KR20170045770A - A compostion comprising Osmanthus matsumuranus extract having anti-oxidation or anti-inflammation activity - Google Patents
A compostion comprising Osmanthus matsumuranus extract having anti-oxidation or anti-inflammation activity Download PDFInfo
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- KR20170045770A KR20170045770A KR1020150145221A KR20150145221A KR20170045770A KR 20170045770 A KR20170045770 A KR 20170045770A KR 1020150145221 A KR1020150145221 A KR 1020150145221A KR 20150145221 A KR20150145221 A KR 20150145221A KR 20170045770 A KR20170045770 A KR 20170045770A
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- extract
- osmanthus
- antioxidant
- matsumuranus
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/92—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof
- A61K8/922—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof of vegetable origin
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- A—HUMAN NECESSITIES
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- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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- A23V2200/00—Function of food ingredients
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
The present invention relates to a process for the preparation of a medicament comprising Osmanthus matsumuranu s) extract as an active ingredient, and to a pharmaceutical composition, a cosmetic composition and a pharmaceutical composition for antioxidant or antiinflammation. Since the composition comprising the Osmanthus matsumuranus extract of the present invention exhibits antioxidant or anti-inflammatory activity, it can be used for prevention, improvement or treatment of diseases caused by oxidative and inflammatory action, or for use in foods, cosmetics and pharmaceuticals .
Description
The present invention relates to a process for the preparation of a medicament comprising Osmanthus The present invention relates to a composition for antioxidant or antiinflammation comprising an extract of Osmanthus matsumuranus as an active ingredient, a cosmetic composition and a pharmaceutical composition.
Although the human body has a balance between the oxidation promoting substance and the oxidation inhibiting substance, due to various factors, if such a balance state is lost and it is inclined to promote oxidation, oxidative stress is induced in the living body. Reactive oxygen species (ROS), which is a direct cause of oxidative stress, are chemically unstable and highly reactive, and can easily react with various biomaterials such as DNA, protein, lipid and carbohydrate. Attack, causing irreversible damage to cells and tissues, or mutation and cytotoxicity.
The deleterious ROS as described above is generally caused by superoxide dismutase, catalase, glutathione peroxidase, glutamine reductase, vitamin C, vitamin E, uric acid, bilirubin (bilirubin) is removed by the action of antioxidant system. However, when oxidative stress occurs, unremoved ROS causes various pathological changes, leading to cellular damage and pathological disease. In fact, it has been confirmed that oxidative stress is increased in many clinical diseases. Various studies on new antioxidants have been made to reduce such oxidative stress.
The macrophages respond to pathogens in the body and induce inflammatory factors such as tumor necrotic factor-α, TNF-α, interleukin-6 (IL-6) and interleukin-1β (IL-1β) (NO) and prostaglandin (prostaglandin E2, PGE2) were synthesized by synthesizing inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) . Physiologically, NO plays a variety of roles such as removing bacteria and tumors, regulating blood pressure, or mediating neurotransmission. However, when the inflammatory reaction occurs, the expression of iNOS is increased in related cells, and a large amount of NO is produced. Excessively produced NO causes tissue damage, gene mutation, nerve damage, etc., And promote the inflammatory response.
On the other hand, Korean Patent Publication No. 10-2015-0075757 entitled "Anticancer composition containing Osmanthus matsumuranus extract" is known about the cancer cell killing effect of Osmanthus matsumuranus extract. However, antioxidant activity of Osmanthus matsumuranus The anti-inflammatory effect was not known.
Under these circumstances, the present inventors have made intensive studies to find natural products having antioxidant or anti-inflammatory effects and less side effects. As a result, it has been found that the extract of Osmanthus matsumuranus, one of the native plants derived from China, The present inventors have confirmed that there is a strong antioxidant and antiinflammatory effect and completed the present invention.
One object of the present invention is to provide a method and apparatus for detecting the presence of < RTI ID = 0.0 > Osmanthus & matsumuranu s) extract as an active ingredient.
Another object of the present invention is to provide an antioxidant or anti-inflammatory cosmetic composition comprising the Osmanthus matsumuranus extract as an active ingredient.
Another object of the present invention is to provide a pharmaceutical composition for antioxidant or antiinflammation comprising the Osmanthus matsumuranus extract as an active ingredient.
In one aspect of the present invention, there is provided an antioxidant or anti-inflammatory food composition containing Osmanthus matsumuranus extract as an active ingredient.
The term " Osmanthus " matsumuranu s) "belongs to the genus Oleaceae. No specific effects have been known yet, and in particular, no antioxidant and anti-inflammatory effects are known at all. Thus, the present inventors have found that the extract of Osmanthus matsumuranus And anti-inflammatory effect, and confirmed that it can be applied to foods or cosmetics, and moreover, to pharmaceuticals.
The term "extract " of the present invention means a preparation which is obtained by squeezing a herbal medicine with an appropriate leaching solution and concentrating the liquid by evaporating the leaching solution, and is not limited thereto. The extract is obtained by drying the extract, diluted solution, Dried products, controlled products thereof or purified products thereof. The extract of the present invention can be prepared by extracting with an extraction solvent or extracting with an extraction solvent and fractionating the extract with a fraction solvent. The extraction solvent may be, but is not limited to, water, an organic solvent or a mixed solvent thereof. The organic solvent may be an alcohol having 1 to 4 carbon atoms, a polar solvent such as ethyl acetate or acetone, hexane or dichloromethane Non-polar solvents or mixed solvents thereof may be used. Specifically, water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof may be used. In one embodiment of the present invention, an extract using 95% ethanol as the solvent was prepared.
The Osmanthus matsumuranus extract can be prepared by using common extraction, isolation and purification methods known in the art. Examples of the extraction method include, but are not limited to, hot-water extraction, hot water extraction, cold extraction, reflux cooling extraction, or ultrasonic extraction.
The term "antioxidant" of the present invention means an antioxidant action. The human body has a balance between prooxidant and antioxidant, but due to various factors, And leaning towards oxidative stimulation leads to oxidative stress leading to potential cellular damage and pathological disease. Reactive oxygen species (ROS), which is a direct cause of oxidative stress, are unstable and highly reactive. They react easily with various biological materials, attack the body's polymers, cause irreversible damage to cells and tissues, And cytotoxicity. Active nitrogen species (RNS) such as NO, HNO 2 , and ONOO - are produced in large quantities due to the immune response of macrophage neutrophils and other immune cells. Such active oxygen oxidizes and destroys cells in the body, thereby exposing them to various diseases. Therefore, when the Osmanthus matsumuranus extract of the present invention is contained in a food, the antioxidative effect can be attained, thereby contributing to health promotion.
In one embodiment of the present invention, it was confirmed that DPPH radical scavenging activity (Table 1) and active oxygen species scavenging ability (Fig. 2) of Osmanthus matsumuranus extract were analyzed to have strong antioxidant ability, And that the antioxidant activity was due to the induction of HO-1 expression through the Nrf-2 signaling pathway (FIG. 3). Thus, the antioxidant effect of the present invention may be achieved by removal of active oxygen species.
When the composition of the present invention is used as a food additive, the Osmanthus matsumuranus extract can be directly added or used in combination with other food or food ingredients, and can be suitably used according to ordinary methods. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment), and may further include a food-acceptable food-aid additive. Since the composition of the present invention contains an extract derived from a natural substance as an active ingredient, there is no problem in terms of stability, so there is no great limitation on the amount of the mixture.
The food composition of the present invention may include all foods having a conventional meaning, and may be mixed with terms known in the art such as functional foods, health functional foods, and the like.
The term "functional food" in the present invention means a food prepared and processed by using a raw material or ingredient having functionality useful to the human body according to Law No. 6727 on health functional foods, and " And function of the nutrient for the purpose of obtaining a beneficial effect in health use such as controlling the nutrient or physiological action.
The term "health functional food" of the present invention refers to a food prepared by processing a specific ingredient as a raw material for the purpose of health assisting or by extracting, concentrating, refining, mixing, or the like a specific ingredient contained in a food raw material, Refers to a food which is designed and processed so that the body control function such as bio-defense, regulation of biorhythm, prevention and recovery of disease and the like can be sufficiently exhibited to the living body by the above components. Recovery, and so on.
There is no limitation on the kind of food in which the composition of the present invention can be used. In addition, the composition comprising the Osmanthus matsumuranus extract of the present invention as an active ingredient can be prepared by admixing other suitable auxiliary ingredients that can be contained in foods and known additives, according to the selection of a person skilled in the art. Examples of foods that can be added include dairy products, such as meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen and other noodles, gums, ice cream, various soups, drinks, tea, Vitamin complex, and the like, and can be prepared by adding to the juice, tea, jelly, and juice prepared from the extract of the present invention as a main component.
Foods that can be applied to the present invention include, for example, special nutritional foods (such as crude oil, infant formula, infant formula, etc.), meat products, fish products, tofu, mackerel, noodles Food, seasoned foods (eg soy sauce, miso, kochujang, mixed potatoes), sauces, confectionery (eg snacks), dairy products (eg fermented milk, cheese), other processed foods, kimchi, pickled foods Etc.), beverages (such as fruit, vegetable beverages, beverages, fermented beverages, etc.), and natural seasonings (eg, ramen soup, etc.).
When the health functional food composition of the present invention is used in the form of a drink, it may contain various sweeteners, flavors, or natural carbohydrates as an additional ingredient such as ordinary beverages. In addition to the above, the health functional food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, , Alcohols, carbonating agents used in carbonated drinks, and the like. In addition, it may contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks.
In another aspect, the present invention provides an antioxidant or anti-inflammatory cosmetic composition containing Osmanthus matsumuranus extract as an active ingredient. The antioxidant and antiinflammatory effects of the Osmanthus matsumuranus extract are as described above. Since the extract of the present invention has antioxidative and antiinflammatory effects, it can be added to the cosmetic composition for the purpose of antioxidant or antiinflammation .
The cosmetic composition of the present invention can be produced in the form of a general emulsified formulation and a solubilized formulation. Examples of the emulsified formulations include nutritive lotions, creams, essences, and the like, and the solubilization formulations include softening longevity. Suitable formulations include, but are not limited to, for example, solutions, gels, solid or kneaded anhydrous products, emulsions obtained by dispersing the oil phase in water, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes) May be in the form of a follicle dispersant of the type, cream, skin, lotion, powder, ointment, spray, or conical stick. It may also be in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.
The cosmetic composition may further contain at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickening and gelling agents, softening agents, antioxidants, suspending agents, stabilizers, foaming agents, perfumes, surfactants, water, ionic or nonionic emulsifiers, Auxiliaries such as ionic sequestering agents, chelating agents, preservatives, vitamins, blocking agents, moisturizers, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredients conventionally used in cosmetic compositions ≪ / RTI >
In another aspect, the present invention provides a pharmaceutical composition for antioxidant or antiinflammation comprising an extract of Osmanthus matsumuranus as an active ingredient. The extract of Osmanthus matsumuranus is as described above. Since the extract of the present invention has an antioxidative and anti-inflammatory effect for removing active oxygen, it can be used as a pharmaceutical composition to treat diseases caused by active oxygen and inflammatory And can be included in medicines for preventing, improving and treating diseases.
The above-mentioned antioxidation is as described above, and thus can be included in a pharmaceutical composition for the purpose of antioxidation, and can have a preventive or therapeutic effect on a disease caused by active oxygen. The diseases caused by the active oxygen include, but are not limited to, degenerative neurological diseases including arteriosclerosis, Lou Gehrig's disease, Parkinson's disease, Alzheimer's disease, proximal axillary sclerosis and Huntington's disease, myocardial infarction, angina pectoris, coronary artery disease, Cardiovascular diseases including diseases, ischemic brain diseases including stroke, and various diseases such as aging, and may be specifically aging caused by active oxygen.
In addition, the anti-inflammatory activity is as described above, and thus can be included in a pharmaceutical composition for the prophylaxis or treatment of anti-inflammatory, i.e., inflammatory diseases. The inflammatory diseases include allergic diseases including allergic asthma, allergic rhinitis, allergic mucositis, urticaria, and anaphylax, although it is not limited thereto. Arthritis, atopic dermatitis, psoriasis, asthma, multiple sclerosis, ssRNA and dsRNA viral infection, sepsis, multiple chondritis, scleroderma, including systemic sclerosis, dermatomyositis and inclusion body myositis , Eczema, gout, periodontal disease, Behcet's syndrome, edema, vasculitis, Kawasaki disease, diabetic retinitis, autoimmune pancreatitis, vasculitis, glomerulonephritis, acute and chronic bronchitis, and influenza infection.
The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable" of the present invention means that it exhibits properties that are not toxic to the cells or humans exposed to the composition. Such carriers may be used without limitation as long as they are known in the art such as buffers, preservatives, wetting agents, solubilizers, isotonic agents, stabilizers, bases, excipients and lubricants.
The pharmaceutical composition of the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols or the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method . Furthermore, it can be used in the form of an external preparation for skin in the form of ointments, lotions, spray agents, patches, creams, powders, suspensions, gels or gels. Examples of carriers, excipients and diluents that can be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used.
Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like. These solid preparations may contain at least one excipient, such as starch, calcium carbonate, etc., in the Osmanthus matsumuranus extract, Sucrose, lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.
Meanwhile, the pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The term "administering" of the present invention means introducing a predetermined substance into an individual by an appropriate method, and the administration route of the composition can be administered through any conventional route so long as it can reach the target tissue. But are not limited to, intraperitoneal, intravenous, intramuscular, subcutaneous, intradermal, oral, topical, intranasal, intrathecal, rectal.
The term "individual" refers to all animals, including humans, including mice, mice, and livestock. Preferably, it may be a mammal, including a human.
The term "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and not causing side effects, and the effective dose level is determined by the patient's sex, Including, but not limited to, medicaments and other medical fields that are used in combination, or in combination with, or in combination with, a pharmaceutically acceptable carrier, excipient, Can be readily determined by those skilled in the art according to known factors. The administration may be such that the recommended dose is administered once a day or divided into several doses.
Since Osmanthus matsumuranus extract of the present invention is a natural material as a raw material, it can be used safely and usefully when it is used as a food composition, a cosmetic composition, or a pharmaceutical composition because its side effects are less than general synthetic compounds.
Since the composition comprising the Osmanthus matsumuranus extract of the present invention exhibits antioxidant or anti-inflammatory activity, it can be used for prevention, improvement or treatment of diseases caused by oxidative and inflammatory action, or for use in foods, cosmetics and pharmaceuticals .
Figure 1 shows the effect of Osmanthus matsumuranus extract on RAW 264.7 cell viability.
Fig. 2 shows the active oxygen species-removing activity of Osmanthus matsumuranus extract.
Figure 3 shows the expression control of antioxidant enzyme HO-1 and its parent transcription factor Nrf-2 protein by Osmanthus matsumuranus extract in RAW 264.7 cells.
Figure 4 shows the effect of Osmanthus matsumuranus extract on LPS-induced RAW 264.7 cell viability.
Figure 5 shows LPS-induced NO production regulation by Osmanthus matsumuranus extract in RAW 264.7 cells.
Hereinafter, the constitution and effects of the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.
Example One. Osmanthus Matsumuranus ( Osmanthus matsumuranus ) Preparation of extract
Osmanthus matsumuranus was extracted using a 95% ethanol solvent. Specifically, 95% ethanol was added to the sample, followed by sonication for 15 minutes and then for 2 hours. The procedure was repeated 10 times a day for 3 days at room temperature. After completion of extraction, the extract was filtered, concentrated, dried and stored in a cold room at 4 ° C until used in the experiment.
Example 2. RAW 264.7 Rats Macrophage culture
A major murine macrophage cell line, RAW 264.7, which is used as an experimental model for antiinflammatory activity, was purchased from the American Type Tissue Collection (ATCC), and 10% fetal bovine serum (FBS) and penicillin / streptomycin Were cultured in DMEM medium.
Example 3. Cytotoxicity analysis
To determine the effect of Osmanthus matsumuranus extract on RAW 264.7 cell viability, cytotoxicity tests were performed.
RAW 264.7 cells were plated at 3.0 × 10 5 cells per well in a 24-well tissue culture vessel and the cytotoxicity of Osmanthus matsumuranus extract was confirmed by WST analysis.
The water soluble tetrazolium (WST) assay is a representative method for measuring cell viability using water-soluble tetrazolium. After 24 hours of sample treatment, the cells were incubated at 37 ° C for 1 hour after replacing with WST medium. Absorbance was measured at 450 nm using a multi-plate reader, and the value was expressed as the average value of three repeated experiments.
As a result of examining the effect of the extract of Osmanthus matsumuranus on the cell viability of RAW 264.7, it was confirmed that it did not induce cytotoxicity up to the concentration of 200 / / mL, but increased the cell survival rate (Fig. 1).
Example 4. DPPH Analysis of radical scavenging activity
The electron donating ability is used as an index of antioxidant activity and is widely used for measuring the antioxidant activity of plant extracts. The electron donating action is used to donate electrons of free radicals generated in the human body, thereby suppressing aging and disease by free radicals. The electron donating ability was measured by the DPPH radical scavenging method.
DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity (DPPH radical scavenging activity) measurement was performed to measure the electron donating ability of the Osmanthus matsumuranus extract prepared by the method of Example 1 above. Specifically, Osmanthus matsumuranus extract was dissolved in methanol at various concentrations (0.1024 to 12.8 ㎍ / mL) and prepared. Then, 40 μl of DPPH dissolved in methanol at a concentration of 1.5 × 10 -4 M in a 96-well plate and 40 μl of The mixed solution containing 160 쨉 l of the prepared Osmanthus matsumuranus extract solution was reacted at room temperature for 30 minutes, and the absorbance was measured at 520 nm using a microplate reader. The free radical scavenging activity was expressed as a percentage of the negative control to which no sample was added, and a 50% inhibitory concentration (IC 50 ) was calculated. As a positive control, ascorbic acid (vitamin C), a representative antioxidant, was used. The formula for calculating the percentage of the free radical scavenging ability is as follows.
DPPH radical scavenging activity (%) = {1- (A-B) / C} 100
A is the absorbance of the sample at 520 nm, B is the color control absorbance at 520 nm, and C is the control absorbance at 520 nm.
The analytical results of the DPPH radical scavenging activity of the Osmanthus matsumuranus extract of the present invention and the calculated IC 50 values are shown in Table 1 below. The calculated IC 50 value for the DPPH radical scavenging activity of Osmanthus matsumuranus extract was 1.87 ㎍ / mL, which showed similar antioxidative activity to ascorbic acid (1.53 ㎍ / mL) used as a positive control. .
Osmanthus matsumuranus ( Osmanthus matsumuranus )
1.87
(Vitamin C, positive control)
1.53
Example 5. Active Oxygen species (reactive oxygen species, ROS ) Scatters analysis
The effects of hydrogen peroxide (H 2 O 2 ) - induced ROS on the antioxidative capacity of Osmanthus matsumuranus extract were investigated. RAW 264.7 cells were pretreated with 50 μM DCFH-DA, a cell permeable fluorescent dye, for 2 hours, removed, and treated with 500 μM H 2 O 2 and concentration-dependent extracts. The inhibition of ROS formation was analyzed by fluorescence measurement using a multiplate reader.
As a result, it was confirmed that the extract of Osmanthus matsumuranus effectively inhibited ROS generation induced by H 2 O 2 in a concentration-dependent manner, and thus it was confirmed that the extract had a strong antioxidant ability (FIG. 2).
Example 6. HO-1 and Nrf -2 expression analysis
In order to examine the antioxidant activity of Osmanthus matsumuranus extract, the expression of HO-1, a representative antioxidant enzyme, and Nrf-2, a transcription factor, was examined by western blotting And analyzed by Western blot hybridization. Proteins were extracted from the sample-treated cultured cells, and the protein concentration was determined by Bradford assay. Then, 50 μg of the protein was electrophoresed on 10% SDS-PAGE, blotted onto a nitrocellulose membrane, Lt; / RTI > After washing the membranes, HRP-labeled secondary antibodies were reacted for 1 hour and protein expression was analyzed using a chemiluminescence detection system.
As a result, HO-1 and Nrf-2 showed significant induction of expression depending on the concentration of Osmanthus matsumuranus extract, and the antioxidant ability of the extract was attributed to the induction of HO-1 expression through the Nrf-2 signaling pathway (Fig. 3).
Example 7. Cytotoxicity and nitrogen monoxide (NO) production Inhibition analysis
In order to confirm the anti-inflammatory activity of the Osmanthus matsumuranus extract prepared in Example 1, the ability to inhibit the production of nitrogen monoxide (NO), which is one of inflammatory substances, was analyzed.
RAW 264.7 cells were adhered to a 24-well tissue culture vessel at a rate of 3.0 X 10 5 cells / well, treated with 1 쨉 g / mL of lipopolysaccharide (LPS) to induce NO production, To confirm the inhibitory effect of NO production on the NO production, the cytotoxicity induction of Osmanthus matsumuranus extract was first confirmed by WST analysis and the inhibition of NO production by Griess reaction was analyzed.
The water soluble tetrazolium (WST) assay is a representative method for measuring cell viability using water-soluble tetrazolium. After 24 hours of sample treatment, the cells were incubated at 37 ° C for 1 hour after replacing with WST medium. Absorbance was measured at 450 nm using a multi-plate reader, and the value was expressed as the average value of three repeated experiments.
The grease reaction is a method for quantitatively analyzing the amount of released NO produced from the cultured cells. After 24 hours of sample treatment, 100 μl of the cell culture medium is dispensed into a 96-well plate, and then the grease reaction solution A (5% (0.1% naphthylenediamine dihydrochloride) buffer solution (1%) was dispensed into a 96-well plate in an amount of 100 쨉 l per well, The absorbance at 540 nm was measured using a plate reader, and the value was expressed as an average value of three repeated experiments.
First, the effect of Osmanthus matsumuranus extract on LPS-induced RAW 264.7 cell survival rate was examined. As shown in Fig. 4, it did not induce cytotoxicity up to 100 ㎍ / mL, In the case of the present invention.
Furthermore, the anti-inflammatory activity of the extract of Osmanthus matsumuranus was analyzed by RAW 264.7 cells in the rat macrophage cell line induced by LPS. As a result, it was confirmed that the extract showed inhibitory effect on NO production as shown in FIG. 5, It was confirmed that the compound had a strong anti-inflammatory activity.
As a result, it was confirmed that the extract of Osmanthus matsumuranus did not induce toxicity to RAW 264.7 cells induced by LPS, but had anti-inflammatory action.
Thus, the above results suggest that Osmanthus matsumuranus extract has strong antioxidant ability by inhibiting the production of reactive oxygen species by inducing DPPH radical scavenging activity and HO-1 expression by Nrf-2 signal transduction pathway , And has a strong anti-inflammatory effect through inhibition of NO production. Therefore, it can be used for prevention, improvement or treatment of diseases caused by oxidative and inflammatory action, or for foods, cosmetics and medicines.
Claims (7)
Wherein the extract is extracted with water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.
Wherein the alcohol is ethanol.
Wherein said antioxidant effect is achieved by removal of reactive oxygen species.
Wherein the antiinflammatory effect is achieved by inhibiting nitric oxide (NO) production.
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