CN114767600A - Antioxidant, anti-inflammatory and anti-pollution composition containing mixed extract of myrtle and sparassis crispa - Google Patents
Antioxidant, anti-inflammatory and anti-pollution composition containing mixed extract of myrtle and sparassis crispa Download PDFInfo
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- CN114767600A CN114767600A CN202210617200.6A CN202210617200A CN114767600A CN 114767600 A CN114767600 A CN 114767600A CN 202210617200 A CN202210617200 A CN 202210617200A CN 114767600 A CN114767600 A CN 114767600A
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- myrtle
- sparassis crispa
- inflammatory
- mixed extract
- antioxidant
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Abstract
The invention relates to an antioxidant or anti-inflammatory composition, which comprises a mixed extract of myrtle and sparassis crispa as an active ingredient, wherein the mixed extract of myrtle and sparassis crispa has DPPH free radical scavenging activity and a remarkable lipid peroxidation inhibition effect, has an excellent inflammation inhibition effect, and has a remarkable anti-inflammatory effect. Therefore, the mixed extract of the myrtle and the sparassis crispa can be used in medicines, foods, cosmetics and the like to achieve the purposes of antioxidation and/or anti-inflammation. In addition, it can be used as an anti-pollution material by utilizing its anti-oxidation and anti-inflammatory effects.
Description
Technical Field
The present invention relates to an antioxidant, anti-inflammatory and anti-pollution composition comprising a mixed extract of myrtle and sparassis crispa.
Background
The fine dust is dust having a particle size of 10 μm or less, abbreviated as PM10(Particulate Matter 10), and the ultrafine dust having a particle size of 2.5 μm or less is classified as PM2.5(Particulate Matter 2.5). It is mainly produced when fossil fuels are burned, and fine dust from china, which relies on coal to a degree of 70%, accounts for a large part. In addition, yellow sand is a phenomenon that fine sand, loess and dust in deserts of the places such as china and mongolian countries in the middle of the continental asia and in loess areas float in the sky, fly to the upper layer of the air and are blown away by wind, and may fall into the ju-zhou island as an air pollutant which can move in a long distance.
In particular, when fine dust comes into direct contact with the skin, which is located at the outermost part of our body, the skin acts as a barrier between the organisms and the external environment, and if it is frequently exposed directly to pollution, the barrier is lowered, thereby causing various problems. In particular, the fine dust is 20 times smaller than pores and easily permeates into human skin. For example, the skin is considered to be a major site for absorbing pollutants in the environment, as well as inhaling toxic substances such as chloroform and the like through the respiratory system.
Fine dust causes oxidative stress and inflammation in the body or skin. Inflammation induced in this way affects the whole body and increases the risk of inflammatory diseases of the skin, alopecia, exacerbation of allergic dermatitis, stroke, inflammatory diseases of the brain (such as alzheimer's disease), respiratory diseases and cardiovascular diseases.
At present, the demand for materials having anti-inflammatory and antioxidant functions, which have skin-protecting effects on inflammatory reactions and oxidative stress caused by fine dust and the like, is rapidly increasing, and thus, there is a need to obtain new materials having good effects and little side effects from natural materials having safety.
Prior application patent documents: (patent document 0001) korean registered patent No. 10-1851603.
Disclosure of Invention
The invention aims to provide an antioxidant and anti-inflammatory composition containing a mixed extract of myrtle and sparassis crispa as an active ingredient, and the composition comprises a food composition, a cosmetic composition, a pharmaceutical composition and a pharmaceutical external composition.
It is another object of the present invention to provide an anti-contamination cosmetic composition comprising a mixed extract of myrtle and sparassis crispa as an active ingredient.
One embodiment of the present invention is an antioxidant or anti-inflammatory food composition comprising a mixed extract of myrtle and sparassis crispa as an active ingredient.
The mixed extract can be extracted by mixing the myrtle and the sparassis crispa in a weight ratio of 1: 0.1-10.
The mixed extract can be hot water extract of Myrtus citriodora and Sparassis crispa.
Another embodiment of the present invention is an antioxidant or anti-inflammatory cosmetic composition comprising a mixed extract of myrtle and sparassis crispa as an active ingredient.
Another embodiment of the present invention is a pharmaceutical composition for preventing or treating inflammatory diseases, comprising a mixed extract of myrtle and sparassis crispa as an active ingredient.
The inflammatory disease may be a brain or neurological inflammatory disease.
Another embodiment of the present invention is an antioxidant or anti-inflammatory pharmaceutical external composition comprising a mixed extract of myrtle and sparassis crispa as an active ingredient.
Another embodiment of the present invention is an anti-contamination cosmetic composition comprising a mixed extract of myrtle and sparassis crispa as an active ingredient.
The anti-pollution effect may be to suppress inflammation or oxidative stress caused by sand dust or fine dust.
The invention relates to an antioxidant or anti-inflammatory composition, which comprises a mixed extract of myrtle and sparassis crispa. The mixed extract of myrtle lemon and sparassis crispa has excellent DPPH free radical scavenging activity and lipid peroxidation inhibiting effect, and has been confirmed to be effective in inhibiting inflammation, and thus can be used for antioxidant or anti-inflammatory use.
In addition, the mixed extract of Myrtus citriodora and Sparassis crispa can be used for developing anti-pollution cosmetics for relieving oxidative stress and inflammation caused by fine dust.
Detailed Description
Hereinafter, specific embodiments of the present invention will be described in detail so that those skilled in the art can easily practice the present invention. The invention may, however, be embodied in several different forms and is not limited to the embodiments described.
In one aspect, the present invention relates to an antioxidant or anti-inflammatory food composition comprising a mixed extract of myrtle and sparassis crispa as an active ingredient.
The antioxidant or anti-inflammatory food composition of the present invention comprises active ingredients of a mixed extract of myrtle and sparassis crispa.
In the present invention, "myrtle (Backhausia citriodora)" mainly refers to shrubs growing in forests in the tropical zone of Queensland, Australia. The leaves of lemon myrtle, which have a lemon flavor, are the most widely used flavoring material in australia, and recently are used as a raw material or a flavoring agent for herbal tea, candies, beverages, nutritional supplements, and the like. In addition, the lemon myrtle essential oil has the strongest fragrance in various oils with lemon fragrance, has the longest fragrance retention time, and is mainly used for manufacturing perfumed soaps, lip balms or perfumes. The lemon aroma of lemon myrtle originates from the citral component, and unlike lemon which contains about 3 to 10% citral, lemon myrtle contains about 90% citral and therefore gives off a stronger taste or aroma than lemon, but reportedly, it is also more astringent and bitter. Citral is a monoterpene aldehyde component known to be insoluble in water, mainly soluble in alcohols or ethers.
In the present invention, "Sparassis crispa" is a mushroom belonging to Aphyllophorales (Aphyllophorales) and Sparassidaceae (Sparassidaceae), is distributed in Korea, Japan, China, Europe, North America, Australia, etc., is longer than near the root of coniferous trees or on stumps of trees for 8 to 9 months, has a fruiting body 10 to 25cm high in flesh quality, has a large number of branches on a thick handle at the bottom, has a wavy upper flat edge, is in the shape of white flower cabbage, emits a light fragrance, has good chewing feeling, and has an edible value. In addition, the content of beta-1, 3-D-glucan is significantly higher than that of other mushrooms, and is known to have various physiological activities. That is, it is known to have anticancer and anti-angiogenic effects, antitumor effects, antibacterial and antifungal effects, and antiallergic effects, even on diabetes. It is also involved in cytokine production in mice and is effective in acute liver injury.
In the present invention, "extract" means an extract obtained by an extraction treatment, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, a preparation or purified product of the extract, or a mixture thereof; it includes all formulations of extracts that can be formed using the extract itself and the extract.
In the present invention, useful components in the dried plant of the extract are not destroyed. This can be carried out by a known method, for example, by natural drying in the shade. In addition, useful components of plants can be sufficiently extracted in a subsequent extraction process by breaking the walls or pulverizing into powder. The drying, pulverizing or wall breaking process may be performed in reverse order or repeatedly as necessary.
The extract can be prepared by conventional preparation method of plant extract, specifically solvent extraction, water immersion extraction, cold immersion extraction, warm immersion extraction or ultrasonic extraction, and existing extraction device, ultrasonic pulverizing extractor, fractionator, etc. can be used.
The extraction solvent of the extract can be water or organic solvent. The organic solvent may be a polar solvent, a non-polar solvent or a mixture thereof, such as alcohol, aqueous alcohol solution, hexane, dichloromethane, acetone, ethyl acetate, diethyl ether, chloroform or a mixture thereof. The alcohol is C1-C5 alcohol, such as methanol, ethanol, propanol, butanol, isopropanol, etc. Further, the alcohol aqueous solution can be obtained by diluting alcohol in water to 50 to 99.9% (v/v).
Further, the extract is extracted with a solvent, and then subjected to a fractional distillation process using any one solvent selected from the group consisting of hexane, dichloromethane, acetone, ethyl acetate, diethyl ether, chloroform, water and a mixture thereof. The fractionation process may be prepared using two or more solvents, and each solvent fraction may be used sequentially or in a mixture depending on the polarity of the solvent.
The prepared extract or the fraction obtained by the fractional distillation process may then be filtered or concentrated or dried to remove the solvent, and may be filtered, concentrated and dried at the same time. Specifically, for example, filter paper or a reduced pressure filter may be used for filtration, concentration may be performed under reduced pressure using a reduced pressure concentrator such as a rotary evaporator, and drying may be performed by a freeze-drying method.
In the present invention, the "mixed extract" refers to an extract prepared by extracting Cimyrtus citriodora and Sparassis crispa, and may be prepared by extracting Cimyrtus citriodora and Sparassis crispa respectively and then mixing them, or may be prepared by extracting Cimyrtus citriodora and Sparassis crispa respectively and then mixing them.
The lemon myrtle may be at least one selected from the group consisting of stem, leaf, root, flower and fruit, preferably lemon myrtle leaf.
In one embodiment of the invention, dried myrtle and sparassis crispa are mixed and then extracted under pressure at 80-120 ℃ for 1-5 hours by using distilled water as a solvent.
Therefore, the mixed extract can be a hot water extract of myrtle and sparassis crispa, and specifically can be a hot water extract at 90-110 ℃, and the extraction time is 2-4 h.
In one embodiment of the present invention, the extract is prepared by mixing myrtle and sparassis crispa in a weight ratio of 1: 0.1-10, and the cytotoxicity of the extract is analyzed according to the mixing ratio of myrtle and sparassis crispa, and it is shown that the extract prepared by mixing myrtle and sparassis crispa in a weight ratio of 1:1 (example 1) does not show cytotoxicity.
Therefore, in the invention, the mixed extract can be prepared by mixing and extracting the myrtle and the sparassis crispa in a weight ratio of 1: 0.1-10, specifically, the weight ratio of the myrtle to the sparassis crispa is 1: 0.5-5, preferably 1:1, and extracting with hot water at 90-110 ℃.
In one example of the present invention, it was demonstrated that the concentration of a mixed extract of myrtle and sparassis crispa (example 1, LB) was positively correlated with an increase in DPPH clearance activity. In order to analyze the antioxidant effect, after LPS induced brain inflammation in mice, a mixed extract (LB) of myrtle and sparassis crispa was administered to analyze the change in the production of lipid peroxide (MDA), which is an index of oxidative damage, and it was shown that LPS increased the production of MDA in the brains of experimental animals by about 170%, indicating that LPS induced oxidative damage. It was confirmed that the amount of MDA production increased by LPS was statistically significantly reduced when a high dose (1000mg/kg) of a mixed extract (LB) of Myrtus citriodora and Sparassis crispa was injected.
That is, it was confirmed that the mixed extract of myrtle lemon and sparassis crispa has DPPH scavenging activity and has excellent antioxidant effect by inhibiting lipid peroxide (MDA) caused by oxidative damage, and the extract can be used as an antioxidant composition.
In the present invention, "antioxidation" means protection of cells from oxidative stress, specifically, inhibition of the production of active oxygen or inhibition of the production of lipid peroxide.
In another example of the present invention, anti-inflammatory effects of myrtle and sparassis crispa (LB) were analyzed, and after inducing brain inflammation in mice using LPS, the increase degree of inflammation-associated cells, namely microglia (microglia) and astrocyte (astrocyte), in brain tissues of mice was analyzed after oral administration of a mixed extract of myrtle and sparassis crispa (LB) for 7 days. The results of the analysis showed that LPS induced an increase in GFAP-positive cells (an astrocyte marker) and Iba-1-positive cells (a microglia marker) in the brains of the experimental animals. It was confirmed that when a mixed extract (LB) of myrtle and sparassis crispa was orally taken, the increase of astrocytes (GFAP-positive cells) and microglia (Iba-1-positive cells) induced by LPS was significantly reduced.
That is, it was confirmed that the mixed extract of myrtle and sparassis crispa has an anti-inflammatory effect, and the mixed extract of myrtle and sparassis crispa of the present invention can be used as an anti-inflammatory composition.
In the present invention, "anti-inflammatory" refers to inhibiting or preventing the occurrence of inflammation, or protecting cells from inflammatory damage.
In the present invention, inflammation may be caused by, but is not limited to, microbial infection, exposure to sand or fine dust, exposure to chemicals, or oxidative stress.
The food composition of the present invention may include a form of pill, powder, granule, injection, tablet, capsule or liquid, and the kind of food to which the inventive myrtle and sparassis crispa mixed extract can be added is not particularly limited, such as various beverages, chewing gum, tea, vitamin complex, health functional food, and the like.
The food composition may further comprise other components in addition to the mixed extract of myrtle and sparassis crispa, and the kind thereof is not particularly limited. For example, it may contain various herbal extracts, food additives allowed in foods, or natural carbohydrates as additional ingredients, as in general foods, but is not limited thereto.
In the present invention, the term "food additive" means an ingredient that can be added to a food, and can be added to a functional health food for producing various dosage forms, and can be appropriately selected and used by those skilled in the art. Examples of the food additive include various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, coloring agents and fillers, pectic acid and its salt, alginic acid and its salt, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, and the like, but the above examples are not limited to the kind of the food additive of the present invention.
Examples of natural carbohydrates include monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; and sugar alcohols such as xylitol, sorbitol and erythritol; in addition to the above, natural flavors (e.g., thaumatin), stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (e.g., saccharin, aspartame, etc.) may be used as appropriate.
The kind of the food is not particularly limited. Examples of foods to which the above substances can be added include meats, sausages, breads, chocolates, candies, snacks, biscuits, pizza, stretched noodles, other noodles, chewing gums, dairy products including ice cream, various soups, beverages, teas, oral liquids, alcoholic beverages, vitamin complexes, and the like.
The food composition may be a health functional food composition.
In the present invention, the "health functional food" refers to a food which is produced and processed in the form of tablets, capsules, powders, granules, liquids and pills using functional materials or ingredients useful for human body. Here, the term "functionality" means obtaining a health-beneficial effect on the structure of the human body and the function of regulating the nutritional or physiological effects. The health functional food of the present invention may be prepared by a method commonly used in the art, and raw materials and ingredients commonly used in the art may be added at the time of manufacture. In addition, unlike common medicine, it is produced with food as material and has no side effect and excellent carrying performance.
The health functional food is taken as a beauty food (inner beauty), and has an advantage of more excellent effect. The inner beauty food (inner beauty) is a food called "eatable cosmetic or beauty food", and means a food in which various ingredients beneficial to the skin are absorbed into the body to make the skin healthy, and as well as selecting a cosmetic suitable for the skin type, the inner beauty food suitable for an individual can be selected for ingestion in consideration of the skin condition and the living style. More preferably, when a cosmetic containing the mixed extract of downy rosemyrtle and sparassis crispa is used together with an interior beauty product containing the mixed extract of downy rosemyrtle and sparassis crispa, the antioxidant or anti-inflammatory effect is remarkably improved compared with the case of using only the cosmetic, and the cosmetic is characterized by having a more effective antioxidant or anti-inflammatory effect.
The blending amount of the active ingredient may be appropriately determined depending on the purpose of use (prevention, health function or treatment). In general, the mixed extract of myrtle and sparassis crispa according to the present invention may be added in an amount of 1 to 50%, preferably 5 to 10% by weight of the raw material composition in the preparation of food, but is not limited thereto. However, in the case of long-term ingestion for health and hygiene purposes, or for health regulation, the amount used may be below the above range.
In another aspect, the present invention relates to an antioxidant or anti-inflammatory cosmetic composition comprising a mixed extract of myrtle and sparassis crispa as an active ingredient.
In the present invention, myrtle, sparassis crispa, mixed extract, antioxidant and anti-inflammatory are described as described above.
The cosmetic composition of the present invention may further include a compound or natural extract known to have an antioxidant or anti-inflammatory effect, in addition to the mixed extract of myrtle and sparassis crispa, to enhance the antioxidant or anti-inflammatory effect of myrtle and sparassis crispa.
In the antioxidant or anti-inflammatory cosmetic composition of the present invention, any amount (effective amount) of the active ingredient may be contained according to use, formulation purpose, etc., as long as it can exhibit antioxidant or anti-inflammatory effects, and the conventional effective amount may be included in the range of 0.001 wt% to 99.99 wt% based on the total weight of the composition. Here, "effective amount" refers to an amount of active substance that is capable of inducing an antioxidant or anti-inflammatory effect. Such effective amounts can be determined experimentally within the ordinary ability of those skilled in the art.
The cosmetic composition of the present invention may be prepared in various forms, for example, the cosmetic composition of the present invention may be prepared in any form conventionally prepared in the art, such as a solution, a suspension, an ointment, a gel, a cream, a lotion, a loose powder, a face cleanser, an essential oil, a foundation, an emulsion foundation, a waxy foundation, a spray, etc., but is not limited thereto. Further, it may be specifically selected from various types among a group of a lotion, a toner, a astringent, a skin lotion, an emulsion, a moisturizing emulsion, a nourishing emulsion, a massage cream, a nourishing cream, a moisturizing cream, a hand cream, an essence, a nourishing essence, a mask, a soap, a shampoo, a cleansing foam, a cleansing milk, a cleansing cream, a body lotion, an emulsion, a lipstick, a makeup base, a foundation, a compact, and a loose powder, but is not limited thereto.
The cosmetic composition of the present invention may contain, in addition to its active ingredient, a cosmetically acceptable carrier. Herein, "cosmetically acceptable carrier" means a compound or composition known and used, which may be included in cosmetics, or which will be developed in the future, which, when in contact with the skin, is not more toxic, unstable or irritating than the human body in the prior art. The carrier is present in the cosmetic composition of the present invention in an amount of about 1% to about 99.99% by weight, preferably about 5% to about 99% by weight. However, since the above ratio varies depending on the formulation of the cosmetic of the present invention and its specific application site (face or hand) or desired applicable amount, the above ratio should not be construed as limiting the scope of the present invention in any way.
On the other hand, the carrier may be, for example, alcohol, oil, surfactant, fatty acid, silicone oil, humectant, viscosity modifier, emulsion, stabilizer, sunscreen agent, developer, perfume, etc. Alcohols, oils, surfactants, fatty acids, silicone oils, humectants, moisturizers, viscosity modifiers, emulsifiers, stabilizers, sunscreens, colorants, fragrances, and the like that may be used as carriers are known in the art and one skilled in the art can select and use the appropriate materials/compositions.
In another aspect, the present invention relates to a pharmaceutical composition for preventing or treating inflammatory diseases, comprising a mixed extract of myrtle and sparassis crispa as an active ingredient.
The description of the myrtle, sparassis crispa and mixed extract in the invention is as described above.
In the present invention, the "inflammatory disease" is a disease caused by inflammation (inflammation) which may be caused by microbial (bacterial, viral, fungal) infection, sandstorm, fine dust, exposure to chemicals or oxidative stress, but is not limited thereto.
The inflammatory disease may be one of allergic disease, inflammatory intestinal disease, atherosclerosis, inflammatory collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, pharyngitis, bronchitis, pneumonia, pancreatitis, septicemia, brain or nerve inflammatory disease, and nephritis.
In the above embodiment of the present invention, the mixed extract (LB) of myrtle and sparassis crispa inhibits the induction of the increase of astrocytes and microglia, cells associated with inflammation, in the brain of experimental animals using LPS. It was confirmed that the mixed extract of myrtle and sparassis crispa can prevent or treat inflammatory diseases by inhibiting inflammation.
In addition, it was confirmed that the mixed extract of myrtle and sparassis crispa inhibits inflammation of brain tissue, and thus, in the present invention, the inflammatory disease may be specifically a brain or nerve inflammatory disease.
The brain or nerve inflammatory disease may be degenerative encephalopathy caused by oxidative stress and excessive inflammatory reaction, and specifically may be one of Alzheimer's disease, Parkinson's disease, Huntington's disease and Amyotrophic Lateral Sclerosis (ALS).
In the present invention, "treatment" refers to alleviation, inhibition, sedation, or eradication of an infectious disease or an inflammatory disease, which is a subject disease of the present invention. In the present invention, "prevention" means any action of suppressing or delaying the onset of an infectious disease or an inflammatory disease which is a target disease of the present invention, and includes a therapeutic meaning.
The pharmaceutical compositions of the present invention may be administered in a pharmaceutically effective dose.
In the present invention, the "pharmaceutically effective dose" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose standard may be determined by factors including the type and severity of a subject, age, sex, type of disease, pharmaceutical activity, sensitivity to drugs, administration time, administration route and excretion rate, duration of treatment, drugs used concurrently, and other factors well known in the medical field. The compositions of the present invention may be used as the sole therapeutic agent or may be used in combination with other therapeutic agents, either sequentially or simultaneously with conventional therapeutic agents. And may be administered in a single or multiple doses. Importantly, in view of all of the above factors, the maximum effect can be obtained by administering the minimum amount without causing side effects, and can be easily determined by one of ordinary skill in the art.
The pharmaceutical composition of the present invention is not particularly limited as long as it is an individual for the purpose of treating or preventing infectious diseases or inflammatory diseases, and any individual may be used. For example, non-human animals such as monkeys, dogs, cats, rabbits, guinea pigs, rats, mice, cows, sheep, pigs, goats, etc., humans, birds, and fish, etc., may be used, and the pharmaceutical compositions may be administered parenterally, subcutaneously, intraperitoneally, intrapulmonary, and intranasally, and if necessary topically by any suitable means including intralesional administration. The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and body weight of an individual, the degree of disease, the form of the drug, the administration route and the period, but can be appropriately selected by those skilled in the art. For example, it can be administered by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine, or intracerebral injection, but is not limited thereto.
The pharmaceutical composition can be selected from any one of tablets, pills, powder, granules, capsules, suspensions, oral liquid preparations, emulsions, syrups, sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilizates and suppositories, and can also be various oral or parenteral dosage forms. When formulated, they may be prepared using conventional diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants and surfactants.
Solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid preparations include at least one excipient, which can be prepared by mixing, for example, starch, calcium carbonate, sucrose (sucrose) or lactose, gelatin and the like. Besides simple excipients, lubricants such as magnesium stearate, talc, etc. may also be used. The liquid preparations for oral administration include suspensions, oral liquid preparations, emulsions, syrups and the like, and may include various excipients such as wetting agents, sweeteners, flavors, preservatives and the like in addition to the common simple diluents water and liquid paraffin. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, synthetic fatty acid ester (witepsol), polyethylene glycol, tween (tween)61, cacao butter, lauric acid glyceride, glycerogelatin, and the like can be used.
The appropriate total daily amount can be determined by the attending physician within the scope of sound medical judgment, and is generally 0.001-1000 mg/kg, preferably 0.05-200 mg/kg, more preferably 0.1-100 mg/kg, and can be administered once to several times per day.
In another aspect, the present invention relates to an antioxidant or anti-inflammatory pharmaceutical topical composition comprising a mixed extract of myrtle and sparassis crispa as an active ingredient.
The description of myrtle, sparassis crispa, mixed extract, antioxidant and anti-inflammatory in the present invention is as described above.
In the present invention, the term "pharmaceutical product" refers to a product that has a milder action than a drug, in terms of diagnosis, treatment, amelioration, remission, treatment, or prevention of a disease in a human or an animal. For example, according to the "medical law", the term "medical foreign product" refers to products other than those used as medicines, including textiles and rubber products for treating or preventing human and animal diseases, articles like non-appliances or machines which have little or no direct effect on the human body, disinfectants and insecticides for preventing it, and the like.
The type or formulation of the external pharmaceutical product of the present invention is not particularly limited, but a disinfectant, a bath lotion, a mouth wash, a wet wipe, a laundry soap, a hand cleanser, a humidifier filler, a mask, an ointment, a filter filler, or the like is preferable.
When the composition of the present invention is included in a pharmaceutical external product for anti-oxidant or anti-inflammatory purposes, the composition may be used as it is, or may be used together with other pharmaceutical external product components, and may be suitably used according to a conventional method, and the mixing amount of the active components may be suitably determined according to the purpose of use, and in the pharmaceutical external product composition of the present invention, the mixed extract of myrtle and sparassis crispa may account for 0.01 to 20% by weight relative to the total weight of the composition.
In another aspect, the present invention relates to a cosmetic composition for anti-pollution, comprising a mixed extract of myrtle and sparassis crispa as an active ingredient.
The description of myrtle, sparassis crispa, mixed extract and cosmetics in the present invention are the same as above.
In the present invention, "anti-contamination" means inhibition of oxidative stress or inflammation caused by sand dust or fine dust.
In the above examples of the present invention, it was confirmed that the mixed extract of myrtle and sparassis crispa has excellent DPPH radical scavenging activity, and it was confirmed that it has excellent antioxidant effect by inhibiting the production of lipid peroxide induced in mice by LPS and that it can protect cells from sand dust or fine dust by inhibiting or reducing the oxidative stress induced in cells by sand dust or fine dust.
In addition, when LPS was used to induce inflammation in the brain tissue of mice, the mixed extract of myrtle and sparassis crispa orally administered to the mice, which inhibited the production of astrocytes (GFAP positive cells) and microglia (Iba-1 positive cells), which are inflammation-related cells, compared to the control group administered with LPS alone, and the mixed extract of myrtle and sparassis crispa of the present invention protected cells by inhibiting or preventing inflammation caused by fine dust.
In addition, it was confirmed that oral administration of a mixed extract of myrtle and sparassis crispa can inhibit inflammation of brain tissue, which may be for the purpose of protecting cells from damage.
In addition, it was confirmed that oral administration of a mixed extract of myrtle and sparassis crispa inhibits inflammation in brain tissue, and the anti-contamination of the present invention can specifically protect cells from nerve or brain cell damage caused by inflammation induced by fine dust.
Hereinafter, the constitution and effect of the present invention will be described in more detail by way of examples. These examples are provided only for illustrating the present invention, and the scope of the present invention is not limited to these examples.
1. Preparation method of mixed extract of myrtle and sparassis crispa
Mixing dried Myrtus citriodora and Sparassis crispa at the weight ratio shown in Table 1, extracting with distilled water at 100 deg.C for 3 hr, filtering, concentrating, freeze drying at-75 deg.C, and collecting the dried product.
The result of cytotoxicity analysis of the prepared mixed extracts showed that example 1 was not cytotoxic.
TABLE 1
2. In vitro (in-vitro) antioxidant Activity assay
1) DPPH free radical scavenging Activity assay
The samples prepared in example 1, comparative examples 1 and 2 were dissolved in distilled water to a concentration of 1000. mu.g/ml and used for experiments. DPPH (1, 1-diphenyl-2-picrylhydrazyl) solution is prepared by dissolving 3mg DPPH in 15ml ethanol, and adding 3ml ethanol and 0.5ml DMSO to 1.5ml solution.
DPPH radical scavenging activity, 50. mu.l of the sample and DPPH solution were mixed and reacted at room temperature for 10 minutes, and then the concentration of residual radicals was measured at 517nm using an ultraviolet spectrophotometer.
DPPH radical scavenging activity electron donating ability (EDA%) was calculated according to the following mathematical formula 1.
[ mathematical formula 1 ]
EDA(％)＝(B-A)/B×100
A is absorbance of sample application group, B is absorbance of sample-free group
TABLE 2
| Distinguishing between | DPPH radical scavenging ability |
| Example (LB) | 88.91 |
| COMPARATIVE EXAMPLE 1(L) | 67.59 |
| COMPARATIVE EXAMPLE 2(B) | 71.44 |
In order to determine the in vitro antioxidant effect of the mixed extract of myrtle and sparassis crispa (example 1, LB), the results of analyzing DPPH radical scavenging activity (%) showed that the mixed extract of myrtle and sparassis crispa (example 1, LB) exhibited more excellent DPPH scavenging activity than the individual extracts of myrtle (comparative examples 1, L) or sparassis crispa (comparative examples 2, B).
3 analysis of antioxidant and anti-inflammatory action in animal models (in vivo)
1) Preparation of laboratory animals
Male ICR mice (6 weeks old) were purchased from Samtaco Bio, and used after 1 week of adaptation. Animals were housed in individual housing units at 22 + -1 deg.C and 50 + -10% humidity, with a day/night set to circulate every 12 hours, with water and food provided as an ad hoc diet. The experimental group consisted of a non-administered control group (sham) to which LPS (lipopolysaccharide) was not added, an LPS control group (control), a mixed extract (LB) of myrtle and Sparassis crispa, a mixed extract low-dose group (100LB, 100mg/kg) to which LPS was added together with a mixed extract (LB) of myrtle and Sparassis crispa, a mixed extract (LB) high-dose group (1000LB, 1000mg/kg) to which myrtle and Sparassis crispa, a single extract (L) of myrtle and a single extract high-dose group (1000L, 1000mg/kg) to which LPS was added together with a single extract (L) of myrtle and Sparassis crispa, a high-dose group (1000B, 1000mg/kg) to which Sparassis crispa, and a positive control group (ferulic acid ).
2) Drug (sample) administration and brain sample collection
The administration dose of the mixed extract of myrtle and sparassis crispa was set to a low dose (100LB, 100mg/kg) and a high dose (1000LB, 1000mg/kg), the administration dose of the single extract of myrtle (L) and the single extract of sparassis crispa (B) was set to a high dose (1000mg/kg), and the positive control group was administered with ferulic acid at 20 mg/kg.
LPS was dissolved in PBS, administered to the third ventricle at a dose of 1 μ g/3 μ l, each sample was dissolved in 10% tween 80 solution and administered orally 3 days before LPS administration.
Samples were taken orally 7 days after LPS administration and brain tissue from experimental animals was collected 7 days later. Half of the brain tissue was ground for western blot and ELISA analysis, and the other half was fixed for tissue immunostaining.
3) Evaluation of antioxidant effect: lipid peroxidation assay (MDA assay)
In order to analyze the antioxidant effects of the mixed extract (LB) of myrtle and sparassis crispa, the single extract (L) of myrtle and the single extract (B) of sparassis crispa, the change in the production of lipid peroxide (MDA) after administration of the mixed extract (LB) of myrtle and sparassis crispa, the single extract (L) of myrtle and the single extract (B) of sparassis crispa was analyzed by using lipid peroxide (MDA), which is an index of oxidative damage caused by LPS after induction of brain inflammation in mice.
To determine changes in lipid peroxides, mouse hippocampus tissues were homogenized at 4 ℃ using Phosphate Buffered Saline (PBS). Then, the mixture was centrifuged at 10,000 rpm for 5 minutes, and the supernatant was extracted and used. Samples were treated with 0.15M potassium chloride and 500. mu.l of each homogenate and 100. mu.l of 20mM ferric chloride was added thereto. Thereafter, after reaction in a 37 ℃ incubator for 30 minutes, 500. mu.l of 0.25N cold HCl containing 15% TCA, 0.38% TBA and 0.5% BHT was added. After heating at 90 ℃ for 60 minutes, the resulting mixture was centrifuged at 5,000rpm at 4 ℃ for 5 minutes to separate the extract, and the thiobarbituric acid-reactive substance (TBARS) in the supernatant was measured at an absorbance of 532 nm.
TABLE 3
| Distinguishing | MDA (percent control) |
| Non-drug control group (sham) | 100 |
| LPS control group (control) | 170 |
| 100LB(100mg/kg) | 160 |
| 1000LB(100mg/kg) | 120 |
| 1000L(100mg/kg) | 140 |
| 1000B(100mg/kg) | 135 |
| Positive control group (FA,20mg/kg) | 115 |
The analysis results showed that LPS increased the production of MDA in the brains of experimental animals by 170%, which means that LPS induced oxidative damage. When the extracts of Cimyrtle and Sparassis crispa (LB), Cimyrtle single extract (L) and Sparassis crispa single extract (B) were administered at high doses (1000mg/kg), it was confirmed that the amount of MDA production increased by LPS was significantly reduced. Compared with the single extract (L) of the myrtle lemon and the single extract (B) of the sparassis crispa, the MDA production amount increased by LPS is statistically and remarkably reduced.
In addition, it was confirmed that Ferulic Acid (FA) used as a positive control also inhibited MDA produced by LPS (table 3).
4) Evaluation of anti-inflammatory Effect: histological immunostaining analysis of inflammatory cells
In order to analyze the anti-inflammatory effects of the mixed extract (LB) of myrtle and sparassis crispa, the single extract (L) of myrtle and sparassis crispa and the single extract (B) of sparassis crispa, the increase degree of inflammatory-related cells, namely microglia (microglia) and astrocyte (astrocyte), in brain tissues of mice was analyzed after the mixed extract (LB) of myrtle and sparassis crispa, the single extract (L) of myrtle and sparassis crispa and the single extract (B) of sparassis crispa were orally administered within 7 days after the mice were induced with LPS to the brain inflammations. Specifically, the degree of increase of each cell was analyzed by tissue immunostaining of microglia using Iba-1 antibody and astrocytes using GFAP antibody.
The immobilized hippocampus portion of brain tissue was selected, washed 3 times with PBS, and then treated with hydrogen peroxide to remove endogenous peroxidase (peroxidase). The primary antibody, rabbit-Iba-1 (1:1000 dilution) or rabbit-GFAP (1:1000 dilution), was allowed to react overnight. The secondary antibody was developed with DAB for 3 minutes by ABC reaction using biotin and rabbit antibody (1:200 dilution).
Wash 3 times with PBS between each procedure. After the staining reaction was completed, the coverslip with hippocampus tissue was covered with mounting solution and dried, and then stored. After placing the brain tissue on a glass slide coated with gelatin, the brain tissue is treated with 70-100% ethanol and xylene, and then the tissue is covered with a cover glass and stored. The stained hippocampus sections were visualized using an optical Microscope (Olympus Microscope System BX 51; Olympus, Tokyo, Japan). The ratio of the area of each antibody positive region to the total hippocampus area is indicated.
TABLE 4
| Distinguishing between | GFAP positive area (%) in CA3 region |
| Non-drug control group (sham) | 0.021 |
| LPS control group (control) | 0.053 |
| 100LB(100mg/kg) | 0.045 |
| 1000LB(100mg/kg) | 0.029 |
| 1000L(100mg/kg) | 0.038 |
| 1000B(100mg/kg) | 0.041 |
| Positive control group (FA,20mg/kg) | 0.027 |
TABLE 5
| Distinguishing | Iba-1 Positive area (%) -in CA3 region |
| Non-drug control group (sham) | 0.012 |
| LPS control group (control) | 0.059 |
| 100LB(100mg/kg) | 0.039 |
| 1000LB(100mg/kg) | 0.019 |
| 1000L(100mg/kg) | 0.034 |
| 1000B(100mg/kg) | 0.037 |
| Positive control group (FA,20mg/kg) | 0.017 |
The results of the analysis showed that LPS induced an increase in astrocyte marker GFAP positive cells and microglia marker Iba-1 positive cells in the brains of the experimental animals. When the mixed extract (LB) of downy rosemyrtle and sparassis crispa, the single extract (L) of downy rosemyrtle and the single extract (B) of sparassis crispa are orally administered, the increase of astrocytes (GFAP-positive cells) and microglia (Iba-1-positive cells) caused by LPS can be reduced, and the increase of astrocytes (GFAP-positive cells) and microglia (Iba-1-positive cells) caused by LPS is remarkably reduced by the mixed extract (LB) of downy rosetle and the sparassis crispa compared with the single extract (L) of downy rosemyrtle and the single extract (B) of sparassis crispa.
The positive control group (ferulic acid, FA) also inhibited the increase of astrocytes by LPS (table 4, table 5).
That is, it was confirmed in animal experiments that the mixed extract (LB) of myrtle and sparassis crispa orally administered has an anti-inflammatory effect, and it was confirmed that it has an excellent anti-inflammatory effect as compared to the single extract (L) of myrtle and the single extract (B) of sparassis crispa.
As described above, according to the results of the present invention, it can be seen that the mixed extract of myrtle and sparassis crispa has more excellent synergistic effect than the case of using myrtle and sparassis crispa alone.
While the preferred embodiments of the present invention have been described in detail, it will be understood by those skilled in the art that the present invention is illustrative only, and can be easily modified into other specific forms without changing the technical idea or essential features of the present invention.
Accordingly, the scope of the present invention is indicated by the appended claims rather than the description, and all changes or modifications that come within the meaning and range and equivalence of the claims are to be construed as being included in the scope of the invention.
Claims (9)
1. An antioxidant or anti-inflammatory food composition, characterized in that the composition is an antioxidant or anti-inflammatory food composition comprising a mixed extract of myrtle and sparassis crispa as an active ingredient.
2. The antioxidant or anti-inflammatory food composition as claimed in claim 1, wherein the mixed extract is obtained by mixing and extracting myrtle and sparassis crispa at a weight ratio of 1: 0.1-10.
3. The antioxidant or anti-inflammatory food composition as set forth in claim 1, wherein the mixed extract is a hot water extract of myrtle and sparassis crispa.
4. An antioxidant or anti-inflammatory cosmetic composition, characterized in that it is an antioxidant or anti-inflammatory cosmetic composition comprising, as an active ingredient, a mixed extract of myrtle and sparassis crispa.
5. A pharmaceutical composition for preventing or treating inflammatory diseases, which comprises a mixed extract of myrtle and sparassis crispa as an active ingredient.
6. The pharmaceutical composition of claim 5, wherein the inflammatory disease is a brain or neuroinflammatory disease.
7. An antioxidant or anti-inflammatory pharmaceutical external composition is characterized by comprising a mixed extract of myrtle and sparassis crispa as an active ingredient.
8. An anti-pollution cosmetic composition, characterized in that the composition is an anti-pollution cosmetic composition comprising a mixed extract of myrtle and sparassis crispa as an active ingredient.
9. The anti-contaminant cosmetic composition of claim 8, wherein said anti-contaminant inhibits inflammation or oxidative stress caused by sand or fine dust.
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| KR20100032713A (en) * | 2008-09-18 | 2010-03-26 | (주)우성바이오 | Extract of sparassis crispa and its use as anti-inflammatory medicine |
| JP2015024982A (en) * | 2013-07-29 | 2015-02-05 | 上野製薬株式会社 | Advanced glycation end products formation inhibitor |
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| KR20190064714A (en) * | 2017-11-30 | 2019-06-11 | 신라대학교 산학협력단 | Anti-inflammatory agent containing backhousia citriodora extract |
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| KR101851603B1 (en) | 2017-04-07 | 2018-04-24 | 부산대학교 산학협력단 | Composition for preventing or treating of parkinson's disease through controlling neuroinflammation |
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