KR20090040631A - Composition for preventing or treating immune disease comprising dandelion water extract as an active ingredient - Google Patents

Abstract

A composition for preventing or treating immune diseases comprising dandelion(Taraxacum offininale) water extract is provided to suppress the generation of nitrogen oxide (NO) and iNOS(inducible nitrogen oxide synthetase) by accelerating the expression of mRNA(messenger RNA) of heme oxygenase-1(HO-1) and protein and increasing the enzyme activity. A composition for preventing or treating immune diseases comprising dandelion(Taraxacum offininale) water extract as an active ingredient. The dandelion water extract is obtained by heating after mixing organic dandelion powder and water, filtering after decompression, freezing and drying them. The one day dose of the active compound is 0.000001-1 mg/kg, preferably 0.001-500 mg/kg, more preferably 10-300 mg/kg. The immunization disease is acute and chronic infection, acute and chronic bronchitis, osteoarthritis, rheumatoid arthritis, sinusitis, gastroenteritis, colitis, cystitis, urethritis, dermatitis, conjunctivitis, pericarditis, peritonitis, synovitis, pleurisy, tendinitis, cholecystitis, vaginitis or uveitis.

Description

COMPOSITION FOR PREVENTING OR TREATING IMMUNE DISEASE COMPRISING DANDELION WATER EXTRACT AS AN ACTIVE INGREDIENT}

The present invention relates to a novel use of dandelion ( Taraxacum offininale ) water extract (TOWE), and more particularly, containing the dandelion hot water extract, which induces the expression and activity of hemeoxygenase-1, as an active ingredient. It relates to a composition for the prevention or treatment of immune diseases.

Dandelion ( Taraxacum offininale ) is a perennial herbaceous plant that is called the genus of the Anzalea , which is also called a wildebeest . Flowers bloom in April-May, and young leaves have been edible by herbs and national streets. Sunghwan Kil, Herbal Medicine and Trees, Chung-Ang University Press, p332, 1997). Dandelion is also called pogongyoung (Kim Won, Botanical Botany, Kyungpook National University Press, p190, 1987), and it has been used in tonic, fever, diuresis, dry stomach, expectorant, and detoxification in oriental medicine. 101 native species, Woongjin Press, 1990, 18). Dandelion taste and properties are bitter and sweet. Outpost has an edema and speeds up the secretion of gastric juice.

Dandelion contains ingredients such as taraxasterol, choline, inulin and pectin, and roots are taraxol, taraxenol, taraxolol, betaamiline, stigmasterol, choline, organic acids, fructose , Sucrose, glucose, glucoside and the like, the leaves contain rutin, violaxanthin, plastoquinone, vitamin C (50-70 mg / 100 g) and vitamin D (5-9 mg / 100 g), Flowers contain alnidiol, rutin and flavoxanthin (Oriental Medicine Dictionary, Kyung Hee University Press, p514, 1999). As a special ingredient, it contains a lot of inulin, sterol, choline, palmitine and serine. Especially, dandelion linoleic acid and choline are known to be effective in adult diseases such as hypertension, heart disease and liver disease (Kim YG et al. , Free radical, YeoMoon Gak, Seoul, 1997).

Component in the study of these plants is in Taraxacum Taraxacum officinale roots ρ - hydroxyphenyl acetic acid (ρ -hydroxyphenyl-acetic acid), 3,4- dimethyl hydroxycinnamic acid (3,4-dihydroxycinnamic acid), pyromellitic seusan (melissic acid), taraxasteryl acetate, taraxol, taraxerol, and ψ -taraxasterol have been reported to have been isolated (Hwanwan, etc.) Phenol component, 25 (3): 209-213, 1994; and Hans-Willi R and Jai-tung H, Phytochemistry , 24: 1557-1562, 1985). In the West, dandelion is used as a medicine for bile secretion, antirheumatic and diuretic (Yang KS and Jeon CM, Korean J. Pharmacogn. , 27: 267-273, 1996). (Jeong JY et al. , Arch. Pharm. Res. , 14: 68-72, 1991), especially chlorophyll and vitamin C, among the polyphenolic compounds such as flavonoids, luteolin, cinnamic acid, coumarin, taraxasterol, etc. (Williams CA et al. , Phytochemistry , 42: 121-127, 1996).

Heme oxygenase (HO) has recently been recognized for its role in antioxidant activity. HO is a rate-limiting enzyme that converts heme into biliverdin, where incidental production of carbon monoxide (CO) and iron (Fe) occurs. Biliburdine is then reduced to bilirubin and used to clean various oxides. HO is known to have three isomers (McCoubrey et al. , 1997) . Unlike HO-2 and HO-3, which are still expressed, HO-1 is an enzyme induced by several stimuli, especially oxidative. It is known to be induced by stress (Tenhumen et al. , 1970; and Choi and Alam, 1996), and it is known to perform protective functions, particularly antioxidant and anti-inflammatory (Otterbein et al. , 2000; and Lee). et al. , 2003).

In addition, bilirubin, the final oxide of HO, protects cells from oxidative stress, and recent studies suggest that carbon monoxide (CO) in addition to bilirubin is also involved in protecting cells. HO is a particularly important enzyme for the protection of the immune system against oxidative stress, as evidenced by reports of increased cell damage by oxidative stress in HO-deficient cells (Akihiro et al. , 1999).

Increasing HO-1 inhibits inducible nitric oxide synthase (iNOS), which is overproduced by the bacterial toxin lipopolysaccharide (LPS). Development of useful substances that induce iNOS inhibition through an increase of -1 is underway (Chen TH et al. , 2007).

NO is known to be a multifunctional mediator produced by various cells and acting on these cells and involved in the destruction of tissues mediated by inflammation and autoimmunity. Such NO is produced by nitric oxide synthase (NOS) in vivo, and NOS is continuously produced (constitutive NOS, cNOS) and induced by stimulation (inducible NOS, iNOS) is present.

The NOS is an enzyme for producing NO from L-arginine, among which nNOS (neuronal NOS) present in the nervous system and eNOS (endothelial NOS) present in the vascular system are always expressed at a constant level in the body. NO produced by them plays an important role in maintaining normal homeostasis, such as inducing neurotransmission and vasodilation. However, iNOS is induced by various cytokines or external stimulants, and it is known to rapidly produce excess NO, causing cytotoxicity and various inflammatory reactions. In particular, chronic inflammation is closely related to the increase of iNOS activity. (Miller MJ et al., Mediators of inflammation , 4: 387-396, 1995; and Appleton L. et al., Adv. Pharmacol. , 35: 27-28, 1996). In addition, gene expression of iNOS is regulated by electronic regulators such as NF-κB and AP-1, which are rarely expressed in normal time, but are rapidly or temporarily expressed when the immune system is activated. It is known to be synthesized.

When the NO is excessively expressed in the cell, it causes an immune system disease such as inflammation or autoimmune disease, resulting in inflammation and autoimmune disease. Therefore, a component having an activity that inhibits NO production will be effective in preventing and treating the inflammation or autoimmune disease which is a disease caused by the overproduction of NO.

Therefore, the present inventors have been conducting research for the development of a therapeutic agent for preventing or treating a disease caused by the overproduction of NO, and the dandelion hot water extract promotes the mRNA and protein expression of HO-1 and increases the enzyme activity. The present invention has been completed by identifying that iNOS can be inhibited and cell damage caused by hydrogen peroxide can be reduced.

An object of the present invention to provide a composition that can prevent or treat immune diseases through the induction of the expression and activity of HO-1.

Another object of the present invention is to provide a food composition that can effectively prevent immune diseases.

In order to achieve the above object, the present invention provides a composition for the prevention or treatment of immune diseases containing dandelion hot water extract as an active ingredient.

In order to achieve the above another object, the present invention provides a food composition for the prevention of immune diseases comprising dandelion hot water extract.

Since the composition of the present invention comprising dandelion hot water extract can increase the gene expression and enzyme activity of HO-1 in immune cells, it can be effectively used for the prevention or treatment of immune diseases.

The pharmaceutical composition containing dandelion hot water extract according to the present invention as an active ingredient not only has the activity of increasing the expression and enzyme activity of HO-1, but also inhibits the production of NO and iNOS induced by overproduction by LPS. In addition, it prevents cell damage induced by hydrogen peroxide, and the dandelion hot water extract does not affect the survival rate of the cells and does not show side effects, and is safe because it is a natural substance derived from an edible plant.

In one embodiment of the present invention, to determine whether the dandelion hot water extract induces the expression of HO-1 treated with dandelion hot water extract in the macrophage line RAW 264.7 cell line of the mouse and cultured, and then isolate the protein from the cells, An enzyme reaction was performed on the HO-1 protein. As a result, it was confirmed that dandelion hot water extract increased the enzyme activity of HO-1, and the increase of HO-1 enzyme activity by dandelion hot water extract was comparable to the effect of hemin, which is a conventional HO-1 activity increasing agent. (See Figure 1).

In another embodiment of the present invention, the dandelion hot water extract alone or mixed and cultured with a bacterial toxin LPS (lipopolysaccharide), which induces an immune response such as an inflammatory reaction, and then cultures the protein from the cells, and HO-1 protein Western blots were performed using antibodies against. As a result, the dandelion hot water extract was confirmed to induce an increase in the protein production of HO-1 (see Fig. 2). Therefore, it can be seen that the increase of HO-1 enzyme activity by dandelion hot water extract is due to the effect of increasing HO-1 protein expression.

In another embodiment of the present invention, after culturing the dandelion hot water extract and LPS-treated cells, RNA was isolated from the cells and subjected to reverse transcription-polymerase chain reaction analysis. As a result, it was confirmed that the dandelion hot water extract induces gene expression of HO-1 at the mRNA level (see FIG. 3).

In addition, in another embodiment of the present invention, after culturing the cells treated with dandelion hot water extract and LPS in order to determine whether inhibition of iNOS production by HO-1 expression induction, and to separate the protein from the cells, for the iNOS protein Western blots were performed using the antibodies. As a result, it was confirmed that the dandelion hot water extract inhibits the synthesis of iNOS at the protein level (see FIG. 4). Therefore, it can be seen that the induction of HO-1 expression by dandelion hot water extract is related to the iNOS enzyme expression inhibitory effect.

In addition, as a result of recovering the medium in which the cells were cultured and measuring the amount of nitrite produced by the cells, the degree of NO production was measured. As a result, dandelion hot water extract effectively inhibited NO production in macrophages in a concentration-dependent manner. At this time, it was confirmed that there was no change in viability of the cells (see FIGS. 5 and 6).

In another embodiment of the present invention, in order to confirm the cellular protective effect of dandelion hot water extract when treated with hydrogen peroxide as oxidative stress inducing cell damage, after treating and cultivating dandelion hot water extract and hydrogen peroxide, the survival rate of the cells was measured. . As a result, it was confirmed that the decrease in cell viability caused by hydrogen peroxide was recovered by the dandelion hot water extract (see FIG. 7). From this, it was found that the dandelion hot water extract prevented cell damage caused by oxidative stress such as hydrogen peroxide. there was.

Therefore, the composition of the present invention comprising the dandelion hot water extract as an active ingredient can induce the expression and activity of HO-1, can be used as a therapeutic agent for immune diseases.

Representative examples of the immune diseases include inflammatory diseases, which are manifested by fever, flushing, swelling, pain and loss of function. Causes of inflammation include factors such as microbial infections (bacterial and fungal infections), physical factors (burns, irradiation and trauma), chemical agents (toxins and triggers), tissue necrosis and various types of immunological reactions, especially NO Enzymes related to the biosynthesis of NOS and prostaglandins, which are enzymes that produce N, are known to play an important role in mediating the inflammatory response, and NO, one of the many reactive products produced in immune and inflammatory responses, is particularly responsible for chronic inflammation. It is known to cause nonspecific tissue destruction.

Diseases caused by the overproduction of NO to which the dandelion hot water extract of the present invention can be applied are, but are not limited to, for example, acute and chronic infections, acute and chronic bronchitis, osteoarthritis, rheumatoid arthritis, sinusitis, gastroenteritis, colitis Cystitis, urethritis, dermatitis, conjunctivitis, pericarditis, peritonitis, synovitis, pleurisy, tendonitis, cholecystitis, vaginitis and uveitis.

Dandelion hot water extract can be used in the present invention can be prepared by using an organic dandelion powder from dandelion obtained from nature or commercially purchased dandelion, for example, dandelion foods of Gyeongnam. The extract can be extracted using a known method, it is preferable to use after filtration and freeze-dried under reduced pressure.

In the pharmaceutical composition of the present invention, the dandelion hot water extract of the present invention may be used alone or formulated in combination with one or more pharmaceutically acceptable carriers according to the selected route of administration.

The route of administration of the pharmaceutical composition of the present invention can be administered orally or parenterally, and the parenteral route of administration is not limited thereto, but may be administered subcutaneously, intravenously, intramuscularly, or intraperitoneally. .

For oral administration of the pharmaceutical compositions of the present invention may be formulated in solid forms such as capsules, tablets, pills, granules and powders or in liquid form such as elixis syrups and suspensions. When formulated in solid form for oral administration, fillers, extenders, binders, wetting agents, disintegrants, surfactants and the like known in the art can be used as suitable carriers. For example, to formulate the pharmaceutical composition of the present invention in capsules, carriers such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid can be used. In addition, it can be prepared in the form of compressed tablets using a diluent and both the tablets and capsules can be prepared in a sustained release according to methods known in the art to allow the continuous release of the drug over several hours. In addition, the compressed tablets may have a sugar coating or a film coating. When formulated in liquid form for oral administration, suspending agents, emulsions, syrups, diluents, wetting agents, sweetening agents, fragrances, preservatives and the like known in the art can be used as suitable carriers.

For parenteral administration of the pharmaceutical compositions of the invention, the form of injectables in ampoules or vials is preferred. Suitable carriers when formulated as injectables include stabilizers, buffers and preservatives known in the art, and suitable stabilizers include sodium bisulfite, sodium sulfite and ascorbic acid, and the like. Benzalkonium, methyl or propyl-paraben and chlorobutanol.

Typical daily dosages of the active compounds of the pharmaceutical compositions of the invention may range from 0.000001 to 1 mg / kg body weight, preferably from 0.001 to 500 mg / kg body weight, preferably from 10 to 300 mg / kg body weight. It can be administered once or divided into several times. However, the actual dosage of the active ingredient is determined by the medical judgment of the patient, including age, health, weight, excretion, diet, severity, activity of the compound used, sex, time of administration, route of administration, duration of treatment and frequency of treatment. It is to be understood that such determinations should be made in accordance with the foregoing, and therefore the above dosages do not limit the scope of the invention in any way.

Dandelion hot water extract of the present invention is secured as a natural drug, 50% lethal dose (LD ₅₀ ) of the intraperitoneal administration to mice is a safe substance of at least 28 g / kg extract (K. Sch ㆌ tz et al. , Journal of Ethnopharmacology , 107: 313-323, 2006).

In addition, the dandelion hot water extract of the present invention may be added to food or beverage for the purpose of prevention of various immune diseases.

The health beverage of the present invention has no particular limitation on the liquid component except for containing the dandelion hot water extract as an essential ingredient, and may contain various flavors or natural carbohydrates as additional components, as in general beverages. As the flavoring agent, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. Examples of the natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And sugars such as conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and xylitol, sorbitol, erythritol.

In addition to the above, the food or beverage composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), colorants and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, and preservatives. , Glycerin, alcohols, carbonation agents used in carbonated drinks, and the like.

Examples of foods to which the dandelion hot water extract of the present invention may be added for the development of dietary supplements include various foods, beverages, gums, mitamine complexes and the like.

Hereinafter, the present invention will be described in more detail based on the following examples. However, the following examples are not intended to limit the invention only.

In the following examples, * and ** indicated on the drawings indicate statistical analysis results. For the group not treated with the dandelion hot water extract (the group having 0 concentration in the figure), * indicates p <0.05. * Indicates p <0.01 (statistical analysis performed Dunnett's two-tailed t-test).

Preparation Example 1 Preparation of Dandelion Hot Water Extract

In order to manufacture the dandelion hot water extract of the present invention, the organic dandelion powder supplied from dandelion food of Gyeongnam Ryeongyeong added 10 times distilled water to 1 kg of the powder sample, and then attached to the reflux cooling tube at 100 ± 5 ℃ for 4 hours Extraction, filtration under reduced pressure using a filter paper (Whatman paper No. 4), and freeze-dried. At this time, the recovery rate of the dandelion extract was 22.5%.

Example 1 Induction of HO-1 Enzyme Activity Increase by Dandelion Hot Water Extract

In order to confirm whether the dandelion hot water extract of the present invention induces the enzyme activity of HO-1, the effect of dandelion hot water extract was measured using RAW 264.7 cells (American Type Culture Collection, Manassas, VA, USA), a mouse macrophage line. .

Specifically, RAW 264.7 cells were treated with 1 × in RPMI 1640 medium (hereinafter referred to as 'RPMI medium' for convenience) containing 100 units / ml penicillin, 100 μg / ml streptomycin and 5% fetal bovine serum. The cells were suspended in a concentration of 10 ⁶ cells / ml, placed in a culture dish, and incubated at 37 ° C. in a 5% CO ₂ incubator. While culturing the RAW264.7 cells, the dandelion hot water extract was treated at concentrations of 100, 500 and 1,000 ㎍ / ml, respectively, and incubated for 18 hours. As a control group, a group treated with 100 μM of hemin known as a HO-1 inducer was used. Thereafter, the cells were collected and subjected to enzymatic reaction using a HO-1 enzyme reaction system. The absorbance of the product, bilirubin, was measured to measure HO-1 enzyme activity, and the results are shown in FIG. 1. In addition, the recovered cells were also used for measuring cell viability, and cell viability was measured according to the MTT assay, and the results are shown in FIG. 6.

As shown in Figures 1 and 6, in the group treated with dandelion hot water extract, the activity of HO-1 HO-1 enzyme increased with increasing dose, and the level could be comparable to that of the control group treated with hemin. there was. In addition, as shown in Figure 5, it was confirmed that there is no change in the cell viability at this time.

Example 2 Measurement of Induction of HO-1 Expression by Dandelion Hot Water Extract

<2-1> Measurement of HO-1 Induction by Western Blot

Whether the dandelion hot water extract of the present invention induces the expression of HO-1 was confirmed by Western blot.

Specifically, the RAW264.7 cells were suspended in a concentration of 1 × 10 ⁶ cells / ml using RPMI medium and placed in a culture dish, followed by culturing at 37 ° C. in a 5% CO ₂ incubator to obtain 100, 100% of dandelion hot water extract. The cells were incubated for 24 hours by treatment at concentrations of 500 and 1,000 μg / ml. In order to measure the effect on the activated cells, after treatment with LPS 200 ng / ㎖ 1 hour after dandelion hot water extract treatment was incubated for another 24 hours. The cells were then harvested and subjected to Western blot using an anti-mouse HO-1 antibody (Santa Cruz Biotechnology, USA; Dilution Ratio 1: 1,000), as a control to show no change in other proteins in the sample. An anti-mouse beta-actin antibody (Cell Signaling Technology, USA; dilution ratio 1: 1,000) was used and the results are shown in FIG. 2.

As shown in Figure 2, whether or not LPS treatment, dandelion hot water extract treatment increased HO-1 protein synthesis, and as the amount of dandelion hot water extract administered increased HO-1 protein synthesis was confirmed to further increase Could.

<2-2> Measurement of HO-1 Synthesis Increase by Reverse Transcription-Polymerase Reaction (RT-PCR)

In order to confirm the increase in the synthesis of HO-1 by the dandelion hot water extract of the present invention at the mRNA level, RT-PCR was performed.

Specifically, after culturing the cells in the same manner as in <2-1>, and treated with dandelion hot water extract at the concentration of 100, 500 and 1,000 ㎍ / ml, and then treated with LPS to activate the cells after 1 hour Incubated for 6 hours. Thereafter, the cells were recovered to separate RNA, and then RT-PCR was performed on the HO-1 gene. RT-PCR was also performed on the beta-actin gene as a control to show that there was no change in other genes in the sample. It was. 1% agarose gel electrophoresis was performed with the reaction product of RT-PCR, and the results are shown in FIG. 3.

As shown in Figure 3, the group treated with the dandelion hot water extract of the present invention was found to increase the amount of mRNA of HO-1 in a concentration-dependent manner.

Example 3 Measurement of iNOS Inhibitory Activity by Dandelion Hot Water Extract

Whether the dandelion hot water extract of the present invention has an activity of inhibiting the expression of iNOS was confirmed by Western blot.

Specifically, the RAW264.7 cells were suspended in a concentration of 1 × 10 ⁶ cells / ml using RPMI medium and placed in a culture dish, followed by culturing at 37 ° C. in a 5% CO ₂ incubator to obtain 100, 100% of dandelion hot water extract. The cells were treated at concentrations of 500 and 1,000 μg / ml, and treated with 200 ng / ml of LPS to activate the cells after 1 hour and then incubated for another 24 hours. As a control group, a group not treated with LPS and a group treated with only LPS were used. Cells were then harvested and subjected to Western blot using an anti-mouse iNOS antibody (Santa Cruz Biotechnology, USA; Dilution Ratio 1: 1,000), and as a control to show no change in other proteins in the sample. Mouse beta-actin antibody (Cell Signaling Technology, USA; dilution ratio 1: 1,000) was used, the results are shown in FIG.

As shown in FIG. 4, iNOS protein synthesis was significantly increased in the LPS-treated group compared with the LPS-treated group, and the iNOS protein was increased as the amount of dandelion hot-water extract increased. It was confirmed that the synthesis was further suppressed.

In addition, since the HO-1 protein concentration-dependently increased when the dandelion hydrothermal extract was treated, it was found that the inhibitory effect of iNOS production of the dandelion hydrothermal extract was related to the increase of HO-1.

Example 4 Measurement of NO Production Inhibition Activity and Cell Viability by Dandelion Hot Water Extract

To confirm whether the dandelion hot water extract of the present invention inhibits NO production, the dandelion using RAW 264.7 cells, a mouse macrophage line known to induce NO production at the time of inactivation, but is activated when activated by LPS. The effect of hot water extract was measured.

Specifically, the RAW264.7 cells were suspended in a concentration of 1 × 10 ⁶ cells / ml using RPMI medium and placed in a culture dish, followed by culturing at 37 ° C. in a 5% CO ₂ incubator to obtain 100, 100% of dandelion hot water extract. The cells were treated at concentrations of 500 and 1,000 μg / ml, and treated with 200 ng / ml of LPS to activate cells after 1 hour and then incubated for another 24 hours. As a control group, a group not treated with LPS and a group treated with only LPS were used. Thereafter, the cell culture was recovered and the amount of NO was measured using a Gries's reaction, and the results are shown in FIG. 5. In addition, cells were recovered and used for measuring cell viability, and cell viability was measured by MTT assay and the results are shown in FIGS. 5 and 6.

As shown in FIG. 5, in the LPS-treated group, excess NO was generated, and when the concentration of the dandelion hot water extract of the present invention was gradually increased and administered together with the LPS, the production of NO was further inhibited as the concentration was increased. It was confirmed that, as shown in Figure 6, it was confirmed that there is no change in the cell viability at this time.

Therefore, it was found that the dandelion hot water extract of the present invention inhibits NO production in a concentration-dependent manner without causing a change in cell viability when immune cells are activated.

Example 5 Recovery of Cell Damage by Hydrogen Peroxide by Dandelion Hot Water Extract

In order to confirm whether the dandelion hot water extract of the present invention restores cell damage caused by oxidative stress, the effect of dandelion hot water extract was measured when the cell damage was induced using hydrogen peroxide.

Specifically, the RAW264.7 cells were suspended in a concentration of 1 × 10 ⁶ cells / ml using RPMI medium and placed in a culture dish, followed by culturing at 37 ° C. in a 5% CO ₂ incubator to obtain 100, 100% of dandelion hot water extract. The cells were treated at concentrations of 500 and 1,000 μg / ml, and treated with 200 μM and 500 μM of hydrogen peroxide to induce cell damage after 1 hour and then incubated for another 18 hours. As a control group, the group without the dandelion hot water extract and the group without the hydrogen peroxide were used. Thereafter, cells were collected and used for measuring cell viability, and cell viability was measured according to the MTT assay, and the results are shown in FIG. 7.

As shown in FIG. 7, in the group treated with hydrogen peroxide only, the cell viability was decreased due to cell damage. When the concentration of dandelion hot water extract of the present invention was gradually increased and administered, the cells with hydrogen peroxide increased as the concentration increased. It was confirmed that the reduction in survival rate was further recovered.

Therefore, the dandelion hot water extract of the present invention was found to have an effect of preventing cell damage by oxidative stress.

Formulation Example 1 : Preparation of a medicament containing dandelion hot water extract as an active ingredient

<1-1> Preparation of Tablet

25 mg of dandelion hot water extract was U-type, with 26 mg of lactose for excipients and 3.5 mg of Avicel (microcrystalline cellulose), 1.5 mg of sodium starch glyconide as a disintegration aid, and 8 mg of low-hydrosyprophylcellulose (L-HPC) for binders. The mixture was put into a mixer and mixed for 20 minutes. After mixing was completed, 1 mg of magnesium stearate was further added as a lubricant and mixed for 3 minutes. Tablets were prepared by tableting and film coating after a quantitative test and a humidity test.

<1-2> Preparation of Syrup

2 g of dandelion hot water extract, 0.8 g of saccharin and 25.4 g of sugar were dissolved in 80 g of warm water. After cooling the solution, 8.0 g of glycerin, 0.04 g of flavor, 4.0 g of ethanol, 0.4 g of sorbic acid, and an appropriate amount of distilled water were mixed, and then water was added to make 100 ml.

<1-3> Preparation of Capsule

A capsule was prepared by mixing dandelion hot water extract 50 mg, lactose 50 mg, starch 46.5 mg, talc 1 mg, and an appropriate amount of magnesium stearate and filling it into a hard gelatin capsule.

<1-4> Preparation of Injection

1 g of dandelion hot water extract, 0.6 g of sodium chloride and 0.1 g of ascorbic acid were dissolved in distilled water to obtain a final volume of 100 ml. The solution was filled in ampoules and heat sterilized.

Figure 1 shows the measurement results of HO-1 enzyme activity according to the dose of dandelion hot water extract.

Figure 2 shows the measurement results of HO-1 protein production induction according to the dose of dandelion hot water extract (TOWE).

Figure 3 shows the induction of mRNA expression of HO-1 according to the dose of dandelion hot water extract (TOWE).

Figure 4 shows the inhibition of protein expression of iNOS according to the dose of dandelion hot water extract (TOWE).

5 and 6 show NO production inhibition and cell viability according to the dose of dandelion hot water extract (TOWE).

Figure 7 shows the recovery of cell damage by hydrogen peroxide according to the dose of dandelion hot water extract (TOWE).

Claims (4)

Dandelion hot water extract (Dandelion ( Taraxacum offininale ) A composition for preventing or treating immune diseases containing water extract, TOWE) as an active ingredient. The method of claim 1, The dandelion hot water extract is characterized in that to promote the mRNA and protein expression of heme oxygenase-1 (heme oxygenase-1, HO-1), a composition for preventing or treating immune diseases. The method of claim 1, These diseases include acute and chronic infections, acute and chronic bronchitis, osteoarthritis, rheumatoid arthritis, sinusitis, gastroenteritis, colitis, cystitis, urethritis, dermatitis, conjunctivitis, pericarditis, peritonitis, peritonitis, pleurisy, tendinitis, cholecystitis, vaginitis and uveitis It is selected from the group consisting of, composition for the prevention or treatment of immune diseases. Claim 1 food composition for the prevention of immune diseases comprising dandelion hot water extract.

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