JP5504405B2 - Food composition effective in preventing vascular disease - Google Patents

Description

  TECHNICAL FIELD The present invention relates to a food composition for preventing vascular diseases, and more specifically, an active ingredient is eicosapentaenoic acid (hereinafter abbreviated as EPA), which is one of unsaturated fatty acids obtained from fish and the like, and vasospasm. The present invention relates to a food composition provided for the prevention of vascular diseases caused by the above.

In this specification, the following terms are defined as follows. Vasospasm refers to abnormal contraction of vascular smooth muscle, particularly Ca ²⁺ -independent contraction that does not depend on an increase in cytoplasmic calcium ion (Ca ²⁺ ) concentration. Membrane raft refers to the structure on the cell membrane rich in cholesterol and sphingolipids formed on the surface of blood vessels. It is important as a signal transduction site for vascular diseases, and is characterized by disappearance by removing cholesterol. all-cis-EPA (cis-type eicosapentaenoic acid) refers to EPA in which all the double bonds contained in the molecule are coordinated to the cis-position, and trans-EPA (trans-type eicosapentaenoic acid) is a molecule. EPA that is not all-cis type, in which at least one or more double bonds are coordinated to the trans position.

  Vasospasm and the accompanying sudden blood flow disturbances cause cerebral infarction, angina pectoris, myocardial infarction, etc., and the number of deaths of sudden death due to these diseases is estimated to be 60,000 every year. It is a problem. In particular, “cerebral vascular spasm” that occurs frequently after subarachnoid hemorrhage is a disease in which most of the blood vessels in the brain contract and widen the infarcted state of the brain, despite the success of subarachnoid hemorrhage surgery. It is causing a serious situation where the patient's life cannot be saved.

The contraction of the blood vessel depending on the Ca ²⁺ "normal shrinkage", not dependent on Ca ²⁺ are known "abnormal contraction (spasm)", for the normal shrinkage have revealed significant portion including its mechanism ing. Therefore, for diseases such as cerebral infarction, angina pectoris, and myocardial infarction, firstly, a therapeutic method and a therapeutic agent based on inhibition of normal contraction with obvious mechanism, particularly inhibition of Ca ²⁺ channel have been proposed (Patent Literature). 1).

  However, little is known about vasospasm, which has a more important association with diseases such as cerebral infarction, angina pectoris, and myocardial infarction. At present, the treatment method for the disease is an apparent treatment method that suppresses the vasoconstriction by suppressing the normal contraction that is less related to the disease. The cause investigation and countermeasures are anxious.

The present inventors have conducted research to solve the above problems, and sphingosylphosphorylcholine (hereinafter abbreviated as SPC), which is a kind of sphingolipid, has an important role in vasospasm, and SPC is Rho. Ca ²⁺ independent manner to contract the vascular smooth muscle through activation of kinase, further activated another protein that Fyn is the SPC, structures coupling which called membrane rafts on vascular smooth muscle cells membrane wall It has been clarified that playing an important role in the activation of Rho kinase. These facts suggested that vasospasm could be treated without inhibiting normal vasoconstriction if the series of Rho kinase activation process by SPC stimulation can be inhibited somewhere (FIG. 1, Non-Patent Documents 1-4).

  Based on the above predictions, the present inventors have advanced research, and eicosapentaenoic acid (EPA), which is a kind of highly unsaturated fatty acid contained in fish, is used as a substance that specifically inhibits vasospasm caused by SPC. I found it.

  EPA is a kind of highly unsaturated fatty acid that is abundant in marine phytoplankton and blue fish, and effects such as suppression of platelet coagulation and thrombus dissolution have been reported so far (Patent Document 2). In addition to this, the present inventors have clarified the fact that EPA inhibits abnormal smooth muscle contraction and does not affect normal contraction of smooth muscle at all (Patent Document 3). That is, the present inventors have established a novel treatment method for circulatory system diseases such as cerebrovascular spasm.

  However, for various diseases caused by vasospasm, prevention thereof is extremely important as well as treatment. Both cerebral vasospasm and coronary vasospasm can lead to serious diseases. Of course, appropriate measures should be taken after symptoms appear, and risks associated with these diseases can be reduced. It is important to prevent the occurrence of disease in advance. However, there has been no report so far on functional foods whose main component is a substance effective in preventing vasospasm.

  In addition, regarding the use of EPA effective for vasospasm, there is a risk of injecting blood with a high concentration of fatty acid, and it is considered that “oral administration” is the main administration form, but as described in the following examples, The absorption efficiency of EPA in the intestinal tract depends greatly on the health condition and age. In particular, in patients whose physical strength is significantly reduced due to diseases etc., there is a problem that even if EPA is administered, it is not sufficiently absorbed in the intestinal tract and the expected effect does not appear, and diseases caused by vasospasm Development of a food composition that can prevent ingestion and can efficiently ingest active ingredients regardless of their health condition has been awaited.

JP, 2004-43374, A Calcium channel inhibitor JP, 9-176005, A L-carnitine or alkanoyl L-carnitine containing pharmaceutical composition which uses together omega-3 system polyunsaturated fatty acid JP, 2001-261556, A An inhibitor of smooth muscle abnormal contraction

Monthly Bioindustry November 2003 "Evasapentaenoic acid (EPA) prevents vasospasm" Nakao F, Kobayashi S. et al. (2002) Circ. Res. 91: 953-960 Shirao S, Kobayashi S. et al. (2002) Circ. Res. 91: 112-119 Somlyo A.V. (2002) Circ.Res. 91: 83-84 (Editorials)

  In view of the above situation, the present invention relates to the prevention of various diseases caused by vasospasm, and an active ingredient is a substance that specifically inhibits a reaction pathway activated by SPC that is a causative agent of vasospasm. And it aims at providing the food composition for the prevention of the various diseases resulting from vasospasm.

  The present inventors have proceeded with research on vasoconstriction of vascular smooth muscle, and found that EPA inhibits the signal transduction action of the SPC-Rho kinase process, and this time, this action has caused the movement of Fyn protein to membrane rafts. The present invention has been completed by newly finding out what is caused by the inhibition. In addition, the effect of EPA discovered by the present inventor is not to act on a diseased site and treat it, but to act on a membrane raft that may cause vasospasm to suppress vasospasm, In other words, it is a preventive and gentle action, and it has been shown that by applying the present invention to a food composition, a food effective for the prevention of vascular disease can be produced.

  In addition, the present invention is intended to solve the “absorption efficiency in the intestinal tract”, which is considered to be a problem when orally administering EPA as an active ingredient, so as to promote secretion of bile and is derived from animals and plants that are not dangerous as a food composition. Provided is a food composition that also includes an auxiliary component, can efficiently take EPA into the body even in patients whose gastrointestinal function is weak due to diseases, etc., and can prevent diseases caused by vasospasm .

  That is, the 1st aspect of this invention provides the food composition for the prevention of the vascular disease which uses EPA as an active ingredient.

  The second aspect of the present invention provides a food composition for preventing vasospasm comprising EPA as an active ingredient.

  The third aspect of the present invention is the first aspect or the first aspect, wherein the EPA consists essentially of all-cis-EPA and a compound selected from at least one of pharmacologically acceptable salts, esters or derivatives thereof. A food composition according to the second aspect is provided.

  According to a fourth aspect of the present invention, there is provided the food composition according to any one of the first to third aspects, comprising EPA extracted from a seafood at a temperature of 100 degrees Celsius or less as an active ingredient. provide.

  According to a fifth aspect of the present invention, all-cis-EPA is extracted from fish and shellfish at a temperature of 100 degrees Celsius or less and is extracted in a state where the three-dimensional structure in the living body is maintained throughout the extraction process. The food composition according to any one of the first to fourth aspects is provided.

  A sixth aspect of the present invention provides a food composition according to the second aspect, which is for prevention of vasospasm induced by sphingosylphosphorylcholine.

  In a seventh aspect of the present invention, the Fyn protein activated by sphingosylphosphorylcholine is used for the prevention of vasospasm caused by moving to a membrane raft on the vascular wall. A food composition according to any of the aspects is provided.

  In an eighth aspect of the present invention, EPA is an active ingredient, and as an absorption auxiliary ingredient, dandelion powder, artichoke powder, curcumin 90% powder, oyster meat extract, Maria thistle extract, ursodeoxycholic acid and turmeric extract The food composition according to any one of the first to seventh aspects, including at least one kind, is provided.

  A ninth aspect of the present invention is the food composition according to the eighth aspect, comprising dandelion powder, artichoke powder, curcumin 90% powder, oyster meat extract, and Maria thistle extract as absorption auxiliary components I will provide a.

  A tenth aspect of the present invention includes EPA as an active ingredient, and contains at least one of ascorbic acid, tocopherol, a salt or ester thereof, vitamin E, and a fermented soybean extract as an auxiliary ingredient for antioxidant prevention of EPA, The food composition according to any one of the first aspect to the ninth aspect is provided.

  The eleventh aspect of the present invention is any one of the first to tenth aspects, comprising EPA as an active ingredient, digestion and absorption promotion, and egg yolk lecithin as an auxiliary component for improving hypercholesterolemia. A food composition is provided.

  A twelfth aspect of the present invention provides a food composition according to the eleventh aspect, further comprising beeswax and / or glycerin fatty acid ester as an auxiliary component for adjustment and stabilization.

  A thirteenth aspect of the present invention provides a functional food for preventing vascular disease, comprising the food composition according to any one of the first to twelfth aspects.

  In a fourteenth aspect of the present invention, at least one absorption aid selected from 70 to 75% by weight of EPA as a main component, dandelion powder, Maria thistle extract, oyster meat extract, ursodeoxycholic acid, and turmeric extract is used. 5-15% of ingredients, 2-15% by weight of at least one antioxidant auxiliary ingredient selected from ascorbic acid, tocopherol, salts or esters thereof, vitamin E, fermented soybean extract, and 0.5% of egg yolk lecithin Provided is a functional food for preventing vascular disease caused by vasospasm, comprising 2% by weight, 10-12% by weight of auxiliary components for adjustment and stabilization, and a total composition of 100% by weight.

  A fifteenth aspect of the present invention is a functional food for preventing vascular disease caused by vasospasm comprising eicosapentaenoic acid (EPA) as an active ingredient, wherein 73.6% by weight of EPA (28% 2% by weight dandelion powder, 2% by weight artichoke powder, 2% by weight curcumin 90% powder, and 2% by weight oyster Contains meat extract and 2% by weight of thistle extract, 2% by weight vitamin E, 1% by weight vitamin C and 1% by weight soy fermentation as auxiliary ingredients to prevent oxidation of EPA As an auxiliary ingredient for promoting digestion and absorption at the time of ingestion and improving hypercholesterolemia, it contains 1% by weight of egg yolk lecithin as an auxiliary ingredient for adjustment and stabilization. Containing 11.4% by weight of beeswax and glycerol fatty acid ester, to provide a functional food according to the thirteenth aspect.

Among them, the present invention includes [1] eicosapentaenoic acid (hereinafter referred to as “EPA”) as an active ingredient, and includes, as an absorption auxiliary ingredient, dandelion powder, artichoke powder, curcumin powder, oyster meat extract, and maria thistle extract . A composition for preventing vasospasm , and [2 ] ascorbic acid, tocopherol, or a salt thereof, vitamin E, and an auxiliary component for antioxidant prevention of EPA selected from a fermented soybean extract wherein, the composition and according to the above [1], [3] and the digestion and absorption promoting, as auxiliary components for hypercholesterolemia improvement, characterized in that it comprises a yolk lecithin, [1] or [2 and compositions described], further adjustments, including beeswax and / or glycerol fatty acid ester as a stabilizing auxiliary component (4) It consists of a composition as described in said [ 3 ] characterized by the above-mentioned.

The present invention also provides [ 5 ] a functional food for preventing vasospasm comprising the composition according to any one of [1] to [ 4 ], and [6 ] eicosapentaenoic acid (EPA ). Ingredients, containing 73.6% by weight of EPA ( 28% solution) as the main component, and 2% by weight of dandelion powder and 2% by weight of artichoke powder as auxiliary ingredients for enhancing the absorption efficiency of EPA in the intestinal tract 2% by weight vitamin E as a supplement to prevent oxidation of EPA, comprising 2% by weight curcumin 90% powder, 2% by weight oyster meat extract, and 2% by weight Maria thistle extract And 1% by weight of vitamin C and 1% by weight of fermented soy bean extract, 1% by weight of yolk resi as an auxiliary ingredient for promoting digestion and absorption at the time of intake and for improving hypercholesterolemia Include emissions, adjustment, as an auxiliary component for stabilizing consists functional food according to 11.4% by weight of beeswax and glycerol fatty acid esters including, in [5].

  By using the food composition of the present invention, it is possible to produce functional foods that have a preventive effect on various diseases caused by vasospasm. The present invention also includes an auxiliary component for increasing the absorption efficiency of EPA, which is an active ingredient, in the intestinal tract, thereby producing a food that can effectively prevent vascular disease even in persons with weak digestive function and the elderly. It becomes possible.

The schematic diagram of the SPC-Fyn-Rho kinase cascade concerning Ca ^<2+> independent vasospasm revealed by the present inventors is shown. (A) shows the molecular formula of EPA. (B) A schematic diagram of the three-dimensional structure of all-cis-EPA is shown. (C) The schematic diagram of the three-dimensional structure of trans-EPA is shown. The effect of EPA on vasospasm induced by SPC was compared by focusing on its three-dimensional structure. The vertical axis represents the degree of vasoconstriction, and the horizontal axis represents time. The bar at the bottom of the graph represents the time when each stimulus (potassium, SPC) was applied. For vasospasm caused by SPC stimulation, all-cis-EPA showed a higher inhibitory effect (faster time and degree of inhibition) than trans-EPA. Recovery from vasospasm induced by SPC stimulation was compared between all-cis-EPA and trans-EPA. The vertical axis represents the rate of recovery (relaxation) from contraction by SPC stimulation. all-cis-EPA had a significantly higher rate of relaxation compared to trans-EPA (t-test, p <0.05). In human coronary artery smooth muscle cells, stimulation with SPC induces migration of Fyn protein to the membrane raft (upper left arrow). On the other hand, in smooth muscle cells pretreated with EPA, Fyn protein migration by SPC stimulation was not observed (lower left), indicating that EPA inhibits Fyn protein migration to membrane rafts. Changes in blood EPA concentration after ingestion of EPA-containing foods were examined for each of the present invention without the auxiliary component (A) and with the auxiliary component (B). Three subjects, Mr. A (50s), Mr. B (40s), and Mr. C (30s), collected blood according to the schedule in Table 2 and measured the EPA concentration. In the absence of an auxiliary component, no significant increase in EPA concentration was observed in all three subjects (A), whereas in the presence of an auxiliary component, all three subjects showed an increase in EPA concentration. In the figure, the vertical axis represents the EPA concentration (μg / ml). Changes in blood arachidonic acid concentration after ingestion of EPA-containing foods were compared in the present invention without the auxiliary component (A) and with the auxiliary component (B). Even when the EPA-containing food was ingested, the blood arachidonic acid concentration did not change significantly. In the figure, the vertical axis represents the arachidonic acid concentration (μg / ml). The results of FIGS. 6 and 7 were expressed as relative values with the concentration at the time of ingesting the EPA-containing food (0 h) being 100. Among the blood concentrations of each component, “EPA concentration with auxiliary components” showed the largest increase rate. In the figure, the vertical axis represents the relative value of fatty acid concentration (0h = 100), and ARA (Solid bar) represents arachidonic acid.

  The present invention is described in detail below. The first aspect of the present invention is a food composition having EPA as a main component and effective in preventing vascular diseases. EPA itself is a known substance and has been known so far to be effective in thrombolysis, but EPA as a substance effective in preventing vascular disease has not been known so far. First revealed by the present invention.

  In the second to fifth aspects of the present invention, a food composition for preventing vascular disease caused by vasospasm, which contains EPA as an active ingredient, is presented. The EPA in the present invention is preferably all-cis-EPA, more preferably all-cis-EPA extracted from fish and shellfish, which is extracted in a state in which a three-dimensional structure is maintained in all extraction steps. All-cis-EPA (FIG. 2). In order to maintain the three-dimensional structure, for example, it is desirable not to be heated to 100 degrees Celsius or higher in the extraction process. However, as described in the examples, what is important is the three-dimensional structure of EPA, and the extraction temperature itself is The invention is not limited in any way. This is because EPA as a health food composition known so far is often exposed to high temperature conditions during its extraction, resulting in at least one of the five double bonds of the EPA molecule. Or two or three or more trans-EPAs replaced with trans-positions, and as shown in the Examples, all-cis-EPA is substantially trans- This is in consideration of the fact that the present inventors have revealed for the first time that it has an effect on vasospasm compared to EPA.

In the sixth and seventh aspects of the present invention, a food composition for preventing vascular diseases caused by vasospasm caused by the following factors is presented, which comprises EPA as an active ingredient. The vascular disease in this embodiment is specifically caused by Ca ²⁺ -independent vasoconstriction induced by sphingosylphosphorylcholine (SPC), and more specifically, the action of EPA on the vascular disease. Is characterized in that it inhibits the migration of Fyn protein activated by SPC to the membrane raft on the vascular wall. The present inventors have revealed that the Fyn protein is activated by SPC stimulation and moves to a membrane raft, and the activated Fyn protein activates Rho kinase to cause vasospasm (FIG. 1). ). Membrane raft is a structure formed by the deposition of cholesterol on the blood vessel wall, and EPA has the effect of inhibiting the migration of Fyn protein to this structure. Therefore, the present invention is a treatment after vasospasm occurs. It has become clear for the first time that it has a so-called “preventive” effect that inhibits the process leading to vasospasm. In the above aspect, food compositions containing the EPA according to the second to fifth aspects are also included in the present invention.

  In the eighth and ninth aspects of the present invention, a food composition for preventing vascular disease caused by vasospasm is provided, which contains EPA as an active ingredient and includes an “auxiliary ingredient that enhances absorption efficiency in the intestinal tract”. . Particularly in elderly people and patients with diseases, the function of the digestive system is lowered, and there is a problem that the effective effect is not sufficiently absorbed and the expected effect does not appear. In view of this point, and in order to guarantee sufficient absorption even in a healthy state, an auxiliary component for increasing the absorption efficiency of EPA in the intestinal tract is shown. The auxiliary component here is preferably at least one of dandelion powder, artichoke powder, curcumin 90% powder, oyster meat extract, and thistle extract, and more preferably a combination of all of the above. These auxiliary ingredients are all derived from natural animals and plants, promote bile secretion, and have been established as food safety. In the practice of the present invention, these may be used in appropriate combination, and the blending ratio and the like do not limit the present invention. Similarly, ursodeoxycholic acid, which is clinically used for the purpose of increasing the absorption efficiency of orally administered drugs, or the present invention is a food composition, the consumer feels resistance as an additive. For this purpose, “turmeric extract” containing ursodeoxycholic acid is also effective as an auxiliary component of this embodiment.

  In a tenth aspect of the present invention, a food composition for preventing vascular disease caused by vasospasm, comprising EPA as an active ingredient and comprising an “auxiliary ingredient for preventing EPA oxidation” is presented. In the present invention, as described above, EPA having a steric structure, preferably all-cis-EPA, is used as an active ingredient, and preventing its oxidation is important for effectively obtaining the effects of the present invention. The auxiliary component is at least one of vitamin E, vitamin C, and fermented soybean extract, preferably all of them. Although these auxiliary ingredients have an effect as an antioxidant ingredient, vitamins are nutritionally important elements themselves, and considering that the embodiment of the present invention is a health food, these auxiliary ingredients It is preferable as food to contain.

  In the eleventh and twelfth aspects of the present invention, a food composition for preventing vascular disease caused by vasospasm, comprising EPA as an active ingredient and containing “other supplementary ingredients” is presented. Other auxiliary components in this embodiment are egg yolk lecithin for promoting digestion and absorption at the time of ingestion and improvement of hypercholesterolemia, beeswax and glycerin fatty acid ester for adjustment and stabilization. Functional foods using the present invention are not necessarily used only by people with good health. In these cases, auxiliary ingredients and adjustment ingredients that are effectively digested and absorbed also bring out the effects of the present invention. Is important for.

  In the thirteenth to fifteenth aspects of the present invention, eicosapentaenoic acid (EPA) is used as an active ingredient, and further, an absorption auxiliary component, an antioxidant component, an auxiliary component for improving digestion and absorption and hypercholesterolemia, and an auxiliary for stabilizing stabilization. Provided is a functional food for preventing vascular disease caused by vasospasm, comprising a food composition containing an ingredient. The aspect of the food is not particularly limited to the present invention. However, since EPA extracted from fish has a so-called “fish odor”, for example, capsule foods, tablets and the like whose outer sides are covered with digestible materials are used. Presented.

Examples of the present invention will be described below.
(Preparation of specimen)
A porcine coronary artery (left anterior descending branch) was cut about 3 cm 1 cm distal from the main trunk bifurcation, and a Krebs solution (123 mM NaCl, 4.7 mM) that had previously been aerated with a mixed gas (95% O ₂ , 5% CO ₂ ) and ice-cooled. KCl, 15.5 mM NaHCO ₃ , 1.2 mM MgCl ₂ , 1.25 mM CaCl ₂ , 11.5 mM D-glucose). Sample preparation was performed while exchanging the Krebs solution every 15 minutes. After removing the fat around the blood vessel, the outer membrane was removed, the endothelium was removed with a cotton swab, and a smooth muscle strip 1 mm × 4 mm was prepared using a razor.

(Load of Fura-2 (Ca ²⁺ indicator) on smooth muscle strip)
First, 2 ml of Krebs solution loaded with the mixed solution was put into a test tube, and 100 μl of fatal calf serum (FCS) was added to perform sufficient pipetting. The 1 ml was then transferred to a 1.5 ml tube. After adding 40 μl of DMSO to 50 μg of Fura-2AM (DOJINDO) in a dark room, pipetting was performed 50-60 times, 10 μl of which was added to the 1.5 ml tube and immediately pipetting thoroughly. Thereafter, the operation of crushing for 5 minutes while cooling with ice in an ultrasonic crushing apparatus protected from light and vigorously stirring was repeated 6 times. In a dark room, the solution in which Fura-2AM was dissolved was returned from the 1.5 ml tube to the original test tube, the sample was put in, kept at 37 ° C., and Fura-2 was loaded for 4 hours. After 4 hours, the specimen was taken out, immersed in Krebs solution for 45 minutes, and after removing Fura-2AM adhering to the cells, a smooth muscle strip was attached to the measuring device.

(apparatus)
The smooth muscle strip loaded with Fura-2 was kept at 37 ° C. in a Magnus tube with a volume of 5 ml. A wire with a smooth strip strip is connected to a transducer (FD pickup: manufactured by Nihon Kohden Co., Ltd.) and passed through an amplifier (amplifier for strain pressure: manufactured by Nihon Koden Co., Ltd.) and a recorder (desktop pen recorder U-228: manufactured by Pantos) ) Detected the tension. For measurement of intracellular Ca ²⁺ concentration, a smooth muscle strip is alternately excited with light of 340 nm and 380 nm using a fluorescence measuring device (CAM230: manufactured by JASCO), and the ratio of the fluorescence intensity (F340 / 380) is determined. Measured by detecting.

(Measuring method)
Depolarization contraction by 118 mM high potassium solution was caused for 15 minutes, and then the operation of relaxing for 15 minutes with Krebs solution was repeated. When the waveform of tension due to high potassium depolarization was stabilized, 40 mM potassium solution was added, and the contraction was plateau When 1 was reached, 1 μM of Bradykinin was added to confirm the presence or absence of endothelium. When smooth muscle was not relaxed by Bradykinin, it was judged that the endothelium was removed. Thereafter, the membrane was relaxed with Krebs solution for 15 minutes and stimulated with 30 μM SPC. Furthermore, when the tension due to SPC stimulation reached a maximum and a plateau, EPA was added to a final concentration of 60 μM. In addition, EPA is the same experiment for all-cis-EPA, which retains the three-dimensional structure in vivo, and trans-EPA, in which one or more of the double bonds in the molecule are coordinated to the trans position by high-temperature treatment or the like. Comparison was made under conditions.

(result)
In the vascular smooth muscle contraction experiment, EPA suppressed contraction due to high potassium depolarization while EPA suppressed contraction due to SPC. In this experiment, the change in intracellular Ca ²⁺ concentration was also measured, but the contraction due to SPC was not accompanied by an increase in intracellular Ca ²⁺ concentration, whereas the contraction due to high potassium depolarization was accompanied by an increase in intracellular Ca ²⁺ concentration. It has been clarified that EPA suppresses abnormal contraction (not accompanied by an increase in intracellular Ca ²⁺ concentration) while not affecting normal contraction (associated with an increase in intracellular Ca ²⁺ concentration).

  The effect of EPA on vasospasm was examined by paying attention to its three-dimensional structure. Using the signal of vasospasm induced by treatment with 30 μM SPC as an index, vasospasm when all-cis-EPA and trans-EPA were treated was compared (FIGS. 3 and 4). As shown in the graph, all-cis-EPA showed significantly higher effects in terms of the speed and degree of vasospasm suppression than trans-EPA.

(Inhibitory action of EPA on Fyn binding to membrane raft)
Human coronary artery smooth muscle (CASMCs, manufactured by Cambrex) was cultured in SmGM-2 medium. Serum was removed 24 hours prior to the experiment, pretreated with 60 μM EPA for 10 minutes, and stimulated with 30 μM SPC for 5 minutes, followed by localization of Fyn and Caveolin-1 protein, which is said to be localized in membrane rafts. The relationship of localization was examined by immunofluorescence staining and confocal microscope observation (LSM-510, manufactured by Carl Zeiss). The results are shown in FIG. In cells not subjected to EPA pretreatment (upper), Fyn protein bound to the plasma membrane from inside the cell by SPC stimulation (left, arrow), and colocalized with Caveolin-1 (middle) (right) ), It was shown that the Fyn protein moves to the membrane raft by SPC stimulation. On the other hand, in EPA-pretreated cells (lower stage), no movement of Fyn protein to the plasma membrane by SPC stimulation was observed (left side), and no co-localization with Caveolin-1 protein (middle) was observed. (A composite image on the right side) shows that EPA suppresses the movement of the Fyn protein to the membrane raft.

(Composition and absorption efficiency of functional food using the present invention)
Since EPA is an oil, it is dispersed and absorbed by bile produced with meals. However, there is no explanation such as “should be taken after meals” in commercially available EPA-containing foods, and data such as the blood concentration of EPA after ingestion is not clear. Therefore, in this example, for the purpose of obtaining an absorption effect of EPA at the time of fasting, “EPA-containing food containing supplemental component of the present invention (EPA-P)” and “EPA-containing food not including supplemental component (EPA-N)”. The absorption efficiency at the time of fasting was examined. The composition of the “EPA-containing food containing supplemental ingredients” is as shown in Table 1, and the EPA content of EPA-N is the same as EPA-P.

  The measurement schedule in this example is shown in Table 2. About 3 people, Mr. A (50s), Mr. B (40s), and Mr. C (30s), about 10 ml of blood was collected at each stage of the schedule, and EPA concentration and arachidonic acid concentration ( Control) was measured. There was an interval of about 1 week between EPA-N and EPA-P measurements.

  The concentration of fatty acid at each stage of the schedule shown in Table 2 is shown in FIG. 6, FIG. 7 and Table 3 (relative values with blood concentration before taking EPA-containing food as 100 are FIG. 7 and Table 4). Of the three blood fatty acids, the arachidonic acid concentration (control) showed an increase of around 10% regardless of the presence or absence of auxiliary components. In EPA, without supplemental ingredients (N), Mr. C (30's) showed a 37% increase after 6 hours, whereas Mr. A (50's) and Mr. B (40's) showed almost an increase. There wasn't. On the other hand, in the case of supplementary component (P), the EPA concentration increased after 4 hours and 6 hours for all three persons, and the increase rate after 6 hours was 59% for A and 13% for B. C was 53%. These results are as follows: (1) In the state where the function of the gastrointestinal tract is weak, such as after fasting, the active ingredient may not reach the blood even if the EPA-containing food is ingested. (2) In these states, the tendency is strong (2) Even in these states, it is suggested that EPA can be taken into blood by adding an auxiliary component in the present invention. Indicates the effectiveness of the present invention in EPA-containing foods.

(Composition and absorption efficiency 2 of functional food using the present invention)
In the said Example 7, the change of the DHA density | concentration in the blood after each ingestion of EPA-P and EPA-N was contrasted with the change of EPA density | concentration (Table 5, Table 6). The blood DHA concentration tended to increase slightly 4-6 hours after ingestion, but the increase rate was lower than that of EPA. It has been shown that the food composition of the present invention specifically increases the blood EPA concentration.

  In the so-called “health food industry”, the present invention provides a food composition for producing a new food that can prevent vascular diseases. Today, the importance of prevention is being recognized, and the food composition provided by the present invention can be used as a composition of “health food”.

Claims (6)

Prevention of vasospasm characterized by comprising eicosapentaenoic acid (hereinafter referred to as “EPA”) as an active ingredient and as an absorption auxiliary ingredient, dandelion powder, artichoke powder, curcumin powder, oyster meat extract, and Maria thistle extract Composition. 2. The composition according to claim 1, comprising at least one EPA antioxidant auxiliary component selected from ascorbic acid, tocopherol, or a salt thereof, vitamin E, and soybean fermented extract. The composition according to claim 1 or 2 , comprising egg yolk lecithin as an auxiliary component for promoting digestion and absorption and improving hypercholesterolemia. The composition according to claim 3 , further comprising beeswax and / or glycerin fatty acid ester as an auxiliary component for adjustment and stabilization. A functional food for preventing vasospasm comprising the composition according to any one of claims 1 to 4 . Eicosapentaenoic acid (EPA ) as an active ingredient, containing 73.6% by weight of EPA ( 28% solution) as a main component, and 2% by weight of dandelion powder as an auxiliary ingredient for enhancing the absorption efficiency of EPA in the intestinal tract And 2% by weight artichoke powder, 2% by weight curcumin 90% powder, 2% by weight oyster meat extract, and 2% by weight maria thistle extract to help prevent oxidation of EPA As an ingredient, it contains 2 wt% vitamin E, 1 wt% vitamin C, and 1 wt% soy fermented extract. As an auxiliary component for promoting digestion and absorption and improving hypercholesterolemia when ingested, comprises 1% by weight of egg yolk lecithin, adjustment, as an auxiliary component for stabilizing, including 11.4% by weight of beeswax and glycerol fatty acid ester, machine of claim 5 Sex food.