KR100883518B1 - The healthy and funtional foods for the xbrassicoraphanus extract to prevent or treat cardiovascular disease including thrombosis - Google Patents

Abstract

A dietary supplement is provided to improve the usefulness for the cardiovascular disease patient by inhibiting the blood clot. A dietary supplement comprises a xBrassicoraphanus extract. Preferably the xBrassicoraphanus extract is extracted by using ethanol. Preferably the xBrassicoraphanus extract extracted with ethanol is concentrated under reduced pressure and the concentrated one is freeze-dried.

Description

The healthy and funtional foods for the xBrassicoraphanus extract to prevent or treat cardiovascular disease including thrombosis}

The present invention relates to a dietary supplement useful for patients with cardiovascular diseases requiring thrombus suppression from xBrassicoraphanus extract.

The blood must be in a liquid state within normal blood vessels, and the blood vessels must coagulate quickly to stop blood loss from damaged blood vessels and maintain homeostasis. In the normal state, when blood vessels in the blood vessels (thrombus) occurs, thrombolytic action is activated to restore fluidity, and the balance between the formation of blood clots and the mechanisms that prevent the formation of blood clots is broken due to various factors. What appears to be a phenomenon is called thrombosis. Thrombosis refers to the formation of solidified mass in the cardiovascular system of a living body, and the blood clot mass is called thrombus.

The spontaneous coagulation of blood when large vessels are ruptured or damaged is an important process that saves lives by preventing further bleeding. However, the formation of unbroken cardiovascular thrombi can be life-threatening. In other words, when a blood clot is formed in an unruptured cardiovascular system, ischemic damage is caused to organs or tissues by blood flow reduction or blockage, and part or all of the blood clot is released, and the broken blood clot causes embolism.

An embolism is a term used to refer to a lump that occludes a vessel (general vessel or lymphatic vessel), and means that a blood clot or foreign substance introduced from the outside blocks the vessel and prevents blood or lymph flow. Embolism moves along the bloodstream and is transported to other places, eventually blocking some of the blood vessels. This is the cause of the blockage of blood vessels by embolism (em: embolism).

Except in accidents, sudden death is mostly due to cardiovascular disease. The most common cause of death worldwide is cardiovascular disease, at 30%. In the past, cardiovascular disease sudden death was referred to collectively as 'heart attack', but these days, depending on the cause of the symptoms are called. The most fatal diseases are caused by blockage of blood vessels.

The heart contains blood vessels called coronary arteries to supply oxygen and nutrients to the cells of the heart itself. The most dangerous cardiovascular disease, acute myocardial infarction, is a disease in which heart cells die because the blood vessels of the coronary artery are blocked by blood clots. Two hours after an acute myocardial infarction, the heart cells begin to die because they do not receive oxygen, and the heart stops causing death.

Angina is similar to myocardial infarction but weak. The heart cells die, but the blood supply to the coronary arteries is not good enough to cause pain in the heart muscle. Angina usually occurs when you move your body and get over your heart. Therefore, pain during exercise is angina, and when resting, myocardial infarction is likely.

'Stroke' (stroke) is also caused by clogged blood vessels, such as myocardial infarction or angina. However, stroke is caused by the blockage of some of the blood vessels that supply blood to the brain, not the heart.

Arrhythmia is a condition in which the normal rhythm of the heart is broken. 'Mac' does not run as 'jeong' is called arrhythmia. At this time, the heart runs faster or slower than 60-100 times per minute, and the running speed is irregular. Arrhythmias are treated by externally applying electrical stimulation to artificially 'reboot' the heart.

Even if the heart structure is wrong, you can get sick. Heart valve disease is a problem in the valve that allows blood to flow in a certain direction in the heart. Because of the backflow of blood, a crowd will pour into the heart and swell.

Ventricular septal defect is a hole in the compartment that separates each part of the heart. A hole in the ventricular wall mixes clean and dirty blood. The blood will not function properly. Because the disease is congenital, it must be detected during infancy and treated surgically.

Therefore, in order to reduce mortality due to cardiovascular disease, it has been required to respond to health foods and therapeutic agents that have the function of inhibiting blood clots and embolism and minimize side effects of existing drugs.

Korean Patent No. 408087 discloses a pharmaceutical composition having an antithrombotic action comprising starfish ethyl acetate extract as an active ingredient, and Korean Patent No. 392175 is produced in the culture of Matsutake mycelium, and the N-terminal amino acid is ATVQGGDAYYINNSA. A thrombolytic enzyme having a molecular weight of 26.5 KDa and fibrin solubility is known.

On the other hand, Korean Patent No. 452518 is known as x Brassica raphanus obtained by crossing the Chinese cabbage "tradition" and the mother "baekkyung" as the father.

The baemuchae (x Brassicaraphanus ) has both the chromosome of the Chinese cabbage and radish, and when grown for more than 90 days, the barrel sits like a Chinese cabbage, and the inner leaves are yellow and have a small sized radish root. 'Baemuchae' is formed in the low-temperature market and is recommended at high-temperature days, so it is difficult to grow mature plants in spring and summer, but it can be grown when used with heat radish, so it can be grown year-round except in winter in non-heated facilities or open field. In addition, the taste of 'Baemuchae' is different from radish, Chinese cabbage and other spicy tastes, and it is known that it can be used as a kimchi material because it is cool and crispy because it has a lot of moisture in the middle part when kimchi is soaked.

The present inventors have completed the present invention by confirming that while studying the usefulness of the natural hybrid of Chinese cabbage and radish, it has an effect of preventing blood clots by inhibiting blood clots in ethanol extract of xBrassicoraphanus.

An object of the present invention is to provide utility as a dietary supplement from the extract of xBrassicoraphanus, a natural hybrid of cabbage and radish.

In order to achieve the above object, the present invention provides a health functional food useful in patients with cardiovascular diseases that require thrombus suppression from xBrassicoraphanus extract.

In particular, Baemuchae extract according to the present invention can inhibit the expression of thromboxane and glycoprotein expression, can be useful in health functional foods that can be ingested by patients with cardiovascular diseases that require inhibition of blood clots because there is no side effect have.

According to the present invention, the extract may be extracted with ethanol, and the extract is concentrated under reduced pressure and freeze-dried to obtain extract powder.

Thromboxane B ₂ is a protein that is secreted when a thrombotic reaction occurs due to aggregation of platelets by collagen. However, Thromboxane B ₂ (TxB ₂ ) can be measured by quantifying the dehydro form thromboxane A ₂ . Baemuchae extract according to the present invention showed a 100% coagulation inhibitory ability was found to be effective in inhibiting the thrombus, as well as very effective in thromboxane expression inhibition test.

In addition, normal platelets have glycoproteins IIb / IIIa (GPIIb / IIIa) on the surface of the platelets and cross-link with fibrinogen or Von Willebrand Factor (VWF) to injure sites or strong antagonists such as collagen. Platelet coagulation occurs. It can be seen that the present invention has an inhibitory effect on glycoprotein expression by abolishing the receptor of Glyciprotein IIb / IIIa.

Therefore, Baemuchae extract according to the present invention can inhibit the expression of thromboxane and glycoprotein expression can be used in health functional foods that can be consumed by patients with cardiovascular diseases that require thrombus inhibition.

Baemuchae extract according to the present invention can be developed as a health functional food by adding to various foods, for example, beverages, tea, vitamin complexes, health supplement foods, etc., Baemuchae extract directly pill, powder, tablets, capsules, etc. It can be prepared and used in the formulation of.

As described above, the Baekmuchae ethanol extract of the present invention has an effect of inhibiting blood clots, so it can be usefully used in health functional foods that can be consumed by cardiovascular patients who need to suppress blood clots.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail. However, the scope of the present invention is not limited by these Examples.

< Example  1> Abandonment  Preparation of Extract

30 g of Baekchae was added to 40 l of ethanol, and ethanol was extracted twice. The extract was concentrated under reduced pressure and lyophilized to obtain Baekchae extract powder (yield: 14%).

< Formulation example  1> Liquid  Produce

The dry powder of Baemuchae extract prepared in Example 1 was diluted with a ratio of DMSO and physiological saline to 1: 1 to prepare a liquid formulation.

< Experimental Example  1> Platelet aggregation Inhibitory ability  Measure

Platelet aggregation inhibition was determined by turbidity measurement using Platelet Aggregometer (Whole Blood Lumi-ionized calcium aggregometer, Chrono-Log Co. USA). Take 400 μl of PRP, incubate at 37 ° C for 3 minutes, and then add 50 μl of sample. Aggregation was induced after 2 minutes by stimulation with 50 µl of platelet aggregation promoter. After the aggregation reaction was measured for 5 minutes, the maximum aggregation point was calculated and the extent of inhibition was calculated. While aggregation was measured, the agitation was 1,000 rpm, and collagen (Chrono-Log co., 2 μg / ml) was used as a platelet aggregation promoter. Inhibition was obtained using the following equation.

The antithrombotic effect is shown in FIG. 1 as a percentage of the experimental group treated with ethanol layer of Baekchae with the platelet aggregation occurring after collagen treatment (channel1, blue) using an aggregometer. Fig. 1 showed no significant effect on the concentration of 25 ㎍ / ㎖, but the concentration of platelet significantly increased in proportion to the concentration of the ethanol extract of the baewoochae by showing 45% at 50 ㎍ / ㎖, 100% coagulation inhibitory ability at 100 ㎍ / ㎖ or more It can be confirmed that aggregation inhibition occurs.

< Experimental Example  2> Glycoprotein  Expression Inhibitory ability  Measure

Normal platelets have glycoproteins IIb / IIIa on the platelet surface and cross-link with fibrinogen or Von Willebrand Factor (VWF), which causes platelet coagulation by potent antagonists such as injured sites or collagen. Get up. This experiment was used to determine whether the ethanol layer of Baekchae was incapacitated to receptors of glycoprotein IIb / IIIa. FACScan was used as an antibody using PAC-1 / FITC (Becton-Dickenson, San Jose, CA, USA). The results of administration of 25 μg / ml, 50 μg / ml, 100 μg / ml, and 200 μg / ml of Baekmuchae extract after normal platelet, collagen addition, and after collagen are shown in FIG. 2. In FIG. 2, M1 represents a percentage of normal platelets in which no protrusions are formed, and M2 means a state in which a receptor is activated by forming a protrusion.

When collagen is added to normal platelets, the glycoprotein receptor is activated, which can be expressed as a percentage by fluorescence immunostaining. As can be seen in FIG. 2, only 2.3% of normal platelets are present, and collagen is added to the platelets. Glycoprotein receptor is activated up to 33.8% when added, but the addition of Baekchae extract increased M2's FITC binding levels from 25 μg / ml to 22.3%, 50 μg / ml to 21.1%, and 200 μg / ml to 19.6%. Baemuchae extract has been shown to have a decrease in glycoprotein expression.

< Experimental Example  3> Thrombox  Expression Inhibitory ability  Measure

400 μl of 3 × 10 ⁸ / mL platelets were taken, each sample was treated by concentration, and incubated with stirring at 37 ° C. incubator for at least 3 minutes. Thromboxane A ₂ was quantified by mixing 2 μg / ml of collagen and incubating with stirring at 37 ° C. incubator for 5 min. The amount of TxA ₂ was measured by Cayman (Ann Arbor, USA) EIA kit and tested according to the user method.

Thromboxane B ₂ is a protein that is secreted when a thrombotic reaction occurs due to aggregation of platelets by collagen. However, Thromboxane B ₂ (TxB ₂ ) can be measured by quantifying the dehydro form thromboxane A ₂ . Therefore, the present invention quantifies the amount of Thromboxane B ₂ (TxB ₂ ) in accordance with the user manual using the TxB ₂ EIA kit of Cayman (Ann Arbor, USA) and the results are shown in FIG.

As can be seen in Figure 3, Baemuchae extract according to the present invention can be seen to reduce the thromboxane, a protein that occurs during platelet aggregation.

< Experimental Example  4> Pulmonary embolism  Suppression measurement

When epinephrine is injected into the veins of mice, the rate of inhibition of lung infarction of the samples is calculated by counting the number of mice immediately killed by pulmonary embolism or paralysis under the cervical spine.

In the present invention, pulmonary embolism was performed by injecting 112 mg of epinephrine into the tail vein of ICR mice and counting the number of rats immediately killed by pulmonary embolism or paralyzed under the cervical spine. Experimental animals were fed from Korea Biolink Co., Ltd. and were adapted for 1 week in a university animal room under certain conditions (temperature: 20 ± 1 ℃, humidity: 55 ± 3%, contrast: 12 hours light / dark cycle). g balb / c female mouse was used. Each sample was orally administered at 5 mg / Kg, and after 90 minutes, 100 μl of epinephrine 10 mM was injected into the tail vein with an antagonist. After intravenous injection, the number of animals died of pulmonary embolism due to coagulation or hind limb paralysis was counted, and the survival rate was calculated by the following formula and is shown in Table 1 below.

As can be seen in Table 1, pulmonary embolism inhibition can be seen to increase the survival rate to 45.5%, 54.5%, 59.1%, 66.7%, respectively, in proportion to the concentration of the broth.

1 is a graph showing the platelet aggregation inhibitory effect of Baekmuchae ethanol extract,

Figure 2 shows the ⒜ steady state, 시 collagen addition and ⒞, ⒟, ⒠ and ⒡ by administration of 25 ㎍ / ㎖, 50 ㎍ / ㎖, 100 ㎍ / ㎖, 200 ㎍ / ㎖ Flow cytometry inhibiting the expression of glycoprotein receptors,

3 is a graph that Baemuchae ethanol extract inhibits the expression of thromboxane.

Claims (2)

Dietary dietary supplement for patients with cardiovascular diseases requiring thrombus suppression, characterized by containing extract of xBrassicoraphanus. The health functional food according to claim 1, wherein the extract of Baengmuchae is extracted by ethanol.

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