KR20160079608A - Composition for anti-inflammation containing an extract of Thymus quinquecostatusvar.japnica Hara - Google Patents
Composition for anti-inflammation containing an extract of Thymus quinquecostatusvar.japnica Hara Download PDFInfo
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- KR20160079608A KR20160079608A KR1020150084523A KR20150084523A KR20160079608A KR 20160079608 A KR20160079608 A KR 20160079608A KR 1020150084523 A KR1020150084523 A KR 1020150084523A KR 20150084523 A KR20150084523 A KR 20150084523A KR 20160079608 A KR20160079608 A KR 20160079608A
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- extract
- inflammatory
- composition
- thymus
- thyme
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- A—HUMAN NECESSITIES
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Abstract
The isoameric extracts described herein can exhibit excellent anti-inflammatory effects without toxicity. For example, inhibition of the production of at least one of nitrite and PGE2, inhibition of the NF-κB signaling pathway, inhibition of mitogen-activated protein kinase (MAPK) signaling, or inhibition of TNF- 6 and < RTI ID = 0.0 > IL-1, < / RTI >
Description
The present disclosure relates to the anti-inflammatory use of isomaltic acid extract.
Inflammation is a complex biological process involving activation of various immune cells such as monocytes / macrophages. Control disorders of the inflammatory immune response cause a variety of pathological conditions, including cancer (Ben-Neriah and Karin, 2011) and metabolic syndrome (Donath and Shoelson, 2011). Macrophages are the major type of cells involved in the inflammatory process by producing a variety of inflammatory mediators such as cytokines / chemokines and nitric oxide (NO) and prostaglandins (PGs) (Zhang and Mosser, 2008). iNOS (inducible NO synthase) is an enzyme that synthesizes NO from L-arginine using NADPH and oxygen molecules (Murakami and Ohigashi, 2007). COX-2 (Cyclooxygenase-2) converts arachidonic acid to prostaglandins such as PGE2 (Aoki and Narumiya, 2012). Control of the reduction of inflammatory mediators in macrophages provides a rationale for the development of therapeutic agents useful in a variety of inflammatory diseases (Murakami and Ohigashi, 2007).
Various transcription factors and cell signaling pathways play a role in the proinflammatory gene expression of macrophages (Guha and Mackman, 2001). Under stimulation by lipopolysaccharides (LPS) or cytokines, NF-κB is activated through activation of the IκB-kinase (IKK) complex. The IKK complex consists of two kinase subunits (IKKα and IKKβ) and the regulatory subunit IKKγ / NEMO. The IKK complex phosphorylates IκBα of Ser32 and Ser36, resulting in ubiquitination followed by proteome degradation (Gloire et al., 2006). Free NF-κB separates from IκBα in the cytoplasm and migrates into the nucleus, activating the transcription of target genes such as proinflammatory genes in the nucleus.
Thyme (Thymus quinquecostatus Celakov) is a perennial deciduous tree belonging to the genus Lepidoptera (Labiatae) originating from the Mediterranean coast of southern Europe and is one of the subtropical and temperate herbivores. In Korea, only thyme thyme ( Thymus quinquecostatus var. Japonica Hara) and two thyme species are native to Japan. In particular, the thyme thyme is native to the Nari Dong Basin, Ulleungdo Island, and is designated as Natural Monument No. 52. It is well- It grows and is known to be easy to cultivate.
Currently, most studies of islet thyme have focused on the study of essential oils and essential oil extracts.
In one aspect, the present invention aims to provide a composition free from toxicity and having excellent anti-inflammatory activity.
In one aspect, the present invention provides an anti-inflammatory composition comprising an isoameric extract as an active ingredient.
In one embodiment, the isthmopharyngeal extract may inhibit the production of at least one of nitrite and PGE2.
In another embodiment, the islet thyme extract may inhibit the NF-kB signaling pathway.
In other embodiments, the islet thyme extract may inhibit mitogen-activated protein kinase (MAPK) signaling.
In another embodiment, the islet thyme extract may inhibit inflammatory cytokines.
In other embodiments, the inflammatory cytokine may be one or more of TNF-a, IL-6, and IL-l [beta].
In other embodiments, the islet thyme extract may inhibit the expression of the inflammatory cytokine.
In one aspect, the composition of the present invention provides a cosmetic, food or pharmaceutical composition.
In one aspect of the present invention, a composition comprising an extract of Ishii thyme as an active ingredient may exhibit an excellent anti-inflammatory effect without toxicity.
The anti-inflammatory composition according to one aspect of the present invention has an effect of inhibiting the production of at least one of nitrite and PGE2.
The anti-inflammatory composition according to one aspect of the present invention has an effect of inhibiting the NF-kB signaling pathway.
The anti-inflammatory composition according to one aspect of the present invention has an effect of inhibiting mitogen-activated protein kinase (MAPK) signaling.
The anti-inflammatory composition according to one aspect of the present invention exhibits an excellent anti-inflammatory effect by inhibiting inflammatory cytokines such as one or more of TNF-a, IL-6 and IL-1 ?.
FIG. 1 is a graph showing the cell survival rate of an extract of isthmus chrysanthemum at Raw 264.7 cell.
Figures 2 and 3 are graphs of the nitrite and PGE2 measurements of the thymus extracts from Raw 264.7 cells.
FIG. 4 shows the results of measuring the expression levels of iNOS and COX-2 in the extracts of the isochrone extracts from Raw 264.7 cells. [(A) Protein (B) mRNA]
FIG. 5 is a photograph showing the effect of an isthmophytic extract on LPS-induced NF-.kappa.B signaling molecule activity in Raw 264.7 cells.
FIG. 6 is a photograph showing the effect of inhibiting the MAPK activity of the isthmophyll extract.
FIG. 7 is a graph showing inhibition of pro-inflammatory cytokine production in the thymus supernatant of Raw 264.7 cells.
In one aspect of the present invention, there is provided a anti-inflammatory composition comprising as an active ingredient an extract of isthmus masca.
In one embodiment, the Thymus extract comprises an anti-inflammatory composition that inhibits the production of at least one of nitrite and PGE2.
In another embodiment, the isoflavone extract provides a anti-inflammatory composition that inhibits the expression of one or more of iNOS and COX-2.
In another embodiment, the isoameric extract provides an anti-inflammatory composition that inhibits the NF-kB signaling pathway.
In another embodiment, the Thymus extracts provide anti-inflammatory compositions that inhibit mitogen-activated protein kinase (MAPK) signaling.
In another embodiment, the isoflavone extract provides a anti-inflammatory composition that inhibits inflammatory cytokines.
In another embodiment, the inflammatory cytokine provides a anti-inflammatory composition that is at least one of TNF-a, IL-6, and IL-l [beta].
In another embodiment, the isoflavone extract provides a anti-inflammatory composition that inhibits the expression of the inflammatory cytokine.
In another aspect of the present invention, the anti-inflammatory composition provides a cosmetic, a food or a pharmaceutical composition.
In one embodiment of the present invention, the present invention is a composition comprising an island thyme extract as an active ingredient.
As used herein, the term " extract " is a broad concept that encompasses all of the substances obtained by extracting components of natural products, regardless of the type of extraction method, extraction solvent, extracted component or extract.
In one aspect of the present invention, a method for preparing an extract of Thymus thaliana is as follows, but a conventional extraction method can be used, and the scope of the present invention is not limited thereto.
A method for preparing an island thyme extract, comprising the steps of: preparing a raw material of island thyme; A step of drying the raw material, and a step of extracting the dried raw material with a solvent,
In one aspect of the present invention, the raw material may be a conventional thyme, preferably Thymus magnus NaKai.
In one aspect of the present invention, the step of drying the raw material comprises a one-stage drying step and may be carried out at a temperature of 40 ° C to 500 ° C for 1 minute to 240 hours.
In one embodiment, the drying step may comprise a primary, secondary, tertiary or quaternary drying step. The drying step may proceed to an appropriate order until the moisture is sufficiently dried.
In one embodiment, the temperature of the drying step is greater than or equal to 40 ° C, greater than or equal to 45 ° C, greater than or equal to 50 ° C, greater than or equal to 60 ° C, greater than or equal to 65 ° C, greater than or equal to 70 ° C, , 95 占 폚 or higher, 100 占 폚 or higher, or 105 占 폚 or higher.
In one embodiment, the temperature of the drying step may be less than 500 ° C, less than 450 ° C, less than 400 ° C, less than 350 ° C, less than 300 ° C, less than 250 ° C, less than 200 ° C, less than 150 ° C, or less than 105 ° C.
When the temperature of the drying step is within the above range, sufficient drying is performed to increase the anti-inflammatory activity in the production of the island thyme extract.
In one embodiment, the drying time is at least 1 minute, at least 5 minutes, at least 10 minutes, at least 20 minutes, at least 30 minutes, at least 40 minutes, at least 50 minutes, at least 1 hour, at least 2 hours, Hour, more than 5 hours, more than 6 hours, more than 12 hours, and more than 24 hours.
In one embodiment, the drying time may be 240 hours or less, 216 hours or less, 192 hours or less, 168 hours or less, 144 hours or less, 120 hours or less, 96 hours or less, 72 hours or 48 hours or less.
When the drying time in the drying step is within the above range, sufficient drying is performed to increase the anti-inflammatory activity in the production of the island thyme extract.
In one aspect of the present invention, the step of extracting the dried raw material may be performed by extracting the dried raw material with one or more selected from the group consisting of water, an organic solvent, and a mixture of water and an organic solvent. The organic solvent includes, but is not limited to, at least one selected from the group consisting of alcohol, acetone, ether, ethyl acetate, diethyl ether, ethyl methyl ketone and chloroform. Wherein the alcohol comprises a C 1 to C 5 lower alcohol and the C 1 to C 5 lower alcohol is selected from the group consisting of methanol, ethanol, isopropyl alcohol, n-propyl alcohol, n-butanol and isobutanol But is not limited thereto.
In one embodiment, the water may be ultra-pure water.
In one embodiment, the organic solvent may be methanol.
In one embodiment, the mixture of water and an organic solvent may be an aqueous solution of methanol.
In one embodiment, the concentration of the aqueous methanol solution is at least 0.01%, at least 0.1%, at least 1%, at least 5%, at least 10%, at least 15%, at least 20% 30 wt% or more, 35 wt% or more, 40 wt% or more, 45 wt% or more, or 50 wt% or more.
In one embodiment, the concentration of the aqueous methanol solution may be less than or equal to 100 wt%, less than or equal to 90 wt%, less than or equal to 80 wt%, less than or equal to 70 wt%, or less than or equal to 60 wt%.
When the concentration of the aqueous methanol solution is within the above-mentioned range, the anti-inflammatory activity is increased in the production of the island thyme extract.
In one aspect of the present invention, the step of extracting the dried raw material may include a step of concentration under reduced pressure and a freeze-drying step.
In one embodiment, the reduced pressure concentration can be reduced to a pressure of 5 to 50 brix at a temperature of 40 to 90 占 폚.
In one embodiment, the temperature of the reduced pressure concentration may be at least 40 ° C, at least 45 ° C, or at least 50 ° C.
In one embodiment, the temperature for the reduced pressure concentration may be 90 ° C or lower, 85 ° C or lower, 80 ° C or lower, 75 ° C or lower, 70 ° C or lower, 65 ° C or lower, 60 ° C or 55 ° C or lower.
When the concentration under reduced pressure is within the above range, the anti-inflammatory activity is increased in the production of the extract of the isomaltic acid.
The isoameric acid extract prepared through the above process has an anti-inflammatory effect.
In one aspect of the present invention, there is provided a composition comprising as an active ingredient an isotactic extract having an anti-inflammatory effect.
In one embodiment, the extract of Thymus thalassemia is at least 10 ug / mL, at least 20 ug / mL, at least 30 ug / mL, at least 40 ug / mL, at least 50 ug / mL, ML or more, 250 μg / mL or more, 300 μg / mL or more, 350 μg / mL or more, 400 μg / mL or more, 450 μg / mL or more, / mL or more, 700 μg / mL or more, 800 μg / mL or more, 900 μg / mL or more, or 1000 μg / mL or more.
In one embodiment, the islet extract may be 100,000 g / mL or less, 50,000 g / mL or less, 10,000 g / mL or less and 5000 g / mL or less based on the total composition.
If the isoparaef extract is contained within the above range, the anti-inflammatory effect can be effectively obtained without toxicity.
In one embodiment, the composition may be a cosmetic, a food or a pharmaceutical composition.
The pharmaceutical compositions according to the present disclosure may be of various oral or parenteral formulations. 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, soft or hard capsules, etc. These solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose, Or lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.
The pharmaceutical dosage forms of the compositions of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in suitable aggregates. The salt is not particularly limited as long as it is pharmaceutically acceptable so long as it is pharmaceutically acceptable and includes, for example, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, formic acid acetic acid, tartaric acid, lactic acid, citric acid, fumaric acid, , Benzenesulfonic acid, toluenesulfonic acid, and naphthalenesulfonic acid.
The composition of the present invention may be administered parenterally or orally, and may be administered in one to several divided doses so as to be administered in an amount of 0.01 to 500 mg, preferably 0.1 to 100 mg, per 1 kg of body weight per day have. The dosage for a particular patient may vary depending on the patient's body weight, age, sex, health condition, diet, time of administration, administration method, excretion rate, severity of disease, and the like.
The pharmaceutical composition according to the present invention can be administered orally or parenterally in the form of powders, granules, tablets, soft or hard capsules, oral preparations such as suspensions, emulsions, syrups and aerosols, external preparations such as ointments and creams, Sterile injection solutions, and the like, and may be formulated in any form suitable for pharmaceutical preparations.
The composition according to the present invention can be administered to mammals such as rats, mice, livestock, humans, and the like by various routes such as parenteral, oral, and the like, and all manner of administration can be expected. For example, Or by intravenous, intramuscular, subcutaneous, intramural or intracerebroventricular injection.
In one aspect of the invention, the food composition may be a health functional food composition.
The formulation of the food composition according to the present specification is not particularly limited, but may be formulated into, for example, tablets, granules, powders, liquid preparations such as a drink, caramels, gels, bars and the like. The food composition of each formulation can be blended with the ingredients commonly used in the field in addition to the active ingredient without difficulty by those skilled in the art depending on the purpose of formulation or use, and synergistic effect can be obtained when the composition is applied simultaneously with other ingredients.
In the food composition according to the present invention, the determination of the dosage of the active ingredient is within the level of those skilled in the art, and its daily dose is, for example, from 0.1 mg / kg / day to 5000 mg / kg / day, mg / kg / day to 500 mg / kg / day, but it is not limited thereto, and may vary depending on various factors such as the age, health condition, and complication of the subject.
The food composition according to the present invention may be used as a food or beverage such as various foods such as chewing gum, caramel product, candy, ice cream, confectionery, beverage such as soft drink, mineral water, alcoholic beverage, healthful food including vitamins and minerals .
In addition to the above, the food composition which is one aspect of the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and enhancers (cheese, chocolate etc.), pectic acid and its salts, Alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the functional food compositions of the present invention may comprise natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not so critical, but is generally included in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.
The formulation of the cosmetic composition is not particularly limited and may be appropriately selected according to the purpose. For example, skin lotion, skin softener, skin toner, astringent, lotion, milky lotion, moisturizing lotion, nutrition lotion, massage cream, nutritional cream, moisturizing cream, hand cream, foundation, essence, nutrition essence, pack, soap, cleansing But the present invention is not limited thereto, and may be manufactured by any one or more formulations selected from the group consisting of a foam, a cleansing lotion, a cleansing cream, a body lotion and a body cleanser.
When the formulation of the present invention is a paste, cream or gel, animal fiber, plant fiber, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .
When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.
In the case of the solution or emulsion of the present invention, a solvent, a solvent or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.
When the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.
When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolenic derivatives or ethoxylated glycerol fatty acid esters.
The content of the active ingredient is not particularly limited, but may be in the range of 0.001 to 100% by weight based on the total weight of the composition. When the active ingredient satisfies the above content, it can exhibit excellent efficacy without side effects.
The cosmetic composition may further contain ingredients included in a functional additive and a general cosmetic composition. The functional additives may include water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymeric polysaccharides, sphingolipids and seaweed extracts.
The cosmetic composition of the present invention may further contain, in addition to the above-described functional additive, components contained in a general cosmetic composition as required. Examples of the other ingredients that can be included in the composition include humectants, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbents, preservatives, bactericides, antioxidants, plant extracts, pH adjusters, alcohols, Accelerators, coolants, antiperspirants, purified water, and the like.
Hereinafter, the constitution and effects of the present invention will be described in more detail with reference to Examples and Experimental Examples. However, these examples and experimental examples are provided for illustrative purposes only in order to facilitate understanding of the present invention, and the scope and scope of the present invention are not limited thereto.
EXAMPLES Preparation of Thyme Thyme Extract
The dried island thyme toppings were ground with a blender and placed in a 2-L round-shaped flask (inlet size 24) in an amount of 200 g, and each was subjected to reflux extraction by adding 1 L of distilled water and 99.9% methanol. Reflux extraction was carried out in a constant-temperature water bath at 90 ° C and a cooler at -10 ° C for 4 hours, then the extract was transferred to another vessel, and then 1 liter of the same solvent was added thereto. And 2 L of the extract was filtered and used for filtering and concentration.
The collected extract was concentrated under reduced pressure to a concentration of 10 to 20 brix at a temperature of 50 to 60 ° C. The concentrate was frozen at -80 DEG C for 2 hours or more and then lyophilized in a freeze dryer. The lyophilized state was checked every 24 hours, and when the lyophilization was not completed, the ice of the trap was removed and the lyophilized was again dried.
[Experimental Example 1] Toxicity of the extract of isthmus maximus extract to RAW 264.7 cells
The cytotoxicity of extract extracts was investigated in the cells used for the experiment to confirm the proper concentration of the extract for the evaluation of anti - inflammatory activity of the extract.
RAW 264.7 cell is ATTC (American type culture collection, Manassas , VA, USA) 10% FBS purchased from and antibiotics (100 U / mL penicillin, 100 ug / mL streptomycin), 5% of the contained DMEM CO 2 and 95 % Air in a humidified incubator at 37 < 0 > C.
RAW 264.7 cells were cultured in a 6-well plate at a density of 1 × 10 6 cells / well. The concentration of thyme extract was 20, 50, 100, 200, 300, 400 ug / mL. And cultured in a humidified incubator containing 5% CO 2 and 95% air for 24 hours at 37 ° C. After 24 hours, the medium was removed and MTT {3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide} was added to each well at a concentration of 1 mg / Lt; / RTI > MTT medium was removed and 0.5 mL of isopropanol was added to each well to dissolve the blue formazan crystals. The absorbance was then measured at 570 nm on a microplate reader.
The results are shown in Fig. Thymus extracts showed cell viability close to 100% because they did not show toxicity to RAW 264.7 cells at concentrations of 20, 50, 100 and 200 ug / mL. However, at 300 and 400 ug / mL, Survival rate was about 70% and 50%, respectively.
[Experimental Example 2] Effect of LPS-induced NO and PGE2 extracts
RAW 264.7 cells were cultured for 24 hours in the culture medium supplemented with the extracts of thyme and LPS, and the supernatant was collected and stored at -80 ° C. until use. The nitrite concentration in the culture medium was measured according to the Griess reaction (Promega, Madison, WI, USA). The PGE2 concentration in the culture medium was measured using a PGE2 ELISA kit (R & D system, Minneapolis, MN, USA).
(1) Nitrite assay: Measurement using a grease reagent
1) A plate was taken out from an incubator for nitrite assay.
2) The culture supernatant was transferred into a new 96-well plate (70 μl).
3) Griess reagent was taken out from the refrigerator and allowed to stand at
4) 70 μl of a sulfanilamide solution was added to a 96-well plate containing 30 μl of the sample.
5) After 30 minutes, 70 μl of NED (N-1-napthylethylenediamine dihydrochloride) solution was added and left in a dark place for 30 minutes.
6) At the end of the color reaction, the absorbance (OD) value was measured at 520 nm with a microplate spectrophotometer.
(2) PGE2 measurement: ELIA assay
1) Capture antibody (capture anti-body (ab)) of ELISA kit (PGE2) was diluted with dilution factor according to the instruction manual in coating buffer and added to 96-well nunc plate (Nunc cat No. 439454) Lt; RTI ID = 0.0 > 4 C. < / RTI >
2) 200 uL each with washing solution containing 0.05
3) 200 μl of 2% skim milk solution was added and the mixture was blocked at room temperature (RT) for 1 hour.
4) 200 uL each with washing solution.
5) The culture supernatant obtained in the above (1) and the standard solution diluted in steps 1) to 4) were dispensed into each well in an amount of 100 μL, and then incubated in a 37 ° C CO 2 incubator for 2 hours .
6) Washing solution was washed 5 times with 200 uL each.
7) detection anti-body (ab) and streptavidin-HRP were diluted with a dilution factor of the protocol in the kit and 100 uL each was dispensed. After incubation at 37 ° C in a CO 2 incubator, For 1 hour.
8) 200 uL aliquots were washed with washing solution 7 times.
9) TMB substrate solution was dispensed in 100 μl aliquots, and the color development was observed in cows.
10) 2N H 2 SO 4 as a stop solution was dispensed in 100 μl aliquots to stop color development.
11) The absorbance (OD) value was measured using a microplate spectrophotometer at a wavelength of 450 nm within 40 minutes after stopping the color development.
The measurement results are shown in FIG. 2 and FIG. The nitrite concentration was measured by the Griess reaction in the cell culture supernatant. The highest nitrite was produced in the LPS (1 ug / mL) - treated group and the thymus thymus As the concentration of extract increased, the nitrite production was decreased statistically. In addition, the concentration of nitrite in the island thyme alone group was similar to that of the untreated group.
PGE2 concentration was measured by ELISA method. As a result, it was found that PGE2 was not produced in the case of the treatment with isothiocyanate alone, but the concentration of PGE2 was increased as the treatment concentration of the island thyme extract was increased LPS alone treatment group.
[Experimental Example 3] Effect of LPS-induced iNOS and COX-2 on Isolation of Thymus Thymus Extract
To investigate the inhibitory effects of the extracts of thymus thrips on the expression of iNOS and COX-2 in RAW 264.7 cells, pretreatment of thymus thalassemia extract with different concentrations of LPS was carried out for 4 hours for mRNA analysis and 18 hours for protein analysis. (RT-PCR) and Western blot were performed. The primer sequences used for RT-PCR are shown in Table 1.
The measurement results are shown in Fig. As a result, it was confirmed that the expression of mRNA and protein of iNOS and COX-2 induced by LPS was significantly inhibited by the isothiocyanate extract in a concentration-dependent manner. As shown in Experimental Example 2, thymus thymus extract significantly decreased NO and PGE 2 produced by LPS, which was confirmed by the inhibition of iNOS and COX-2 mRNA and protein expression.
[Experimental Example 4] Effect of LPS-induced NF-κB signaling molecule activity on the activity of island thyme extract
In order to confirm whether NF-κB signal transduction pathway is inhibited, LPS was treated for 20 minutes at the concentration of the extract of thymus thymus, and Western blot was performed by extracting the nucleus and cytosol extract.
The results are shown in Fig. As shown in FIG. 5, phosphorylation of NF-κB p65 and p65 migrated into the nucleus when LPS treatment was performed, and it was confirmed that IκBα was degraded. In the case of treatment with the extract of Thymus japonicus at different concentrations, the migration of NF-κB p65 to the nucleus was inhibited and it was observed to inhibit the phosphorylation of p65. In addition, it was confirmed that decomposition of IκBα was remarkably suppressed at a concentration of 20 μM. Thus, the extract of the islet thymus inhibits the expression of iNOS and COX-2 by inhibiting the activity of NF-κB.
[Experimental Example 5] Effect of LPS-induced MAPK activity on the activity of the island thyme extract
Mitogen-activated protein kinase (MAPK) signaling is activated by several signals such as LPS, which is known to affect the activity of NF-kB and AP-1. There are three kinds of MAPK, ERK, p38, and JNK, which are activated by phosphorylation.
Therefore, the activity of LPS - induced MAPK was tested for the efficacy of the extract. When LPS was treated, the phosphorylation of ERK, p38, and JNK was increased by Western blot. The results are shown in Fig. When thymus extracts were treated in the same manner, LPS - induced phosphorylation of p38 and JNK was inhibited in a concentration - dependent manner. However, phosphorylation of ERK was not inhibited.
Experimental Example 6 Effect of LPS-Induced Thyme Extract on the Production of Pro-inflammatory Cytokine
TNF-α, one of the cytokines, plays an important role in triggering inflammatory responses by activating T cells and macrophages and by increasing other pro-inflammatory cytokines. Similarly, IL-6, IL-1β is one of the major inflammatory cytokines secreted by macrophages by LPS and is known to always exhibit high levels of inflammation. Finally, we investigated whether thyme extract inhibits TNF-α, IL-6 and IL-1β, which are inflammatory or inflammatory cytokines, in RAW 264.7 cells using ELISA kit and RT-PCR Protein production and mRNA expression were analyzed. The primer sequences of RT-PCR are shown in Table 1.
The results are shown in Fig. It was confirmed by ELISA assay that the protein production of TNF-α, IL-6 and IL-1β was reduced in a concentration-dependent manner in the case of the extract of thymus thymus (FIG. 7A) As shown in FIG. 7B, the expression of TNF-α, IL-6 and IL-1β mRNA was inhibited when the extract of Ishimaki thymus was treated at different concentrations as in the case of protein.
Reverse: TGGTCAAACTCTTGGGGTTC
Reverse: GCTCGGCTTCCAGTATTGAG
Reverse: CATTCGAGGCTCCAGTGAAT
Reverse: TGGATGGTCTTGGTCCTTAGCC
Reverse: TACCAGTTGGGGAACTCTGC
Reverse: GGACAGTGAGGCCAGGATGG
Hereinafter, as described above, a formulation example of a composition having anti-inflammatory effect according to an aspect of the present invention will be described below. However, the present invention can be applied to various other formulations, It is.
[Formulation Example 1] Health food
Island Thyme Extract ................... 1000 mg
Vitamin mixture
Vitamin A Acetate ............... 70 ㎍
Vitamin E ....................... 1.0 mg
Vitamin B1 ...................... 0.13 mg
Vitamin B2 ...................... 0.15 mg
Vitamin B6 ........................ 0.5 mg
Vitamin B12 ....................... 0.2 g
Vitamin C .......................... 10 mg
Biotin ............................. 10 μg
Nicotinic acid amide ... 1.7 mg
Folic acid ............................... 50 ㎍
Calcium pantothenate ..................... 0.5 mg
Mineral mixture
Ferrous sulfate ........................ 1.75 mg
Zinc oxide .......................... 0.82 mg
Magnesium carbonate ...................... 25.3 mg
Potassium Phosphate ......................... 15 mg
Secondary Calcium Phosphate ..................... 55 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 the ingredient suitable for health food as a preferred embodiment, the compounding ratio thereof may be arbitrarily modified.
[Formulation Example 2] Health drinks
Island Thyme Extract .......................... 1000 mg
Citric acid ................................ 1000 mg
Oligosaccharides ................................ 100 g
Taurine .................................... 1 g
Purified water .................................. Remaining amount
The above components are mixed according to a conventional health drink manufacturing method, and the mixture is stirred and heated at 85 DEG C for about 1 hour, and then the solution is sterilized by filtration.
[Formulation Example 3] Tablets
The granules were prepared by mixing 100 mg of islet extract, 50 mg of soybean extract, 100 mg of glucose, 50 mg of red ginseng extract, 96 mg of starch and 4 mg of magnesium stearate and 40 mg of 30% ethanol, And tableted using a tablet machine.
[Formulation Example 4]
100 mg of thyme extract, 50 mg of soybean extract, 100 mg of glucose, and 600 mg of starch are mixed and 100 mg of 30% ethanol is added to form granules, which are then dried at 60 ° C. to form granules,
<110> HONGCHEON INSTITUTE OF MEDICINAL HERB <120> Composition for anti-inflammation containing an extract of Thymus quinquecostatusvar.japnica Hara <130> 14P744 <150> KR 10-2014-0190740 <151> 2014-12-26 <160> 12 <170> KoPatentin 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer sequence for RT PCR of iNOS (Forward) <400> 1 ttccagaatc cctggacaag 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer sequence for RT PCR of iNOS (reverse) <400> 2 tggtcaaact cttggggttc 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer sequence for RT PCR of COX-2 (forward) <400> 3 agaaggaaat ggctgcagaa 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> Primer sequence for RT PCR of COX-2 (reverse) <400> 4 gctcggcttc cagtattgag 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer sequence for RT PCR of TNF-a (forward) <400> 5 agcccccagt ctgtatcctt 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> Primer sequence for RT PCR of TNF-a (reverse) <400> 6 cattcgaggc tccagtgaat 20 <210> 7 <211> 25 <212> DNA <213> Artificial Sequence <220> Primer sequence for RT PCR of IL-6 (forward) <400> 7 caagaaagac aaagccagag tcctt 25 <210> 8 <211> 22 <212> DNA <213> Artificial Sequence <220> Primer sequence for RT PCR of IL-6 (reverse) <400> 8 tggatggtct tggtccttag cc 22 <210> 9 <211> 20 <212> DNA <213> Artificial Sequence <220> Primer sequence for RT PCR of IL-1B (forward) <400> 9 gggcctcaaa ggaaagaatc 20 <210> 10 <211> 20 <212> DNA <213> Artificial Sequence <220> Primer sequence for RT PCR of IL-1B (reverse) <400> 10 taccagttgg ggaactctgc 20 <210> 11 <211> 25 <212> DNA <213> Artificial Sequence <220> Primer sequence for RT PCR of B-actin (forward) <400> 11 agtgtgacgt tgacatccgt aaaga 25 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> Primer sequence for RT PCR of B-actin (reverse) <400> 12 ggacagtgag gccaggatgg 20
Claims (9)
Wherein said Thyme Thyroid Extract inhibits the production of at least one of nitrite and PGE2.
Wherein said isoflavone extract inhibits the expression of at least one of iNOS and COX-2.
The isoflavone extract inhibits the NF-κB signaling pathway.
The islet thyme extract inhibits mitogen-activated protein kinase (MAPK) signaling.
The isoflavone extract inhibits inflammatory cytokines.
Wherein said inflammatory cytokine is at least one of TNF-a, IL-6 and IL-l [beta].
Wherein said isoflavone extract inhibits the expression of said inflammatory cytokine.
The anti-inflammatory composition is a cosmetic, a food or a pharmaceutical composition.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020140190740 | 2014-12-26 | ||
| KR20140190740 | 2014-12-26 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| KR20160079608A true KR20160079608A (en) | 2016-07-06 |
Family
ID=56502535
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020150084523A KR20160079608A (en) | 2014-12-26 | 2015-06-15 | Composition for anti-inflammation containing an extract of Thymus quinquecostatusvar.japnica Hara |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR20160079608A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20200002567A (en) * | 2018-06-29 | 2020-01-08 | 주식회사 인투바이오 | Cosmetic composition containing botanical extract complex |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20050089452A (en) | 2004-03-05 | 2005-09-08 | 경상북도농업기술원생물자원연구소 | Perfume using essential oil of tymus quinquecostatus var.japonica hara and manufacturing process of the same |
-
2015
- 2015-06-15 KR KR1020150084523A patent/KR20160079608A/en not_active Application Discontinuation
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20050089452A (en) | 2004-03-05 | 2005-09-08 | 경상북도농업기술원생물자원연구소 | Perfume using essential oil of tymus quinquecostatus var.japonica hara and manufacturing process of the same |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20200002567A (en) * | 2018-06-29 | 2020-01-08 | 주식회사 인투바이오 | Cosmetic composition containing botanical extract complex |
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