KR20200048057A - Antiinflammatory composition comprising Locusta migratoria extract - Google Patents

Abstract

The present invention relates to an anti-inflammatory composition including a Locusta migratoria extract. The Locusta migratoria extract has an excellent effect of inhibiting production of inflammation-related factors, such as NO (nitric oxide), iNOS (inducible nitric oxide synthase), COX-2 (cyclooxygenase-2) and TNF-α (tumor necrosis factor-α), and thus can be used advantageously for preventing, alleviating or treating inflammatory bowel disease, inflammatory collagen blood vessel disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, laryngopharyngitis, bronchitis, pneumonia, pancreatitis, sepsis, cystitis, nephritis, neuritis, or the like.

Description

Anti-inflammatory composition comprising Locusta migratoria extract

The present invention relates to a composition for anti-inflammatory containing the extract of Locusta migratoria . More specifically, the present invention inhibits the production of inflammation-related factors such as NO (Nitric oxide), iNOS (Inducible Nitric oxide synthase), COX-2 (cyclooxygenase-2), TNF-α (tumor necrosis factor-α). It contains the extract of Locusta migratoria , which is excellent in inflammatory bowel disease, inflammatory collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, pharyngitis, bronchitis, pneumonia, pancreatitis, The present invention relates to an anti-inflammatory composition having excellent prevention, improvement or treatment efficacy such as sepsis, cystitis, nephritis, and neuritis.

Inflammation is one of the biological tissue's defense responses to a certain stimulus, and refers to a complex lesion that combines tissue degeneration, circulatory disorder and exudation, and tissue proliferation. The causes are mechanical injuries, physical factors such as temperature and radiation, chemical factors such as poisons, and parasitic organisms such as bacterial infections. In addition to these main causes, its occurrence is complicated by several minor factors and the predisposition or immunity of the individual.

In addition, the inflammatory reaction can be said to be one of the most important pathological mechanisms according to the host's defense mechanism against wounds, microbial infections, and the like. Macrophages are the most representative immune cells that regulate this inflammatory response, and activated macrophages are TNF-α (tumor necrosis factor-α), IL-6 (interleukin-6), PGE2 (prostaglandin E2), NO (nitric oxide) ), ROS (reactive oxygen species) and secretes various inflammatory mediators (Laskin, DL et al., 2011).

Accordingly, the present inventors were able to complete the present invention while studying various physiological activities using the extract of Pulmuchi, confirming that the Pulmuchi extract has anti-inflammatory efficacy and easily using it as a therapeutic agent for various inflammatory diseases.

Republic of Korea Patent Publication No. 10-2016-0001880 (Name of the invention: rice locust extract with anti-inflammatory activity and composition containing the same, Applicant: Republic of Korea, Publication date: 2016.01.07) Republic of Korea Registered Patent No. 10-1730065 (Invention name: Full-color flower plain extract with anti-inflammatory activity and composition containing same, Applicant: Republic of Korea, Registration date: 2017.04.19)

An object of the present invention is to provide a composition for anti-inflammatory containing the extract of Locusta migratoria . More specifically, the object of the present invention is the production of inflammation-related factors such as NO (Nitric oxide), iNOS (Inducible Nitric oxide synthase), COX-2 (cyclooxygenase-2), TNF-α (tumor necrosis factor-α). Contains the extract of Locusta migratoria with excellent inhibitory effect, inflammatory bowel disease, inflammatory collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, sore throat, bronchitis, pneumonia, It is to provide an anti-inflammatory composition excellent in prevention, improvement or treatment efficacy of pancreatitis, sepsis, cystitis, nephritis, neuritis, and the like.

The present invention relates to a composition for anti-inflammatory containing the extract of Locusta migratoria .

The extract of the beetroot can be extracted with water, C1 ~ C4 alcohol or a mixed solution thereof as a solvent.

The C1 ~ C4 alcohol may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol and isobutanol.

The Pulmuchi extract inhibits the expression of inflammatory response factors selected from the group consisting of NO (Nitric oxide), iNOS (Inducible Nitric oxide synthase), COX-2 (cyclooxygenase-2) and TNF-α (tumor necrosis factor-α). It is characterized by.

The present invention can also provide a pharmaceutical composition for the prevention or treatment of inflammatory diseases containing Pulmuchi extract.

The inflammatory diseases include inflammatory bowel disease, inflammatory collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, sore throat, bronchitis, pneumonia, pancreatitis, sepsis, cystitis, nephritis and neuritis. It can be selected from the group consisting of.

In another aspect, the present invention also relates to a health functional food for the prevention or improvement of inflammatory diseases containing beetroot extract.

In addition, the present invention provides an anti-inflammatory cosmetic composition comprising the extract of the beetroot as an active ingredient.

Hereinafter, the present invention will be described in detail.

Water, C1 ~ C4 alcohol or a mixed solution used in the preparation of the beetroot extract may be used in a volume of 1-40 times (1-40% based on 1kg) by weight, and preferably 5-40 times the volume. Can be used. The extraction conditions of the Pulmuchi extract may be 1 minute to 48 hours at 20 to 100 ° C. The above process can be repeated 1 to 4 times.

In addition, as a conventional method in the art, after dissolving the extract of water, C1 ~ C4 alcohol or a mixed solution thereof in water, among the group consisting of n-hexane, methylene chloride, acetone, chloroform, ethyl acetate and n-butanol It may be prepared as a fraction by further fractionation using one or more solvents selected.

The preparation temperature of the extract or its fraction may be 20 to 100 ° C, but is not limited thereto. The extraction or fractionation time is not particularly limited, but it is preferable to extract within 10 minutes to 2 days, and an ordinary extraction device, an ultrasonic pulverization extractor, or a fractionator may be used as the extraction device. The thus prepared Pulmuchi extract or fraction can be removed by hot air drying, vacuum drying or freeze drying. In addition, the extract or fractions of Pulmuchi can be used by purification using column chromatography.

In addition, when preparing the extract of the beetroot of the present invention, after mixing the dried powder of the beetle with a solvent, after crushing 1 to 3 times for 5 to 30 seconds at a strength of 200 to 250 J, extraction may be started. In addition, extraction may be performed by standing at 4 to 30 ° C. for 10 to 60 minutes after ultrasonic treatment. The dried powder of the beetle may be obtained by pulverizing the dried beetle through freeze drying or a natural drying method at 4 to 30 ° C.

According to the commercial method, the beetroot extract is a single or suitable known method used for separation and extraction of plant components, such as extraction with an organic solvent (alcohol, ether, acetone, etc.), distribution of hexane and water, and method by column chromatography. It can be used by fractionation or purification using the combined method.

The chromatography is silica gel column chromatography, LH-20 column chromatography, ion exchange resin chromatography, medium pressure liquid chromatography chromatography, thin layer chromatography (TLC), silica gel vacuum liquid chromatography, and high performance liquid chromatography.

In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of inflammatory diseases, including the Pulmuchi extract as an active ingredient. The Pulmuchi extract may be added as 0.001 to 100% by weight to the pharmaceutical composition of the present invention.

More specifically, the pharmaceutical composition is formulated in the form of an oral dosage form such as a powder, a granule, a tablet, a capsule, a suspension, an emulsion, a syrup, an aerosol, an external preparation, a suppository, and a sterile injection solution, respectively, according to a conventional method. Can be used interchangeably. Carriers, excipients, and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose , Methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include at least one excipient in the extract of the present invention, for example, starch, calcium carbonate, sucrose or lactose, It is prepared by mixing gelatin. In addition, lubricants such as magnesium stearate and talc are used in addition to simple excipients. Liquid preparations for oral use include suspensions, intravenous solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used diluents, various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, can be included. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, 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 a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin may be used.

The dosage of the pharmaceutical composition of the present invention will vary depending on the age, gender, weight of the subject to be treated, the specific disease or pathology to be treated, the severity of the disease or pathology, the route of administration, and the judgment of the prescriber. Dosage determination based on these factors is within the level of those skilled in the art, and dosages generally range from 0.01 mg / kg / day to approximately 2000 mg / kg / day. A more preferred dosage is 1 mg / kg / day to 500 mg / kg / day. The administration may be administered once a day, or may be divided into several times. The above dosage does not limit the scope of the present invention in any way.

The pharmaceutical composition of the present invention can be administered to various mammals, such as rats, livestock, and humans. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dura mater or intracranial injection. Since the extract of the present invention has little toxicity and side effects, it is a drug that can be used safely even for a long period of time for prevention purposes.

The present invention can provide a health functional food for the prevention or improvement of inflammatory diseases, including the Pulmuchi extract as an active ingredient.

The health functional food may include a food supplement that is food acceptable. The health functional food of the present invention includes tablets, capsules, pills or liquids, and foods to which the extract of the present invention can be added, for example, various drinks, meat, sausage, bread, candy, There are snacks, noodles, ice cream, dairy products, soups, drinks, alcoholic beverages, chewing gum, tea and vitamin complexes.

In another aspect, the present invention can provide a cosmetic composition for anti-inflammatory comprising the extract of the beetroot as an active ingredient. The formulation of the cosmetic composition is not particularly limited, but is preferably a skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, foundation , Essence, Nutrition Essence, Pack, Soap, Cleansing Foam, Cleansing Lotion, Cleansing Cream, Body Lotion and Body Cleanser. The components included in the cosmetic composition of the present invention may include all of the ingredients commonly used in cosmetics in addition to the berry extract and horse oil of the present invention as active ingredients. For example, it may include general auxiliary ingredients such as emulsifiers, thickeners, emulsions, surfactants, lubricants, alcohols, water-soluble polymers, gelling agents, stabilizers, vitamins, and fragrances.

The present invention relates to a composition for anti-inflammatory containing the extract of Locusta migratoria . The extract is excellent in inhibiting the production of inflammation-related factors such as NO (Nitric oxide), iNOS (Inducible Nitric oxide synthase), COX-2 (cyclooxygenase-2), TNF-α (tumor necrosis factor-α). Prevention of inflammatory bowel disease, inflammatory collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, pharyngitis, bronchitis, pneumonia, pancreatitis, sepsis, cystitis, nephritis, neuritis, It can be easily used for improvement or treatment.

FIG. 1 shows the results of confirming the cytotoxicity of the Pulmuchi extract of the present invention with respect to Raw 264.7, a macrophage of a mouse, through an MTS assay.
Figure 2 is a result of confirming the activity of various inflammation-related factors of the extract of the present invention in the raw 264.7 LPS (Lipopolysaccaride) treated mouse macrophages, Figure 2A is a result of reducing the amount of NO (Nitric oxide) secretion, Figure 2B Shows the results of mRNA inhibition of iNOS (inducible Nitric oxide synthase), and FIG. 2C shows the results of confirming the protein expression inhibitory effect of iNOS and COX-2 (cyclooxygenase-2) (left) and numerically graphing them (right) Indicates.
3 is a result of confirming the effect of inhibiting the expression of mRNA (FIG. 3A) and protein (FIG. 3B) of TNF-α (tumor necrosis factor-α) possessed by the extract of the present invention in Raw 264.7, LPS-treated mouse macrophages. Shows.
Figure 4 shows the results of confirming the change of the signaling factors of the NF-κB and MAPK related to the extract of the present invention in Raw 264.7 LPS-treated mouse macrophages.

Hereinafter, the present invention will be described in more detail through examples. The following examples are presented only to illustrate the present invention, and the present invention is not limited by the following examples.

<Example 1. Preparation of Pulmuchi extract>

Locusta migratoria used six generations of Haenam that were bred indoors by the Insect Industry Division of the National Academy of Agricultural Sciences. Experimental insects were used for feeding wheat and were raised under conditions of 30 ° C, 65% RH, 9L / 15D, and 1,800 Lux. After washing the beetle (adult) twice with water, drying it using a freeze dryer (Eyela, Japan) to remove moisture, and then pulverizing it with a multifunctional grinder (Korea Medi, Korea) to obtain a beetle powder. The thus obtained Pulmuchi powder was dissolved in a 70% (v / v) ethanol aqueous solution (100 g / 1 L) and then crushed twice for 10 seconds at a strength of 230 J using an ultrasonic crusher (LabaX, MAm USA) at 25 ° C. for 30 minutes. After standing and centrifuging at 4500 rpm for 10 minutes, the supernatant was obtained, filtered with a 0.25 μm syringe filter (Whatman, ND, USA), and then completely dried using a centrifugal evaporator (CVE-3100, Tokyo, Japan). Pulmuchi extract ( Locusta migratoria ethanol extract, LME) was prepared.

<Example 2. Confirmation of cytotoxicity of Pulmuchi extract>

RAW264.7 cells were cultured using Dublecco's Modified Eagle Medium (Gibco) medium containing Penicillin-streptomycin 100 unit / ml and 10% fetal bovine serum (Gibco, MD, USA). C., cultured in a 5% CO ² incubator (Incubator; Thermo Scientific, IL, USA), and subcultured at intervals of 2 days, and in subsequent experiments, RAW264.7 cells cultured as above were used in the experiment.

Cytotoxicity was confirmed by measuring cell viability using MTS assay. To this end, RAW264.7 cells are cultured by dispensing them into 96 well plates at 4 × 10 ⁴ cells / well, and treating the extract of Pulmuchi by concentration (100, 500, 1000, 2000 μg / ml), 37 ℃, 5% Incubated in a CO ₂ incubator for 24 hours. Subsequently, 10 μl of MTS reagent CellTiter 96 ^® AQueous One Solution Reagent (Promega, USA) was added and reacted for 4 hours to confirm the change in color and using a microplate reader (Beckman Coulter, CA, USA) at 590 nm. Cell viability was measured by measuring absorbance, and the results are shown in FIG. 1.

Referring to FIG. 1, the extract of Pulmuchi (Locusta migratoria ethanol extract, LME) showed an effect of promoting cell growth up to a concentration of 2000 µg / ml and did not show toxicity, but little by little from a concentration exceeding 2000 µg / ml. It can be confirmed that it appears. Therefore, the highest concentration of locust extract (Locusta migratoria ethanol extract, LME) used in subsequent experiments was used at a concentration of 2000 µg / ml or less, which does not show toxicity to RAW 264.7 cells.

<Example 3. Anti-inflammatory activity assay of Pulmuchi extract-I>

Example 3-1. NO (Nitric oxide) production inhibitory effect confirmed

Normal NO (Nitric oxide) is known to be very important for neuroprotection or brain development, but it is excessively produced from microglia activated by LPS (Lipopolysaccharide), interferon-gamma (IFN-γ), β-amyloid, etc. It is known to induce cytotoxicity and inflammatory reactions.

Thus, the raw 264.7 macrophages were treated with extracts of Pulmuchi and the amount of NO (Nitric oxide) produced by lipopolysaccharide (LPS) was measured. Raw264.7 macrophages were divided into 8 × 10 ⁴ cells / well in 96 well plates and cultured for 24 hours, followed by pretreatment of Pulmuchi extract (LME) for each hour for each concentration. After that, to induce inflammation, LPS was treated at a concentration of 100 ng / ml and cultured for 24 hours. 100 µl of the supernatant of the cultured cells was used in the experiment, and a microplate reader (Beckman Coulter) showed how much the amount of NO secreted by LPS was reduced by the Pulmuchi extract using the NO detection kit (Intron, Korea). CA, USA) to confirm by absorbance measurement at 550nm wavelength. The results for this are shown in Figure 2A.

Referring to Figure 2A, it can be seen that in the experimental group treated with Pulmuchi extract, the concentration-dependent NO secretion decreases.

Example 3-2. Confirmation of iNOS mRNA expression inhibitory effect

iNOS does not normally exist in cells, but once induced by external stimulation, it produces a large amount of NO for a long time, and the generated NO causes harmful effects on living organisms such as pathological vasodilation, cytotoxicity, and tissue damage. It is known that in the inflammatory state, not only iNOS is produced, but also COX is activated to promote the biosynthesis of inflammatory mediators such as prostaglandin to deepen inflammation.

Therefore, the mRNA expression level of iNOS (inducible Nitric oxide synthase) produced by LPS (Lipopolysaccharide) was measured by using Quantitative RT-PCR (qRT-PCR) after treatment of Pulmuchi extract on Raw264.7 macrophages.

To this end, Raw264.7 cells were dispensed into 6 well plates at 2 × 10 ⁵ cells / well each, and then Pulmuchi extract (LME) was treated for each hour for each concentration. After that, to induce inflammation, LPS was treated at a concentration of 100 ng / ml and cultured for 24 hours. Thereafter, the supernatant was removed, washed with PBS, and the cells were collected. 1 ml of TRIZOL solution is mixed with the collected cells and reacted at room temperature for 5 minutes. 200 μl chloroform was added and mixed, followed by centrifugation at 4 ° C. for 12,000 × g for 15 minutes, supernatant was taken, 500 μl isopropanol was added, reaction was performed at room temperature for 10 minutes, and then centrifugation at 4 ° C for 12,000 × g for 10 minutes. do. The precipitated RNA was washed 3 times with 75% (v / v) ethanol aqueous solution, dried and then dissolved in sterile water. Next, for the synthesis of cDNA, cDNA was synthesized with High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster city, CA) using 1 μg of total RNA, and qRT-PCR experiment was conducted using the primers below. The results for this are shown in Figure 2B.

i-NOS Forward: 5'-CAGCACAGGAAATGTTTCAGC-3 '

i-NOS Reverse: 5'-TAGCCAGCGTACCGGATGA-3 '

GAPDH Forward: 5'-AAGGTCATCCCAGAGCTGAA-3 '

GAPDH Reverse: 5'-CTGCTTCACCACCTTCTTGA-3 '

Referring to FIG. 2B, it can be confirmed that the gene expression amount of iNOS is decreased in a concentration-dependent manner similar to NO, depending on the concentration of Pulmuchi extract.

Example 3-3. Confirmation of protein-inhibiting effect of i-NOS and COX-2

Next, the effect of inhibiting protein expression of i-NOS and COX-2 possessed by Pulmuchi extract was confirmed through Western blotting. For this, each 2 × 10 ⁵ cells / well cell was prepared in 6 wells of Raw264.7 cells, and then treated with Pulmuchi extract (LME) for 1 hour at each concentration. After that, to induce inflammation, LPS was treated at a concentration of 100 ng / ml and cultured for 24 hours. Thereafter, the supernatant was removed, washed with PBS, and then the protein was extracted using RIPA lysis buffer. Protein was quantified using a BCA kit (Thermo Fisher), and developed using an SDS-PAGE gel (Sodium dodecyl sulphate polyacrylamide gel). After the developed proteins are transferred to the PVDF membrane, they are blocked with 5% (w / v) skim milk for 1 hour, diluted with primary antibody (1: 1000), overnight at 4 ° C for 18 hours, and then 0.05%. Washed three times at 10 minute intervals with TBST (TBST; 20 mM Tris [pH 7.5], 145 mM NaCl, 0.05% (w / v) Tween-20), and diluted each secondary antibody to 1: 10,000 at room temperature 1 The reaction was carried out at room temperature for an hour. After washing with 0.05% TBST for 3 times for 10 minutes, reacting with ECL solution and confirming the signal was measured using an image analyzer FluorChem (Alpha Innotech, USA), and the results are shown in FIG. 2C. The left side of FIG. 2C is a result of confirming the protein expression inhibitory effect of iNOS and COX-2 (cyclooxygenase-2), and the right side of FIG. 2C shows the result of digitization and graphing.

Referring to Figure 2C, it can be seen that the inflammatory mediators COX-2 and iNOS are concentration-dependently reduced in the Pulmuchi extract.

<Example 4. Anti-inflammatory activity assay of prunus extract-II>

Quantitative RT-PCR of mRNA and protein expression of TNF-α (tumor necrosis factor-α), an inflammatory factor cytokine produced by LPS (Lipopolysaccharide) after treatment of Pulmuchi extract on Raw264.7 macrophages 3-2) and ELISAELISA (Enzyme-linked immunosorbent assay), and the results are shown in FIG. 3.

At this time, the primer sequence used for Quantitative RT-PCR is as follows.

TNF-α Forward: 5'-ATGAGAAGTTCCCAAATGGC-3 '

TNF-α Reverse: 5'-CTCCACTTGGTGGTTTGCTA-3 '

GAPDH Forward: 5'-AAGGTCATCCCAGAGCTGAA-3 '

GAPDH Reverse: 5'-CTGCTTCACCACCTTCTTGA-3 '

Also, for ELISA, Raw264.7 cells were prepared in 96 well plates, each 8 × 10 ⁴ cells / well cell, and then treated with Pulmuchi extract (LME) for 1 hour at each concentration. After that, to induce inflammation, LPS was treated at a concentration of 100 ng / ml and cultured for 24 hours. The supernatant of the cultured cells was collected and the secretion amount of TNF-α was measured using an ELISA kit (ThermoFisher, Waltham, MA).

Referring to FIG. 3, it was confirmed that the expression level of TNF-α was decreased in a concentration-dependent manner by the Pulmuchi extract, and these results demonstrate that the Pulmuchi extract can exhibit excellent effects on inflammatory diseases.

<Example 5. Activity test of anti-inflammatory related signaling substances>

Using the Western blotting method used in Example 3-3, changes in NF-κB and MAPK-related signaling substances due to the treatment of Pulmuchi extract were confirmed.

Nuclear factor (NF) -κB plays a very important role in the inflammatory response by regulating the expression of genes such as inflammatory cytokine, growth factor, COX-2 and iNOS. NF-kB is present in an inactivated state in the cytoplasm by binding to an inhibitory κB (IBK) protein that inhibits NF-κB activity in most cells. The active pathway of NF-κB has been reported to separate NF-κB from the inhibitory protein IκB and the cytoplasm, pass through the nuclear membrane, move into the nucleus, and regulate gene expression. At this time, the separated IκB is decomposed in the proteasome through phosphorylation and polyubiquitination.

For this, each 1.6 × 10 ⁶ cells / well cells were prepared in ⁶ wells of Raw264.7 cells, and then treated with Pulmuchi extract (LME) for 1 hour at each concentration. After that, to induce inflammation, LPS was treated with 100 ng / ml concentration for 30 minutes, washed with PBS, and extracted with protein using RIPA lysis buffer to observe the expression and phosphorylation changes of NF-kB and MAP kinases.

In addition, in the inflammatory reaction, mitogen-activated protein kinase (MAPK) plays an important role in the production of immune response mediators, so it is necessary to examine the phosphorylation process of MAPKs. Pulmuchi extract was pre-treated for 1 hour to induce an inflammatory reaction with LPS for 40 minutes to confirm the phosphorylation process of each MAPK.

The results of each experiment are shown in FIG. 4. Referring to FIG. 4A, it can be confirmed that phosphorylation of IkB increased by LPS is decreased depending on the concentration of Pulmuchi extract, and the amount of IkB protein significantly reduced by LPS is also Pulmuchi extract It can be seen that is increased by.

In MAPK signaling, extracellular signal-regulated kinases (ERK) and c-Jun N-terminal Kinase (JNK) in the LPS-only treatment group, when inducing an inflammatory response with LPS for 40 minutes after 1 hour of pre-treatment of the extract of Pulmuchi as shown in FIG. It was confirmed that phosphorylation of was significantly increased, but phosphorylation of the two proteins in the group treated with prunus extract was decreased depending on the concentration of prunus extract. However, the phosphorylation of p38 MAP kinase increased by LPS did not decrease in the group treated with prunus extract. These results indicate that Pulmuchi extract regulates the inflammatory response through inhibition of phosphorylation of ERK and JNK, which are the main mechanisms of the inflammatory response.

<Formulation Example 1. Pharmaceutical preparation>

200 g of the beetle ethanol extract of the present invention was mixed with 175.9 g of lactose, 180 g of potato starch and 32 g of colloidal silicic acid. After adding 10% gelatin solution to this mixture, it was ground and passed through a 14 mesh sieve. This mixture was dried, and a mixture obtained by adding potato starch 160g, talc 50g, and magnesium stearate 5g was purified.

<Formulation Example 2. Food manufacturing>

Formulation Example 2-1. Cooking seasoning

Pulmuchi ethanol extract of the present invention was added to the cooking seasoning in 1% by weight to prepare a cooking seasoning for health promotion.

Formulation Example 2-2. Production of flour food

Pulmuone ethanol extract of the present invention was added to wheat flour at 0.1% by weight, and bread, cake, cookies, crackers, and noodles were prepared using this mixture to prepare foods for health promotion.

Formulation Example 2-3. Preparation of soup and gravy

Pulmuchi ethanol extract of the present invention was added to soups and broths at 0.1% by weight to prepare health promotion soups and gravy.

Formulation Example 2-4. Manufacturing dairy products

The beetle ethanol extract of the present invention was added to milk at 0.1% by weight, and various dairy products such as butter and ice cream were prepared using the milk.

Formulation Example 2-5. Vegetable juice production

0.5 g of the beetle ethanol extract of the present invention was added to 1,000 ml of tomato juice or carrot juice to prepare a vegetable juice for health promotion.

Formulation Example 2-6. Fruit juice manufacturing

0.1 g of beetle ethanol extract of the present invention was added to 1,000 ml of apple juice or grape juice to prepare a fruit juice for health promotion.

<Cosmetic formulation example 1. Preparation of flexible lotion>

As shown in the composition of Table 1, the flexible lotion (skin, 100 g) containing the extract of the beetle ethanol of the present invention was prepared according to a conventional method.

Raw material Content (g) Beet ethanol extract 0.3 glycerin 3.0 Butylene glycol 2.0 Propylene glycol 2.0 Polyoxyethylene (60) hardened castor oil 1.0 ethanol 10.0 Triethanolamine 0.1 antiseptic a very small amount Coloring a very small amount Spices a very small amount Purified water Balance

<Cosmetic formulation example 2. Preparation of nutritional lotion>

As shown in the composition in Table 2, a nutritional lotion (lotion, 100 g) containing the extract of the beet ethanol of the present invention was prepared according to a conventional method.

Raw material Content (g) Beet ethanol extract 0.1 Citosterol 1.7 Polyglyceryl 2-oleate 1.5 Seteares 1.2 cholesterol 1.5 Dicetyl phosphate 0.4 Concentrated glycerin 5.0 Sunflower Oil 10.0 Carboxyvinyl polymer 0.2 Shotgun 0.3 antiseptic a very small amount Spices a very small amount Purified water Balance

Claims (8)

Anti-inflammatory composition, characterized in that it contains an extract of Locusta migratoria . According to claim 1,
The Pulmuchi extract is an anti-inflammatory composition, characterized in that the extract is extracted with water, C1 ~ C4 alcohol or a mixed solution thereof as a solvent. According to claim 1,
The Pulmuchi extract inhibits the expression of inflammatory response factors selected from the group consisting of NO (Nitric oxide), iNOS (Inducible Nitric oxide synthase), COX-2 (cyclooxygenase-2) and TNF-α (tumor necrosis factor-α). Anti-inflammatory composition, characterized in that. A pharmaceutical composition for the prevention or treatment of inflammatory diseases, comprising the composition of claim 1. The method of claim 4,
The inflammatory diseases include inflammatory bowel disease, inflammatory collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, sore throat, bronchitis, pneumonia, pancreatitis, sepsis, cystitis, nephritis and neuritis. Pharmaceutical composition for the prevention or treatment of inflammatory diseases, characterized in that selected from the group consisting of. A health functional food for the prevention or improvement of inflammatory diseases, comprising the composition of claim 1. The method of claim 6,
The inflammatory diseases include inflammatory bowel disease, inflammatory collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, sore throat, bronchitis, pneumonia, pancreatitis, sepsis, cystitis, nephritis and neuritis. Health functional food for preventing or improving inflammatory diseases, characterized in that selected from the group consisting of. An anti-inflammatory cosmetic composition comprising the composition of claim 1.

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