externalizes early boils and skin rashes, chronic ulcers, cervical erosions, eczema, fire or hot water. Known. In addition, it has an inhibitory effect on Staphylococcus aureus, E. coli, influenza bacteria, dysentery bacteria, and skin fungi, exhibits an inhibitory effect on immune responses, exhibits anti-inflammatory and mild antipyretic effects, and significantly inhibits the menstrual cycle and development of the uterus. It has been known to act as a contraceptive action, exhibit an antimicrobial effect when reacting with physiological blood, and suppresses the oxidative environment by inhibiting the reproduction of microorganisms to eliminate the smell of odor during menstruation. Republic of Korea Patent No. 661014, Republic of Korea Patent Korean Patent No. 299916 and Korean Patent Publication No. 388668 disclose that self-inflamation has an anti-inflammatory effect.

Baekgulchae is a herb that uses the outpost of Chelidonium majusLinne (Papiaceae). It has been reported to have analgesic effects on pain, ulcers, antitumor, blood pressure lowering, antibacterial, central nervous system suppression, and isoquinoline. Chelidonine, hemoelidonine, chelirubine, protopine, alpha-allocryptopine, beta-allocryptopine, β-allocryptopine, Stylopine and benzophenanthridine alkaloids, sanguinarine and chelerythrine, have been reported as main ingredients of pharmacological action. In No. 503926, analgesic, anticancer, immunopotentiating and hematopoietic effects of Baekchulchae extract are known.

Cinnamomum cassia A young branch of cassia (or cinnamon or radish), an evergreen arboretum of the berry family, spicy, sweet and warm in nature, acting on the heart, lungs and bladder. It is known to strengthen the stomach, suppress paralysis, have analgesic and cardiovascular effects, expand the blood vessels of the skin and stimulate sweat glands to cause sweating, antipyretic effect, and inhibit the virus. Chills, fever, headache, body pain In the case of sweating or palpitations, it has been used, and Korean Laid-Open Patent Publication No. 2005-97013 and Korean Patent Publication No. 778078 are known that gyeji extract has an effect on inflammation and pain.

However, the inventors of the present invention, while searching for herbal medicines having anti-inflammatory and anti-pain effects, iNOS, which are two important enzymes that mediate the inflammatory response in the case of single administration of licorice, oleander, and gage described in the prior art, respectively, There is no significant effect on COX-2 enzyme inhibitory activity and inhibitory activity on the production of cytokines such as IL-8, IL-1β, IL-6, and MCP-1, which cause inflammation. Could.

However, surprisingly, the present inventors exhibited significantly more synergistic effect when using the herbal medicine alone in anti-inflammatory analgesic action and anti-inflammatory action by essentially administering the herbaceous, white oyster and gyeji together to effectively have anti-inflammatory analgesic action and anti-inflammatory action. As well as confirming that it does not show side effects such as gastrointestinal disorders, gastritis, gastric ulcer, and extracts containing essentially the herbaceous, white bran and gyeji, the present invention can be usefully used in dietary supplements for preventing and improving arthritis Completed.

The present invention exhibits a significant synergistic effect on anti-inflammatory analgesic and anti-inflammatory action, effectively have anti-inflammatory analgesic and anti-inflammatory action, and health functional food for arthritis prevention and improvement that does not exhibit side effects such as gastrointestinal disorder, gastritis, gastric ulcer, etc. To provide.

In order to achieve the above object, the present invention is to prevent and improve arthritis having anti-inflammatory analgesic action and anti-inflammatory action from extracts containing essentially Lipospermum erythrorhizon, Chelidonium majusLinne and Cinnamomum cassia Provide dietary supplements for

Specifically, the health functional food according to the present invention essentially contains the herb, white oyster and gyeji, the composition ratio of the herb: white oyster: weight ratio of wedge (w / w) 1-5: 0.5-2: 0.5-2 And preferably 1: 1: 0.5: 0.5.

In the present invention, the soybeans, white oysters and cabbages contained in the health functional foods are solvent-soluble extracts selected from water or lower alcohol solvents such as methanol and ethanol or a mixture thereof, preferably water and ethanol mixed solvent soluble extracts, More preferably 60-90% ethanol soluble extract.

The present invention washes and then chops licorice, white oysters and ginseng and about 2 to 20 times its weight, preferably about 5 to 10 times by volume of 85% ethanol at about 80 to 100 ℃ about 1 to 3 days Extraction methods, such as hot water extraction, reflux cooling extraction, ultrasonic extraction, preferably repeated extraction 1 to 3 times by hot water extraction method to obtain the extract, and then the extract is left to cool at room temperature and filtered and concentrated under reduced pressure with a rotary vacuum concentrator. Then freeze-dried can be obtained a dietary supplement containing essentially the liquorice, white oysters and gyeji of the present invention.

Herbal herbicide, white buckwheat, and basal gland used in the present invention were known to have anti-inflammatory or anti-pain effects, but the anti-inflammatory effect and mouse model in carrageenan-induced acute arthritis model and serotonin-induced acute arthritis model in real rats. Analgesic Effect and Hot Plate Method by Acetic Acid wirthing Method As a result of measuring the analgesic effect by the hot plate method, in contrast to the known literature, anti-inflammatory and analgesic effects are not only weak, but also in the anti-inflammatory action test using mouse macrophages, NO production amount, COX-2, prostaglandin 2 (PGE2) ), And cytokines (IL-8, IL-1β, IL-6) were not found to be significant in the dietary supplement for the prevention and improvement of arthritis.

Surprisingly, however, the composition essentially containing the moths, moths and moss in accordance with the present invention, unlike the results shown in the moss, moths and moss, respectively, has anti-inflammatory effect in the carrageenan-induced acute arthritis model and serotonin-induced acute arthritis model of rats. Analgesic Effect and Hot Plate Method by Acetic Acid wirthing Method in Korean and Mouse Models As a result of measuring the analgesic effect by the hot plate method, it showed the anti-inflammatory effect equivalent to or higher than the control drugs ibuprofen and indomethacin, and the present invention also showed analgesic effect and hot plate method by Acetic acid wirthing method in mouse model. As a result of measuring the analgesic effect by the hot plate method, it showed an analgesic effect equivalent to or higher than that of the control drugs aspirin and morphine.

In addition, the present invention inhibited the synthesis of nitric oxide at the cellular level, inhibited two important enzymes, iNOS and COX-2, which mediate the inflammatory response, and induce the inflammation of IL-8, IL-1β, IL-6, It was confirmed that the inhibitory activity on the production of cytokines such as MCP-1 is excellent and has an excellent effect on anti-inflammatory activity.

In particular, the present invention is very excellent PGE2 inhibitory effect does not show side effects such as gastrointestinal disorders, gastritis, gastric ulcers can be used as a health functional food for the purpose of preventing and improving arthritis having anti-inflammatory analgesic and anti-inflammatory action.

The present invention can be developed as a health functional food by adding to various foods, for example, beverages, teas, vitamin complexes, health supplements, etc., and can be prepared and used in the form of direct pills, powders, tablets, capsules and the like. In addition, when formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants that are generally used are prepared.

As described above, the present invention not only exhibits side effects such as gastrointestinal disorder, gastritis, and gastric ulcer by essentially administering the herbaceous, baekryechae, and gyeji, but also exhibits a synergistic effect in anti-inflammatory and anti-inflammatory action. When using the herbal medicine alone can be used more effectively in health functional food for the prevention and improvement of arthritis having anti-inflammatory analgesic action and anti-inflammatory action.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail. However, the scope of the present invention is not limited by these Examples and Experimental Examples.

<Example 1> Preparation of the extract according to the present invention

1 g of ethanol was added to a mixture of 50 g of vinegar, 25 g of white oysters and 25 g of ethanol, and ethanol was extracted at 80 ° C. for 3 days, and ethanol extraction was repeated twice. The extract was concentrated under reduced pressure in a rotary vacuum evaporator and lyophilized to prepare 2.21 g of powder.

Comparative Example 1 Preparation of Licorice Extract

50 g of licorice was added to 1 L of ethanol for ethanol extraction for 3 days, and ethanol extraction was repeated twice, and the extract was concentrated under reduced pressure and freeze-dried in the same manner as in Example 1 to prepare 1.32 g of powder.

Comparative Example 2 Preparation of Baekgulchae Extract

25g of white oysters were extracted in the same manner as in Comparative Example 1 to prepare 0.85g of powder.

Comparative Example 3 Preparation of Cinnamon Extract

25 g of gage was extracted in the same manner as in Comparative Example 1 to prepare 0.78 g of powder.

Experimental Example 1 Anti-inflammatory Effects of Rats Carrageenan-Induced Acute Arthritis Model 1

Sprague-Dawley's 180-200 g of rats (Dae Biolink, Korea) were used, and as a prophylactic agent for inducing arthritis, 1% carrageenin was used. 1g each of the freeze-dried powder extracted in Example 1, Comparative Examples 1 to 3 according to the present invention was dissolved in 10ml of physiological saline, orally administered 200 mg / kg for 2 weeks, and then 0.1 ml of 1% carrageenan solution was added to rats. Injection into the right hind paw. Edema that occurred 1 hour and 2 hours after injection was measured by volume method to the volume of the right hind paw of the rat up to the malleous lateral area using a Plethysmometer (UGO Basile, Italy). Table 1 shows. As a control drug, ibuprofen was administered at 100 mg / kg.

As can be seen in Table 1, in the group administered orally administered the extract of Example 1 of the present invention showed an edema inhibitory effect compared to the control group not administered, in particular, it has an acute anti-inflammatory effect equivalent to or greater than ibuprofen as a control drug. Although it can be confirmed, it can be seen that the anti-inflammatory effect is very weak in the administration group administered the same amount of the licorice, baekryechae and gyeji in Comparative Examples 1 to 3, respectively.

Therefore, although the anti-inflammatory action is very weak anti-inflammatory effect unlike the conventionally known, soybean, baekryokchae and gyeji extract, it can be seen that the present invention, which contains essentially the soybean, cabbage and gyeji is very excellent anti-inflammatory action.

Experimental Example 2 Determination of Anti-inflammatory Effects of Rats in Serotonin-induced Acute Arthritis Model 2

In order to reconfirm the anti-inflammatory effect according to the present invention, the anti-inflammatory effect of serotonin-induced rats was performed. As in Experimental Example 1, 1g each of the freeze-dried powder extracted in Example 1, Comparative Examples 1 to 3 according to the present invention was dissolved in 10ml of physiological saline and orally administered 200 mg / kg two weeks before serotonin ( 1mg / ml) was injected into the right sub plantar region of the rat by 0.1ml to induce edema, and the pedal volume was measured in 18 minutes, 24 minutes and 30 minutes immediately before the injection. Indomethacin (10 mg / kg) was used as a comparative drug.

From Table 2, it can be seen that the anti-inflammatory effect is very weak in the administration group administered the same amount of licorice, baekryechae and gyeji, Comparative Examples 1 to 3, but the control drug in the oral administration of the extract of Example 1 of the present invention It can be seen that there is an acute anti-inflammatory effect equivalent to that of phosphorus indomethacin, the present invention containing essentially the herbaceous, baekryokchae and gyeji is very excellent anti-inflammatory action.

Experimental Example 3 Analgesic effect measurement in mouse model 1

As in Experiment 1 by Acetic acid wirthing method, 1 g of each freeze-dried powder extracted in Example 1 and Comparative Examples 1 to 3 according to the present invention was dissolved in 10 ml of physiological saline and 200 mg / kg of ICR mouse for 7 days. (20 g) were administered orally, respectively. One hour after the last oral administration on day 7, 0.1 ml / 10 g of 0.6% acetic acid-physiological saline solution was administered intraperitoneally, followed by writhing syndrome (acetic acid intraperitoneally in animals) When you hit a person to stretch the waist and stretch the hind legs, such as stretching, which means that when you feel the pain takes this type of action) and the frequency is shown in Table 3. Aspirin (100 mg / kg) was used as a comparative drug.

As shown in Table 3, the experimental results showed only a slight analgesic effect in the same administration groups of Comparative Examples 1 to 3, the same dose of Licorice, White Oysters and Gyeji, but in the group orally administered the extract of Example 1 of the present invention. It can be confirmed that the drug has an analgesic effect equivalent to that of aspirin, and the present invention, which essentially contains licorice, white oyster and gyeji, can confirm that the analgesic effect is very excellent.

Experimental Example 4 Analyzing Analgesic Effects in a Mouse Model 2

Hot plate method to reconfirm analgesic effect The analgesic effect by the hot plate method was measured. Using a hot plate, as in Experiment 1, each sample was orally administered to an ICR mouse (20 g), and after one hour was placed in a hot plate (70 ° C.), the time until the mouse jumped and jumped at the same time was measured. Table 4 shows. Morphine (10 mg / kg) was used as a comparative drug.

As can be seen in Table 4, the experimental results of comparative examples 1 to 3, the administration of the same amount of licorice, baekryechae and gyeji respectively, the effect of alleviating the pain, but the oral administration of the extract of Example 1 of the present invention In the group, it can be confirmed that the effect of alleviating pain equivalent to that of the control drug morphine.

Therefore, it can be seen that the present invention is very excellent in the pain relief effect in the composition containing essentially the herbaceous, baekryechae and gyeji.

Experimental Example 5 Anti-inflammatory Effect Using Mouse Macrophages: Measurement of NO Production

After dispensing Murine RAW264.7 macrophages into 6 well plates so as to have a number of ^{6 6} cells per well, after 24 hours, 50 μg / ml of each of Examples 1 and 3 according to the present invention was taken and 10 ng / ml of LPS. and IFN-γ by simultaneously processing a 100U / ㎖ for 48 hours and then cultured in a CO ₂ incubator and separated for 5 minutes using a centrifugal by 2000rpm NO ₂ to the amount present in the cell culture medium of the nitric oxide (nitric oxide) produced from supernatant-for It was measured using Griess reagent as the form. Each supernatant was dispensed into a 96 well plate and 100 μl of Griess reagent (0.8% sulfanilamide / 0.75% N- (naphthyethylene) diamine in 0.5 N HCl, sigma) was added. After standing at room temperature for 15 minutes, the nitrite concentration was measured using a microplate reader (Molecular Devices, Sunnyvale, CA, USA) at 540 nm wavelength is shown in FIG. Sodium nitrite (0.5-100M) was used as the nitrite standard.

As can be seen in Figure 1, Example 1 according to the present invention inhibited NO synthesis of LPS and IFN-γ-induced cells by 50 ㎍ / ㎖ by 79%, Comparative Examples 1 to 3 NO of the cells The degree of synthesis inhibition was very weak.

Experimental Example 6 Anti-inflammatory effect using mouse macrophages; Cyclooxygenase (COX-2) Determination

After dispensing Murine RAW264.7 macrophages in 6 well plates to 10 ⁶ cells per well, Example 1 according to the present invention 1, 10 and 50 ㎍ / ㎖ take LPS 10 ng / ㎖ and IFN-γ was treated with 100U / ml at the same time and cultured in a CO ₂ incubator for 48 hours, and then the cells were collected and RNA was extracted by the method using Trizol Reagent, and cDNA was synthesized with 2ug RNA, COD-1, PCR was performed with each primer of COX-2. PCR conditions were 30 cycles of Pre-denaturation 94 ℃ 5 minutes, Denaturation 94 ℃, Annealing 58-65 ℃ 30 seconds, polymeration 72 ℃ 30 seconds, Extend polymeration 72 ℃ 5 minutes. As a comparative drug, cecoxibe, a cox-2 inhibitor, was used.

As shown in Figure 2, 1, 10, 50 ㎍ / ㎖ of Example 1 according to the present invention showed the effect of inhibiting COX-2 induced by LPS and IFN-γ. In particular, 10 ㎍ / ㎖ of Example 1 according to the present invention was inhibited to the same extent as the positive control group selecoxib, 50 ㎍ / ㎖ showed a more inhibitory effect than celecoxib.

Experimental Example 7 Anti-inflammatory Effect Using Mouse Macrophages: Determination of Prostaglandin 2 (PGE2) Content

After dispensing Murine RAW264.7 macrophages into 6 well plates so as to have a number of ^{6 6} cells per well, after 24 hours, 50 μg / ml of each of Examples 1 and 3 according to the present invention was taken and 10 ng / ml of LPS. And IFN-γ were treated with 100U / ml simultaneously and incubated in a CO ₂ incubator for 48 hours, followed by centrifugation at 2000 rpm for 5 minutes, and the amount of prostaglandin 2 (PGE2) was measured by enzyme-immunoassay (FIG. 3). Kits, R & D system, USA). As a comparative drug, cecoxibe, a cox-2 inhibitor, was used.

As can be seen in Figure 3, Example 1 according to the present invention showed that the LPS and IFN-γ-induced PGE2 production in RAW264.7 macrophage main cells significantly reduced compared to the comparable drug, but compared Examples 1-3 rarely reduced PGE2 production.

Experimental Example 8 Anti-inflammatory Effects Using Mouse Macrophages

Murine RAW264.7 macrophages were dispensed into 6 well plates to have 10 ⁶ cells per well, and after 24 hours, LPS 10 ng / ml and IFN-γ 100U / ml, and Examples 1 and 1 according to the present invention. 50 μg / ml each of 3 was co-treated and incubated for 48 hours in a CO ₂ incubator using celecoxib (50 μg / ml) as a positive control. After incubation, the plate was centrifuged at 2000 rpm for 5 minutes to collect the culture supernatant. The amounts of MCP-1, IL-8, IL-1β and IL-6 were measured by enzyme-immunoassay and shown in FIGS. 4A, 4B, 4C and 4D (Kits, BD Pharmigen, USA). As a comparative drug, cecoxibe, a cox-2 inhibitor, was used.

As can be seen in Figures 4a, 4b, 4c and 4d, Example 1 according to the present invention is chemokine (MCP-1) and cytokines (IL-) induced by LPS and IFN-γ in RAW264.7 macrophage line cells 8, IL-1β, IL-6) was shown to significantly reduce the production of the same as the comparative drug, Comparative Examples 1 to 3 almost did not show an effect of reducing significantly.

Experimental Example 9 Inhibitory Activity of Gastritis Model

100 mg, 200 mg and 300 mg of the freeze-dried powder of each extract prepared in Example 1 according to the present invention were dissolved in 0.1% CMC, respectively, and orally administered to SD rat t (male, SPF, 150 200 g, purchased from Orient Bio Company). It was. Indomethacin was orally administered to SD rats at a dose of 10 mg / rat as a control. After 5 hours, the stomach was excised, ground with a polytron homogenizer, and centrifuged to obtain a supernatant. Then extracted with ethyl acetate and dried. After dissolving in methanol, Park (Eur. J. Pharmacol. 425, 153-157, 2001) was added to Sep-Pak C18 column, and after distillation, it was dried with N2 and PGE, which is a parameter of gastric inflammation, by ELISA method. The amount of 2 was quantified and the results are shown in Table 5.

As can be seen in Table 5, Example 1 according to the present invention almost did not cause inflammation of the stomach irrespective of the dose, but the control drug is a high risk of gastritis, the present invention is very effective in inhibiting PGE2 gastrointestinal It does not show side effects such as disorders, gastritis and gastric ulcers and can be used as a health functional food for the purpose of preventing and improving arthritis having anti-inflammatory and anti-inflammatory effects.

1 is a graph showing the inhibitory effect of NO production in the macrophages of the lyophilized powder prepared in Example 1, Comparative Examples 1 to 3 according to the present invention,

2 is a graph showing the effect of inhibiting COX-2 production in mouse macrophages by concentration (1, 10 and 50 ㎍ / ㎖) of the lyophilized powder prepared in Example 1 according to the present invention,

3 is a graph showing the effect of inhibiting the production of prostaglandin 2 in mouse macrophages of the lyophilized powder prepared in Example 1, Comparative Examples 1 to 3 according to the present invention,

Figure 4 is a graph showing the inhibitory effect of inflammatory cytokines and chemokines in mouse macrophages of the lyophilized powder prepared in Example 1, Comparative Examples 1 to 3 according to the present invention,

    4A is a graph showing the inhibitory effect of MCP-1 production;

    Figure 4b is a graph showing the inhibitory effect of IL-8 production,

    Figure 4c is a graph showing the inhibitory effect of the production of IL-1β and

    Figure 4d is a graph showing the inhibitory effect of IL-6 production.

Claims (4)

A dietary supplement for the prevention and improvement of arthritis with anti-inflammatory and anti-inflammatory effects, which contains essential extracts of Lithospermum erythrorhizon, Chelidonium majusLinne and Cinnamomum cassia. The dietary supplement according to claim 1, wherein the weight ratio (w / w) of the moths: white buckwheat: gye is contained in a weight ratio of 1: 0.5: 0.5. According to claim 1, wherein the health functional food powder, tablets, granules, Health functional food, characterized in that selected from pills, hard capsules, soft capsules or liquid formulations. According to claim 1, wherein the health functional food is a juice, pouch, Health functional food characterized in that the drink or tea.

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