KR20160001880A - An Extract of Oxya chinensis sinuosa Having Anti-inflammatory and Immuno-modulating Activity and Composition Comprising the Same - Google Patents
An Extract of Oxya chinensis sinuosa Having Anti-inflammatory and Immuno-modulating Activity and Composition Comprising the Same Download PDFInfo
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Abstract
The present invention relates to an extract of rice plant ( Oxya chinensis sinuosa ) having anti-inflammatory activity and a composition containing the same.
The present invention relates to a pharmaceutical composition for preventing and treating inflammatory diseases, which comprises the extract of rice plant ( Oxya chinensis sinuosa ) obtained by extracting with ethanol, There is provided a food composition for improving an inflammatory disease.
The rice-grasshopper extract having anti-inflammatory activity according to the present invention and a composition comprising the same are natural insect-derived natural resources and have excellent safety because they are not cytotoxic. In addition, they inhibit the expression of inflammation-related factors and have anti-inflammatory effects, Which can be used as a prophylactic and therapeutic agent for inflammation.
Description
The present invention relates to an extract of Oxya chinensis sinuosa having anti-inflammatory activity and a composition containing the same, and more particularly, to a plant extract obtained by extracting oxya chinensis sinuosa with ethanol and a composition containing the extract will be.
The inflammatory reaction is the first innate immunity reaction to protect the living body from injury and external foreign material infiltration (Non-Patent Document 8), and can be divided into acute inflammation and chronic inflammatory reaction according to the action and duration of inflammation .
Acute inflammation is caused by inflammatory mediators such as inflammatory cytokines, chemokines, nitric oxide (NO) and prostaglandin E2 (PGE2), which are produced by macrophage activation by external stimuli. (Fig. 1), and then regenerated by regeneration of tissues after removing foreign substances by macrophages and leukocytes activated by the microorganisms (Non-Patent Document 1).
Acute inflammation is an essential reaction to defend the body, but excessive inflammatory mediators in this reaction can cause acute septic shock due to excessive vasodilation and cause continuous survival and activation of immune cells, resulting in chronic inflammation or immune Inflammatory diseases such as rheumatoid arthritis can develop into autoimmune diseases (Non-Patent
Immune cells include macrophages, monocytes, leukocytes, and mast cells. Among them, macrophages are present in all tissues. They first recognize the cause of inflammation in the body and then produce an inflammatory mediator to activate and activate other immune cells. It plays an essential role in the inflammation process, such as inducing chemotaxis, eliminating the cause of inflammation, and restoring the inflammatory site.
TLR-4 (toll like receptor-4) on the surface of macrophages during LPS treatment, which is a cell wall component of Gram-negative bacteria belonging to pathogen-associated molecular patterns (PAMPS) ) And MYD88 (myeloid differentiation primary response gene 88), activating MYD88 protein binds IRR-4 (IL-1 receptor associated kinase-4) to TLR-4 and TRAF-6 (TNF receptor associated factor- 6).
TRAF-6 activates IκK (I kappaB kinase), which degrades IκBα bound to NF-κB p50 / p65, a cytoplasmic transcription factor, resulting in the transfer of NF-κB p50 / p65 to the nucleus. Nuclear translocated NF-κB p50 / p65 transcription factors express the inflammatory cytokines TNF-α and IL-6 and inflammatory enzymes iNOS and COX-2 (
TNF-α binds to TNF-α receptor (TNFR) of its own and other macrophages via endocrine and exocrine ligands and activates NF-κB p50 / p65 to induce inflammatory cytokines TNF-α and IL- And iNOS and COX-2 (
IL-6 activates the JAK-STAT (Janus kinase-signal transducer and activator of transcription) signal transduction and stimulates T cells and B cells, which are lymphocytes involved in white blood cells and acquired immunity, to convert acute inflammation into chronic inflammation (Non-Patent
NO is produced by iNOS and normally acts as a defense mechanism, neurotransmitter, and vascular regulation. However, NO produced during inflammation can cause acute septic shock and has a strong toxicity, Causing chronic inflammation and autoimmune diseases (Non-Patent Documents 12 and 14).
COX-2 plays an important role in the inflammatory response by converting arachidonic acid derived from the phospholipid of the cell membrane into PGE2, increasing vasodilation and permeability, and acting on sensory neurons to produce pain Non-Patent
Oxya chinensis sinuosa is an incomplete metamorphic insect in the grasshopper and grasshopper, also called our rice-grasshopper. Its body length is 21-35 mm and appears in August-October, and has been used for a long time in Korea for domestic use (Non-Patent Document 18).
However, a study on the function of the rice-grasshopper has not been conducted except for the antioxidative activity against SH-SY5Y, which is an oxidative stress-inducing neuroblastoma cell line, and the suppression of COX-2 transcription (Non-Patent Document 16).
Accordingly, the inventors of the present invention have completed the present invention by confirming the anti-inflammatory effect of rice hogwort by treating the RAW 264.7 cells with LPS-induced RAW 264.7 cells as a part of a functional study on the rice-grasshopper used for food.
It is an object of the present invention to provide a rice-grasshopper extract having anti-inflammatory activity and a composition containing the same.
It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not intended to limit the invention to the particular embodiments that are described. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not restrictive of the invention, There will be.
In order to attain the above object, the present invention is characterized in that the extract of rice plant having anti-inflammatory activity is obtained by extracting the rice plant ( Oxya chinensis sinuosa ) with ethanol.
The rice grasshopper extract inhibits the inflammatory cytokines TNF-a and IL-6, inhibits the expression of inflammatory proteins such as cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) It is characterized by inhibiting the activation of NF-kB p65.
The pharmaceutical composition for preventing and treating inflammatory diseases according to the present invention is characterized by containing the above-mentioned rice plant extract ( Oxya chinensis sinuosa ) as an active ingredient.
Characterized by further comprising a carrier, an adduct or a diluent conventionally used in the production of the pharmaceutical composition.
The rice grasshopper extract of the composition is contained in an amount of 1000 to 2000 占 퐂 / ml.
The food composition for improving inflammation according to the present invention is characterized by containing the above-mentioned Oxya chinensis sinuosa extract as an active ingredient, and the above-mentioned rice grasshopper extract is contained in an amount of 1000 to 2000 占 퐂 / ml.
The composition containing the rice-grasshopper extract having anti-inflammatory activity according to the present invention is a natural resource derived from insects, which is not cytotoxic and has excellent safety. In addition, the composition inhibits the expression of inflammation-related factors and has an anti-inflammatory effect, It can be used as a prophylactic and therapeutic agent for inflammation.
FIG. 1 is a graph showing the cell survival rate after treatment of RAW 264.7 cells with a concentration of a rice plant extract. FIG.
FIG. 2 is a graph showing the results of the relative quantitative analysis of the inhibitory effect of the inflammatory cytokines TNF-.alpha. And IL-6 after the real-time PCR and the ddCT method.
(A) TNF-α mRNA
(B) IL-6 mRNA
FIG. 3 is a graph showing the inhibitory effect of the inflammatory cytokines TNF-.alpha. And IL-6 treated with rice plant extracts by ELISA analysis.
(A) TNF-a protein
(B) IL-6 protein
FIG. 4 is a graph showing the inhibitory effect of NF-κB p65 on the activation of rice plant extracts.
FIG. 5 is a graph showing the inhibitory effect of iNOS and COX-2 by treating a rice plant extract. FIG.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail, and a detailed description of known functions and configurations that may unnecessarily obscure the gist of the present invention will be omitted.
The invention comprises byeomettugi (Oxya chinensis sinuosa) having anti-inflammatory activity extract and this by checking the efficacy of the inflammation which may cause various diseases to find the new value of byeomettugi (Oxya chinensis sinuosa) which have been used only as a food Thereby providing a composition.
First, the rice-grasshopper extract having anti-inflammatory activity can be prepared as follows.
For example, Oxya chinensis sinuosa was lyophilized to remove moisture and then pulverized into powder form. The powder was then dissolved in 70% ethanol at a volume of 2 to 10 times the weight of the powder and dissolved by ultrasonic extraction And then centrifuged to obtain a rice hull extract.
The extract of the rice plant of the present invention obtained by the above-mentioned method of the present invention was used to examine the inhibitory effect of inflammatory cytokine (TNF-α, IL-6) and the inflammatory expression protein (cyclooxygenase- 2) and inducible nitric oxide synthase (iNOS)) and inhibition of NF-kB p65 activation. As a result, it was confirmed that the anti-inflammatory effect was excellent.
Accordingly, the present invention provides a pharmaceutical composition for preventing or treating an inflammatory disease or a food composition for improving an inflammatory disease, which contains the rice plant extract having anti-inflammatory activity as an active ingredient as described above.
At this time, the rice plant extract of the composition is contained at 1000 to 2000 占 퐂 / ml, and preferably the rice plant extract of 2000 占 퐂 / ml is included in the composition.
When the composition is contained at less than 1000 μg / ml, the inhibitory effect of inflammatory cytokines (TNF-α, IL-6) is low and it is insufficient to obtain the desired anti-inflammatory effect. When the composition contains more than 2000 μg / ml There is a fear that the added synergistic effect will not be obtained as compared with the case where the concentration is 2000 占 퐂 / ml.
Thus, when the composition of the present invention is used as an active ingredient of a pharmaceutical composition for the purpose of prevention and treatment of an inflammatory disease, it can be used for the treatment of inflammation such as rheumatoid arthritis, asthma, acute pain, pain such as chronic pain, migraine and joint pain, It becomes possible to prevent and treat related diseases. In this case, according to a method which can be easily carried out by a person having ordinary skill in the art to which the present invention belongs, the preparation is carried out in the form of a unit dose form by using a pharmaceutically acceptable carrier or excipient, .
The formulations may be in the form of tablets, capsules, powders, granules, liquids or rings, and may additionally contain dispersing or stabilizing agents. That is, it can be selected without difficulty by those skilled in the art depending on the purpose of use, and the addition amount thereof can be selected within a range that does not impair the purpose and effect of the present invention.
Examples of the pharmaceutically acceptable carriers include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline Cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and also pharmaceutically acceptable excipients Include, but are not limited to, lubricants, wetting agents, sweeteners, flavoring agents, emulsifying agents, suspending agents, preservatives and the like.
In another aspect, when the composition of the present invention is used as an active ingredient of a food composition for the purpose of improving inflammatory diseases, the composition may be used as it is or may be used in combination with components of other conventional food compositions, Can be suitably used. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). In general, when the food composition is prepared, it is added in an amount of 0.01 to 10 parts by weight, preferably 0.05 to 1 part by weight, based on the raw material of the active ingredient.
However, in the case of long-term consumption intended for health or hygiene purposes or for health control purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.
Such a food composition may be contained in a health food for the purpose of improving and preventing inflammation, and the kind thereof is not particularly limited. Examples of the food to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy foods in a conventional sense.
In addition to the above, the food composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, Alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the food composition of the present invention may be contained as pulp for the production of natural fruit juice, fruit juice drink and vegetable drink. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for illustrating the present invention and that the scope of the present invention is not construed as being limited by these embodiments.
< Example 1> Manufacture of rice-grasshopper extract
Byeomettugi was used as the byeomettugi (Oxya sinuosa chinensis) collected in Kangwon Province Yeongwol youngwoleup youngheungri.
The rice grasshopper was washed twice with running water, dried using a freeze dryer (Eyela, Japan) to remove water, and then pulverized with a multifunction grinder (Korea Medi, Korea) to obtain a rice grasshopper powder.
After dissolving in the 70% ethanol, the thus obtained rice flour was dissolved in 70% ethanol and pulverized with an ultrasonic wave shaker (LabaX, MA, USA) for 20 seconds at 230 joules. After centrifugation at 4500 rpm for 10 minutes, the supernatant was obtained and 0.25 μm syringe After filtration with a filter (Whatman, ND, USA) and drying, the rice grasshopper extract of the present invention ( Oxya chinensis sinuosa ethanol extract, OCE) were prepared.
< Experimental Example 1> Confirmation of the cytotoxicity of the grasshopper extract
The cell survival rate of the rice plant extract ( Oxya chinensis sinuosa ethanol extract, OCE) of Example 1 was measured by RAW 264.7 macrophages (100, 500, 1000, 2000 μg / ml).
1) Experimental method
RAW 264.7 cells were cultured in medium supplemented with penicillin-streptomycin (100 units / ml) and 10% fetal bovine serum (Gibco, MD, USA) Using a 37 ° C, 5% CO 2 incubator (incubator; Theromo Scientific, IL, USA) and subcultured at 2-day intervals.
To measure the cell survival rate, RAW 264.7 cells were seeded in 96-well plates at 2 × 10 5 cells / ml, and treated with the concentrations of 100, 500, 1000 and 2000 μg / ml of the grasshopper extract prepared in Example 1 , a control group using the well-known turmeric extract as anti-inflammatory substance (obtained by extraction in the same manner as in example 1), 37 ℃, 5
(Microplate Reader, Beckman Coulter, CA, USA) at 37 ° C in a 5% CO 2 incubator for 4 hours after adding CellTiter 96 ® aqueous non-radioactive cell proliferation assay reagent (Promega, And the absorbance was measured at 450 nm to measure cell viability.
2) Experimental results
As shown in FIG. 1, it was confirmed that the extract of rice plant ( Oxya chinensis sinuosa ethanol extract, OCE) did not affect the cell growth up to the concentration of 2000 μg / ml and was not toxic.
In the present experiment, C. longa , which has been reported to have anti-inflammatory activity, was used as a positive control. The ethanol extract (CLE) was prepared in the same manner as in the case of the turmeric sample used in the present experiment and 1000 μg / And no cytotoxicity was observed up to the concentration. Therefore, the highest concentrations of oxya chinensis sinuosa ethanol extract (OCE) and turmeric extract (CLE) used in the following experiments were 2000 and 1000 μg / ml, respectively, which do not show toxicity to RAW 264.7 cells.
< Experimental Example 2> Influence on the expression of inflammatory cytokines in rice plant extracts
To investigate the anti-inflammatory effect of rice plant extracts ( Oxya chinensis sinuosa ethanol extract, OCE), RAW 264.7 cells were treated with LPS for inducing inflammation, and the expression of inflammatory cytokines was measured by treatment with Oxye chinensis sinuosa ethanol extract (OCE) And protein levels, respectively, by real-time PCR and ELISA
1) Experimental method
① Real-time PCR analysis
Real-time PCR was performed to verify the effect of rice plant extracts on the transcription of inflammatory cytokine genes. RAW 264.7 cells were stimulated with 100 ng / ml of LPS (Sigma, MO, USA) and the rice plant extracts of Example 1 were treated with concentrations (100, 500, 1000, 2000 / / ml) The extract of turmeric was used at a concentration of 1000 / / ml, which does not show cytotoxicity, at 37 ° C, 5% CO 2 incubator for 24 hours.
After washing the cells twice with PBS, total DNA was extracted from the cells using trizol reagent (Invitrogen, CA, USA) and cDNA was synthesized using high capacity cDNA reverse transcription kit (Applied Biosystems, IL, USA).
The synthesized cDNA is stylized amplifying the TNF-α and IL-6 gene using the Power SYBR ® GreenMasenMix (Applied Biosystems, IL, USA) in 7500 real-time Thermal Cycler (Applied Biosystems, IL, USA) using, this time with The primers are shown in Table 1 below.
Using GAPDH (Glyceraldehyde 3-phospate dehydrogenase) as an endogenous control, the amount of TNF-α and IL-6 mRNA expression relative to GAPDH was analyzed by ddCT method.
② ELISA (Enzyme-linked immunosorbent assay) analysis
RAW 264.7 cells were stimulated with 100 ng / ml of LPS to examine the effect of the rice plant extract on the production of inflammatory cytokines such as TNF-a and IL-6, and the rice plant extracts of Example 1 , 1000, was treated with 2000 ㎍ / ㎖), as a control group was treated with
TNF-α and IL-6 cytokine production was quantitated using a microplate reader (Beckman Coulter, CA, USA) using an ELISA kit (Thermo Scientific, IL, USA) And measured at a wavelength of 450 nm.
2) Experimental results
As shown in FIGS. 2 and 3, TLR-4 present on the surface of macrophages during LPS treatment, which is a kind of PAMPS, recognizes and activates TLR-4 to express inflammatory mediators. Therefore, TNF-α and IL-6, which are inflammatory cytokines, The level of expression was confirmed at the transcript level.
That is, in the case of TNF-α was increased about 7.5 times the amount of expression when compared to the LPS treated LPS non-treatment group, with
IL-6 was approximately 21.4 times the gene expression compared to non-treatment group in the LPS-only treatment group increased, LPS and 2000 ㎍ / ㎖ byeomettugi extract (Oxya chinensis sinuosa ethanol extract, OCE) was inhibited by about 10.4-fold over the LPS-treated group (FIGS. 2B and 3B).
TNF-α and IL-6 were compared with LPS alone at a concentration of 1000 ㎍ / ㎖, which did not show cytotoxicity. , And the same concentration of rice plant extract ( Oxya chinensis sinuosa ethanol extract, OCE) with LPS co-processed with about 3.1 times the TNF-α, IL-6 is about 2.4 times because the expression is reduced byeomettugi in macrophage inflammation induced extracts (Oxya chinensis sinuosa ethanol extract, OCE ) is It was confirmed that the inhibition of inflammatory cytokine expression is more strongly inhibited than that of the turmeric extract (CLE).
<Experimental Example 3> Inhibition of activation of NF-κB p65 by rice plant extracts
1) Experimental method
RAW 264.7 cells were stimulated with LPS (Sigma, MO, USA) at a concentration of 100 ng / ml, and the rice plant extracts were added to the culture medium to determine the efficacy of rice plant extracts against the inflammatory-related transcription factor NF- and then treated with (100, 500, 1000, 2000 ㎍ / ㎖) 37 ℃, 5
Cultured cells were washed twice with PBS (phosphate buffered saline, Sigma, MO, USA) and fixed with formaldehyde (Sigma, MO, USA) for 25 min at room temperature.
The immobilized cells were incubated with 5% horse serum (Vector Labs, CA, USA) at room temperature using 0.01% Triton X-100 (Sigma, MO, USA) For 1 hour.
NF-κB p65 primary antibody (Santa Cruz, CA, USA) was diluted at a ratio of 1: 1000 and incubated at 4 ° C overnight. The cells were washed four times with PBS for 10 minutes at room temperature and diluted 1: And reacted with antibody alexa fluor 488 (Molecular Probes, MD, USA) for 2 hours at room temperature.
The reacted cells were washed four times with PBS for 10 minutes at room temperature, and then attached to a slide glass with a VECTA SHIELD (Vector Labs, CA, USA) solution and confirmed with a DMI6000B fluorescence microscope (Leica, NJ, USA).
2) Experimental results
Rice grasshopper extract ( Oxya chinensis sinuosa ethanol extract, OCE) and inhibited the activation of NF-κB p65 upstream of the inflammatory mediator during activation of LPS and TNF-α. In RAW 264.7 cells induced by LPS (100, 2000 ㎍ / ㎖), and the intracellular location of NF-κB p65 was confirmed by immunofluorescence staining.
When NF-κB p65 is not induced by inflammation, it is present in the cytoplasm in association with IκBα and can not activate inflammatory mediator-related gene transcription. However, when signaling by LPS and TNF-α is activated, IκBα is degraded NF-kB p65 translocates to the nucleus and binds to the NF-kB element present in the promoter of the inflammatory mediator gene to activate transcription.
Thus, as shown in Fig. 4, the RAW 264.7 cells induced with inflammation were injected with 2000 占 퐂 / ml rice plant extract ( Oxya chinensis sinuosa NF-κB p65 was detected in NF-κB p65 by Oxycholesterol extract (OCE), which was present in the cytoplasm without migration to the nucleus. Activation was suppressed.
In addition, since NF-κB p65 activity is inhibited, it is expected to further block the production of other inflammatory cytokines such as IL-1β and chemokines that increase the chemotaxis of immune cells.
< Experimental Example 4> Confirmation of inhibition of iNOS and COX-2 protein expression by rice plant extracts
1) Experimental method
Byeomettugi extract on the production of the inflammatory enzymes iNOS and COS-2 (Oxya chinensis sinuosa ethanol extract, OCE) were treated with 100, 500, 1000 and 2000 ㎍ / ㎖ of RAW 264.7 cells with 100 ng / ㎖ of LPS at a concentration of 100 ng / 2 After incubation in an incubator for 24 hours, proteins were separated from the cells and subjected to Western blot analysis.
Cells were washed twice with PBS and collected in a cell scraper (SPL Life Science, Korea). The cells were then pulverized using Cytobuster ™ Protein Extraction Reagent (Novagen, ND, USA) and centrifuged at 12000 rpm for 15 minutes. Respectively.
The recovered supernatant was measured by Bio Rad Protein Assay (Bio Rad, CA, USA), adjusted to a concentration of 10 μg / ml and subjected to 10% SDS-PAGE. After electrophoresis, the PVDF membrane (GE Healthcare, NJ , USA) and blotting grade blocker (Bio Rad, CA, USA) was dissolved in TBST to a concentration of 5%. Then, the PVDF membrane was blocked for 1 hour at room temperature.
Actin (Sigma, MO, USA) was diluted at a ratio of 1: 10000 to 1: 1000 and the primary antibody COX-2 (Santa Cruz, CA, USA) and iNOS After overnight incubation, the cells were washed four times with TBST for 15 minutes each day. The secondary antibody HRP-conjugated secondary antibody (Promega, WI, USA) was reacted at a ratio of 1: 3000 for 40 minutes at room temperature. Lt; / RTI >
The Western Lightning ® Plus ECL (Perkin Elmer, MA, USA) by the X-ray photosensitive film (Fujifilm, Japan) in the dark and then treated in the PVDF membrane was confirmed that the protein expression of iNOS and COX-2.
2) Experimental results
In order to confirm the anti-inflammatory effect by inhibiting the expression of iNOS and COX-2, the inflammatory enzymes activated by TLR-4 (toll like receptor-4) on the surface of macrophages during LPS treatment, byeomettugi the RAW 264.7 cell extracts (oxya chinensis sinuosa ethanol extract, OCE) at 100, 500, 1000 and 2000 ㎍ / ㎖, respectively, and the amount of iNOS and COX-2 protein expression was confirmed by Western blot analysis.
As a result, as shown in Fig. 5, both of the iNOS and COX-2 proteins were treated with 2000 / / ml rice plant extract ( Oxya chinensis sinuosa ethanol extract, OCE) significantly reduced protein expression.
NO plays a protective role against vasodilation and external foreign substances in the inflammatory reaction, but NO excessively secreted by inflammatory cytokines causes DNA damage, inflammation amplification through inflammatory mediator hyperplasia, septic shock due to excessive vasodilation, Cell and tissue necrosis due to strong cytotoxicity, dysfunction, and the like, and may cause chronic inflammation and autoimmune diseases.
It has also been reported that NO is closely related to tumorigenesis through VEGF (angiogenesis effect) with COX-2.
In this experiment, the expression of iNOS protein was confirmed by Western blot analysis after treatment of rice-derived RAW 264.7 cells with Oxya chinensis sinuosa ethanol extract (OCE), since NO produced by inflammation reaction can be indirectly confirmed by iNOS production Respectively. As a result, 2000 / / ml rice plant extract ( Oxya chinensis sinuosa The expression of iNOS was induced by the activation of NF-kB p65 by LPS. Therefore, the activity of NF-kB p65 was blocked at this concentration and iNOS (Fig. 5).
NF-kB p65 and NO-produced COX-2 are enzymes that produce PGE2 in the arachidonic acid produced by phospholipid metabolism during inflammation, while PGE2 produced by COX-2 acts on vasodilatation, pain and fever .
Irradiation- induced RAW 264.7 cells were injected with 2000 占 퐂 / ml rice plant extract ( Oxya The reduction of COX-2 in chinensis sinuosa ethanol extract (OCE) treatment seems to be due to the marked decrease in the expression of TNF-α and NO-producing iNOS protein through inhibition of NF-kB p65 activation at this concentration .
A number of NSAIDs have been invented since the discovery that aspirin and indomethacin, a class of non-steroidal anti-inflammatory drugs (NSAIDs), have anti-inflammatory effects through the inhibition of COX-2 expression Have been used as anti-inflammatory drugs.
However, recent studies have shown that long-term administration of NSAIDs can cause gastrointestinal and renal dysfunction, and in particular, several side effects related to cardiovascular events have been reported by inhibiting the production of PGEs and prostaglandins (prostacyclin) that expand blood vessels.
Therefore, recently, COX-2 inhibitors derived from natural products are more absorbed and metabolized in the living body than NSAIDs, and therefore, the side effects are considered to be less. Thus, it has been reported that a combination of silymarin derived from milk thistle and six natural herbs Efforts are being made to find COX-2 inhibitors in natural products such as shinbaro.
Therefore, the extract of rice plant ( Oxya chinensis sinuosa ethanol extract, OCE) used in the present invention is a natural product derived from insects and inhibits the expression of inflammatory cytokines such as TNF-α and IL-6, iNOS and COX-2 and activates NF- It is anticipated that it can be developed as an anti-inflammatory substance derived from natural materials.
It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the inventions. Examples and experiments may be implemented. The scope of the present invention is defined by the appended claims, and all differences within the scope of the claims are to be construed as being included in the present invention.
Claims (7)
Mushroom extracts with anti-inflammatory activity.
The rice grasshopper extract inhibits the inflammatory cytokines TNF-a and IL-6, inhibits the expression of inflammatory proteins such as cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) Lt; RTI ID = 0.0 > NF-kB < / RTI > p65,
Mushroom extracts with anti-inflammatory activity.
A pharmaceutical composition for the prevention and treatment of inflammatory diseases.
A pharmaceutical composition for the prevention and treatment of inflammatory diseases.
Wherein said rice plant extract is contained in an amount of 1000 to 2000 占 퐂 / ml.
A pharmaceutical composition for the prevention and treatment of inflammatory diseases.
A food composition for improving inflammatory diseases.
Wherein said rice plant extract is contained in an amount of 1000 to 2000 占 퐂 / ml.
A food composition for improving inflammatory diseases.
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| KR102050287B1 (en) * | 2018-07-20 | 2019-11-29 | 재단법인 전라북도생물산업진흥원 | Composition for relieving skin itching using grasshopper powder, preparation method thereof, cosmetics and feed containing the same |
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| KR20200048057A (en) | 2018-10-29 | 2020-05-08 | 대한민국(농촌진흥청장) | Antiinflammatory composition comprising Locusta migratoria extract |
| KR20210011051A (en) | 2018-10-25 | 2021-01-29 | 대한민국(농촌진흥청장) | Composition comprising Locusta migratoria extract for preventing or treating Neuritis |
| KR20210060233A (en) * | 2019-11-18 | 2021-05-26 | 세종대학교산학협력단 | Pharmaceutical composition for treating or preventing leaky gut syndrome |
-
2014
- 2014-06-27 KR KR1020140079824A patent/KR20160001880A/en active Search and Examination
Non-Patent Citations (18)
| Title |
|---|
| .2005.Advanced glycation in health and disease role of the modern environment. Ann N Y Acad Sci 1043, 452-460. |
| Baldwin, A.S. Jr. 1996. The NF-kappa B and I kappa B proteins: new discoveries and insights. Annu Rev Immunol 14, 649-683. |
| Cerella, C., Sobolewski, C., Dicato, M. and Diederich, M. 2010. Targeting COX-2 expression by natural compounds: a promising alternative strategy to synthetic COX-2 inhibitors for cancer chemoprevention and therapy. Biochem Pharmachol 15, 1801-1815. |
| Ferrero-Miliani, L., Nielsen, O.H., Andersen, P.S. and Girardin, S.E. 2007. Chronic inflammation: Importance of NOD2 and NALP3 in interleukin-1beta generation. 147, 227-235. |
| Gabay, C. 2006. Interleukin-6 and chronic inflammation. Arthritis Research & Therapy 8, S3. |
| Giuliani, C., Napolitano, G., Bucci, I., Montani, V. and Monaco, F. 2001. NF-κB transcription factor: role in the pathogenesis of inflammatory, autoimmune, and neoplastic diseases and therapy implications. Clin Ter 152, 249-253. |
| Hink, U. and M, T. 2006. COX-2, another important player in the nitric oxide-endothelin cross-talk: good news for COX-2 inhibitors? Circ Res 98, 1344-1346. |
| Iwalewa, E.O., McGaw, L.J., Naidoo, V. and Eloff, J.N. 2007. Inflammation: the foundation of diseases and disorders. A review of phytomedicines of south african origin used to treat pain and inflammatory conditions. 6, 2868-2885. |
| Kang, C.H., Choi, Y.H., Choi, I.W., Lee, J.D. and Kim, G.Y. 2011. Inhibition of lipopolysaccharide-induced iNOS, COX-2, and TNF-α expression by aqueous extract of Orixa Japonica in RAW 264.7 cells via suppression of NF-κB activity. Trop J Pharm Res10, 161-168. |
| Lee, S.Y., Kwon, H.K. and Lee, S.M. 2011. SHINBARO, a new herbal medicine with multifunctional mechanism for joint disease: first therapeutic application for the treatment of osteoarthritis. 34, 1773-1777. |
| Medzhitov, R. 2008. Origin and physiological roles of inflammation. Nature 454, 24. |
| Miyataka, M., Rich, K.A., Ingram, M., Yamamoto, T. and Bing, R.J. 2002. Nitric oxide, anti-inflammatory drugs on renal prostaglandins and cyclooxygenase-2. Hypertension 39, 785-789. |
| Nakanishi, M. and Rosenberg, D.W. 2013. Multifaceted roles of PGE2 in inflammation and cancer. Semin Immunopathol 35, 123-137. |
| Paek, M.K., Hwang, J.M., Jung, K.S., Kim, T.W., Kim, M.C., Lee, Y.J., Cho, Y.B., Park, S.W., Lee, H.S., Ku, D.S., Jeong, J.C., Kim, K.G., Choi, D.S., Shin, E.H., Jwang, J.H., Lee, J.S., Kim, S.S. and Bea, Y.S. 2010. Checklist of Korean insects. Nature & Ecology pp. 36. |
| Park, J.Y., Heo, J.C., Woo, S.U., Yun, C.Y., Kang S.W., Hwang, J.S. and Lee, S.H. 2006. Anti-inflammatory and cellular protective effects on hydrogen peroxide-induced cytotoxicity of grasshopper extracts. Preserv 13, 796-802. |
| Tak, P.P. and Firestein, G.S. 2001. NF-kappaB: a key role in inflammatory diseases. J Clin Invest 107, 7-11. |
| Tracey, D., Klareskog, L., Sasso, E.H., Salfeld, J.G. and Tak, P.P. 2008. Tumor necrosis factor antagonist mechanisms of action: a comprehensive review. Pharmacology & Therapeutics 117, 244-279. |
| Wang, X., Luo, Y., Liao, W.B., Zhang, J. and Chen, T.M. 2013. Effect of osteoprotegerin in combination with interleukin-6 on inhibition of osteoclast differentiation. Chin J Traumatol 16, 277-280. |
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| KR20200046772A (en) | 2018-10-25 | 2020-05-07 | 대한민국(농촌진흥청장) | Composition comprising Locusta migratoria extract for preventing or treating Neuritis |
| KR20210011051A (en) | 2018-10-25 | 2021-01-29 | 대한민국(농촌진흥청장) | Composition comprising Locusta migratoria extract for preventing or treating Neuritis |
| KR20200048057A (en) | 2018-10-29 | 2020-05-08 | 대한민국(농촌진흥청장) | Antiinflammatory composition comprising Locusta migratoria extract |
| KR102095265B1 (en) * | 2019-02-28 | 2020-04-01 | 건국대학교 글로컬산학협력단 | Composition for preventing, treating or improving of atopic dermatitis comprising edible insects based oil as an effective ingredient |
| KR20210060233A (en) * | 2019-11-18 | 2021-05-26 | 세종대학교산학협력단 | Pharmaceutical composition for treating or preventing leaky gut syndrome |
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