JPH0913076A - Oil and fat controlling thrombocyte agglutination - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an oil and fat serving to control thrombocyte agglutination by using a triglyceride which contains a specified unsaturated fatty acid as the constituent of glyceride and in which less than a specified amount of the unsaturated fatty acid is bound to the 2-position of the glyceride. SOLUTION: This oil and fat comprises a triglyceride which contains an n-3 series long-chain polybasic unsaturated fatty acid as the constituent of glyceride and in which less than 40mol% of the total of the unsaturated fatty acid is bound to the 2-position of the glyceride. The oil and fat preferably contains the triglyceride in an amount of at least 5wt.%. The n-3 series long- chain polybasic unsaturated fatty acid preferably comprises at least one acid selected from α-linolenic, arachidonic, eicosapentaenoic, decosapentaenoic and docosahexaenoic acids.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to oils and fats having an action of suppressing platelet aggregation ability.

[0002]

BACKGROUND ART Arteriosclerosis is pathologically divided into atherosclerosis, medial sclerosis and arteriosclerosis. Atherosclerosis is the formation of fat deposits and plaque formation on the intima of coronary arteries, basilar arteries, renal arteries, thoracic and abdominal aortas, while medial sclerosis is of moderate limbs such as femoral arteries. Arteriosclerosis is a sclerosing change found in arterioles such as kidney / adrenal gland, spleen, ovary, and pancreas. It is considered that these arteriosclerosis are caused by unrestricted proliferation of medial smooth muscle cells in the intima of blood vessels and abnormal accumulation of intracellular cholesterol. To promote adhesion and aggregation of platelets and progress to arterial thrombus.

Eicosanoids such as prostaglandins and thromboxane synthesized in vivo from polyunsaturated fatty acids have a platelet aggregation action and a vasoconstriction action, and conversely have a platelet aggregation inhibition action and a vasodilation action. It is known that these eicosanoids are closely associated with arteriosclerosis. Further, among eicosanoids, prostaglandin I ₂ (hereinafter sometimes abbreviated as PGI ₂ ) has a platelet aggregation inhibitory action, a vasodilatory action and a blood pressure lowering action, and is a trovoxane A ₂ (hereinafter,
It may be abbreviated as TXA ₂ . ) Is known to have a platelet aggregation-inducing action and a vasoconstrictor action.

By the way, according to the conventional epidemiological survey, Eskimos who live on Greenland Island, which normally eats fish and sea animals, have arteriosclerosis as compared with Danish people who have almost the same high fat diet. It has been revealed that the incidence of ischemic heart disease, such as erythrocyte, is low, and that dietary polyunsaturated fatty acids are involved in the development of such disease (Dyerberg, T. et al., Am. J. . Clin. Nutritio
n, 28, 958, 1975). In addition, further epidemiological studies have shown that eicosapentaenoic acid (all cis
-5,8,11,14,17-eicosapentaenoic acid, EPA
Abbreviated. The numbers after C _{20: 5} and C represent the total number of carbons: the number of double bonds, and the same shall apply hereinafter. ) And docosahexaenoic acid (al
l cis -4,7,10,13,16,19-docosahexaenoic acid, hereinafter abbreviated as DHA. There is a significant correlation between ingestion of n-3 long-chain polyunsaturated fatty acids such as C _{22: 6} ) and suppression of platelet aggregation ability and decrease in whole blood viscosity, and cardiovascular diseases and brain It has been reported that there is an inverse correlation with the mortality rate due to vascular diseases or a reduction in the mortality rate due to the diseases (for example, Hirai, A. et al., Lancet, Volume 2, 1132).
P., 1980). As a mechanism for reducing the mortality rate from ischemic heart disease due to the ingestion of n-3 long-chain polyunsaturated fatty acids, antiplatelet aggregation action and plasma lipid improving action are considered.

For the treatment of arterial thrombosis using drugs, antiplatelet therapy mainly consisting of platelet aggregation inhibitors (eg warfarin, aspirin etc.) and thrombolytic agents (eg urokinase, etc.) for dissolving the resulting thrombus and embolus. Streptokinase) is widely used. On the other hand, EPA
As described above, intake of n-3 long-chain polyunsaturated fatty acids such as DHA and DHA is effective for prevention and treatment of ischemic heart disease such as arteriosclerosis as described above. Has been revealed.

Therefore, for the purpose of preventing or improving arteriosclerosis, a food containing a large amount of fish containing EPA or DHA is intentionally ingested, or a fish oil containing EPA or DHA or a fish oil concentrate is used as a material. Healthy foods that are available are commercially available.
However, it was necessary to ingest or administer these in large amounts and over a long period of time. Fish oils containing EPA and DHA are mainly sardine oil, cod liver oil, herring oil, squid oil,
Tuna orbital oil or the like is used, but the chemical structure of these fats and oils is such that 50 mol% or more of the total amount of the n-3 long-chain polyunsaturated fatty acid present by ester bond to glyceride is at the 2-position of triglyceride. In other words, the n-3 long-chain polyunsaturated fatty acid has a triglyceride structure in which the 2nd-position is more ester-bonded than the 1st-position and the 3rd-position.

On the other hand, EPA and DHA have the effect of preventing or improving arteriosclerosis as described above, but have a large number of double bonds in the molecule as compared with the usual fatty acids constituting edible vegetable oils and fats. Therefore, it is known that it is easily oxidized and, if ingested excessively, has a harmful effect on the living body. It is presumed that when the peroxidation reaction of lipids progresses in the living body, the membranes of the living body are damaged and cause lesions such as ischemic disease, arteriosclerosis, cataract, cancer, Alzheimer's disease, collagen disease and amyloidosis.

[0008]

SUMMARY OF THE INVENTION The present invention has been made in view of such a situation as described above, and an object of the present invention is to have no side effects on humans and animals and to obtain a conventional n-3 system. An object of the present invention is to provide an oil and fat having an action of suppressing the platelet aggregation ability and eventually facilitating the prevention and amelioration of arteriosclerosis by ingesting a smaller amount than a long-chain polyunsaturated fatty acid source.

[0009]

Means for Solving the Problems As a result of intensive studies aimed at achieving the above object, the present inventors have found that n-3 long-chain polyunsaturated fatty acids at the 1-position and / or 3-position of a glyceride structure. Oils and fats having a large amount of platelets are used as a source of n-3 long-chain polyunsaturated fatty acids, and have a higher platelet aggregation ability than fish oils having a large amount of n-3 long-chain polyunsaturated fatty acids at the 2-position. It was found that the above-mentioned object can be achieved, because the effect of suppressing erythrocyte is remarkably high, and the effect of preventing and improving arteriosclerosis can be expected.
The present invention has been completed based on such findings.

That is, the gist of the present invention is to include an n-3 long chain polyunsaturated fatty acid as a constituent fatty acid of glyceride,
Less than 40 mol% of the total amount of n-3 long-chain polyunsaturated fatty acids consists of triglycerides bonded to the 2-position of glycerides, or contains said triglycerides, which suppresses platelet aggregation ability It is a fat and oil.

The triglyceride featured in the present invention is n
In a triglyceride composed of a fatty acid containing a -3 long-chain polyunsaturated fatty acid and glycerin, n-3
When the total amount of the system long-chain polyunsaturated fatty acids is 100 mol%, less than 40 mol% thereof and any fatty acid other than the n-3 system long-chain polyunsaturated fatty acids form an ester bond at the 2-position of the triglyceride. And at least 60 mol% of the n-3 long-chain polyunsaturated fatty acid and any fatty acid other than the n-3 long-chain polyunsaturated fatty acid randomly or at the 1st and 3rd positions of the triglyceride. It is non-randomly distributed and has an ester bond.

The n-3 long-chain polyunsaturated fatty acid is an n-3 having 18 or more carbon atoms and 3 or more double bonds.
System linear unsaturated fatty acid, specifically, α-linolenic acid (C _{18: 3} ), octadecatetraenoic acid (C _{18: 4} , 6,9,
12,15-octadecatetraenoicacid), arachidonic acid (C _{20: 4} ), EPA (C _{20: 5} ), docosapentaenoic acid (C _{22: 5} , 7,10,13,16,19-docosapentaenoic acid)
), DHA (C _{22: 6} ) and the like. In the present invention, among these, one or two or more kinds of mixed fatty acids selected from the group consisting of α-linolenic acid, arachidonic acid, EPA, docosapentaenoic acid and DHA are preferable, and further EPA and / or Or D
HA is more preferred.

As the fatty acids other than the n-3 long-chain polyunsaturated fatty acids, any of short-chain, medium-chain and long-chain fatty acids and saturated and unsaturated fatty acids can be used. A straight-chain, medium- or long-chain saturated or unsaturated fatty acid having 6 or more carbon atoms is preferable. Such fatty acids include caproic acid (C _{6: 0} ), caprylic acid (C _{8: 0} ), capric acid (C _{10: 0} ), lauric acid (C _{12: 0} ), myristic acid (C _{14: 0} ), palmitic acid. (C _{16: 0} ), palmitooleic acid (C _{16: 1} ), stearic acid (C _{18: 0} ), oleic acid (C _{18: 1} ), elaidic acid (C _{18: 1} ), linoleic acid (C _{18 : 2} ), α'-linolenic acid (C _{18: 3} , 5,8,11-octadecatrienoic acid), γ-linolenic acid (C _{18: 3} , 6,9,12-octadecatrienoic acid), Rheostearic acid (C _{18: 3} , 9,11,13-octadecatrienoic acid), arachidic acid (C _{20: 0} ), gadoleic acid (C
_{20: 1} ), behenic acid (C20 _{: 0} ), erucic acid (C22 _{: 1} ), brassic acid (C22 _{: 1} ) and the like. These fatty acids may be used alone or may be used as mixed fatty acids in any proportion. Of these, myristic acid, palmitic acid, stearic acid, oleic acid, and linoleic acid are preferable.

To produce the triglyceride of the present invention composed of the above-mentioned n-3 long-chain polyunsaturated fatty acid and other fatty acids, a chemical synthesis method, a transesterification method, or an extraction method from a natural product is used. The technology such as the above may be used.
Examples of the chemical synthesis method include a desired amount and composition of a fatty acid, a fatty acid anhydride or a fatty acid halide (fatty acid chloride) and glycerin, an acidic substance (hydrochloric acid, sulfuric acid, paratoluenesulfonic acid, etc.), an alkaline substance (sodium hydroxide). Esterification of metals (zinc, tin, titanium, nickel, etc.), metal oxides (zinc oxide, alumina, ferrous oxide, etc.), metal halides (aluminum chloride, tin chloride, etc.), etc. 100 to 250 ° C. in a nitrogen gas stream in the presence or absence of a catalyst
It is preferable that the esterification reaction is carried out for 1 to 25 hours while heating to room temperature and removing the produced water.

The esterification product may be subjected to alkali deoxidation treatment, activated carbon, activated clay, alumina, silica gel, if necessary.
Free fatty acids, monoglycerides, diglycerides, which are subjected to adsorption / fractionation treatment using an ion exchange resin or the like, solvent separation treatment using a hydrophilic organic solvent such as methanol or ethanol and / or a lipophilic organic solvent such as n-hexane or xylene, Impurities such as coloring substances and odorous components are removed, and these treatments are appropriately combined to obtain triglyceride
The content of the n-3 long-chain polyunsaturated fatty acid residues bonded to the position is the total of the n-3 long-chain polyunsaturated fatty acid residues bonded to the 1st, 2nd and 3rd positions of the triglyceride. 40 of content
The triglyceride component may be fractionated or concentrated so as to be less than mol%. The triglyceride of the present invention is
For example, it is desirable to blow steam under heating and decompression to perform deodorizing treatment.

To obtain the triglyceride of the present invention using the transesterification method, for example, triglycerides of fatty acids (component a-1) containing a large amount of n-3 long-chain polyunsaturated fatty acids as raw materials and n- Fatty acid (component a-2) which does not substantially contain or contains a small amount of 3 type long chain polyunsaturated fatty acid, lower alcohol ester (methyl ester, component a-2)
Ethyl ester and the like. The same applies hereinafter. Or a component a-2 triglyceride in a desired ratio, or a fatty acid triglyceride (component b-1) substantially containing no or a small amount of n-3 long-chain polyunsaturated fatty acid and n-. Fatty acids containing a large amount of 3 long-chain polyunsaturated fatty acids (component b-
2) or a lower alcohol ester of component b-2 is mixed in a required amount, and an alkaline substance such as sodium hydroxide and potassium hydroxide as a catalyst, a metal alcoholate (metal alkoxide) such as sodium methylate, sodium ethylate and lithium butyrate. ), An anion exchange resin such as a basic anion exchange resin or an acidic cation exchange resin, or a lipase, it is convenient to carry out the transesterification reaction. When transesterification is carried out using a specific lipase as a catalyst, a new fatty acid group can be selectively introduced into the 1-position and 3-position of the triglyceride as described later, which is desirable as a method for producing the triglyceride of the present invention. .

The raw materials for transesterification include vegetable oils such as linseed oil, sesame oil, perilla oil, etc. as component a-1, fish oils such as sardine oil, cod liver oil, herring oil, squid oil, tuna orbital oil, whales, seals, Pressed or extracted oil obtained from the origin of marine mammals such as fur seals, milk fat of the animals, chlorella, spirulina, donariella, etc.Also genus Nannochloropsis (for example, Nannochloropsis oculata), genus Trustchitrium (for example, Thraustochytrium aureum)
), Crypthecodinium (eg Crypthecodinium
cohnii), isochrysis (eg Isochrysis galban
a) fats and oils extracted from microalgae belonging to, for example, fats and oils derived from microorganisms of the genus Mortierella,
Further, triglycerides of n-3 long-chain polyunsaturated fatty acids or mixed fatty acids with the above-mentioned various fatty acids containing the fatty acids at an arbitrary ratio (see paragraph 0013) can be used. As the component a-2, various fatty acids or derivatives thereof described in paragraph 0013 can be used.

As the component b-1, there are triglycerides obtained from plants, animals, microorganisms, microalgae, etc., soybean oil,
Rapeseed oil, cottonseed oil, corn oil, palm oil, palm oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, olive oil, peanut oil, cacao butter, Chinese tallow, sal butter, shea butter, beef tallow,
Lard, hydrogenated oils and fats thereof, fractionated oils and fats, and the aforementioned component a
-2 triglyceride, medium chain fatty acid triglyceride and the like can be exemplified, and as the component b-2, there is a fatty acid obtained by the hydrolysis treatment of the component a-1.

In the transesterification reaction, for example, the above raw materials are mixed in a molar ratio of component a-1: component a-2 = 1: 0.1.
5, component b-1: component b-2 = 1: 2 to 10 are mixed, and when an alkali or a metal alcoholate is used as a catalyst, it is in an anhydrous state and is 0.5 to 0.5 at 80 to 120 ° C. Allow transesterification for 3 hours. Similarly, when an ion exchange resin is used, it is also in an anhydrous state, but it is preferable to circulate and contact the raw materials in a column system at room temperature to 40 ° C. When using lipase as a catalyst, the water content in the raw material is set to 1% by weight or less, and commercially available lipase powder or a known carrier such as celite, diatomaceous earth, activated carbon, porous glass, ion exchange resin, chitosan, high Immobilized lipase immobilized on molecular gel, cellulose powder or the like is added, and transesterification reaction is performed at 20 to 80 ° C. for 0.5 to 20 hours.

As the lipase, those originating from the following microorganisms or originating from animal organs can be used. That is, the genus Aspergillus (eg Aspergillus niger), the genus Mucor (eg Mucor miehei), the genus Candida (eg Candida cyrindracea), the genus Pseudomonas (eg Pseudomonas fragi), the genus Alcaligenes sp (eg Japanese Patent Publication No. 58-36953). .
), A lipase of microbial origin belonging to the genus Rhizopus delemar (eg Rhizopus delemar), a genus Geotrichum (eg Geotrichum candidum), and a pancreatic lipase of pigs, cattle, etc. Of these, lipases originating from microorganisms of the genus Aspergillus, genus Mucor, genus Alcaligenes and genus Rhizopus, and porcine pancreatic lipase are the glyceride positions 1 and 3
Since it acts specifically on the position, it is suitable for producing the triglyceride of the present invention.

The transesterification reaction product obtained by the above-mentioned various transesterification processes can be the triglyceride used in the present invention depending on the kind of the raw material selected, but it can be obtained by the above-mentioned chemical synthesis method. As in the case of the esterification product, if necessary, alkali deoxidation treatment, adsorption / fractionation treatment, solvent fractionation treatment or solvent-free fractionation (wintering) treatment, etc. are appropriately combined and subjected to the transesterification reaction product, The triglyceride used in the present invention can be obtained by removing impurities or fractionating or concentrating the glyceride component. The triglyceride is preferably deodorized.

The triglyceride according to the present invention can also be obtained by a method of extracting fats and oils from natural products. That is, among those described as the raw materials for transesterification (component a-1), whales, harbor seals,
harp seal etc.), body tissues of marine mammals such as fur seals, milk secreted from the animals, cells of microalgae such as chlorella, spirulina, and donariella, or cultured cells thereof, genus Nannochloropsis, Trustokis Microalgae belonging to the genus Thraustochytrium, the genus Crypthecodinium and the genus Isochrysis, for example Nannochloropsisoculata
), Trustchytrium aureum (Thraustochytr
ium aureum), Crypthecodinium cornie (Cry
pthecodinium cohnii), Isocrisis galvana (Isoch
rysis galbana) etc. or these cultured cells are used as raw materials. In addition, when it originates from a microorganism, it does not matter if the triglyceride obtained therefrom satisfies the glyceride structure of the present invention.

These are squeezed or n-hexane,
Chloroform, benzene, diethyl ether, an organic solvent such as methanol is extracted or fractionated to obtain an oil, which is subjected to degumming, alkaline deoxidation, decolorization, deodorization, and other free fatty acid, phospholipid, The glyceride fraction can be obtained by removing impurities such as glycolipids, unsaponifiable substances, coloring substances and odorous components. This glyceride fraction can be used as the triglyceride used in the present invention, but the glyceride fraction is further fractionated by solventless low temperature fractionation, solvent fractionation or a silica gel column etc. to bind to the 2-position of triglyceride. It is also possible to produce triglycerides with even fewer long-chain polyunsaturated fatty acid residues.

The triglyceride of the present invention produced by the chemical synthesis method, the transesterification method, the extraction method from a natural product, etc. as described above has an n-3 long-chain polyunsaturation as its constituent fatty acid. Less than 40 mol% of the total amount of fatty acids form an ester bond at the 2-position of the triglyceride, but more preferably less than 20 mol%. If it is 40 mol% or more, the desired effect of the present invention becomes small. The triglyceride of the present invention can be used as it is as fats and oils,
It can also be used as a fat or oil by mixing with a common edible fat or oil, for example, the animal or vegetable fat or oil as described as the component b-1. At this time, the content of the triglyceride of the present invention is preferably 5 to 100% by weight of the total fat and oil, and further 10 to
100% by weight is even more preferred. Most preferably 20
１００100% by weight. If it is less than 5% by weight, the desired effect of the present invention is small.

The fats and oils according to the present invention can be ingested in the form of capsules such as soft capsules and microcapsules together with ordinary edible animal and vegetable fats and oils, vitamin E, β-carotene and the like, and in the same manner as ordinary edible fats and oils. As a food material, it can be used by being used as a raw material for various processed foods and a material for cooking. Further, the oil and fat according to the present invention can be expected to be used for the prevention and treatment of arteriosclerosis.

[0026]

【Example】

Example 1 1 kg of triolein and fish oil (manufactured by Tama Biochemical Co., Ltd., trade name: EPA-18) were hydrolyzed mixed fatty acids at low temperature fractionated fish oil hydrolyzed fatty acid concentrate (C _{20: 5} : 3 in total fatty acids).
7.4 mol%, C22 _{: 5} : 5.4 mol%, C22 _{: 6} : 25.
2 mol%. 72. as an n-3 long-chain polyunsaturated fatty acid.
5 mol%. Add 0.01 wt% BHT. ) Is mixed at a molar ratio of 1: 5 and the water content is adjusted to 0.2% by weight, and then Lipozyme IM20 (trade name; lipase derived from Mucor miehei, manufactured by Novo Nordisk) is used. Packed glass column (10cmφ × 60c
m) was allowed to undergo a selective transesterification reaction at 40 ° C.

After removing free fatty acids from the transesterification reaction product by steam distillation and washing with water, a stainless steel column filled with silica gel (Wako Pharmaceutical Co., Ltd., trade name: Wakogel C100) infiltrated with n-hexane. The resulting mixture was eluted with n-hexane to remove the diglyceride to obtain 720 g of the triglyceride of the present invention. Total fatty acid composition constituting the triglyceride, 1st and 3rd of glyceride
The fatty acid composition of each of the 2nd and 3rd positions was determined by GLC analysis.
Table 1 shows the results. C that constitutes this triglyceride
90 mol% of _{20: 5} . 95 mol% or more of C _{22: 6} and 93.5 mol% of the total amount of the n-3 long-chain polyunsaturated fatty acids were distributed in the 1st and 3rd positions of the triglyceride. That is, 6.5 mol% of the total amount of the n-3 long-chain polyunsaturated fatty acid was distributed at the 2-position of this triglyceride. This triglyceride was used as a test oil for the following animal experiments.

0.1% by weight of sodium methoxide was added to a part of the present triglyceride, and a random transesterification reaction was carried out at 100 ° C. under reduced pressure, followed by filtration using Celite to obtain a random transesterification product of the present triglyceride. Got The total fatty acid composition, the respective fatty acid compositions at the 1st, 3rd and 2nd positions were determined in the same manner as described above (see Table 1). At the 2-position of this triglyceride, 50.6 mol% of the total amount of the n-3 long-chain polyunsaturated fatty acid was distributed. This random transesterification product was used as a control oil for animal experiments.

[0029]

[Table 1] * Total carbon number: Displayed as the number of double bonds. (N-3) represents an n-3 fatty acid.

Seven 4-week-old SD male rats were used as one test group, and a breeding experiment was carried out using a feed (see Table 2) containing 5% by weight of each of the test oil and the control oil. During this period, the feed was prepared and fed daily in order to prevent oxidative deterioration of the feed components. After allowing water and each of the above-mentioned feeds to be freely ingested and breeding for 3 weeks, each amount of PGI _{2 in} the aorta of each test group rat and TXA ₂ in blood was measured. Table 3 shows the results.
No significant difference was observed in feed intake, body weight gain and liver weight in each test plot. From these experimental results, the triglyceride (test oil) according to the present invention has no side effect on rats, and in the group to which the test oil is added, the production amount of PGI ₂ is remarkably increased (that is, the inhibitory effect on the platelet aggregation ability and the arteries). It is revealed that the relaxation effect is increased) and the production amount of TXA ₂ is extremely decreased (that is, the inducing effect of the platelet aggregation ability and the decrease of the arterial contraction effect), and therefore the triglyceride according to the present invention prevents the arteriosclerosis. And the potential for treatment.

[0031]

[Table 2] * 1 Nippon Claire Co., Ltd., AIN-93G-MX * 2 Nippon Claire Co., Ltd., AIN-93-VX * 3 t-Butylhydroquinone

[0032]

[Table 3] * There is a significant difference with a risk rate of 5% or less with respect to the value of the control oil added section.

Example 2 A test fat (fat containing a triglyceride of the present invention) and a control fat were prepared as follows. That is, the test fat is ha
The rp seal (seal) oil and fat was cooled with a dry ice / acetone refrigerant at −80 ° C. for 1 hour, and the precipitated crystal part was filtered off with a filter paper to prepare. The control fats and oils are two types of fish oils with different fatty acid composition (mixed oil of cod liver oil and miscellaneous fish oil, tuna orbital oil), cooled with dry ice / acetone refrigerant in the same manner, and the concentrated fractions are blended to obtain the total fatty acid composition. It is almost similar to that of the test fat. Table 4 shows these fatty acid compositions.

[0034]

[Table 4] *: Same as the notes in Table 1.

Seven 4-week-old SD male rats were used as one test group, and oils and fats containing 20% by weight of each of the above-mentioned test oils and control oils (20 weight parts of test oils or control oils, 50 weight parts of palm oil, A mixed fat containing 5 parts by weight of oleic safflower oil and 25 parts by weight of high linoleum safflower oil (see Table 5 for the composition of fatty acids), each containing 10% by weight (the composition of the feed is 5% by weight of fat and 10% by weight). , The same as in Example 1 except that 41.7% by weight of corn starch was changed to 36.7% by weight). During this time,
Feed was prepared and fed daily to prevent oxidative degradation of feed ingredients. After allowing the animals to freely ingest water and the above-mentioned feeds for 3 weeks, the amount of PGI _{2 in} the aorta and TXA _{2 in} the blood of each test group rat was measured (see Table 6). No significant difference was observed in feed intake, body weight gain and liver weight in each test group. From this experimental result, in the group to which the test fat and oil (fat and fat containing the triglyceride according to the present invention) was added, no side effect was observed in rats,
Further, it was revealed that the increase in PGI ₂ production and the decrease in TXA ₂ production, that is, the inhibitory effect on the platelet aggregation ability and the arterial vasodilatory effect are enhanced. Therefore, it was confirmed that the fats and oils containing the triglyceride according to the present invention can be used for prevention and treatment of arteriosclerosis.

[0036]

[Table 5] *: Same as the notes in Table 1. (N-6) shows an n-6 fatty acid. * *: Ratio of each fatty acid among saturated fatty acids, monounsaturated fatty acids, n-6 fatty acids and n-3 fatty acids.

[0037]

[Table 7] *: Same as the notes in Table 3.

Example 3 A rat breeding experiment was carried out in the same manner as in Example 2 except that fats and oils having different mixing ratios of the test fat and oil and the control fat used in Example 2 were added to the feed. That is, seven 4-week-old SD male rats were used as one test group, and the fats and oils containing 10 wt% of the test fat and oil or the control fat and oil described in Example 2 (10 parts by weight of the test fat and oil and the control oil and fat, 50 parts by weight of palm oil) were used. , A mixed oil of 10 parts by weight of high oleic safflower oil and 30 parts by weight of high linoleum safflower oil. See Table 7 for the fatty acid composition. The same as the above). During this time, the feed was prepared daily to prevent oxidative degradation of the feed components. After allowing water and each of the above-mentioned feeds to be freely ingested and breeding for 3 weeks, the amounts of PGI _{2 in} the aorta and TXA _{2 in} the blood of each test group rat were measured (see Table 8). No significant difference was observed in feed intake, body weight gain and liver weight in each test group. From the results of this experiment and the results of Example 2, the fats and oils containing the triglyceride according to the present invention include the fats and oils in which a small amount of the test fats and oils were mixed as compared with the control fats and oils without causing side effects on rats. As a result, it was revealed that the PGI ₂ production amount was increased and the TXA ₂ production amount was decreased, thereby enhancing the inhibitory action on the platelet aggregation ability and the arterial vasodilatory action. From this, it was confirmed that the fats and oils containing the triglyceride according to the present invention can be used for the prevention and treatment of arteriosclerosis.

[0039]

[Table 7] * And **: Same as the notes in Table 5.

[0040]

[Table 8] *: Same as the notes in Table 3.

Example 4 Microalga Crypthecod
inium cohnii, ATCC30336) was planted in 30 liters of the medium shown in Table 9, aerobically cultivated at 30 ° C. for 100 hours in a jar fermenter, and cultured alga cells were collected by centrifugation and freeze-dried. (Yield 6
25 g). Chloroform: methanol =
Hyscotron (trade name) in a mixed solvent of 1: 1 (weight ratio).
Cell disruption and extraction by Nissin Medical Science Instruments)
520 g of oil was obtained. Silica gel dispersed in n-hexane (manufactured by Wako Pure Chemical Industries, Ltd., trade name: Wakogel C10)
0) was filled with a stainless steel column,
Triglyceride 250 according to the present invention was eluted with diethyl ether: n-hexane = 10: 90 (volume ratio).
g was obtained. This triglyceride (this was used as a test fat)
Was determined in the same manner as in Example 1 (see Table 10).

[0042]

[Table 9] (PH: 6.8) * 1: Vitamin mix aqueous solution (unit: weight in 1 liter of the aqueous solution) Biotin: 0.003 g Thiamine: 1.000 g * 2: Metal mix aqueous solution (unit: in 1 liter of the aqueous solution) Weight) Na ₂ EDTA: 1.00 g FeCl ₃ .6H ₂ O: 0.05 g H ₃ BO ₃ : 1.00 g MnCl ₂ .4H ₂ O: 0.15 g ZnCl ₂ : 0.01 g CoCl ₂ · 6H ₂ O: 0.005g

[0043]

[Table 10] *: Same as the notes in Table 1.

The thus-obtained triglyceride derived from microalgae (test fat and oil) and the fat and oil containing 10 wt% of the control fat and oil described in Example 2 (10 wt parts of test fat and oil or control fat and oil, 50 parts by weight of palm oil, Hiole) 10 parts by weight of Ick safflower oil and 3 of high linoleum safflower oil
0 parts by weight of mixed fats and oils. See Table 11 for fatty acid composition. 10% by weight of each feed was prepared (the feed composition is the same as in Example 3 except for fat content), and the same breeding test as in Example 3 was performed. Table 12 shows the results of analysis of the amount of PGI _{2 in} the aorta and the amount of TXA ₂ in blood of each test group rat. No significant difference was observed in feed intake, body weight gain and liver weight in each test plot. Results of this experiment and Example 2
From the results, the fats and oils according to the present invention have no adverse effects on rats, and increase the PGI ₂ production amount and the TXA ₂ production amount, including the case of the fats and oils mixed with a small amount of the test fats and oils as compared with the control fats and oils. It was revealed that the inhibitory effect on platelet aggregation and the effect on dilating arterial blood vessels were enhanced by decreasing the blood pressure. From this, it was confirmed that the fats and oils containing the triglyceride according to the present invention can be used for the prevention and treatment of arteriosclerosis.

[0045]

[Table 11] * And **: Same as the notes in Table 5.

[0046]

[Table 12] *: Same as the notes in Table 3.

[0047]

INDUSTRIAL APPLICABILITY According to the present invention, PGI ₂ production is increased and TXA ₂ production is decreased as compared with the conventional n-3 long-chain polyunsaturated fatty acid source without causing side effects on animals. It is highly effective in producing eicosanoids that suppress platelet aggregation and expand arterial blood vessels by ingesting a smaller amount than conventional n-3 long-chain polyunsaturated fatty acid sources such as fish oil. Therefore, it is possible to provide an oil or fat having an action of facilitating prevention and improvement of arteriosclerosis.

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[Procedure amendment]

[Submission date] August 12, 1996

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] 0017

[Correction method] Change

[Correction contents]

The raw materials for transesterification include vegetable oils such as linseed oil, sesame oil, perilla oil, etc. as component a-1, fish oils such as sardine oil, cod liver oil, herring oil, squid oil, tuna orbital oil, whales, seals, Compressed or extracted oil obtained from marine mammals such as fur seals, milk fat of the animals, chlorella, spirulina, donaliella , etc., and also genus Nannochloropsis (eg Nannochloropsis oculata , UTEX
LB 2164 etc. ), Trust chytrium (eg Thrausto
chytrium aureum , ATCC 28211, 34304 etc. ), Crypthecodinium cohnii , AT
CC 30021, 30334, 30336, 50052, etc. ), genus Isochrysis (for example, Isochrysis galbana , CCAP927 / 1, UTE
X LB 987, etc. ) and other oils and fats extracted from microalgae belonging to the genus Mortierella (M.
isabellina, IFO 6336, 6739, 7873, 7884, ATCC
44853) or the like, or a triglyceride of an n-3 long-chain polyunsaturated fatty acid or a mixed fatty acid with the above-mentioned various fatty acids (see paragraph 0013) containing this at an arbitrary ratio. ATCC: American Type Cul
ture collection (USA), CCAP: Culture collection
of Algae and Protozoa (UK), UTEX: Culture Colle
ction of Algae at the University of Texas (US
Country), IFO: Osaka Fermentation Research Institute. Ingredient a-2
The various fatty acids or their derivatives described in paragraph 0013 can be used.

[Procedure amendment 2]

[Document name to be amended] Statement

[Correction target item name] 0030

[Correction method] Change

[Correction contents]

Seven 4-week-old SD male rats were used as one test group, and a breeding experiment was carried out using a test oil and a control oil and 5% by weight of each feed (see Table 2). During this time,
Feed was prepared and fed daily to prevent oxidative degradation of feed ingredients. After allowing water and each of the above-mentioned feeds to be freely ingested and breeding for 3 weeks, each amount of PGI _{2 in} the aorta of each test group rat and TXA ₂ in blood was measured. Table 3 shows the results. No significant difference was observed in feed intake, body weight gain and liver weight in each test plot. From this experimental result, the triglyceride (test oil) according to the present invention has no side effect on rats, and PGI was not observed in the group to which the test oil was added.
₂ significantly increases (ie, suppresses platelet aggregation and increases arterial relaxation) and TXA ₂ production extremely decreases (ie, induces platelet aggregation and decreases arterial contraction) Therefore, it was confirmed that the triglyceride according to the present invention can be used for the prevention and treatment of arteriosclerosis.

[Procedure 3]

[Document name to be amended] Statement

[Correction target item name] 0035

[Correction method] Change

[Correction contents]

Seven 4-week-old SD male rats were used as one test group, and the above-mentioned test fats and control fats and oils containing 20% by weight each (test fats or control fats or oils 20 parts by weight, palm oil 50 parts by weight) were used. , A mixed fat of 5 parts by weight of high oleic safflower oil and 25 parts by weight of high linoleum safflower oil. See Table 5 for the fatty acid composition. % And the same as in Example 1 except that 41.7% by weight of corn starch was 36.7% by weight).
During this period, the feed was prepared and fed daily in order to prevent oxidative deterioration of the feed components. Free intake of water and each of the above feeds 3
After breeding for a week, PGI _{2 in} the aorta of each test group rat
And the amount of TXA ₂ in blood was measured (see Table 6). No significant difference was observed in feed intake, body weight gain and liver weight in each test group. From this experimental result, in the group to which the test fat and oil (fat and fat containing the triglyceride according to the present invention) was added, no side effect was observed in the rat, and PGI ₂ production was increased and TXA ₂ production was decreased, that is, platelet aggregation ability. It was clarified that the inhibitory effect of erythrocyte and the arterial vasodilatory effect are enhanced. Therefore, it was confirmed that the fats and oils containing the triglyceride according to the present invention can be used for prevention and treatment of arteriosclerosis.

[Procedure amendment 4]

[Document name to be amended] Statement

[Correction target item name] 0044

[Correction method] Change

[Correction contents]

The triglyceride derived from microalgae (test fat and oil) thus obtained and the control fat and oil described in Example 2 were used.
Used, the mixed oil and fat. The fatty acid composition of each 10 wt% including fat (test oils or control oil 10 parts by weight, palm oil 50 parts by weight, high-oleic safflower oil 10 parts by weight of high-linoleic safflower oil 30 parts by weight Table 11 (see 11.) was mixed with each 10% by weight (the composition of the feed was the same as in Example 3 except for the fat content), and the same breeding test as in Example 3 was performed. PGI ₂ of aorta of each test group rat
Table 12 shows the results of analysis of the amount of TXA ₂ produced in blood. No significant difference was observed in feed intake, body weight gain and liver weight in each test plot. From the results of this experiment and the results of Example 2, the fats and oils according to the present invention had no adverse effects on rats, and the PGI ₂ production amount including the fats and oils containing a small amount of test fats and oils compared to the control fats and oils was included. It was clarified that the inhibitory effect on the platelet aggregation ability and the arterial vasodilatory effect are enhanced by increasing the amount of the protein and the amount of TXA ₂ produced. From this, it was confirmed that the fats and oils containing the triglyceride according to the present invention can be used for the prevention and treatment of arteriosclerosis.

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Claims (8)

[Claims] 1. An n-3 long-chain polyunsaturated fatty acid is contained as a constituent fatty acid of the glyceride, and less than 40 mol% of the total amount of the n-3 long-chain polyunsaturated fatty acid is bonded to the 2-position of the glyceride. An oil and fat having an action of suppressing platelet aggregation ability, which is characterized by comprising triglyceride. 2. An oil and fat containing 5% by weight or more of the triglyceride according to claim 1, which has an action of suppressing the platelet aggregation ability. 3. The n-3 long-chain polyunsaturated fatty acid is one or more selected from the group consisting of α-linolenic acid, arachidonic acid, eicosapentaenoic acid, docosapentaenoic acid and docosahexaenoic acid. The oil and fat according to claim 1 or 2. 4. The oil or fat according to claim 1, wherein the n-3 long-chain polyunsaturated fatty acid is eicosapentaenoic acid and / or docosahexaenoic acid. 5. The oil or fat according to any one of claims 1 to 4, wherein the triglyceride is obtained from a marine mammal or a microalgae, is concentrated, or is transesterified. 6. The oil or fat according to claim 5, wherein the marine mammal is a whale or a seal. 7. The oil or fat according to claim 5, wherein the microalgae belongs to the genus Nannochloropsis, the genus Trustchitrium, the genus Isochrysis or the genus Crypthecodinium. 8. The oil or fat according to claim 1, wherein the triglyceride is produced by a transesterification reaction using a lipase having specificity at the 1- and 3-positions of the glyceride.