KR101284772B1 - Functional food composition with the effects of anti-inflammation and pain-relieving - Google Patents

Abstract

The present invention relates to a functional food composition having an anti-inflammatory and analgesic effect obtained by extracting an active ingredient (anthocyanine, quercetin, curcumin) from grains and fruits, and selectively combining the obtained extracts, and a method for producing the same.
Functional food composition of the present invention for extracting red cabbage, apple peel and turmeric, respectively, and then mix them in an appropriate ratio of antioxidant effect, inflammatory edema alleviation effect, acute and chronic inflammation inhibitory effect, inflammation Inhibitory effects of inducing enzymes Lipoxygenase, Cyclooxygenase-2 and inducible Nitric oxide synthase (iNOS), Inhibitory activity of COX-2 and Type I collagenase, TNF-α (tumor necrosis factor-alpha) and IL- 1β (interleukin 1-beta), IL-6 (interleukin 6) and Prostaglandins production of excellent anti-inflammatory analgesic and functional food composition that is effective in preventing and improving symptoms of arthritis.

Description

Functional food composition with the effects of anti-inflammation and pain-relieving}

The present invention relates to the establishment of an optimal method of extracting active ingredients effective in the prevention of inflammation and pain and improvement of symptoms using leafy vegetables, root vegetables and fruits, and to develop a mixed composition containing the extract in an optimum ratio. In more detail, red cabbage, apple peel and turmeric extract were extracted, respectively, and then mixed at an appropriate ratio. (lipoxygenase, LOs), Cyclooxygenase-2 (COX-2) and Inducible Nitric Oxide Synthase (iNOS) Inhibitory Effects, COX-2 and Type 1 Collagen Hydrolase Inhibitory activity of Type I collagenase, nitric oxide (NO), an inflammatory trigger, tumor necrosis factor-alpha (TNF-α), interleukin 1-beta (interl) anti-inflammatory and analgesic effects such as eukin 1-beta, IL-1β), interleukin-6 (IL-6) and prostaglandins (PGs) production inhibitory effect, morphological and histological knee joint inhibitory effect It relates to a functional food composition and a method for producing the same.

Red Cabbage ( Brassica) oleracea L. var . capitata f. rubra . ) Is a type of cabbage with a reddish leaf color and is called red vegetable, red cabbage, and purple cabbage. Red cabbage contains 90% of water, 1.5 ~ 2.3% of crude protein, 6.4% of sugar, 0.6% of fat.The main nutrients include β-carotene 15.0μg, vitamin A 3.0μg, vitamin B2 0.05mg, vitamin B6 0.25mg per 100g. , Vitamin C 51.0mg, Vitamin E 0.11mg, Dietary Fiber 1.82g, Zinc 0.16mg, Phosphorus 41.0mg, Iron 0.5mg, Potassium 282.0mg, Calcium 31.0mg and the fat component contains a lot of linolenic acid. In addition, the main pigment of red cabbage is a flavonoid-based anthocyanin (anthocyanin), known as a powerful antioxidant, often has four hydroxyl groups. In particular, red cabbage contains a lot of methyl-methionine-sulfonate and vitamin K called vitamin U. It is effective in preventing gastritis, gastric ulcer and duodenal ulcer by inhibiting gastric secretion and promoting regeneration of ulcer tissue. have. [Reference: 1. Doosan Encyclopaedia encyber Doosan Co., Ltd. 2. Food Hygiene, University Seorim, pp138 (1993)]

Apple ( Malus pumila Mill . ) Is classified as fruit in the classification of raw materials of food industry and is edible. The varieties include early varieties (honggu, hongro, axis) and mesothelioma (uk, ruby, stararching) and late varieties (gwanggwang, gourmet Delicious, India, Fuji). Depending on the varieties, the water content is about 86%, the sugar content is 11-14%, and the composition of these sugars is 52% fructose, 26% glucose and 20% sucrose. Organic acids are mainly malic acid and citric acid, and there are small amounts of kinasan, succinic acid, glycolic acid, citramal acid, α-ketoglutamic acid and iso citric acid. Inorganic ingredients include potassium 110mg, calcium 3mg, phosphorus 8mg, iron 0.1mg per 100g, potassium is the most contained in combination with the pectin component to release sodium to the outside by lowering blood pressure, effective in treating and preventing hypertension. The amino acid composition is aspartic acid and asparagine, and a small amount of threonine, serine, glutamic acid, glutamine, alanine, isoleucine, and phenylalanine. Aromatic ingredients include methyl acetate, ethyl acetate, pyrophyll acetate, butyl acetate, hexyl acetate, butyl propionate, ethyl 2-methylbutylate, isobutyl alcohol, butyl alcohol, 2-methylbutyl alcohol, hexyl alcohol, t-2 -Hexenyl alcohol, 4-methoxylbenzene and the like, and the aroma consists of a combination of these materials. Pectin, a water-soluble plant fiber contained in apples, is effective in intestinal action, promotes the intestinal lactic acid bacteria, and absorbs moisture to increase volume, which is effective for constipation and helps prevent obesity due to satiety.

In addition, the apple peel contains most of the vitamin C of the apple, contains a large amount of bioactive ingredients such as quercetin and camphorol, and contains antioxidants such as caproic acid and chlorogenic acid. It has been reported that it contains a large amount of cardiovascular disease, cerebrovascular disease, arteriosclerosis prevention, anti-inflammatory action and anti-cancer effect. [Reference: 1. Primary Korean Plant Book, Kyohaksa (1996) 2. Doosan World Daebaek and encyber Doosan Corporation 3. Chosun Medicinal Plants, Korean Cultural History, pp197 (1999)]

Turmeric is steamed and dried after removing the bark of Curcuma longa L. , which is widely used for edible and medicinal purposes. The main ingredient is about 15% curcuminoids (curcumin, diferuloylmethane) is the most contained among these, in addition to dicaffeoylmethane (dicaffeoylmethane) and caffeoylferuloylmethane (cafferoylferuloymethane). In addition, the essential oil is about 15%, including sesquiterpene 56.5%, sesquiterpene alcohol 22%, d-camphor 2.5% and d-camphene. It is contained. Fat-soluble components include campesterol, stigmasterol, β-sitosterol, cholesterol, and the like. Curcumin, the ingredient of turmeric, is the No. 1 candidate for cancer prevention in the joint project of Ministry of Health, Welfare, Ministry of Science and Technology and Ministry of Science and Technology. Scientific evidence has been reported, including prevention of diabetes complications, suppression of hypertension, and prevention of atherosclerosis. In 2001, according to the American Journal of Neurology, Indians reported only 25% of Alzheimer's incidence compared to Americans.In 2002, the World Health Organization (WHO) found that Indian cancers were one of the most common cancers in the United States. / 7.

Inflammation can include acute inflammation (immediate response, nonspecific response, days to weeks), chronic inflammation, delayed response, specific response, several weeks or more, and acute inflammation. Middle stage of chronic inflammation, characterized by the mixed product of multinucleated and monocytes).

In addition to the peptide-inducing factor, inflammatory factors include prostaglandins, leukotrienes, lipid factors such as platelet activators, inflammatory synthase, free radicals such as NO (nitric oxide), various cell adhesion molecules, Immune system, coagulation factor is involved.

The mechanisms of inflammation known to date are cellular damages caused by external biological causes (bacteria, viruses, parasites), physical causes (mechanical stimulation, heat, radiation, electricity), chemical causes, etc. Vasodilation, increased capillary permeability, and aggregation of macrophages occur in inflammatory sites, resulting in increased blood flow, edema, immune cell and antibody migration, pain, and fever.

Currently used therapeutic agents for inflammation include synthetic drugs such as ibuprofen, antihistamines, steroids, cortisones, immunosuppressants, and immunosuppressive drugs, but there are many side effects such as temporary treatment, simple symptoms, hypersensitivity reactions, and immune system deterioration. The underlying treatment of inflammation is difficult.

The pain may be acute pain, subacute pain, recurrent pain, persistent cancer pain, chronic pain, or chronic pain syndrome depending on the period. The mechanism of pain known to date is an unpleasant sensation of stimulation of the cerebral cortex and systemic region through the afferent neuropathy from the nociceptors to the nerve fibers. Pain-related substances include prostaglandin, leukotriene, and thromboxane produced by cyclooxygenese and lipooxigenase enzymes using arachidonic acid as a substrate. Serotonin, catecholamine, protons, histamine, potassium ions, and the like.

In addition, pain treatment includes medication (painkillers, non-narcotic analgesics, nonsteroidal anti-inflammatory drugs, antidepressants), injection therapy (mainly local injection into muscle pain), physical therapy (cold therapy, heat therapy, high voltage direct current therapy, Percutaneous electrical neurostimulation therapy, exercise therapy, ultrasound therapy, iontophoresis and sonic electrophoresis), but these treatments usually have side effects, are resistant to repetitive treatments, and their effects are often temporary, so they are expected to be realistic and appropriate. Difficult to do

Degenerative arthritis, also called osteoarthritis (degenerative joint disease), is the most common type of arthritis. In some cases, trauma, disease, and joint malformations are the major causes of inflammation and pain, mainly due to degenerative damage to cartilage that protects joints. Although aging generally increases the incidence of degenerative arthritis, sex, genetic factors, obesity, and lifestyle are also associated. In severe cases of degenerative arthritis, surgical treatment such as vitreous removal of the vitreous and synovial membranes, synovectomy, curettage of the knee, multiple perforation, and arthroplasty is performed.

The cause of rheumatoid arthritis is not yet clear, but autoimmunity is known as a major mechanism. Inflammation of the synoviium begins to spread to surrounding cartilage and bones, leading to deformation of the joints (primarily the fingers, wrists, toes, and joints of the elbows, shoulders, ankles, and knees as the disease progresses). In addition, complications such as subcutaneous nodules, vasculitis, pulmonary fibrosis, anemia and skin ulcers occur. Various drugs are used to treat the disease, but no drug has been developed to cure rheumatoid arthritis.

Drugs used for degenerative arthritis and rheumatoid arthritis are mainly included in anti-inflammatory analgesic drugs such as aspirin and acetaminophen, non-steroidal anti-inflammatory drugs (NSAIDs), such as cerebrex and viox, and TNF blockers. Rheumatoid arthritis) is used, but the underlying treatment of the disease is impossible, and gastrointestinal side effects (gastritis, vomiting, peptic ulcer, gastrointestinal bleeding, ulcerative colitis) and circulatory side effects (hypertension, palpitations) and blood coagulation Serious side effects have been reported in mechanisms, etc., so caution is required.

In addition, products such as joins tablets, adroda capsules, imoton tablets, seimi tablets, atrene tablets, glucosamine, and chondroitin sulfate are currently used in natural medicines, but the products are insignificant or have undergone rigorous clinical research to prove their effectiveness. rare. Indeed, taking glucosamine for the treatment of arthritis in Korea has become a social epidemic, but a study conducted by the US FDA to investigate the therapeutic effect of a large number of patients showed no effect at all.

In view of the above-mentioned problems of inflammatory diseases, painful diseases and arthritis symptom improving agents in use, the present invention has no concern about side effects compared to other synthetic drugs by using natural plant extracts. It is an object of the present invention to provide a functional food composition and a method for producing the functional food composition having an excellent effect on the improvement and prevention of the underlying symptoms.

In addition, if most of the anti-inflammatory and analgesics developed so far focused on inhibiting the activity of proinflammatory enzymes such as COX-2, Lipoxygenase, iNOS, the present invention not only inhibits the activity of these enzymes, but also in vivo It is an object of the present invention to provide a functional food composition that aims at improving the root symptom of inflammation and pain by inventing a natural extract that also inhibits biosynthesis (expression).

In addition, the present invention develops a plant composition that prevents the destruction of articular chondrocytes after the onset of the disease as well as before the onset by playing an important antioxidant action in the joint health, the development of degenerative arthritis of the elderly in the high risk group of the disease treatment and disease development It is another object to provide a composite natural food functional composition that prevents.

In addition, after exploring a variety of natural products having the effect of anti-inflammatory and analgesic, to find a natural plant extract that can replace a synthetic drug, and in particular, to provide a functional food composition consisting of a combination of these natural plant extracts having a synergistic effect For the purpose of

Functional food composition of the present invention for achieving the above object, containing the mixed plant composition of red cabbage extract, apple peel extract and turmeric extract as a main component, to improve the prevention and symptoms of inflammatory diseases and pain diseases and arthritis It is characterized by.

The mixed plant composition is preferably red pepper and cabbage extract and apple peel extract and turmeric extract in a weight ratio of 1: (0.1 ~ 5): (0.1 ~ 10), respectively.

Functional food composition of the present invention for achieving the above object, containing a plant composition mixed with red cabbage extract and apple peel extract as a main component, characterized in that to improve the prevention and symptoms of inflammatory diseases, painful diseases and arthritis do.

The mixed plant composition is preferably to be mixed red pepper cabbage extract and apple peel in a weight ratio of 1: (0.1 to 5).

Functional food composition of the present invention for achieving the above object, containing the plant composition mixed red cabbage extract and turmeric extract as a main component, it characterized in that the prevention and symptoms of inflammation, pain, arthritis.

The mixed plant composition is preferably to be mixed red pepper cabbage extract and turmeric extract in a weight ratio of 1: (0.1 to 10).

In addition, the functional food composition of the present invention for achieving the above object is characterized by containing a plant composition of the apple peel extract and turmeric extract as a main component, to prevent inflammation and pain, arthritis and improve symptoms.

The mixed plant composition is preferably such that the apple peel extract and turmeric extract is mixed in a weight ratio of 1: (0.1 to 10).

The functional food to which the functional food composition is administered is preferably one of capsules, pills, granules, powders, candy, gum, tablets, beverages, and syrups.

The manufacturing method for producing red cabbage extract of the extract contained in the functional food composition of the present invention, washed red cabbage two times with purified water and then crushed 30 ~ 100 ℃ with 0 ~ 99% ethanol aqueous solution of 4 to 10 times by weight 2 to 8 hours, 1 to 4 times extraction, cooling and filtration with the first step; A second step of concentrating the filtrate obtained in the first step at 50 to 100 ° C. under reduced pressure; And a third step of lyophilizing the extract obtained in the second step and preparing a powder extract, characterized in that an anthocyanin, which is an indicator substance extracted through the process, is 0.1 to 20%.

The method for producing an apple peel extract among the extracts contained in the functional food composition of the present invention, the apple peel is 4 to 10 times by weight of 0 to 100% alcoholic aqueous solution at 30 to 100 ℃ for 2 to 8 hours, 1 ~ A first step of extracting four times, cooling and filtering; A second step of concentrating the filtrate obtained in the first step at 30 to 80 ° C. under reduced pressure; And a third step of lyophilizing the extract obtained in the second step and preparing a powder extract, wherein the total quercetin, which is an indicator material extracted from the process, is 0.2 to 10%. .

The manufacturing method for preparing turmeric extract among the extracts contained in the functional food composition of the present invention, washing the turmeric with purified water twice and pulverizing and then 2 to 60 to 100 ℃ with 0 to 100% alcoholic aqueous solution of 4 to 10 times the weight ratio A first step of extracting, cooling and filtering for 1 to 4 times for 8 hours; A second step of concentrating the filtrate obtained in the first step at 30 to 80 ° C. under reduced pressure; And a third step of lyophilizing the extract obtained in the second step and preparing a powder extract, wherein the curcumin, which is an indicator material extracted from the process, is 0.1 to 40%.

Antioxidant activity (DPPH radical scavenging activity, SOD-like activity, H ₂ O ₂ scavenging activity), inflammatory enzyme (COX-2, Lipoxygenase, iNOS, Type I collagenase) using functional food composition using the composition of natural plant extract of the present invention Inflammatory and painful diseases with excellent effects on inhibition of activity and inhibition of expression in vivo, inhibition of in vivo production of inflammatory mediators (nitric oxide, prostaglandin, TNF-α, IL-1β, IL-6), and inflammatory edema It is useful as a prophylactic and symptomatic agent for preventing arthritis.

1 is a graph showing the SC ₅₀ value in ppm concentration of the mixed extract prepared by the present invention and the comparative substance scavenged DPPH radicals by 50%.
Figure 2 is a graph showing the SC ₅₀ value in ppm concentration showing the SOD-like activity of the mixed extract prepared by the present invention.
Figure 3 is a graph showing the SC ₅₀ value in ppm concentration of the mixed extract prepared in accordance with the present invention and the comparator removes H ₂ O ₂ by 50%.
Figure 4 is a graph showing that the mixed extract prepared by the present invention and the comparative substance inhibits the production of nitrogen monoxide induced by LPS in mouse macrophages.
Figure 5 is a graph showing the IC ₅₀ value in ppm concentration of the mixed extract prepared by the present invention and the comparative material 50% inhibition of the COX-2 enzyme.
6 is a ratio of the mixed extract prepared by the present invention and the comparative substance inhibits the activity of lipooxygenase enzyme in the blood of chronic inflammation animal model by MIA (mono sodium iodoacetate) compared to the untreated group of the sample. Graph shown.
Figure 7 is a graph showing the amount of production of TNF-α in the foot portion in which the mixed extract prepared by the present invention and the comparative material edema of the acute inflammatory animal model caused by carrageenan.
8 is a graph showing the amount of production of TNF-α in the knee joint site where the mixed extract prepared by the present invention and the comparative material inflamed chronic animal model by MIA.
Figure 9 is a graph showing the amount of production of IL-1β in the foot portion of the mixed extract prepared by the present invention and the comparative material edema of acute inflammation animal model caused by carrageenan.
10 is a graph showing the amount of production of IL-1β in the knee joint site where the mixed extract prepared by the present invention and the comparative material is inflammation of the chronic inflammation animal model by MIA.
FIG. 11 is a graph showing the amount of IL-6 production in the foot region in which the mixed extract prepared by the present invention and the comparative substance caused swelling of an acute inflammatory animal model caused by carrageenan.
Figure 12 is a graph showing the amount of IL-6 produced in the knee joint site where the mixed extract prepared by the present invention and the comparative material inflamed chronic animal model by MIA.
Figure 13 is a graph showing that the mixed extract prepared by the present invention and the comparative substance inhibits the production of prostaglandin induced by LPS in mouse macrophages.
14 is a graph showing that the mixed extract prepared by the present invention and the comparative substance inhibits the production of prostaglandin in the knee joint site where inflammation of the chronic inflammation animal model by MIA occurred.
Figure 15 is a graph showing the IC ₅₀ value in ppm concentration of the mixed extract prepared by the present invention and the comparative substance inhibits Type I collagenase 50%.
FIG. 16 is a diagram illustrating the effect of inhibiting paw edema in acute-inflammatory animal model induced by carrageenan by the mixed extract prepared by the present invention.
FIG. 17 is a diagram showing the effect of inhibiting paw edema in an animal model of acute inflammation caused by carrageenan with the mixed extract prepared by the present invention after 6 hours.
FIG. 18 is a magnification of 1000 times with an optical microscope after immunocytochemical staining to inhibit the expression of 5-lipooxygenase induced by LPS in mouse macrophages in a mixed extract prepared by the present invention. to be.
19 is an enlarged view 1000 times with an optical microscope after immunocytochemical staining that the mixed extract prepared by the present invention and the comparative substance inhibit the expression of iNOS induced in LPS in mouse macrophages.
FIG. 20 is an enlarged scale 1000 times with an optical microscope after immunocytochemical staining to inhibit the expression of COX-2 induced by LPS in mouse macrophages in a mixed extract prepared by the present invention.
Figure 21 is a mixed extract prepared by the present invention and stained histological changes of the knee cartilage of the knee cartilage of chronic inflammation animal model by MIA using hematoxylin and eosin 400 times magnified by optical microscope The figure was observed.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will now be described in detail with reference to the accompanying drawings.

In the present invention, red cabbage extract, apple peel extract and turmeric extract are mixed at an appropriate ratio, and optimized in a predetermined amount, thereby inhibiting inflammatory edema superior to each extract, Type I collagenase, lipoxygenase, and COX- 2 inhibitory effect, inflammatory edema alleviation effect, inhibitory effect of the production of inflammatory triggers nitric oxide (NO) and prostaglandins, iNOS, 5-lipoxygenase and COX-2 inhibitory effects in vivo, inflammation Inflammatory disease that is more prominent by inhibiting the production of the induced cytokines TNF-α, IL-1β and IL-6, as a potential inducer of arthritis, and having a radical free radical scavenging effect and an antioxidant effect that are considered as a serious factor of the disease The present invention relates to a complex functional food composition useful for the prevention and improvement of symptoms of painful diseases and arthritis.

Such functional food composition was used to extract red cabbage extract, apple peel extract and turmeric extract with water or alcoholic aqueous solution to prepare the composition, respectively.

1) First, the method of producing a composition by extracting red cabbage is as follows.

After washing red cabbage two times with purified water, use a food grinder to grind for 5 to 10 minutes at 300 to 500 rpm, and then use 4 to 10 times of 0 to 99% ethanol aqueous solution at 30 to 100 ℃ for 2 to 8 hours, 1 to 4 Extract and cool once and perform a first step of filtration with a stainless steel strainer of 0.125mm (80mesh) size. The second step of concentrating the filtrate obtained in the first step under reduced pressure to 15brix at 50 ~ 100 ℃. A third step of lyophilizing the extract obtained in the second step and producing a powdery extract is carried out.

In particular, the red cabbage powder extract obtained by the above method contains 0.1 to 20% by weight of anthocyanin.

2) Next, the method of preparing a composition by extracting the apple peel is as follows.

The apple peel is extracted with 4 to 10 times of 0 to 100% ethanol aqueous solution at 30 to 100 ° C for 2 to 8 hours, 1 to 4 times, cooled, and filtered using a 0.125 mm (80 mesh) stainless steel strainer. Perform step 1.

The second step of concentrating the filtrate obtained in the first step under reduced pressure to 20brix at 50 ~ 100 ℃. A third step of lyophilizing the extract obtained in the second step and producing a powdery extract is carried out.

The apple peel powder extract obtained by the above method contains, in particular, quercetin in an amount of 0.2 to 10% by weight.

3) Next, the method of preparing the composition by extracting turmeric is as follows.

Wash the turmeric twice with purified water and then grind for 5 to 10 minutes at 500 to 700rpm using a food grinder and then 4 to 10 times water or 0 to 100% aqueous ethanol solution at 60 to 100 ℃ for 2 to 8 hours, 1 Extract 4 times, and after cooling, perform the first step of filtering with a stainless steel sieve of 0.074mm (200 mesh).

The second step of concentrating the filtrate obtained in the first step under reduced pressure to 15brix at 30 ~ 80 ℃. The extract obtained in the second step is lyophilized and a third step of preparing a powder extract is performed.

In particular, the turmeric powder extract obtained by the above method usually contains 0.2 to 30% by weight of curcumin.

The red cabbage extract, apple bark extract and turmeric extract are mixed at a weight ratio of 1: 0.2 to 1 through the process of 1) to 3) above and contain 0.1 to 30% by weight of anthocyanin.

For reference, the main production area of red cabbage used in the present invention was Jeju Island, which was harvested in May-July, and the main production area of apple peel was Gyeongsangbuk-do, and the mixed species and Manjong species were mixed. Harvest was used in mid-December. However, the scope of the present invention is not limited because of the origin or variety of each plant.

The plant extract may be used as such, but food-acceptable moisture absorbents, excipients, forming agents, diluents, carriers, and the like, for preparing powders, tablets, granules, or gels may be used. Can be mixed together.

In addition, the present invention has a combination selected from the group consisting of red cabbage extract and apple peel extract and turmeric extract obtained by the above method, and comprises a combination selected from the group consisting of vitamin group and quercetin as an additional ingredient It provides a food containing other food ingredients in addition to the functional composition for inflammatory diseases, pain diseases and arthritis.

The food includes beverages, special nutritional supplements, health supplements, functional foods and other foods.

Meanwhile, the form of the food includes powder, granule, tablet, capsule, liquid or beverage form.

In addition, the present invention, the food additives in the food, such as fungicides, spices, seasonings, various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, such as colorants and neutralizing agents (cheese, chocolate, etc.) ), Pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, or the like as essential raw materials for food materials It provides a method of using a food additive characterized in that. At this time, the food additive may be added to the food by dipping, spraying or mixing the food, the ratio of these additives is not so important but selected from the range of 0 to about 20% by weight per 100% by weight of the functional composition of the present invention Is common.

It may also be added to foods or beverages for the purpose of preventing and improving symptoms of inflammatory diseases, painful diseases and arthritis. In this case, the amount of the composition in the food or beverage is generally 10 to 80% by weight, preferably 20 to 70% by weight of the total food weight in the case of the health functional food composition of the present invention, It can be added in the ratio of 0.1-30 g, Preferably it is 1-30 g based on 100 ml.

Hereinafter, the present invention will be described in detail with reference to the following examples, but the scope of the present invention is not limited by the following examples.

Example  1: Preparation of Mixed Extract

Red cabbage extract, apple peel extract and turmeric extract extracted by the above method were mixed using a powder mixer in a 5: 2: 2 weight ratio.

Experimental Example  One : DPPH Radical Scatters  Experiment

DPPH (2,2-Diphenyl-1-picrylhydrazyl, Sigma, MO, USA), a very stable free radical, was used to determine the electron donating ability of each sample by using a hydrogen proton-radical scavenger that discolors purple to yellow diphenylpicrylhydrazine. It was. The results are as shown in FIG. 1 and Table 1.

1 is a graph showing the SC ₅₀ value in ppm concentration of the mixed extract prepared by the present invention and the comparative substance eliminates DPPH radicals by 50%.

Experimental Method

After dissolving each sample by concentration, 75 μl of each sample is mixed with 750 μl of DPPH (0.2 mM in ethanol) and 675 μl of tertiary purified water. After reacting for 30 minutes at room temperature, 200 μl of the reaction solution is transferred to a 96 well plate, and then absorbance is measured at a wavelength of 520 nm with an ELISA reader device (Molecular Devices, CA, USA). The DPPH radical scavenging activity (%) was calculated from the formula (1-absorbance of sample 1-added sample / absorbance of sample-free sample) X 100. SC ₅₀ The value represents the minimum concentration, in ppm, required to eliminate 50% of the generated radicals.

TABLE 1

As shown in Table 1 and Figure 1, the scavenging ability of DPPH radicals for Example 1 prepared by the present invention is the case of using each extract except the apple peel extract or when using the SK tablets It can be seen that better. Vitamin C was used as a positive control.

Experimental Example  2 : SOD Similar activity  Experiment

The scavenging activity of oxygen radicals liberated from xanthine by xanthine oxidase was compared to determine the scavenging activity of the superoxide radical of each plant extract composition. As shown in FIG. 2 and <Table 2> which were attached.

Figure 2 is a graph showing the SC ₅₀ value in ppm concentration showing the SOD-like activity of the mixed extract prepared by the present invention.

Experimental Method

Coloring reagent consisting of xanthine solution (0.4mM) and NBT solution (0.24mM, nitro blue tetrazolium) in 500µl of xanthine oxidase solution (0.045U / ml) dissolved in buffer solution (0.1M phosphate buffer, pH 8.0) at 37 ℃ Add 500 μl, add 50 μl of sample solution dissolved at each concentration, and react for 20 minutes. The reaction was stopped by the addition of 950 μl of SDS (70 mM) solution. The degree of discoloration due to oxidation of cytochrome-c at 560 nm was stopped. Was measured.

<Table 2>

As shown in Table 2 and Figure 2, the SOD-like activity for Example 1 prepared by the present invention than when using each extract except the apple peel extract or using the joining tablet of SK Pharmaceuticals It is excellent. Vitamin C was used as a positive control.

Experimental Example  3: H ₂ O ₂ Scatters  Experiment

In general, the Fenton reaction is used to measure the antioxidant activity of converting hydrogen peroxide, a product generated after the decomposition of superoxide dismutase (SOD) into safer water and oxygen. The hydroxyl radical was measured by the degree of absorption of MDA (malondialdehyde) generated by oxidizing 2-deoxyribose at 520nm and the results are shown in Figure 3 and Table 3.

Figure 3 is a graph showing the mixed extract prepared by the present invention and the comparative substance SC ₅₀ value of 50% scavenging H ₂ O ₂ in a concentration of ppm.

Experimental Method

To the solution of 150 μl of 2-deoxyribose (50mM) solution in 100 μl of FeSO _{4 · 7} H ₂ O (50mM) -EDTA (50mM) solution, add 150 μl of the sample dissolved in each concentration, and then add buffer solution (0.2 300 ml of M sodium phosphate buffer, pH 7.0) and 10 ml of H ₂ O ₂ (50 mM) and 200 µl of distilled water were reacted at 37 ° C. for 4 hours, and 1% 2 dissolved in 1000 µl of 2.8% trichloroacetic acid and NaOH (50 mM) solution. 1000 μl of -thiobarbituric acid was added, heated at 100 ° C. for 15 minutes, rapidly cooled, and the absorbance was measured at 530 nm using a spectrophotometer. Hydroxy radical scavenging activity (%) was calculated by the formula (1-absorbance of sample added / absorbance of sample free) X 100.

<Table 3>

As shown in Table 3 and Figure 3, H ₂ O ₂ for Example 1 prepared by the present invention It can be seen that the scavenging ability is much superior to the case of using red cabbage extract, apple peel extract and turmeric extract, respectively, or using the joining tablet of SK Pharmaceuticals. Vitamin C was used as a positive control.

Experimental Example  4 : LPS  Judo NO  Production Suppression Experiment

The effect of inhibiting the production of NO (nitric oxide), which is an inflammation-causing substance, was measured by Griess reaction of nitric oxide concentration in the culture medium produced by treating LPS (lipopolysaccharide) on mouse macrophages. Is as shown in FIG. 4 and <Table 4>.

Figure 4 is a graph showing that the mixed extract prepared by the present invention and the comparative material inhibits the production of nitric oxide induced in LPS in mouse macrophages.

Experimental Method

NO concentration was measured by Griess reaction of nitric oxide concentration in the culture. First, 1 × 10 ⁴ cells / well of mouse macrophage-derived RAW 264.7 cells were dispensed into 96 well culture plates using DMEM medium containing FBS and antibiotics, followed by 37 ° C., 5% CO ₂ incubator. Incubated for 24 hours at. After 24 hours, the medium used for the previous culture was removed and fresh DMEM medium containing no FBS and antibiotics was dispensed and the plant composition prepared in Example 1 was treated by concentration (mg / mL). After 1 hour, 1 mg / mL LPS was treated and incubated for 24 hours. NO produced during the cultivation was measured by the concentration of total NO ₂ ⁻ present in the cell culture solution using the Griess reagent, which was mixed with 50 µl of the cell culture supernatant and 50 µl of the Griess reagent for 10 minutes in a 96 well plate. After the reaction, the absorbance was measured at 540 nm using an ELISA reader. Nitric oxide scavenging ability (%) was expressed using the formula ((absorbance of the reaction group without the 1-sample / absorbance of the control group without the sample) × 100.

TABLE 4

As shown in Table 4 and Figure 4, the use of three plant extracts prepared by the present invention was found to significantly inhibit the production of NO, one of LPS-induced inflammatory substances in mouse macrophages. In particular, when Example 1 was treated, NO production was found to be excellent.

Experimental Example  5: COX -2 inhibition experiment

In order to determine the efficacy of each sample to inhibit the activity of the proinflammatory enzyme COX-2 (cyclooxygenase-2), it is used to induce the discoloration by inducing the oxidation of hematin using arachidonic acid, which is the substrate of the enzyme. The results are as shown in FIGS. 5 to 6 and <Table 5>.

Figure 5 is a graph showing the IC ₅₀ value in ppm concentration of the mixed extract prepared by the invention and the comparative substance inhibits 50% of COX-2 enzyme.

Experimental Method

For COX-2 inhibitory effect, put 40 units of 30 units / ml of COX-2 per well in a 96 well plate, 90 μl of 100 mM Tris-HCl buffer (pH 8.0), 20 μl of 30 μM EDTA, 20 μl of 150 μM hematin and 20 μl of sample were mixed for 5 minutes at 25 ° C., 5 μl of 5 mM TMPD and 5 μl of 20 mM arachidonic acid were reacted at 25 ° C. for 5 minutes, and then absorbance was measured at 590 nm using an ELISA reader device. It was. The inhibition rate (%) of COX-2 was expressed as 50% inhibitory concentration (IC ₅₀ ) using the following equation. COX-2 inhibition rate (%) = [(absorbance of the control group-absorbance of the experimental group) / absorbance of the control group] × 100

<Table 5>

As shown in Table 5 and Figure 5, when using the plant extract composition prepared according to the present invention it was found that the activity of COX-2, an inflammation-related enzyme is significantly inhibited, especially in Example 1 In red cabbage extract, apple peel extract and turmeric extract, respectively, COX-2 showed much better activity inhibition.

Experimental Example  6: MIA ( Mono sodium iodoacetate Induced Degenerative Arthritis Animal Model

Intra-articular injection of MIA into SD rats inhibits metabolism of chondrocytes, causing cartilage, ligament, and tendon damage to osteoarthritis Was used to generate the principle. The results are as shown in Table 6.

Experimental Method

50 μl of MIA (40mg / ml) was injected into the joint cavity of SD rats weighing 300g or more using a 27-gauge needle. Each sample or saline or Ibuprofen Was administered orally at daily doses for 14 days.

<Table 6>

Experimental Example  7: Lipooxygenase ( LOs , Lipoxygenases Deactivation experiment

Lipogenasease using Lipoxygenase Inhibitor Screening Assay Kit (Cayman Chemical Com, MI, USA) to measure the efficacy of each sample to inhibit lipooxygenase, an inflammation and pain-causing enzyme, in a degenerative arthritis animal model using MIA. The principle of measuring the amount of hydrogen peroxide as a result of the lipoxygenation reaction was used, and the results are shown in FIG. 6 and <Table 7>.

7 is a graph showing the ratio of the mixed extract prepared by the present invention and the comparative substance inhibits the Lipoxygenase enzyme activity in the blood of the chronic inflammation animal model by MIA compared to the untreated group of the sample.

Experimental Method

To 10 μl of the serum sample of the degenerative arthritis animal model, 90 ml of 5-LO (220 units / ml) and 10 ml of 1 mM linoleic acid were added and reacted at room temperature for 5 minutes, and then 100 ml of chromogen was added for 5 minutes at room temperature. The absorbance was measured at 490 nm with ELISA reader. The results were expressed as 50% inhibitory concentration (IC ₅₀ ) and 5-LO inhibition (%) was expressed as IC ₅₀ value using [(absorbance of control-absorbance of control group) / absorbance of control group] × 100 equation. .

<Table 7>

As shown in Table 7 and Figure 6, it was found that the activity of the lipooxygenase, an inflammation-related enzyme, was inhibited when the plant extract composition prepared according to the present invention was used. Showed very good inhibition of the lipoxygenase activity. In addition, Ibuprofen, which was used as a positive control, did not appear to affect the inhibition of lipooxygenase activity.

Experimental Example  8 : TNF -α production inhibition experiment

To evaluate the efficacy of each sample in reducing the expression of TNF-α, an inflammation-inducing factor, the TNF-α sandwich ELISA kit (eBiosience, Vienna, Austria) was used in animal models of carrageenan-induced edema and MIA-induced degenerative arthritis. ), And the results are shown in FIGS. 7 to 8 and <Table 8 to 9>.

7 is a graph showing the amount of production of TNF-α in the serum of the acute inflammatory animal model of carrageenan by the mixed extract prepared by the present invention,

8 is a graph showing the amount of production of TNF-α in the serum of an animal model of chronic inflammation caused by MIA of the mixed extract prepared by the present invention.

Experimental Method

Each sample was sacrificed by carrageenan-induced edema animal model and MIA-induced degenerative arthritis animal model, which were orally administered once daily at the period and dose shown in Tables 8 to 9, and blood and plasma were separated. The concentration of -α was measured by the method suggested by the manufacturer of the ELISA kit. First, TNF-α in plasma was bound to anti-rat TNF-α antibody coated on a 96 well plate, and each well was washed five times, followed by biotin-conjugated anti-rat TNF-α. After the reaction of the antibody, each well was washed five times, treated with streptavidin-HRP, and finally colored with tetramethyl-benzidine, and the absorbance was measured at 450 nm using an ELISA reader. The amount of TNF- [alpha] was calculated in an arithmetic proportion using the standard curve of the standard recombinant TNF- [alpha] contained in the ELISA kit.

<Table 8>

<Table 9>

As shown in Tables 8 to 9 and 7 to 8, TNF is an inflammation-related factor when a plant extract composition prepared by the present invention is used in an acute inflammatory animal model by carrageenan and a chronic inflammatory animal model by MIA. In vivo production of -α was suppressed, and in particular, in the case of Example 1, the inhibition of in vivo production of TNF-α was much better than that of red cabbage extract, apple peel extract, or turmeric extract, respectively. appear. In addition, Ibuprofen used as a positive control was found to have only a slight effect on the in vivo production of TNF-α.

Experimental Example  9: IL -1β production inhibition experiment

To evaluate the efficacy of each sample to reduce the in vivo expression of IL-1β, an inflammation-inducing factor, the IL-1β sandwich ELISA kit (eBiosience, Vienna, Austria) was used in animal models of carrageenan-induced edema and MIA-induced degenerative arthritis. ), And the results are shown in FIGS. 9 to 10 and <Table 10 to 11>.

Figure 9 is a graph showing the amount of IL-1β production in the serum of the acute inflammation animal model of the mixed extract prepared by the present invention and carrageenan,

10 is a graph showing the amount of IL-1β production in the serum of the animal model of chronic inflammation caused by MIA of the mixed extract prepared by the present invention.

 Experimental Method

IL-1β present in plasma after sacrificing blood collection and plasma separation after sacrificing carrageenan-induced edema animal model and MIA-induced degenerative arthritis animal model orally administered once daily with the periods and doses shown in Tables 10-11. The concentration of was measured by the method suggested by the manufacturer of the ELISA kit. First, IL-1β in plasma is bound to anti-rat IL-1β antibody coated on a 96 well plate, and each well is washed five times, followed by biotin-conjugated anti-rat IL-1β. After the reaction of the antibody, each well was washed five times, treated with streptavidin-HRP, and finally colored with tetramethyl-benzidine, and the absorbance was measured at 450 nm using an ELISA reader. The amount of IL-1β was calculated in an arithmetic proportion using the standard curve of the standard recombinant IL-1β contained in the ELISA kit.

<Table 10>

<Table 11>

As shown in Tables 10 to 11 and 9 to 10, IL-related inflammation factor when the plant extract composition prepared by the present invention was used in an acute-inflammatory animal model by carrageenan and a chronic-inflammatory animal model by MIA In vivo production of -1β was suppressed, and especially in Example 1, the inhibition of in vivo production of IL-1β was much superior to that of red cabbage extract, apple peel extract, or turmeric extract, respectively. appear. In addition, Ibuprofen, used as a positive control, was found to have only a minor effect on the in vivo production of IL-1β.

Experimental Example  10: IL -6 generation inhibition experiment

IL-6 sandwich ELISA kit (eBiosience, Vienna, Austria) for carrageenan-induced edema and MIA-induced degenerative arthritis animal models ), And the results are shown in FIGS. 11 to 12 and Tables 12 to 13.

11 is a graph showing the production amount of IL-6 in the serum of the acute inflammatory animal model of carrageenan by the mixed extract prepared by the present invention,

FIG. 12 is a graph showing the amount of IL-6 production in the serum of an animal model of chronic inflammation caused by MIA of the mixed extract prepared by the present invention.

Experimental Method

IL was present in plasma after sacrificing blood collection and plasma separation at the expense of carrageenan-induced edema animal model and MIA-induced degenerative arthritis animal model, which were orally administered once daily at the periods and doses shown in Tables 12-13. The concentration of -6 was measured by the method suggested by the manufacturer of the ELISA kit.

First, IL-6 in plasma was bound to anti-rat IL-6 antibody coated on a 96 well plate, and each well was washed five times, followed by biotin-conjugated anti-rat IL-6. After reacting the antibody, streptavidin-HRP was treated, and finally, tetramethyl-benzidine was developed and the absorbance was measured at 450 nm using an ELISA reader. The amount of IL-6 was calculated arithmetic proportionally using the standard reaction curve of the standard recombinant IL-6 included in the ELISA kit.

<Table 12>

<Table 13>

As shown in Tables 12 to 13 and 11 to 12, IL is an inflammation-related factor when a plant extract composition prepared by the present invention is used in an acute inflammatory animal model by carrageenan and a chronic inflammatory animal model by MIA. In vivo production of -6 was inhibited, especially in Example 1 treated with red cabbage extract or turmeric extract, except that the inhibition of in vivo production of IL-6 treated apple peel extract It was found to be much better than one case. In addition, Ibuprofen, used as a positive control, was found to have only a minor effect on the in vivo production of IL-6.

Experimental Example  11: Prostaglandin ( PGs , Prostaglandins ) Generation inhibition experiment

To evaluate the efficacy of each sample to inhibit the production of prostaglandins, inflammation and pain-causing factors, in inflammatory cell models using LPS and degenerative arthritis animal models using MIA, Ptostaglandin Screening EIa Kit (Cayman Chemical Co., MI, USA) using the principle of prostaglandin and tracer (prostaglandin artificially coupled to acetylcholinesterase) competitively binds to the anti-PGs-antibody, the results are shown in Figures 13-14 and Tables 14-15 As shown.

FIG. 13 is a graph showing that the mixed extract prepared by the present invention and the comparative substance inhibit the production of prostaglandin induced by LPS in mouse macrophages,

14 is a graph showing that the mixed extract prepared by the present invention and the comparative substance inhibit the production of prostaglandin in the serum of animal models of chronic inflammation by MIA.

Experimental Method

RAW 264.7 cells derived from mouse macrophages were cultured in 96 well culture plates using FMEM and DMEM medium containing antibiotics, and then dispensed at 1 × 10 ⁴ cells / well at 37 ° C. and 5% CO ₂ incubator. Incubated for hours. After 24 hours, the medium used for the previous culture was removed and a new DMEM medium containing no FBS and antibiotics was dispensed and each sample was treated by concentration. After 1 hour, 1 mg / mL LPS was treated and incubated for 24 hours. Prostaglandin AChE Tracer and antiserum were added to a 96-well plate coated with anti-PGs antibody, and 50 μl of standard prostaglandin or each sample-treated medium was added and reacted at room temperature for 18 hours. Wash the wells five times, add 200 μl of Ellman's Reagent, and develop for 70 minutes. Measure the absorbance at a wavelength of 420 nm using an ELISA reader. The amount of prostaglandin was arithmetic proportionally calculated using the standard curve included in the ELISA kit.

TABLE 14

<Table 15>

As shown in Tables 14 to 15 and FIGS. 13 to 14, when the plant extract composition prepared according to the present invention is used, the production of prostaglandin, which is one of LPS-induced inflammatory substances, is inhibited in mouse macrophages. In particular, in the case of treating Example 1, it was shown that the prostaglandin production inhibition rate was superior to that of the red cabbage extract, the apple peel extract, or the turmeric extract, respectively.

Experimental Example  12: Type  I Collagenase  Deactivation experiment

PZ-peptide (4-phenylazobenzyloxycarbonyl-Pro-Lue-Gly-), a substrate of recombinant collagenase enzyme (Sigma, MO, USA), was used to examine the effects of each sample on the activity of Type I collagenase, which has a major adverse effect on articular cartilage health. Pro-D-Arg, Fluka Chemie GmbH, Buchs, Switzerland) was measured using the discoloration phenomenon when decomposing the results are shown in Figure 15 and Table 16.

Figure 15 is a graph showing the IC ₅₀ value in ppm concentration, the concentration of the mixed extract prepared by the present invention and the comparative substance inhibits Type I collagenase 50%.

Experimental Method

After dissolving collagenase enzyme at the concentration of 0.2mg / ml and diluting PZ-peptide in the buffer solution at the concentration of 0.3mg / ml, vortex by mixing 100μl of sample according to concentration, 150μg of collagenase and 250μl of PZ-peptide After reaction at 37 ° C. for 30 minutes, 500 μl of citric acid (25mM) was added, ethyl acetate 1.5ml was added, followed by reaction at room temperature for 10 minutes, and the absorbance was measured at 320 nm using a quartz cuvette.

<Table 16>

As shown in Table 16 and Figure 15, it can be seen that Type I collagenase inhibitory activity for Example 1 prepared by the present invention is superior to the case of using each extract or the joining tablet of SK Pharmaceuticals. .

Experimental Example  13: Carrageenan ( carrageenan Edema for induced acute inflammation edema Suppression experiment

In the carrageenan-induced acute inflammatory response by the plant composition prepared in Example 1, the degree of inhibition of foot edema was observed for each time period compared to the control group, and the results are shown in Table 17 and FIGS. 16 to 17.

FIG. 16 is a diagram illustrating the edema-inhibiting effect of carrageenan-induced acute inflammation animal model experimented with the mixed extract prepared in accordance with the present invention by the method of <Table 17> according to time zones.

FIG. 17 is a photograph showing the effect of inhibiting the edema of an animal model of acute inflammation caused by carrageenan experimented with the mixed extract prepared in accordance with the present invention by the method of <Table 18> after 6 hours of edema.

Experimental Method

The number of populations in each group was set to 10 in SD (Spraque-Dqwley) rats and administered orally at the concentrations shown in the following <Table 19>, and 100 ml of 1% carrageenan-dissolved saline was injected into the left plantar foot to prevent foot edema. After induction, the degree of foot edema was measured at each hour for 6 hours using a Digimatic micrometer (Mitutoyo Corporation, Japan).

<Table 17>

<Table 18>

As shown in FIGS. 16 to 17 and Tables 17 to 18, it can be seen that the edema of the acute inflammatory animal model is suppressed in proportion to the concentration of Example 1 prepared by the present invention. In addition, it is slightly less inhibited than ibuprofen, a synthetic drug, but it can be seen that edema is more suppressed than when each extract is used or when the SK tablets are used as a joining tablet.

Experimental Example  14: inflammation-related enzyme (5- LO , iNOS , COX -2) expression suppression experiment

Immunocytochemistry was performed for each sample to measure the efficacy of inhibiting the expression of 5-LO, iNOS, and COX-2 in vivo in inflammatory cell model using LPS. As shown in 18-20.

FIG. 18 is a diagram showing the number of cells expressing the 5-LO enzyme reduced in the inflammatory cell model of LPS-induced mixed extract prepared by the present invention by 1000-fold magnification using an optical microscope,

FIG. 19 is a diagram showing the number of cells expressing iNOS enzyme in the inflammatory cell model of LPS-induced mixed extract prepared by the present invention at 1000-fold magnification using an optical microscope.

FIG. 20 is a graph showing that the number of cells expressing COX-2 enzyme in the inflammatory cell model of LPS-induced mixed extract prepared by the present invention is increased by 1000 times using an optical microscope.

Experimental Method

24 well culture plates containing 12 mm circular optical microscope glass coverslides specially coated to allow cell proliferation after attachment of mouse macrophage-derived RAW 264.7 cells using FMEM and DMEM medium containing antibiotics Culture plate) was incubated for 24 hours in 37 ℃, 5% CO ₂ incubator after dispensing 2 × 10 ⁴ cells / well. After 24 hours, the medium used for the previous culture was removed and a new DMEM medium containing no FBS and antibiotics was dispensed and each sample was treated by concentration. After 1 hour, 1 mg / mL LPS was treated and incubated for 24 hours. After 24 hours, cells were fixed with acetone, treated with 0.3% H ₂ O ₂ for 10 minutes for inactivation of endogenous peroxidase, washed three times with PBS, and treated with goat serum to prevent nonspecific binding of primary antibodies. The primary antibody is reacted at 4 ° C. for 24 hours. After 24 hours, washed three times with PBS, amplified the signal using VECTASTAIN elite ABC kit (VECTOR laboratories, CA, USA) and stained with DAB (Sigma, MO, USA). The cell-covered cover slides were separated from the 24-well culture plate using forceps, dehydrated, and attached to slide glass using Canada balsam (Sigma, MO, USA). Diluted concentrations of primary antibodies, 5-LO (Santa Cruz Biotechnology, CA, USA), iNOS (Santa Cruz Biotechnology, CA, USA), and COX-2 (Cell Signaling Technology, MA, USA) were set to 200: 1. The slide thus prepared was observed with an optical microscope at a magnification of 1000 times.

<Table 19>

As shown in Table 19 and FIGS. 18 to 20, the expression in the macrophages of 5-LO, iNOS, and COX-2, which are inflammation-related enzymes, is proportional to the concentration of Example 1 prepared by the present invention. It can be seen that it is inhibited. Ibuprofen also slightly inhibited iNOS expression but did not affect the expression of 5-LO and COX-2.

Experimental Example  15: Chronic inflammation of animal models Knee tube insulation goal  Histomorphological changes (H & E staining)

As in Experimental Example 6, MIA was injected intra-articular injection into SD rats to prepare a chronic inflammation animal model, sacrifice the animal, separate the knee joint, and observe changes in the knee joint. The results are as shown in Table 19 and FIG.

FIG. 21 shows that the mixed extract prepared by the present invention and the comparative substance were changed 400 times by using an optical microscope to change the thickness of the knee cartilage in the animal model of chronic inflammation caused by MIA.

Experimental Method

Inject 50 ml of MIA (40mg / ml) using a 1ml syringe with a 27 gauge needle into the joint cavity of SD rats weighing more than 300g. Each sample, 10 saline, or Ibuprofen It was administered orally at the dosage as indicated in Table 6 daily for 14 days. After 14 days, the animals were sacrificed, the knee joints were separated, the tissues were fixed, calcium was removed using 4% formic acid, dehydration and transparent processes using xylene, and paraffin was embedded. It was cut to 6 mu m using a rotary cutting machine and attached to a gelatin-coated slide glass. The paraffin between the tissues xylene-derived was removed again and hydrated with staged ethanol and purified water, and the nucleus and cytoplasm were stained with hematoxylin and Eosin. This was again dehydrated using staged ethanol and xylene and sealed by attaching to a cover glass using Canada balsam (Sigma, MO, USA). The slide thus produced was magnified 400 times using an optical microscope.

<Table 20>

As shown in Table 20 and FIG. 21, it can be seen that, according to Example 1 prepared in the present invention, damage of the chondrocyte layer caused by MIA in the knee joint of the chronic inflammation animal model was reduced in proportion to the concentration of Example 1. have.

Preparation Example : Although a manufacturing example of a food or health functional food containing the extract of the present invention will be described, the preparation example is for explaining the present invention in more detail, and the present invention is not limited to the preparation example and various other Subject to change.

Manufacturing example  One: Sanje  Produce

300 mg of the functional composition of Example 1, 100 mg of lactose, 10 mg of talc

The above components are mixed and filled in airtight bags to prepare powders.

Manufacturing example  2: Preparation of tablets

50 mg of the functional composition of Example 1, 100 mg of corn starch, 100 mg of lactose, 2 mg of magnesium stearate

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

Manufacturing example  3: Manufacture of capsule

50 mg of the functional composition of Example 1, 100 mg of corn starch, 100 mg of lactose, 2 mg of magnesium stearate

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Manufacturing example  4: Liquid  Produce

100 mg of the functional composition of Example 1, 10 g of isomerized sugar, 5 g of mannitol,

According to the conventional method of preparing a liquid solution, each component is added and dissolved in purified water, lemon flavor is added to the appropriate amount, the above components are mixed, purified water is added, and the whole is adjusted to 100 ml by adding purified water to a brown bottle. Fill and sterilize to prepare a liquid.

Manufacturing example  5: Manufacture of dietary supplements

1000 mg of the functional composition of Example 1, a proper amount of vitamin A, 70 g of vitamin A acetate, 1.0 mg of vitamin E, 13 mg of vitamin B10, 15 mg of vitamin B20, 0.5 mg of vitamin B6, 0.2 g of vitamin B12, 10 mg of niacinamide, 1.7 mg of nicotinic acid amide, 50 g of folic acid, 0.5 mg of calcium pantothenate, an appropriate amount of inorganic mixture, 1.75 mg of ferrous sulfate, 0.82 mg of zinc oxide, 25.3 mg of magnesium carbonate, 15 mg of potassium phosphate monobasic Mg, potassium citrate 90 mg, calcium carbonate 100 mg, magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

Manufacturing example  6: manufacture of health drinks

1,000 mg of the functional composition of Example 1, 1000 mg of citric acid, 100 g of oligosaccharide, 2 g of a plum concentrate, 1 g of taurine and purified water were added to adjust the total amount to 900 ml.

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was heated at 85 DEG C for about 1 hour with stirring, and the solution thus prepared was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, &Lt; / RTI &gt;

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Claims (12)

delete Red cabbage extract, apple peel extract and turmeric extract 1: (0.1 ~ 5): functional food composition having an anti-inflammatory, analgesic effect, characterized in that they are mixed in a weight ratio of (0.1 to 50), respectively. delete delete delete delete delete delete The method according to claim 2,
The functional food composition is a functional food composition having an anti-inflammatory, analgesic effect is administered in any one of capsules, pills, granules, powder, candy, gum, tablets, beverages, syrup form.
delete delete delete

Images