KR20190035973A - Compositon containing extract of Platycodon grandiflorum and Glycyrrhiza uralensis Fischer - Google Patents

Abstract

The present invention relates to a use of a Platycodon grandiflorum soup containing a Platycodon grandiflorum extract and a Glycyrrhiza uralensis extract or crude saponin separated therefrom for lowering blood pressure. The Platycodon grandiflorum soup and crude saponin separated therefrom can be usefully used in preventing and treating hypertension by relaxing blood vessels. In addition, since extracts of the present invention are natural products, it is possible to be used stably without side effects, thereby being able to be provided as medicine or food for preventing, alleviating, or treating hypertension.

Description

Compositions Containing Gakyung Extracts and Licorice Extracts and Their Use {Compositon containing extracts of Platycodon grandiflorum and Glycyrrhiza uralensis Fischer}

The present invention relates to a composition comprising Gakyung extract and licorice extract; Or a composition for preventing, improving or treating hypertension comprising crude saponin isolated therefrom as an active ingredient.

Hypertension is a risk factor for cardiovascular disease worldwide, with 15-20% of adults suffering from it, but the precise cause of hypertension has not been identified. Family history, race, salt intake, insulin resistance, excessive drinking and aging.

Renin-angiotensin system (RAS) plays an important role among various factors of blood pressure rise. It has been known that general physiological activity causes renin-angiotensin system (ROS) by blood flow, sodium reduction and sympathetic activity increase RAS), and renin degrades angiotensin into angiotensin I, which is then converted to angiotensin II, which is a vasoconstrictor of angiotensin I-converting enzyme (ACE) Re-uptake, increased potassium ion release, aldosterone production to increase blood flow, vasoconstriction, and blood pressure.

Currently known ACE inhibitors include ramipril, D-3-mer-capto-2methylpropanoyl-L-proline, enarapil, risinopril, fosinoril, There is spirapril, which is widely used as a treatment for hypertension. However, since these compounds are easily decomposed in a pharmaceutical dosage form, they are not only inferior in stability but also have other side effects such as vigorous vomiting, coughing, headache, anorexia and taste abnormalities, . Therefore, it is required to develop an ACE inhibitor excellent in ACE inhibitory activity and stability and to develop an antihypertensive agent having another pharmacological mechanism.

Accordingly, the present inventors have conducted research to develop a composition for prevention, improvement or treatment of hypertension using natural substances having less side effects, such as plant extracts, rather than chemical synthetic products.

Kim, Jeong-Sook et al., 4, ACE inhibitory activity and blood pressure lowering effect of essential hypertensive rats (SHR) administered with Mulberry japonica extract, Journal of the Korean Society of Food Science and Nutrition 37, No. 1, 2008.1, 27-34

The object of the present invention is to provide a ginseng root extract comprising ginseng extract and licorice extract; Or a crude saponin isolated therefrom as an effective ingredient.

The inventors of the present invention have confirmed that Gwangyang extract and licorice extract have an effect of preventing or treating hypertension by inducing vasodilatory action during research on natural products against hypertension, and completed the present invention.

The present invention relates to a composition comprising Gakyung extract and licorice extract; Or a new use for lowering the blood pressure of crude saponin isolated therefrom, Gilgyeongtang consisting of Gyunggyeong extract and licorice extract; Or a pharmaceutical composition for lowering blood pressure comprising crude saponin isolated therefrom as an active ingredient, a therapeutically effective amount of Gakkyeong extract and licorice extract; Or a crude saponin isolated therefrom to a subject.

In the following examples, rabbit blood vessel relaxation effect was observed after administration of crude saponin extract and crude saponin isolated from licorice root extract to rabbits inducing the contraction of brain basal artery with endodelin. Therefore, Gwangyang extract and licorice extract; Or crude saponin separated therefrom can be used as an active ingredient of a composition for lowering blood pressure.

In the present invention, Gyeonggyang-tang means a herbal medicine product containing Gyungyang and licorice, which is a medicinal herb, and can be prepared by mixing Gyungyang extract and licorice extract with extracting Gyungyang and licorice extracts, ≪ / RTI >

In the present invention, "endothelin" is the most potent vasoconstrictor secreted in endothelial cells, and is considered to be a target of pulmonary hypertension, diabetes and inflammatory diseases. Gilgyeong tang of the present invention; Or crude saponin isolated therefrom showed vasoconstriction effect specifically on endothelial - induced vasoconstriction.

In the present invention, "pulmonary hypertension" or "pulmonary hypertension" is a disease in which the blood pressure of the pulmonary artery rises due to a problem in blood vessels supplying blood to the lungs unlike systemic hypertensive diseases. The pulmonary artery pressure is 25 mmHg or more, When exercising, it means 30mmHg or more.

In the present invention, " crude saponin " may mean saponin of macromolecular weight separated from Gilgyeongtang mixed with Gakyung extract and licorice extract.

In one embodiment, the composition according to the present invention comprises, as an active ingredient, Gil-gyeong-tang, which consists of Ganoderma lucidum extract and licorice extract.

In the present invention, the extract of Ganoderma lucidum and Licorice extract contains all the extracts obtained by the compression method or the solvent extraction method.

In one embodiment of the present invention, Ganoderma lucidum extract and licorice extract may be respectively extracted from Gakugei and licorice extracts using water, an organic solvent or a mixture thereof as an extraction solvent. The kind of the organic solvent used and the mixing ratio of water and the organic solvent are not particularly limited.

For example, the organic solvent may be one or more solvents selected from the group consisting of lower alcohols, hexane, acetone, ethyl acetate, chloroform and diethyl ether. The lower alcohol may be an alcohol having 1 to 6 carbon atoms. For example, as the lower alcohol, methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol, 2-butoxyethanol or ethylene glycol can be used. The organic solvent may be a polar solvent such as acetic acid, dimethyl-formamide (DMFO), or dimethylsulfoxide (DMSO), acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, 2,2,4- Butene, 1-chloropentane, o-xylene, diisopropyl ether, 2-chloropropane, toluene, 1-chloropropane, chlorobenzene, A nonpolar solvent such as benzene, diethyl ether, diethyl sulfide, chloroform, dichloromethane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride and THF (tetrahydrofuran) may be used.

In one embodiment, the ginseng extract and the licorice extract may be hot water extracts of Gakuen and licorice or a lower alcohol extract. Preferably ethanol extracts of Ganoderma lucidum and licorice root. In another embodiment, the ginseng extract and the licorice extract may be hexane extract of licorice and licorice. In the present specification, the extract of organic solvent includes organic solvent alone or extract obtained by using a mixed solvent of organic solvent and water as extraction solvent. For example, the ethanol extracts of Gyunggi and licorice include not only extracts using ethanol alone but also extracts obtained by using a mixture of water and ethanol.

For example, the ginseng extract and the licorice extract of the present invention may be prepared by mixing ginseng and licorice at a ratio of 1: 9 to 4: 6, and mixing the ginseng at a ratio of 3 to 10 times, preferably 5 times water extraction, heat extraction, ultrasonic extraction, and reflux extraction, in water or in a mixed solvent of these, preferably water or 5 to 95% ethanol, in a solvent such as water, ethanol, methanol, Preferably 70 to 120 ° C, preferably 80 to 110 ° C, for 1 to 12 hours, preferably 4 to 10 hours by a heating method, preferably a heating method; A second step of filtrating and concentrating the extract obtained in the above step under reduced pressure; And then lyophilized at 50 ° C to 20 ° C, preferably at 45 ° C to 20 ° C for 12 to 72 hours, preferably for 12 to 50 hours. An extract can be obtained. In this case, in the first step, the process of obtaining extract from Gyeonggyeong and licorice may include mixing gyeonggyeong and licorice extract to extract gyeonggyeong and licorice, or extracting gakyung and licorice with organic solvent, respectively, have.

Also, in one embodiment, the Ganoderma lucidum extract and Licorice root extract are blanched for about 5 minutes at about 100 < 0 > C before breaking down the Gakgye and licorice to obtain an effective preventive or therapeutic extract of hypertension Followed by inactivation of the intracellular enzyme.

As used herein, the term " extract " also includes fractions obtained by further fractionating the extract. That is, the extract of Ganoderma lucidum and licorice extract includes not only those obtained by using the above-mentioned extraction solvent, but also those obtained by further applying a purification process thereto. Further, fractions obtained by passing the above extract or fractions through an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity), and the like The fractions obtained through various purification methods were also included in the Gakyung extract and licorice extract of the present invention.

In one embodiment, the ginseng extract and the licorice extract may be fractions obtained by fractionating an organic solvent extract of Gakugei and licorice with a second organic solvent. In another embodiment, the ginseng extract and the licorice extract may be fractions obtained by fractionating a lower alcohol extract of Gakugei and cinnamon into a second organic solvent. For example, the extract of Ganoderma lucidum and licorice extract may be a fraction obtained by fractionating hexane extract of licorice and licorice with chloroform.

In the present specification, 'Gyunggyeong extract' includes not only crude extract obtained by treating the extracting solvent in Gyeonggi, but also a processed product of Gyungyang extract. For example, Ganoderma lucidum extract can be prepared in powder form by an additional process such as vacuum distillation and lyophilization or spray drying.

The 'Gyeonggyeong' means bellflower, which is a perennial plant that grows frequently on mountains and is 40 to 100 centimeters in height. It has a thick root and a stem. Gakgyeong has been used as a medication for neuralgia and tonsillitis, and it has been used as an anti-inflammatory analgesic and an anti-inflammatory agent because it has a genomic DNA, Jinhae, analgesic, sedative and antipyretic action.

In the present specification, 'licorice extract' includes not only crude extract obtained by treating an extractive solvent with licorice but also a licorice extract. For example, licorice extracts can be prepared in powder form by additional processes such as vacuum distillation and lyophilization or spray drying.

The 'Licorice' stands for a plant with a height of about 1.5m and bushy straight root. Licorice can be used as a treatment for sweeteners, stomach ulcers, and duodenal ulcers in roots and stem bottoms, and it can be used for various purposes in oriental medicine. In general, licorice contains glycyrrhizin, glabric acid, sucrose, glucose, liquiritin and lycoricidine. Glycyrrhizin has been used as a therapeutic agent for tetanus due to the detoxification effect of diphtheria toxin, snake venom and blowfish poison.

The composition of the present invention may contain the ginseng extract and the licorice extract in an amount of 0.01 to 50 parts by weight based on the total weight of the composition. The composition of the present invention is not limited thereto, and may vary depending on, for example, the concentration of Ganoderma lucidum extract and licorice extract, the condition of the subject to which the composition of the present invention is administered, the type of disease,

In one embodiment, the ginseng extract and licorice extract may be included in the composition in a ratio of 1: 9 to 4: 6. By incorporating the composition in the above proportions, the effect as GilYanggang can be realized, and the ratio can be, for example, from 2: 8 to 4: 6, but is not limited thereto.

As used herein, the term " comprising as an active ingredient " means to include an amount sufficient to achieve the potency or activity of Ganoderma lucidum extract and licorice extract. In one embodiment of the present invention, the Ganoderma extract and licorice extract in the composition of the present invention have a concentration of, for example, not less than 0.01 mg / ml, preferably not less than 0.05 mg / ml, more preferably not less than 0.1 mg / More preferably 1 mg / ml or more. The quantitative upper limit of the extract of Ganoderma lucidum and Licorice root extract contained in the composition of the present invention can be selected by a person skilled in the art within a suitable range.

In another embodiment, the composition according to the present invention contains crude saponin isolated from Gilgyeongtang consisting of the Gypsum gypsum extract and licorice root extract as an active ingredient.

In the present invention, the crude saponin may be contained at a concentration of 1 to 10 μg / ml. When the composition according to the present invention contains crude saponin within the above-mentioned concentration range, a significant amount of crude saponin can exhibit significant blood vessel relaxation effect.

The composition according to the present invention can be utilized as a pharmaceutical composition or a food composition.

In one embodiment, the composition may be a pharmaceutical composition for lowering blood pressure.

The blood pressure lowering pharmaceutical composition preferably relaxes endothelial-induced vasoconstriction, but is not limited thereto. The crude saponin isolated from Gilgyeong-tang of the present invention inhibits endocardial contraction induced by endodelin and relaxes the blood vessel, so that it can be deduced that it has endodelin inhibitory effect. Accordingly, the pharmaceutical composition for lowering blood pressure of the present invention can be used for prevention or treatment of endothelial-induced pulmonary arterial hypertension, but is not limited thereto.

The pulmonary arterial hypertension may be different from systemic hypertension due to depolarization induced by increased potassium concentration.

In the following examples, crude saponin isolated from Gilgyeongtang or the present invention had no vasoconstriction effect on high-concentration potassium-related vasoconstriction related to general system hypertension. However, it has been reported that endospermine induced by pulmonary arterial hypertension, diabetes, A blood vessel relaxation effect was specifically observed in the vasoconstriction, so that the composition could be used as a blood pressure lowering composition. Also, through the above experimental results, the composition of the present invention is useful as a renal disease such as chronic renal failure and glomerulosclerosis induced by abnormal hyperactivity of endodermal system; Respiratory diseases such as idiopathic pulmonary fibrosis and chronic obstructive pulmonary disease; Diabetic neuropathy, retinopathy, and necrosis, and cancer diseases such as prostate cancer and colorectal cancer.

The pharmaceutical composition of the present invention may be prepared by using pharmaceutically acceptable and physiologically acceptable adjuvants in addition to the above-mentioned active ingredients. Examples of the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, Or a flavoring agent.

The pharmaceutical composition may be formulated into a pharmaceutical composition containing at least one pharmaceutically acceptable carrier in addition to the active ingredient described above for administration.

The pharmaceutical form of the pharmaceutical composition may be a granule, a powder, a tablet, a coated tablet, a capsule, a suppository, a liquid, a syrup, a juice, a suspension, an emulsion, a drip agent or an injectable liquid agent. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthan gum and the like.

Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile water and sterile water suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants can be added to formulate into injectable formulations, pills, capsules, granules or tablets, such as aqueous solutions, suspensions, emulsions, and the like.

Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

The pharmaceutical composition of the present invention can be administered orally or parenterally. In the case of parenteral administration, the composition can be administered by intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection, transdermal administration, etc., .

The appropriate dosage of the pharmaceutical composition of the present invention varies depending on factors such as the formulation method, the manner of administration, the age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, , The ordinarily skilled physician can readily determine and prescribe dosages effective for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dose of the pharmaceutical composition of the present invention is 0.0001 to 10 g / kg.

The pharmaceutical composition of the present invention may be prepared in a unit dosage form by formulating it with a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by those having ordinary skill in the art to which the present invention belongs Or into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

The present invention also provides a food composition for supplementing blood pressure lowering which comprises Ganoderma lucidum extract and licorice extract as an active ingredient.

The food composition according to the present invention can be formulated in the same manner as the above-mentioned pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectioneries, diet bars, dairy products, meat, chocolates, pizza, ram noodles, other noodles, gums, ice cream, .

The food composition of the present invention may contain, as an active ingredient, Gakyung extract and licorice extract, as well as ingredients that are ordinarily added in the course of food production, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavors do. Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavors (saccharin, aspartame, etc.) may be used as flavorings. For example, when the food composition of the present invention is prepared from a drink and beverage, it may further contain citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, various plant extracts, etc., .

The present invention provides a health functional food comprising the Gypsum gypsum extract and the licorice extract as an active ingredient and a food composition for prevention and improvement of hypertension. Health functional foods are foods prepared by adding Gyungyang extract and licorice extract to food materials such as beverages, tea, spices, gum, confectionery, or encapsulated, powdered, or suspension. However, unlike general medicine, there is an advantage that there is no side effect that can occur when a food is used as a raw material for a long period of taking the medicine. The health functional food of the present invention thus obtained is very useful because it can be ingested routinely. The amount of Gyungyang extract and licorice extract added in such a health functional food can not be uniformly defined depending on the kind of the health functional food to be added. However, it can be added within a range that does not deteriorate the original taste of the food, And usually ranges from 0.01 to 50 parts by weight, preferably from 0.1 to 20 parts by weight. In the case of health functional foods in the form of pills, granules, tablets or capsules, they may be added usually in the range of 0.1 to 100 parts by weight, preferably 0.5 to 80 parts by weight. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule or beverage.

The present invention also provides the use of Ganoderma extract and licorice extract for the manufacture of a medicament or food for the prevention, amelioration and treatment of hypertension. As described above, Gwangyang extract and licorice extract can be used for prevention, improvement and treatment of hypertension.

The present invention also provides a method of preventing, ameliorating and treating hypertension comprising administering to a mammal an effective amount of Ganoderma extract and licorice extract.

As used herein, the term " mammal " refers to a mammal that is the subject of treatment, observation, or experimentation, preferably a human.

As used herein, the term " effective amount " refers to the amount of active ingredient or pharmaceutical composition that elicits a biological or medical response in a tissue, animal, or human subject as contemplated by a researcher, veterinarian, physician or other clinician, ≪ / RTI > inducing a reduction of the symptoms of the disease or disorder. It will be apparent to those skilled in the art that the effective amount and the administration frequency of the active ingredient of the present invention will vary depending on the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art and will vary with the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation and the age, weight, , Sex and diet, time of administration, route of administration and rate of administration of the composition, duration of treatment, concurrent administration of the drug, and the like. In the prevention, treatment, or improvement of the present invention, it is preferable to administer the extract of Ganoderma lucidum and the licorice extract at a dose of 0.001 mg / kg to 10 g / kg once or several times a day.

The composition comprising Ganoderma lucidum extract and Licorice root extract as an active ingredient in the treatment method of the present invention can be administered orally, rectally, intravenously, intraarterally, intraperitoneally, intramuscularly, intrasternally, transdermally, topically, Lt; / RTI >

Accordingly, the composition according to the present invention can be applied to the prevention, improvement, and treatment of diseases caused by endothelial-induced contraction of blood vessels. In one embodiment, the composition of the present invention can be applied to the prevention, improvement, and treatment of hypertension, but the present invention is not limited thereto. For example, the composition may include diabetes, kidney failure, foot necrosis, and peripheral circulatory disorder.

The Gakyung extract and licorice extract according to the present invention may be useful for preventing and treating hypertension by relaxing blood vessels. In addition, the extracts of the present invention can be used as a natural product and can be used stably without side effects, and can be provided as medicines or foods for lowering blood pressure.

FIG. 1 shows the results of blood vessel relaxation after treatment of crude saponin of the present invention in the basal artery of the rabbit inducing vasoconstriction with high concentration of potassium.
FIG. 2 shows the result of confirming the degree of vasorelaxation after treating crude saponin of the present invention in the basal artery of the rabbit which induced vasoconstriction by endodelin treatment.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

≪ Example 1 > Preparation of Ganoderma lucidum extract and licorice root extract

 Gyeonggyung and licorice were purchased at the Yakyong-dong Yakju market. Three grated licorice of 300g in dry weight and three grated licorice in 700g were mixed with each other and immersed in 3L of ethanol. The mixture was heated and extracted with a reflux extractor (TOPS, USA) at 80 ° C for 8 hours, and the vacuum filtered supernatant was recovered. This procedure was repeated three times and then concentrated under reduced pressure using a rotary vacuum-decompression vacuum rinse machine (Tokyo Ridkakikai Co., JP) to obtain 205.53 g of an ethanol extract of Gakugei and licorice.

<Example 2> Preparation of fractions of Gwangyang extract and licorice extract

1) Preparation of fraction

Each of the extracts prepared in Example 1 was suspended in 2 L of distilled water, and the fractions were fractionated using polar and nonpolar solvents, and then each sample was obtained in a vacuum vacuum concentrator. The fractions of Gilchwang extract and licorice extract were obtained as shown in Table 1 below.

[Fraction sequence]

The fractions were fractionated using separate funnel in the order of n-hexane> chloroform> ethyl acetate> n-butanol.

Fraction Gakyung extract and licorice extract (unit: g) chloroform 59.34 Ethyl acetate 72.65 n-butanol 95.63

2) Separation and basis weight of crude saponin

To separate crude saponin from the extracts of Gilchwang extract and licorice root, 1) was fractionated by n-butanol and then concentrated under reduced pressure. The fractions were suspended in 1000 ml of distilled water and passed through a column packed with 200 g of activated carbon. Thereafter, the eluate was concentrated, then developed with 10 L of distilled water in Diaion HP-20 (1 kg, 5 x 100 cm) to remove the sugar fraction, and then eluted with 3 L of ethanol and concentrated to give crude saponin fraction. And weighed. As a result, 9.6 g of crude saponin was obtained from Ganoderma lucidum extract and licorice extract.

<Experimental Example 1> Vascular relaxation effect of crude saponin isolated from Gwangyang extract and licorice extract

EXPERIMENTAL EXAMPLE 1-1 Vascular Smooth Muscle Cell Culture and Calcium Mobilization Regulation

1) Cell culture

The vascular smooth muscle cell line A7r5 was cultured in DMEM culture medium supplemented with 10% FBS, 10 units / ml penicillin, 100 μg / ml streptomycin and 2 mM L-glutamine at 37 ° C in a 5% CO ₂ -95% air incubator. The cultured cells cultured in monolayer culture flasks were washed twice with the basic medium, treated with 2.5% trypsin solution to desorb cells, and centrifuged by neutralizing the suspended cells with serum. The cell pellet obtained by centrifugation was washed three times with a cell culture medium. Next, the number of cells was measured with a hemocyte counter to adjust the number of cells to 5 × 10 ⁴ cells / ml, and 200 μl of each was added to a 96-well plate (Corning, dark-clear bottom) for 24 hours.

2) Calcium mobilization control of vascular smooth muscle

The following experiment was conducted to confirm whether calcium mobilization in vascular smooth muscle cells, which is a major factor of vasoconstriction, can be controlled. 96 well-calcium ion specificity of Fura-2 / AM (molecular probe ) in the dye-cultured cells in the plate by treatment with 5μM concentration for one hour the mixture was kept at 37 ℃ CO ₂ -incubator with Ca ^{2 +} -free PSS 3 Lt; / RTI &gt; After the addition of 190㎕ containing a 1.5mM Ca ^{2 +} PSS to each well, and then scoring the crude saponin isolated from the extract and the licorice extract at a concentration of gilgyeong 10㎍ / ml in Flexstation 3 Assay plate (assay plate) Was inserted. To the compound plate, a 5-fold concentrated K ⁺ solution was added to maintain a final concentration of 50 mM K ⁺ when added, and K ⁺ was automatically added using SoftMax Pro 5.1, followed by a wavelength of 340/380 nm The fluorescence intensity of the 510 nm emission wavelength emitted when excited was measured. The results are shown in Table 2 below.

Calcium mobilization (F _340/380 ratio) Highly concentrated potassium-treated cell line Untreated group 1.85 ± 0.11 Crude saponin treated group 1.62 ± 0.09 Endodelin treated cell line Untreated group 1.65 + 0.13 Crude saponin treated group 0.51 + - 0.11

As shown in the above Table 2, treatment of high-concentration potassium-treated cell line and endodelin-treated cell line treated with crude saponin isolated from Gilchung extract and licorice root extract showed that calcium mobilization inhibitory effect was observed in both cell lines there was. Thus, crude saponin isolated from Gwangyang extract and licorice extract according to the present invention can be expected to inhibit vasoconstriction.

<Experimental Example 1-2> Vascular relaxation efficacy in basal artery

1) Isolation of basilar artery

The separation of the basilar artery was performed by surgical methods. First, 2 to 2.5 kg white rabbits were anesthetized by inhalation with an animal anesthetic, enflurane, and then the basal artery of the brain bottom was excised with the skull cleaved. The extracted basilar artery was dipped in a Ca ^{2 +} -free tyrode solution saturated with 95% O ₂ and 5% CO ₂ to remove surrounding connective tissue and fat. The Ca ^{2 +} -free tyrode solution used for the separation of blood vessels was composed of 137 mM NaCl, 5.4 mM KCl, 1 mM MgCl ₂ , 23.8 mM NaHCO ₃ and 5.5 mM glucose, and the osmolarity was 300 mOsm / kg H ₂ O, respectively.

2) Tension measurement in vessels contracted by high concentration of potassium

Separated blood vessels were stored in a Ca ^{2 +} -free tyrode solution saturated with 95% O ₂ and 5% CO ₂ . First, the basal artery was cut into 5 mm size and fixed to the end of the force transducer and the latch ring under the organ bath maintained at 37 ° C using a chrome ring. Were measured. This experiment was performed by measuring the shrinkage force with both ends fixed. The inside of the organ bath was measured by isometric contraction in the state of continuous saturation with 95% O ₂ and 5% CO ₂ Respectively. By replacing the blood vessels in a normal tyrode state with a tyrode containing high-concentration potassium (50 mM) ions, depolarization of the membrane voltage up to 20 mV according to the Nernst equation results in the passage of ions through the closed channel, By activating the channel, the concentration of calcium in the vascular smooth muscle cells was increased to induce contraction. Such contraction and relaxation were repeated three times. At this time, the tyrode solution containing high concentration of potassium ions is composed of 92.4 mM NaCl, 50 mM KCl, 1.5 mM CaCl ₂ , 1 mM MgCl ₂ , 23.8 mM NaHCO ₃ and 5.5 mM glucose, and the osmotic pressure is maintained at 300 mOsm / kg H ₂ O And saturated with 95% O ₂ and 5% CO ₂ to adjust the pH to 7.4.

In the present study, we investigated the effect of acetylcholine 1 μM on blood vessel relaxation by determining whether the contraction of blood vessels was reduced when the contraction was stabilized by treatment with tyrode solution containing high concentration of potassium ions in the basal artery isolated from 1) Respectively. The results are shown in Fig.

The basic tension of the blood vessel or the tension of the blood vessel, which is changed by a certain condition, is caused by the contraction and relaxation of the vascular smooth muscle. Especially, the contraction of the blood vessel occurs from the outside of the vascular smooth muscle through the membrane voltage- (Ca ²⁺ channel), which is related to the increase of intracellular Ca ^{2 +} ions by the influx of calcium ions. When the vascular smooth muscle is stimulated by electrical stimuli such as the excitation wave propagation of the nerves that control the blood vessels, the vascular smooth muscle is electrically depolarized and the L-type calcium channel opened by the membrane potential Calcium ion is introduced from the outside of the cell into the inside by the difference in the concentration of calcium ions to induce contraction of the vascular smooth muscle, and thus the tension or tension of the blood vessel increases. In FIG. 1, crude saponin isolated from Gilchwang extract and licorice extract was treated with artificial depolarization induced by depolarization using high-concentration potassium. The crude saponin thus obtained showed 15.7 + 4.0% basal artery relaxation rate, And showed significant blood vessel relaxation effect.

3) Tension measurement in vessels contracted by endodelin treatment

The blood vessel relaxation efficacy in the vessels contracted by endodelin treatment was performed in the same manner except that the tyrode solution containing endodelin was used instead of the tyrode solution containing the high concentration potassium ion in 2) Respectively. The endothelin-1 was treated with 3nM of endodelein. The endogenous tyrode solution was composed of 137 mM NaCl, 5.4 mM KCl, 1.5 mM CaCl ₂ , 1 mM MgCl ₂ , 23.8 mM NaHCO ₃ and 5.5 mM glucose, The osmotic pressure was maintained at 300 mOsm / kg H ₂ O and saturated with 95% O ₂ and 5% CO ₂ to adjust the pH to 7.4.

Endodelin is the most potent cause of vasoconstriction in the human body, and is a major target of diseases such as diabetes, hypertension, and renal failure. The basal artery was shrunk with 3 nM endodelin and treated with crude saponin isolated from Gilchung extract and licorice root extract (Fig. 2). The relaxation rate of the basal artery was 92.4 ± 3.1%, indicating a very strong vascular relaxation efficacy I could. In the case of endodelin, the serum concentration of the diabetic patient is increased by 100 times or more to cause hypertension, renal failure, foot necrosis, peripheral circulatory disorder, etc. Therefore, crude saponin according to the present invention may be a good candidate as an endodelin receptor inhibitor .

Claims (14)

Gil - gyeong - tang consisting of Gilchung extract and licorice extract; Or crude saponin isolated therefrom as an effective ingredient. The method according to claim 1,
Wherein the Gakyung extract and licorice extract are extracted with a solvent of water, an organic solvent or a mixture thereof. 3. The method of claim 2,
Wherein the organic solvent is at least one solvent selected from the group consisting of alcohols having 1 to 6 carbon atoms, hexane, acetone, ethyl acetate, chloroform and diethyl ether. The method according to claim 1,
Gakyung extract and licorice extract are alcohol extracts having 1 to 6 carbon atoms of Gakuin and licorice. 5. The method of claim 4,
Gakyung extract and licorice extract are ethanol extracts of Gakgye and licorice. The method according to claim 1,
Wherein the fraction is obtained by fractionating Ganoderma lucidum and licorice extract in one or more solvents selected from the group consisting of water, hexane, chloroform, ethyl acetate, butanol and mixtures thereof. The method according to claim 1,
Wherein the crude saponin is contained at a concentration of 1 to 10 mu g / ml. Gil - gyeong - tang consisting of Gilchung extract and licorice extract; Or crude saponin isolated therefrom as an active ingredient. 9. The method of claim 8,
Wherein the Gakyung extract and the licorice extract are extracted with a solvent of water, an organic solvent or a mixture thereof. 10. The method of claim 9,
Wherein the organic solvent is at least one solvent selected from the group consisting of alcohols having 1 to 6 carbon atoms, hexane, acetone, ethyl acetate, chloroform and diethyl ether. 9. The method of claim 8,
Gakyung extract and licorice extract are alcohol extracts having 1 to 6 carbon atoms in Gilchung and licorice. 12. The method of claim 11,
Gakyung extract and licorice extract are ethanol extracts of Gakyung and licorice. 9. The method of claim 8,
Wherein the fraction is obtained by fractionating Ganoderma lucidum and licorice extract in one or more solvents selected from the group consisting of water, hexane, chloroform, ethyl acetate, butanol and mixtures thereof. 9. The method of claim 8,
And crude saponin is contained at a concentration of 1 to 10 μg / ml.

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