KR20220062258A - Composition of improvement, prevention and treatment in atherosclerosis with radish seed extract - Google Patents

Abstract

The present invention relates to a composition for alleviating, preventing, or treating arteriosclerosis, containing a Raphanus sativus L. seed extract as an active ingredient, and more specifically, to a composition for alleviating, preventing, or treating arteriosclerosis, containing a Raphanus sativus L. seed extract as an active ingredient, and configured to suppress adhesion of mononuclear cells to vascular endothelial cells, induced from an inflammatory response, thereby suppressing the development of arteriosclerosis at an early stage. The present invention can provide a composition containing a Raphanus sativus L. seed extract as an active ingredient, which is capable of alleviating, preventing, or treating arteriosclerosis by suppressing adhesion of mononuclear cells to vascular endothelial cells.

Description

Composition for improvement, prevention or treatment of arteriosclerosis containing the extract of Naitakeja as an active ingredient {Composition of improvement, prevention and treatment in atherosclerosis with radish seed extract}

The present invention relates to a composition for improving, preventing or treating arteriosclerosis containing radish seed extract as an active ingredient, and more particularly, arteriosclerosis by inhibiting mononuclear adhesion to vascular endothelial cells induced from an inflammatory response. It relates to a composition for improving, preventing, or treating arteriosclerosis, which contains an extract of Naebokja as an active ingredient, which exhibits the effect of inhibiting the occurrence of the disease at an early stage.

Cardiovascular disease, including arteriosclerosis and myocardial infarction, is one of the leading causes of death worldwide and accounts for about 40% of all deaths (Seymour, G.J., Clinical Microbiology and Infection, 13:4, 3-10, 2007). Among them, arteriosclerosis has received worldwide attention as an important factor in cardiovascular disease. LDL cholesterol has been known as the most important risk factor for atherosclerosis, but as many studies have been conducted recently, the inflammatory response has been gradually recognized as the main cause of atherosclerosis (Blankenberg, S. et al., Atherosclerosis, 170(2), 191- 203, 2003).

Atherosclerosis is a multifactorial disease that is known to be related to smoking, hypertension, diabetes, and obesity. It is known that an inflammatory response is caused by disorders of vascular endothelial cell function and increased cholesterol, oxidized LDL, and free radicals in the blood vessel wall, which leads to the development of atherosclerosis (Kong, L. et al., Lipids in Health and Disease, 12:115, 2013).

Vascular endothelial cells, a single cell layer in the arterial wall that come into contact with blood, generally resist the adhesion of white blood cells, but when the function of endothelial cells is reduced, the expression of adhesion molecules increases along with an inflammatory response. When the expression of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) is increased, monocytes, T lymphocytes, etc. Atherosclerosis is induced as it adheres and flows into the tissue. Thereafter, monocytes differentiate into macrophages and secrete inflammatory cytokines such as tumor necrosis factor-alpha (TNF-α), oxidizing substances, etc. to further oxidize lipids and cause a continuous inflammatory reaction to cause coronary artery lesions. This causes acute coronary syndromes such as angina pectoris and acute myocardial infarction, leading to death (Libby, P., Nature, 420(6917), 868-874, 2002).

As cardiovascular disease ranks as the number one cause of death worldwide, the cardiovascular disease treatment market is growing steadily every year, and food companies are showing interest in developing products related to arteriosclerosis prevention. The number and sales of functional ingredients such as blood circulation improvement and blood triglyceride and cholesterol improvement are also increasing. However, the problem of side effects is being raised with statins, which are currently used to treat arteriosclerosis, and representative health functional foods for improving vascular health, such as maintaining omega-3, have a disadvantage in that their digestibility is poor. Therefore, there is a need to discover new food and drug materials that are effective and safe for the treatment of arteriosclerosis.

Republic of Korea Patent No. 10-1835397 (registration date: 2018.02.28) describes a composition for the prevention and treatment of inflammation and allergic diseases, which contains a complex extract of Naitake jasmine, ragweed, and sirloin root as an active ingredient.

The present invention exhibits the effect of inhibiting the occurrence of arteriosclerosis in the early stage by inhibiting the adhesion of monocytes to vascular endothelial cells induced from an inflammatory reaction compared to Ginko leaf extract, which is a material widely used in the prior art. An object of the present invention is to provide a composition for improving, preventing, or treating arteriosclerosis containing the extract of Naebokja as an active ingredient.

The present invention provides a food composition for improving arteriosclerosis due to vascular endothelial cell dysfunction, characterized in that it contains radish seed extract as an active ingredient to inhibit mononuclear adhesion in vascular endothelial cells.

In addition, the present invention provides a pharmaceutical composition for preventing or treating arteriosclerosis due to vascular endothelial cell dysfunction, characterized in that it contains radish seed extract as an active ingredient to inhibit mononuclear adhesion in vascular endothelial cells. do.

In addition, the present invention provides a food composition for improving arteriosclerosis independent of blood cholesterol, characterized in that it contains a radish seed extract as an active ingredient, and inhibits monocyte adhesion in vascular endothelial cells.

In addition, the present invention provides a pharmaceutical composition for preventing or treating arteriosclerosis independent of blood cholesterol, characterized in that it contains a radish seed extract as an active ingredient, and inhibits monocyte adhesion in vascular endothelial cells.

The present invention can provide a composition containing an extract of spleen cyst, which can improve, prevent, or treat arteriosclerosis by inhibiting monocyte cell adhesion in human umbilical vein endothelial cells as an active ingredient.

1 is a graph showing the degree of inhibition of mononuclear cell adhesion in vascular endothelial cells of Radish seed extract of the present invention compared with Ginko leaf extract. A) is the result of treating each extract at 80 μg/ml, and B) is the result of treating each extract at 20 ~ 80 μg/ml.

Cardiovascular disease, including arteriosclerosis and myocardial infarction, is considered the leading cause of death worldwide and accounts for about 40% of all deaths. Atherosclerosis is a multifactorial disease, including cholesterol, oxidized LDL, and increased free radicals as causes, which leads to cell dysfunction and inflammatory response.

Vascular endothelial cells, a single cell layer in the arterial wall that come into contact with blood, normally resist the adhesion of white blood cells, but when the endothelial cell function is reduced, an inflammatory response occurs along with vascular cell adhesion molecule-1 (VCAM). -1) and intercellular adhesion molecule-1 (hereinafter, ICAM-1) expression is increased, and monocytes and T lymphocytes attach and flow into the tissue, causing arteriosclerosis. Monocytes differentiate into macrophages and secrete inflammatory cytokines such as tumor necrosis factor-alpha (TNF-α), oxidizing substances, etc. to further oxidize lipids and cause a continuous inflammatory reaction to prevent coronary artery lesions. It causes acute coronary syndromes such as angina pectoris and acute myocardial infarction, leading to death.

Although research to treat such arteriosclerosis continues, there are problems such as side effects and reduced digestibility, so it is necessary to discover new food and drug materials that are effective and safe. Accordingly, the present invention selected radish seed extract as a new material for treating arteriosclerosis.

The present invention provides a food composition for improving arteriosclerosis, characterized in that it contains radish seed extract as an active ingredient. In addition, the present invention provides a food composition for animals for improving arteriosclerosis, characterized in that it contains radish seed extract as an active ingredient. In addition, the present invention provides a pharmaceutical composition for preventing or treating arteriosclerosis, characterized in that it contains radish seed extract as an active ingredient. In addition, the present invention provides an animal pharmaceutical composition for preventing or treating arteriosclerosis, characterized in that it contains radish seed extract as an active ingredient.

Raphanus sativus seed ( Raphanus sativus seed) is the seed of radish, a cruciferous plant. It is warm in nature and has no poison, so it has been used as a medicine since ancient times. To date, radish seed extract has been reported to have physiological functions such as anti-inflammatory and anticancer, and when treated with radish seed extract in a colitis model, intestinal inflammation induced by TNBS or DSS was reduced. It has been reported to recover (Choi KC. et al., J Ethnopharmacol, 179, 55-65, 2016).

According to the present invention, extract by adding 5-15% (v/v) ethanol as an extraction solvent to radish seeds can be used to obtain an extract containing a large amount of sulforaffine. The ethanol may include absolute ethanol and fermented alcohol, and when preparing an edible extract, ethanol is preferably fermented alcohol.

On the other hand, the extraction is preferably chimchi extraction, and when hot water is extracted, an extraction method without an increase in temperature is used because of the thermal stability of sulforaffine in the insider. In general, when preparing an edible extract, a refluxing extraction method is mainly used, but the refluxing extraction method may increase the temperature of the solvent during extraction, and the process is more complicated than that of immersion extraction.

In the food composition or pharmaceutical composition of the present invention, the extract of Naitakeja may be one that preferably inhibits adhesion of monocytes in vascular endothelial cells.

According to the following experiment, it was confirmed that the Naebokja extract of the present invention inhibited TNF-α-induced monocyte adhesion in vascular endothelial cells in a concentration-dependent manner superior to that of the ginkgo leaf extract at a concentration ranging from 20 to 80 μg/ml. . Therefore, it can be said that the extract of the present invention shows efficacy in improving, preventing or treating arteriosclerosis by initially inhibiting the occurrence of arteriosclerosis.

On the other hand, the food composition of the present invention is, for example, meat, grains, caffeinated beverages, general drinks, chocolate, breads, snacks, confectionery, pizza, jelly, noodles, gums, ice cream, alcoholic beverages, alcohol, vitamin complexes and other health It may be any one selected from supplements, but is not necessarily limited thereto.

On the other hand, the food composition for animals of the present invention is an oral preparation for pets, specifically tablets, capsules, liquids, gels, pastes, oral sprays, oral tablets, powders and chewable treats for pets or animals. It may be formulated as feed, but is not limited thereto. Diluents or excipients such as fillers, extenders, binders, moisture agents, disintegrants, and surfactants commonly used in formulation can be used, and starch, calcium carbonate, sucrose, lactose or gelatin, etc. can be used as excipients, In addition, a lubricant such as magnesium stearate talc may be used.

On the other hand, the pharmaceutical composition or veterinary pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier, diluent or excipient in addition to the active ingredient. Usable carriers, excipients or diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, There are microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil, and at least one selected from among them may be used. In addition, when the therapeutic and prophylactic agent is a pharmaceutical, a filler, an anti-aggregant, a lubricant, a wetting agent, a fragrance, an emulsifier, or a preservative may be additionally included.

On the other hand, the pharmaceutical composition or the formulation of the pharmaceutical composition for animals of the present invention can be prepared in a preferred form according to the method of use, and in particular, in order to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal It is better to formulate it by adopting a known method. Examples of specific formulations include PLASTERS, GRANULES, LOTIONS, LINIMENTS, LEMONADES, AROMATIC WATERS, POWDERS, syrups ( SYRUPS, OPHTALMIC OINTMENTS, LIQUIDS AND SOLUTIONS, AEROSOLS, EXTRACTS, ELIXIRS, OINTMENTS, FLUIDEXTRACTS, EMULSIONS ), suspensions (SUSPESIONS), preparations (DECOCTIONS), infusions (INFUSIONS), eye drops (OPHTHALMIC SOLUTIONS), tablets (TABLETS), suppositories (SUPPOSITIORIES), injections (INJECTIONS), alcohol tablets (SPIRITS), CATAPLS ), capsules (CAPSULES), creams (CREAMS), troches (TROCHES), tinctures (TINCTURES), pastas (PASTES), pills (PILLS), it may be any one selected from soft or hard gelatin capsules.

On the other hand, the dosage of the pharmaceutical composition or veterinary pharmaceutical composition of the present invention is preferably determined in consideration of the administration method, the age, sex and weight of the user (animal), and the severity of the disease. For example, the pharmaceutical composition for preventing or treating arteriosclerosis of the present invention can be orally administered at least once per day at 0.000001 to 100 mg/kg (body weight) based on the active ingredient. However, the above dosage is only an example for illustration, and may be changed according to the condition of the user (animal) and the prescription of the doctor.

On the other hand, in the present invention, the meaning of 'contained as an active ingredient' means that the arteriosclerosis improvement, prevention or treatment efficacy required in the present invention is generated from the Naebokja extract, which is a component of the present invention, and other ingredients. It means that it can be included as an auxiliary ingredient.

Hereinafter, the content of the present invention will be described in more detail through the following Examples and Experimental Examples. However, the scope of the present invention is not limited only to the following examples and experimental examples, and includes modifications of technical ideas equivalent thereto.

[Example: Preparation of extract of Naitakeja containing a large amount of sulforaffine]

In order to extract sulforaffine in an optimal yield from the ingestion, a 10% (v/v) fermented alcohol solvent was prepared using ethanol (fermented alcohol). 800 ml of each solvent was added to 100 g of Naesokja finely pulverized using a mixer, followed by immersion extraction at a temperature of 40° C. for 12 hours to obtain an extract of Naesokja. Thereafter, the extract of Naitakeja was concentrated and lyophilized to form a powder.

[Experimental Example: Confirmation of the inhibitory efficacy of monocyte cell adhesion of Naitake extracts]

In this experimental example, an experiment was performed to confirm the efficacy of the radish seed extract of the above example for inhibiting mononuclear cell adhesion in vascular endothelial cells. For comparative analysis, Ginkgo leaf extract, which is widely used in the industry, was used.

THP stained with calcein-AM (calcein-AM, obtained from Sigma-Aldrich, St. Louis, MO, USA) on human umbilical vein endothelial cells (Lonza, Walkersville, MD, USA) The -1 cell line (THP-1 monocytic leukemic cells, obtained from: Korea Cell Line Bank) was co-cultured and washed, and then the degree of adhesion of THP-1 monocytic leukemic cells to vascular endothelial cells was measured using a fluorescence microscope.

Vascular endothelial cells were prepared in 10% (v/v) fetal bovine serum in Medium 199 medium containing 2 mM L-glutamine and HEPES (from Corning, NY, USA) from Gipco, Grand Island, NY, USA), 1 ng/ml recombinant human epidermal growth factor (obtained from Gipco, Grand Island, NY, USA), 2 ng/ml basic fibroblast growth factor (obtained) Source: Gipco, Grand Island, NY, USA), 1 ng/ml hydrocortisone (Sigma-Aldrich, St. Louis, MO, USA), 1% (v/v) streptomycin/penicillin ( Confluent at 37 °C, 5% CO2 incubator (obtained from Forma Scientific Co., Marietta, OH, USA) using medium supplemented with streptomycin/penicillin, available from Corning, Corning, NY, USA). ) and incubated until they grow in the same state.

The THP-1 cell line contains 10% (v/v) fetal bovine serum, obtained from Sigma-Aldrich, St. Louis, MO, USA, and 0.05 mM 2-mercaptoethanol, obtained from : Sigma-Aldrich, St. Louis, MO, USA), 1% (v/v) streptomycin/penicillin added in RPMI-1640 medium (obtained from: Welgene, Daegu, Korea) 2 - 20 x 10 ⁵ pieces/ incubated in ml concentration.

Vascular endothelial cells grown in a confluent state were seeded in a 96-well plate and cultured for 24 hours, followed by 20 radish seed extract and 20 Ginkgo leaf extract, respectively. , 40, 80 μg/ml concentration was pre-treated in the culture medium for 1 hour. After 1 hour of pretreatment, 10 ng/ml TNF-α was treated and activated for 5 hours.

THP-1 monocytes cultured in RPMI-1640 medium were transferred to sterile phosphate buffered saline, 2 μM calcein-AM was added, and placed in an incubator at 37°C, 5% CO2 for 15 minutes for staining. The stained THP-1 was transferred to Medium 199 medium and co-cultured with 5 x 10 ⁵ per 96 well of TNF-α-activated vascular endothelial cells for 5 hours. After 1 hour, they were washed with phosphate buffered saline, and the degree of adhesion of THP-1 monocytes to vascular endothelial cells was measured with Infinite 200 PRO (obtained from Tecan group Ltd., Mannedorf, Switzerland) at 485 nm excitation and 538 nm emission wavelength. .

As a result, as shown in FIG. 1 , it was confirmed that the Naebokja extract significantly inhibited TNF-α-induced monocyte adhesion in vascular endothelial cells at a concentration in the range of 20 to 80 μg/ml in a concentration-dependent manner compared to the ginkgo leaf extract.

Claims (4)

A food composition for improving arteriosclerosis due to vascular endothelial cell dysfunction, characterized in that it contains radish seed extract as an active ingredient to inhibit mononuclear adhesion in vascular endothelial cells.
A pharmaceutical composition for preventing or treating arteriosclerosis due to vascular endothelial cell dysfunction, characterized in that it contains radish seed extract as an active ingredient to inhibit mononuclear adhesion in vascular endothelial cells.
A food composition for improving arteriosclerosis independent of blood cholesterol, characterized in that it contains radish seed extract as an active ingredient to inhibit mononuclear adhesion in vascular endothelial cells.
A pharmaceutical composition for preventing or treating arteriosclerosis independent of blood cholesterol, characterized in that it contains radish seed extract as an active ingredient to inhibit mononuclear adhesion in vascular endothelial cells.

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