KR20240025754A - Anti-inflammatory composition comprising extract of centella asiatica callus and method of preparing the same - Google Patents

Abstract

The present invention provides an anti-inflammatory composition containing Centella asiatica callus extract as an active ingredient and a method for producing the composition. The anti-inflammatory composition specifically includes as an active ingredient a Centella asiatica callus extract cultured in an environment irradiated with LED light of 400 nm to 760 nm, thereby providing significantly improved anti-inflammatory properties. The anti-inflammatory composition effectively inhibits inflammatory mediators without cytotoxicity and provides the effect of preventing, improving, or treating inflammatory diseases.

Description

Anti-inflammatory composition containing centella callus extract and method for manufacturing same {ANTI-INFLAMMATORY COMPOSITION COMPRISING EXTRACT OF CENTELLA ASIATICA CALLUS AND METHOD OF PREPARING THE SAME}

The present invention relates to an anti-inflammatory composition containing Centella asiatica callus extract and a method for producing the same.

In modern society, interest in health is increasing at a high level among various age groups due to the improvement of the economic level and the continued increase in the elderly population. Many medicines, health functional foods, and functional cosmetics are being developed with the goal of improving health and delaying the aging process. In particular, natural materials are used to solve safety problems, suppress inflammation with fewer side effects, and achieve skin improvement effects. Research is actively underway.

Natural products have long been a source of various natural physiologically active substances that serve as active ingredients for medicines, foods, cosmetics raw materials, fragrances, and pigments. These natural physiologically active substances are high value-added substances that exhibit remarkable activity in very small amounts, and numerous types are currently being usefully used.

Various materials derived from natural products with excellent anti-inflammatory activity are being introduced. Inflammatory response is a physiological response to protect the living body from harmful surrounding environments such as invasion of foreign substances such as bacteria or physical and chemical damage. Immune cells are activated by recognizing foreign substances in the living body, and the activated immune cells mediate inflammation. It begins by secreting many factors that This inflammatory phenomenon causes an increase in various types of white blood cells and immune substances, and the increased cells secrete various types of proteolytic enzymes and cytokines, which are inflammatory cell products, enabling treatment and defense. In particular, nitric oxide (NO) is a powerful inflammatory mediator with high reactivity involved in neurotransmission, vascular relaxation, and cell-mediated immune responses, and is produced from L-argine by NO synthase. Nitric oxide (NO) activates macrophages to produce interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor (TNF)-α and The same pro-inflammatory cytokines are produced.

Inflammation is one type of immune response, but if the irritant that stimulates inflammation does not disappear or continues to be created, chronic inflammation occurs, resulting in serious tissue damage, which becomes the cause of various diseases. Chronic inflammation or hyperinflammation causes damage to cells and connective tissues due to proteolytic enzymes, and also has a negative effect on cell regeneration and proliferation. In particular, damage to connective tissue not only reduces the elasticity of the skin, causing wrinkles, but also causes the loss of normal tissue structure and function, resulting in a serious condition.

Accordingly, as a material derived from natural products that is guaranteed to be safe and non-irritating to the human body, it is not only non-toxic, but also has excellent anti-inflammatory activity compared to existing natural materials, so the development of a new material that can be directly applied to medicine, food, or cosmetics is needed. It is being done actively.

Centella asiatica is a perennial creeping herb belonging to the Umbrella family, which mainly grows in hot and humid areas. In traditional medicine, it has been used as a medicinal herb for skin diseases, fever, hemoptysis, diuretic, tonic, euphoria, enlargement, arthritis, etc. Youngno Lee, 2006). The main known components of Centella asiatica are triterpenic acid sugar esters, and asiaticoside, madecassoside, brahmoside, and brahminoside have been reported. It is known that when hydrolyzed, asiatic acid and madecassic acid are produced. The main components of Centella asiatica include asiaticoside and madecassoside, which are pentacyclic triterpene glycosides belonging to the α-amyrin-ursonic acid group. madecassoside) has long been used to treat skin wounds and chronic ulcers, and has been reported to be effective in treating leprosy, antibacterial effects, wounds, stomach ulcers, various skin diseases, mental disorders, tuberculosis, venous diseases, dementia, etc. there is. Due to these effects, Centella asiatica is used for various purposes such as edible, skin treatment, wound treatment, memory enhancer, and tonic. In Korea, Centella asiatica is also used in fields such as medicine and cosmetics.

However, considering the potential of Centella asiatica as a medicinal ingredient, efforts to increase the activity of the active ingredients contained in Centella asiatica are insufficient. In particular, there appears to be insufficient research to increase anti-inflammatory activity.

Republic of Korea Patent Publication No. 10-2022-0014765

The present inventors completed the present invention by discovering that an extract with significantly increased anti-inflammatory activity could be obtained by irradiating LED light when culturing Centella callus.

Therefore, the purpose of the present invention is to provide an anti-inflammatory composition containing Centella asiatica callus extract with significantly increased anti-inflammatory activity.

Additionally, the purpose of the present invention is to provide a method for producing an anti-inflammatory composition that can significantly increase anti-inflammatory activity by irradiating LED light when culturing Centella callus.

In order to achieve the above object, the present invention,

Provided is an anti-inflammatory composition containing as an active ingredient a Centella asiatica callus extract cultured in an environment irradiated with LED light of 400 nm to 760 nm.

In addition, the present invention

(a) cultivating Centella asiatica calli in an environment where LED light of 400 nm to 760 nm is irradiated; and

(b) extracting the cultured callus with a solvent; providing a method for producing an anti-inflammatory composition comprising a.

The anti-inflammatory composition of the present invention contains an extract obtained from Centella asiatica callus, which has increased anti-inflammatory activity by irradiating LED light during cultivation, and thus has excellent anti-inflammatory activity, and therefore provides an excellent effect in preventing, improving or treating anti-inflammatory diseases. .

In addition, the method for producing an anti-inflammatory composition of the present invention provides an anti-inflammatory composition with significantly increased anti-inflammatory activity by a simple method by further performing an LED light irradiation process when culturing Centella callus.

Figure 1 is a graph showing cell survival rate by administration of LED-treated Centella asiatica callus extract during culture;
Figure 2 is a graph showing the NO production inhibition effect of LED-treated Centella asiatica callus extract during culture;
Figure 3 is a graph showing the inhibitory effect of PGE ₂ production of LED-treated Centella asiatica callus extract during culture;
Figure 4 is a graph showing the inhibitory activity of IL-6 production, a pro-inflammatory cytokine, of the LED-treated Centella asiatica callus extract during culture;
Figure 5 is a graph showing the inhibitory activity of IL-1β production, a pro-inflammatory cytokine, of the LED-treated Centella asiatica callus extract during culture;
Figure 6 is a graph showing the inhibitory activity of the LED-treated Centella asiatica callus extract on the production of TNF-α, a pro-inflammatory cytokine, during culture.

The present invention will be described in detail below.

The anti-inflammatory composition of the present invention is characterized by containing as an active ingredient a Centella asiatica callus extract cultured in an environment irradiated with LED light of 400 nm to 760 nm.

In the anti-inflammatory composition, Centella callus extract may be included in an amount of 0.1 to 100% by weight, and various additives known in the art may be included.

The Centella asiatica callus extract may be a water or ethanol extract, and the water extract may be more preferably used. Distilled water may be used as the water. The extraction can be performed by adding Centella asiatica callus extract to a solvent and heating it. At this time, the heating temperature may be a temperature between the boiling point of -30°C and the boiling point.

It may be more preferable that the Centella asiatica callus extract is cultured in an environment irradiated with one or more types selected from LED light of 400 nm to 500 nm and LED light of 610 nm to 760 nm.

The LED light of 400 nm to 500 nm is BLUE light, and the LED light of 610 nm to 760 nm is RED light. In particular, when the Centella asiatica callus extract is cultured in an environment irradiated with LED light (BLUE light) of 400 nm to 500 nm, it provides more improved anti-inflammatory activity.

The LED light may be irradiated at a light quantity of 0.5 to 10 μmol/m ² S, preferably 1 to 5 μmol/m ² S, and more preferably 2 to 3 μmol/m ² S. It is not limited.

The Centella asiatica callus culture may be performed at a temperature of 20 to 30° C. and 50 to 90% humidity, but is not limited thereto.

For example, the centella asiatica callus culture may be performed in an incubator for 1 day to 2 months, 15 days to 45 days, or 20 days to 40 days, but is not limited thereto.

In one embodiment of the present invention, the anti-inflammatory composition may be a pharmaceutical composition for preventing or treating inflammatory diseases.

The pharmaceutical composition can be prepared in a conventional formulation by selecting an effective amount of the Centella asiatica extract and one or two or more conventional pharmaceutically acceptable carriers or one or two or more additives.

The carrier may be one or two or more selected from the group consisting of diluents, lubricants, binders, disintegrants, sweeteners, stabilizers and preservatives, and the additives may include one or two or more of the following: flavorings, vitamins and antioxidants. You can select and use it.

In the present invention, the carrier and additives are not limited to their types, and specifically, diluents include lactose monohydrate, trehalose, corn starch, soybean oil, and microcrystalline cellulose ( microcrystalline cellulose or mannitol (D-mannitol), magnesium stearate or talc as a lubricant, and polyvinylpyrrolidone (PVP) or hydroxypropylcellulose (HPC) as a binder. You can choose. In addition, disintegrants include carboxymethylcellulose calcium (Ca-CMC), sodium starchglycolate, polacrylin potassium or cross-linked polyvinylpyrrolidone, and sweeteners include: White sugar, fructose, sorbitol, or aspartame; stabilizers include carboxymethylcellulosesodium (Na-CMC), beta-cyclodextrin, white bee's wax, or xanthan gum ( xanthan gum), and as a preservative, you can choose from methyl p-hydroxy benzoate (methlparaben), propyl p-hydroxybenzoate (propylparaben), or potassium sorbate.

The amount of the pharmaceutical composition used may vary depending on the age, sex, and weight of the patient or animal to be treated, and, above all, depending on the condition of the subject to be treated, the specific category or type of the disease to be treated, the route of administration, and the properties of the therapeutic agent used. It will be dependent. The pharmaceutical composition is appropriately selected depending on the absorption rate of the active ingredient in the body, excretion rate, age and weight, sex and condition of the patient or animal to be treated, and the severity of the disease to be treated, but is generally administered at 0.1 to 1,000 mg/day. It is preferable to administer in kg. The unit dosage form formulated in this way can be administered several times at regular time intervals as needed.

The pharmaceutical composition may be administered individually as a preventive or therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents.

The pharmaceutical compositions can be used by formulating them into oral dosage forms such as powders, granules, tablets, capsules, troches, suspensions, emulsions, syrups, aerosols, etc. according to conventional methods. When formulating, it can be prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, troches, etc. Liquid preparations for oral use include suspensions, oral solutions, emulsions, syrups, etc. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. .

In one embodiment of the present invention, the anti-inflammatory composition may be a cosmetic composition for preventing or improving inflammatory diseases.

The cosmetic composition has excellent anti-inflammatory activity and can additionally provide excellent effects in anti-aging, skin moisturizing, and skin barrier strengthening.

The cosmetic composition can be prepared in any formulation commonly prepared in the art, for example, solution, suspension, emulsion, paste, lotion, gel, water-soluble liquid, cream, essence, surfactant-containing cleansing, oil. , a basic cosmetic formulation selected from oil-in-water (O/W) type and water-in-oil (W/O) type; skin; Lotion; eye cream; soothing gel; Ointment; Formulation for mask pack; Formulations for body wash; peeling gel; Oil-in-water and water-in-oil makeup bases; foundation; skin cover; A color cosmetic formulation selected from lipstick, lip gloss, face powder, two-way cake, eye shadow, cheek color, and eyebrow pencil; Formulations for scalp use; It may be any one selected from among.

In addition, the cosmetic composition may contain auxiliaries commonly used in the cosmetic field, such as hydrophilic or lipophilic activators, preservatives, antioxidants, solvents, fragrances, fillers, blocking agents, pigments, odorants, dyes, etc. The amounts of these various auxiliaries are those commonly used in the art, for example, 0.001 to 30% by weight relative to the total weight of the composition. However, in any case, the auxiliaries and their ratios will be selected so as not to adversely affect the desirable properties of the cosmetic composition according to the present invention.

In the cosmetic composition, when the formulation is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide are used as carrier ingredients. etc. can be used.

In addition, when the formulation is a solution or emulsion, a solvent, solubilizing agent, or emulsifying agent is used as a carrier component, examples of which include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzoate, propylene glycol, 1 , 3-butylene glycol, glycerol aliphatic ester, polyethylene glycol, or fatty acid ester of sorbitan can be used.

In addition, when the dosage form is a suspension, the carrier ingredients include 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, and miso. Crystalline cellulose, aluminum metahydroxide, bentonite, agar, or tragacanth can be used.

In addition, when the formulation is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder can be used as the carrier ingredient. In particular, when the formulation is a spray, chlorofluorohydrocarbon and protane can be used as carrier ingredients. /May contain propellants such as butane or dimethyl ether.

In addition, when the formulation is a surfactant-containing cleanser, the carrier ingredients include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, sarcosinate, fatty acid amide ether sulfate, and alkylamido. Betaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester can be used.

Additionally, if the formulation is a surfactant-containing cleanser formulation or a surfactant-free cleanser formulation or soap, it can be applied to the skin and then wiped off, removed, or washed with water. For example, the soap is liquid soap, powdered soap, solid soap, and oil soap, the surfactant-containing cleansing formulation is cleansing foam, cleansing water, cleansing towel, and cleansing pack, and the surfactant-free cleansing formulation is cleansing cream. , cleansing lotion, cleansing water and cleansing gel, but are not limited thereto.

In one embodiment of the present invention, the anti-inflammatory composition may be a food composition for preventing or improving inflammatory diseases. The food composition may be a health functional food.

The food composition may be tea, jelly, juice, extract, beverage, etc. containing Centella asiatica extract as an active ingredient, and there is no limitation on the processed form as long as it contains the Centella asiatica extract. In addition, the food composition includes general foods, dairy products including ice cream, milk, soy milk, and cheese, and soy milk products, beverages, meat, sausages, bread, chocolate candies, snacks, confectionery, pizza, ramen, and other noodles, It may be selected from the group consisting of gums, various soups, beverages, tea, drinks, alcoholic beverages, and lactic acid bacteria preparations in capsule or stick packaging.

The beverage composition may contain various flavoring agents, sweeteners, or natural carbohydrates as additional ingredients, like ordinary beverages. The above-mentioned natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. The sweetener may be a natural sweetener such as thaumatin or stevia extract, or a synthetic sweetener such as saccharin or aspartame.

In the present invention, the inflammatory disease includes inflammatory bowel disease, non-alcoholic chronic hepatitis, chronic hepatitis, Crohn's disease, pancreatitis, esophagitis, gastritis, colitis, ulcerative colitis, pneumonia, bronchitis, pharyngitis, myocardial infarction, heart failure, arthritis, and psoriasis. One selected from the group consisting of arthritis, rheumatoid arthritis, renal failure, psoriasis, anemia, diabetes, fibrosis, multiple sclerosis, systemic lupus erythematosus, ankylosing spondylitis, asthma, chronic obstructive pulmonary disease, periodontitis, neuropathic pain, and idiopathic inflammatory myopathy. It may be one of the following diseases.

In addition, the present invention

(a) cultivating Centella asiatica calli in an environment where LED light of 400 nm to 760 nm is irradiated; and

(b) extracting the cultured callus with a solvent. It relates to a method for producing an anti-inflammatory composition comprising a.

In step (a), the Centella callus extract may be a water or ethanol extract, and the water extract may be more preferably used. Distilled water may be used as the water.

It may be more preferable that the Centella asiatica callus extract in step (a) is a Centella asiatica callus extract cultured in an environment irradiated with one or more types selected from LED light of 400 nm to 500 nm and LED light of 610 nm to 760 nm.

The LED light of 400 nm to 500 nm is BLUE light, and the LED light of 610 nm to 760 nm is RED light. In particular, when the Centella asiatica callus extract is cultured in an environment irradiated with LED light (BLUE light) of 400 nm to 500 nm, it provides more improved anti-inflammatory activity.

In another embodiment of the present invention, the anti-inflammatory composition contains as active ingredients a Centella asiatica callus extract and an ethanol extract of Eriophorum scheuchzeri ssp. arcticum cultured in an environment irradiated with LED light of 400 nm to 500 nm in 6: It may be included in a weight ratio of 4 to 7:3.

The Arctic scheuchzeri (Eriophorum scheuchzeri ssp. arcticum) is a grass that is often seen in Arctic tundra wetlands, and has seeds that look like cotton. The Arctic stork grass extract may be an extract extracted from white cotton, which is a seed.

The Arctic stork grass extract is extracted from white cotton, which is a dried seed, with ethanol. In the above, extraction may be performed at 50 to 60°C. After obtaining the extract from the Arctic stork plant, it is filtered, concentrated, and dried to obtain the Arctic stork plant extract.

Specifically, for example, the manufacturing method of Arctic stork grass extract is as follows:

After chopping 1 kg of dried Arctic stork grass cotton, add it to 2 L of ethanol (concentration 40% (v/v)) and extract while stirring at 50 to 60°C for 4 hours. The extract is filtered under reduced pressure using a pressure reduction pump and Whatman No. 2 filter paper, and the filtered extract is concentrated using a vacuum rotary concentrator and then spray-dried to obtain a powdered extract of Arctic dorado.

In the present invention, when the active ingredients include an extract of Centella asiatica callus cultured in an environment irradiated with LED light of 400 nm to 500 nm and an ethanol extract of Eriophorum scheuchzeri ssp. arcticum (weight ratio of 7:3), It was confirmed by the same method as the “Test Example” described below that the anti-inflammatory effect increased compared to the case containing only the extract.

In other words, the composition containing more of the Arctic stork extract has improved cell viability and increased NO, as shown in the graph below, than the composition containing only the Centella asiatica callus extract cultured in an environment irradiated with 400 nm to 500 nm LED light. Inhibitory activity: Improved inhibitory activity was shown in pro-inflammatory cytokines IL-6, IL-1β, and TNF-α.

Below, the present invention will be described in more detail through examples. However, the following examples are intended to illustrate the present invention in more detail, and the scope of the present invention is not limited by the following examples. The following examples can be appropriately modified and changed by those skilled in the art within the scope of the present invention.

Example 1: Culture of Centella asiatica callus and preparation of water extract

Centella asiatica (CA) callus was provided by the Korea Research Institute of Bioscience and Biotechnology, Biological Resources Center. For efficient growth control, 0.43% (w/v) murashige & skoog medium, 100 mg/L myo-inositol, 3% (w/v) sucrose, 0.4% (w/v) gelrite (Duchefa Biochemie, Haarlem, Netherlands). , a medium supplemented with 1 mg/L 2,4-Dichlorophenoxyacetic acid (Sigma-Aldrich, St. Louis, MO, USA) was used. After adjusting the moisture of the medium to around 60 ± 2%, 5 g were placed in bottles and subcultured every month. LED incubation was performed with a light intensity of 2.10 ± 0.5 μmol/m ² S in an LED incubator under conditions of 25 ± 1°C and 70% humidity. Red (660 nm) and LED blue (450 nm) monochromatic light were used. Additionally, as a comparative example, callus under dark culture conditions was used. Afterwards, 10 times distilled water was added to 1 g of callus for hot water extraction, and the extract was filtered through a paper filter (ADVANTEC, Tokyo, Japan), concentrated under reduced pressure, freeze-dried at -110°C, and powdered.

Experiment example.

1) Experimental method

(1) Experimental materials and cell culture

LPS used in the present invention was purchased from Sigma-Aldrich (St. Louis, MO, USA), and RAW 264.7 cells were purchased from the Korea Cell Line Bank and supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg. /mL streptomycin was added to Dulbecco's Modified Eagle Medium (DMEM, Gibco, NY, USA), cultured in a 37°C, 5% CO ₂ incubator, and subcultured every 2 days.

(2) Cytotoxicity evaluation

To measure ^cytotoxicity _, RAW 264.7 cells were dispensed at 8.0 ) were treated simultaneously and cultured for 24 hours. Afterwards, MTT reagent was added and reacted at 37°C for 4 hours. After removing the supernatant, the formed formazan blue was dissolved with DMSO and the absorbance was measured at 570 nm. The average absorbance for each sample group was measured, and cytotoxicity was evaluated by comparing it with the absorbance measurement of the control group.

(3) Measurement of NO production inhibition activity

RAW 264.7 _cells were distributed to ²⁴ -well plates at 8.0 Afterwards, the sample and LPS (1 μg/mL) were simultaneously treated and cultured for 24 hours, and then 100 μL of the supernatant and Griess reagent [1% (w/v) sulfanilamide, 0.1% (w/v) naphylethylenediamine in 2.5% (v/ v) phosphoric acid] 100 μL was mixed in equal amounts in 96-well plates, reacted in the dark for 10 minutes, and absorbance was measured at 540 nm.

(4) Prostaglandin E ₂ (PGE ₂ ) Measurement of production inhibition activity

RAW 264.7 cells were distributed on 24 ^- well plates at 8.0 Afterwards, the culture medium was centrifuged at 10,000 rpm for 3 minutes to remove precipitates, and the supernatant was recovered. The content of PGE ₂ in the recovered supernatant was measured using a mouse enzyme-linked immnunosorbent assay (ELISA) kit (R&D Systems Inc., Minneapolis, MN, USA).

(5) Measurement of inhibitory activity on the production of pro-inflammatory cytokines (IL-6, IL-1β, TNF-α)

RAW 264.7 cells were dispensed onto ²⁴ -well plates at a _{concentration} of 8.0 They were treated simultaneously and cultured for 24 hours. Afterwards, the culture medium was centrifuged (10,000 rpm, 3 min) to remove precipitates, and the supernatant was recovered to measure the amount of pro-inflammatory cytokines produced. The amount of pro-inflammatory cytokines produced in the supernatant was measured using the Mouse TNF-α ELISA Kit (Invitrogen, California, USA), Mouse IL-6 ELISA Kit (BD Biosciences, California, USA), and Mouse IL-1β ELISA Kit (R&D Systems Inc., Minneapolis). , MN, USA).

(6) Statistical processing

All experiments were repeated three times and the results were expressed as mean ± standard deviation. In addition, statistical analysis was conducted using analysis of variance (ANOVA) and multiple comparisons using student's t-test to verify the significance of each treatment section (*p<0.05; **p<0.01: ***p<0.001). did.

2) Experiment results

(1) Cell survival rate

Centella asiatica (CA) callus hot water extract (comparative example) cultured under dark culture conditions and callus hot water extract (Examples R and B) cultured under LED (Red, Blue) light irradiation conditions were 50, 100 , As a result of treatment at a concentration of 200 μg/mL, the groups administered extracts of Examples R and B showed a cell survival rate of more than 80% at all concentrations. As shown in Figure 1, cancer culture was performed at the highest concentration of 200 μg/mL. Centella callus extract showed slightly improved cell viability compared to callus (comparative example).

(2) NO production inhibitory activity

Based on the experimental results of (1) above, an experiment was conducted to determine the effect on NO production in RAW264.7 cells. As a result, the callus extract cultured by LED (Red, Blue) light irradiation (Example R, B) was confirmed to exhibit increased NO inhibitory activity compared to the dark-cultured Centella asiatica callus extract (comparative example). In addition, Blue LED culture (Example B) showed the most increased activity and showed a similar level of NO reduction activity as that of the LPS-free group at the highest concentration of 200 μg/mL.

(3) Prostaglandin E ₂ (PGE ₂ ) production inhibitory activity

To determine the effect of dark-cultured Centella asiatica callus extract (comparative example) and Blue LED-cultured Centella asiatica callus extract (Example B) on the amount of PGE ₂ produced in RAW264.7 cells, LPS (1 μg/ml) and each sample were As a result of treatment at concentrations of 50, 100, and 200 μg/mL, as shown in Figure 3, Blue LED cultured Centella asiatica callus extract (Example B) showed a concentration-dependent increase in PGE ₂ inhibitory activity, and cancer inhibition at all concentrations. It showed PGE ₂ inhibitory activity at a higher level than that of the cultured Centella callus extract (comparative example).

(4) Inhibitory activity on the production of pro-inflammatory cytokines (IL-6, IL-1β, TNF-α)

Based on the results of the cytotoxicity test, the production of pro-inflammatory cytokines IL-6, IL-1β, and TNF-α was measured by treating cancer-cultured Centella Asiatica callus extract (comparative example) and Blue LED-cultured Centella Asiatica callus extract (Example B). Measured. Blue LED cultured Centella Asiatica callus extract (Example B) showed an increased tendency to decrease TNF-α in IL-6 and IL-1β cytokines compared to the dark-cultured Centella Asiatica callus extract (comparative example) (Figures 4-6), and IL-6 In the case of , the highest concentration of 200 μg/mL showed IL-6 reducing activity at a level similar to that of the LPS-free group (control group) (Figure 4). In addition, unlike the dark-cultured Centella asiatica callus extract (comparative example), which did not show TNF-α reducing activity, RAW264.7 cells treated with Blue LED-cultured Centella asiatica callus extract (Example B) showed significant TNF-α reducing activity ( Figure 6).

(5) Conclusion

In the present invention, the effect of changes in growth conditions on the anti-inflammatory activity of Centella callus was confirmed. RAW264.7 cells stimulated with LPS were used for the experiment. LED (Red, Blue) cultured Centella asiatica callus hot water extract (Example R and Example B) showed slightly improved cell survival rate compared to dark-cultured Centella asiatica callus hot water extract (comparative example), at a non-toxic concentration. It showed inhibitory activity against NO, which was increased by LPS stimulation. In particular, the Blue LED-treated Centella callus hot water extract (Example B) showed increased activity compared to the Red LED-treated Centella callus hot water extract (Example R), and the pro-inflammatory cytokines IL-6, IL-1β, and TNF- α also showed increased inhibitory activity compared to the dark-cultured Centella Asiatica callus hot water extract (comparative example). In the case of IL-6, at the highest concentration of 200 μg/mL, it showed a similar level of NO-reducing activity as the LPS-free group (control group), and dark-cultured Centella Asiatica callus hot water extract did not show TNF-α reducing activity (comparative example) ), unlike RAW264.7 cells treated with Blue LED-treated Centella asiatica callus hot water extract (Example B), showed significant reduction activity. This suggests that efficacy was improved due to changes in various components present in Centella asiatica callus during the culturing process using LED.

Claims (12)

An anti-inflammatory composition comprising as an active ingredient a Centella asiatica callus extract cultured in an environment irradiated with LED light of 400 nm to 760 nm. According to paragraph 1,
An anti-inflammatory composition, characterized in that the Centella asiatica callus extract is a water or ethanol extract. According to paragraph 2,
An anti-inflammatory composition, characterized in that the Centella asiatica callus extract is an extract of Centella asiatica callus cultured in an environment irradiated with at least one type selected from LED light of 400 nm to 500 nm and LED light of 610 nm to 760 nm. According to paragraph 2,
An anti-inflammatory composition, characterized in that the Centella asiatica callus extract is a Centella asiatica callus extract cultured in an environment irradiated with LED light of 400 nm to 500 nm. According to paragraph 1,
The anti-inflammatory composition is a pharmaceutical composition for preventing or treating inflammatory diseases. According to paragraph 1,
The anti-inflammatory composition is a cosmetic composition for preventing or improving inflammatory diseases. According to paragraph 1,
The anti-inflammatory composition is a food composition for preventing or improving inflammatory diseases. According to any one of claims 5 to 7,
The inflammatory diseases include inflammatory bowel disease, non-alcoholic chronic hepatitis, chronic hepatitis, Crohn's disease, pancreatitis, esophagitis, gastritis, colitis, ulcerative colitis, pneumonia, bronchitis, pharyngitis, myocardial infarction, heart failure, arthritis, psoriatic arthritis, and rheumatism. One or more diseases selected from the group consisting of arthritis, renal failure, psoriasis, anemia, diabetes, fibrosis, multiple sclerosis, systemic lupus erythematosus, ankylosing spondylitis, asthma, chronic obstructive pulmonary disease, periodontitis, neuropathic pain, and idiopathic inflammatory myopathy. Characterized by an anti-inflammatory composition. (a) cultivating Centella asiatica calli in an environment where LED light of 400 nm to 760 nm is irradiated; and
(b) extracting the cultured callus with a solvent. According to clause 9,
A method for producing an anti-inflammatory composition, characterized in that the solvent in step (b) is water or ethanol. According to clause 9,
The step (a) is a method for producing an anti-inflammatory composition, characterized in that the centella asiatica callus is cultured in an environment where one or more types selected from 400 nm to 500 nm LED light and 610 nm to 760 nm LED light are irradiated. According to clause 9,
The step (a) is a method of producing an anti-inflammatory composition, characterized in that the centella asiatica callus is cultured in an environment where LED light of 400 nm to 500 nm is irradiated.