JP2006306866A - Adiponectin-increasing agent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an adiponectin-increasing agent capable of keeping a biological factor closely involving so-called life-style related diseases, e.g. hypertension, obesity, hyperlipemia and diabetes mellitus to a normal value while usually taking a lipid into the human body. <P>SOLUTION: The adiponectin-increasing agent or a food for increasing adiponectin comprises docosahexaenoic acid or its derivative as an active ingredient. In the adiponectin-increasing agent or the food for increasing adiponectin, the derivative of docosahexaenoic acid is a docosahexaenoic acid ethyl ester or a docosahexaenoic acid-binding type phospholipid. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to an adiponectin elevating agent containing docosahexaenoic acid or a derivative thereof as an active ingredient.

  In modern Japan, the introduction of excessive lipids into the diet brought about by the westernization of dietary habits has caused various lifestyle-related diseases such as obesity, and is now serious not only in the medical field but also in the socioeconomic context. Is causing problems. As dietary habits are reviewed as an improvement measure, not only the amount of fat in the diet, but also the importance of quality is recognized again. For a long time, docosahexaenoic acid (DHA) is abundant in fish oil and has attracted attention as a functional lipid with various nutritional physiology functions such as improvement of hyperlipidemia and prevention of arteriosclerosis. In the present situation that 20 to 30% of patients with lifestyle-related diseases have developed pathology due to obesity, it is considered that functional evaluation of foods using obese model animals is significant. Obese model Otsuka Long-Evans Tokushima Fatty (OLETF) rats overeating due to a receptor mutation of cholecystokinin, an appetite regulating hormone, and develop obesity, hyperlipidemia, and diabetes.

  By the way, the idea that fat cells are not just cells that store energy, but a kind of endocrine organ, has recently been generalized, and is produced by regulating "adipocytokines" at the gene transcription level as needed. Have been reported to be released.

  Among these adipocytokines, adiponectin was found as an expression product of a gene that is specifically expressed in adipose tissue and highly expressed. Although the concentration in blood is secreted specifically in adipocytes, it decreases when obesity, that is, fat is accumulated, and increases when the amount is reduced (Non-patent Document 1). In particular, the concentration in the blood decreases due to an increase in visceral fat, and becomes low even in patients with coronary artery disease or diabetes (Non-Patent Document 2). Moreover, the low value of blood adiponectin is the best index for the future development of diabetes (Non-patent Document 3).

  Adiponectin is considered to be a candidate for adipose tissue-derived insulin sensitivity factor together with leptin. Insulin resistance and lipid metabolism abnormalities in lipotrophic diabetes were partially improved by physiological leptin administration, but almost completely improved by simultaneous administration of adiponectin and leptin. Then, by supplementing adiponectin and increasing the blood concentration, the triglyceride content in tissue that causes insulin resistance can be reduced (Non-patent Document 4).

  It has also been reported that adiponectin exhibits an anti-arteriosclerotic action directly on arteriosclerotic lesions. The mechanism of action is considered to be the action of reducing the accumulation of fat through the regulation of scavenger receptor expression and controlling the expression of molecules involved in inflammation such as TNFα (Non-patent Document 5).

  Thus, adiponectin is closely related to so-called lifestyle-related diseases such as hypertension, obesity, hyperlipidemia, diabetes, and in particular, genetic and acquired deficiency of adiponectin is a major cause of lifestyle-related diseases in Japanese . Therefore, an increase in the blood level is desired in patients, and limiting intake of lipid is an effective method.

In order to increase the blood level of adiponectin, intake of fermented tea extract is effective as food and drink (Patent Document 1). However, limiting the intake of lipid as an energy source is not preferable in maintaining life activity. Furthermore, restricting intake of lipids causes vitamin B deficiency. In addition, absorption of fat-soluble vitamins in intestinal absorption occurs, and night blindness and bleeding tendency are shown due to lack of vitamin A and vitamin K. The nerve treetop is very rich in unsaturated fatty acids,
Vitamin E deficiency can cause neuropathy due to its lipid peroxides. And, fermented tea extract as a food and drink itself is not an energy source, and it is advantageous if the blood adiponectin concentration can be increased while ingesting lipid as an energy source.
JP 2004-315379 A "Biochem. Biophys. Res. Commun." 1999, 257, p.79-83 "Circulation" 1999, 100, p.2437-2476 Ichiro Shimomura: Adipocytokines and diabetes. Edited by Takashi Kadowaki. Diabetes mellitus. Tokyo: Nanzan Hall; 2004, p.71-80 "Nature Med." 2001, Volume 7, p.941-946 "J. Biol. Chem." 2003, 278, p.2461-2468

  The present invention uses docosahexaenoic acid or a derivative thereof as an active ingredient, which can maintain normal values of biological factors that are closely related to so-called lifestyle-related diseases such as hypertension, obesity, hyperlipidemia, and diabetes while usually ingesting lipids. An adiponectin elevating agent is provided.

  In order to solve the above problems, the present inventors have intensively studied a substance that increases the concentration of blood adiponectin while being a lipid, and as a result, found that docosahexaenoic acid or a derivative thereof increases the concentration of blood adiponectin. The invention has been completed.

That is, the present invention is as follows.
(1) An adiponectin increasing agent containing docosahexaenoic acid or a derivative thereof as an active ingredient.
(2) A food for increasing adiponectin comprising docosahexaenoic acid or a derivative thereof as an active ingredient.

  Examples of the derivative of docosahexaenoic acid include docosahexaenoic acid ethyl ester or docosahexaenoic acid-binding phospholipid.

  The adiponectin elevating agent of the present invention can elevate adiponectin in blood without interfering with lipid absorption. Furthermore, the adiponectin elevating agent of the present invention can lower molecular biological plasma parameters, specifically insulin, PAI-1, and CRP, which are indicators of deterioration of obesity and arteriosclerosis. The lipid profile, specifically triacylglycerol and cholesterol, can be lowered, so that biological factors closely related to so-called lifestyle-related diseases such as hypertension, obesity, hyperlipidemia, and diabetes can be maintained at normal levels. it can.

  The present invention is described in detail below.

  The present invention relates to an adiponectin elevating agent containing docosahexaenoic acid or a derivative thereof as an active ingredient, and a food that can increase adiponectin.

<Docosahexaenoic acid>
Docosahexaenoic acid (hereinafter also referred to as “DHA”) has a chemical formula of C ₂₂ H ₃₂ O ₂ , a molecular weight of 328.50, a carbon number of 22, having a cis double bond at positions 4, 7, 10, 13, 16, 19 and n -3 series linear unsaturated fatty acids. DHA is abundant in fish such as sardines and horse mackerel, but is localized in the brain and retina in vivo. DHA is an eicosapentaenoic acid (EPA) that is derived from α-linolenic acid (18: 3 n-3) by using two chain length extensions and two desaturation reactions. ), And further, it can be industrially synthesized via an enzyme system of chain length extension, desaturation, and chain length reduction. DHA has antithrombotic, anti-arteriosclerotic, anti-inflammatory and anti-carcinogenic effects, but according to the present invention, it has been shown to increase blood adiponectin levels.

  The DHA of the present invention may be a free fatty acid, or acts effectively even in the form of a salt such as sodium, potassium, calcium or magnesium.

<DHA derivative>
Derivatives of DHA include derivatives derived from substitution of the carboxyl group of DHA, for example, ester derivatives (acylated amino acids, acylated saccharides, alcohol esters, tocopherol esters, etc.), phospholipid derivatives, amide derivatives (for example, , Ceramide derivatives), salts (for example, alkali metal salts and alkaline earth metal salts, ammonium salts) and the like, but are not limited thereto. In the present invention, the derivatives of DHA are preferably ester derivatives and phospholipid derivatives.

  Examples of ester derivatives of DHA include, but are not limited to, methyl ester derivatives, ethyl ester derivatives, glycerol ester derivatives and the like.

  DHA glycerol esters include triglycerin-containing sardine oil, tuna oil, bonito oil and other fish oils, cod and squid liver oils, or synthetic triglycerides synthesized from DHA free fatty acids, and DHA ethyl esters. Or the synthetic triglyceride synthesize | combined from methyl ester is mentioned. DHA diglycerin or monoglycerin obtained by hydrolyzing or enzymatically degrading the triglyceride can also be used. The fatty acid, methyl ester and ethyl ester of DHA should be purified and concentrated using HPLC (high performance liquid chromatography) or a column after hydrolyzing the fish oil, liver oil, phospholipid, etc. it can.

  As phospholipid derivatives of DHA, DHA-linked phosphatidylcholine, DHA-bound phosphatidylethanolamine, DHA-bound phosphatidylserine, DHA-bound phosphatidylglycerol, in which DHA derived from nature is bound as an acyl group of phospholipid, Examples include DHA-bound inositol and DHA-bound plasmalogen. Furthermore, phospholipids synthesized using glycerophosphatidylcholine and free fatty acid or ethyl ester of DHA are used. In addition, lysophospholipid obtained by hydrolyzing the phospholipid obtained as described above with lipase or phospholipase can also be used.

  In addition, phosphatidylserine obtained by base exchange of DHA-bound phosphatidylcholine with phospholipase D or a phospholipid derivative having a primary hydroxyl group can be used. In addition, a derivative in which DHA is ester-bonded to a primary hydroxyl group of an antioxidant function substance typified by tocopherol or ascorbic acid can also be used.

  The structural formula of a general DHA-binding phospholipid is shown in the following formula (1).

(In the formula, R ¹ and R ² each may be the same or different and indicate a docosahexaenoic acid residue or a group selected from saturated or monoenoic fatty acid residue having a carbon number of 14 to 24,, R ¹ And at least one of R ² is a docosahexaenoic acid residue, and R ³ mainly represents a phosphorylcholine group or an ethanolamine group.
In the present invention, R ¹ and R ² are preferably all docosahexaenoic acid residues, but R ² is a docosahexaenoic acid residue, and R ¹ is a palmitic acid residue, an oleic acid residue or a stearic acid residue. The DHA-binding phospholipids can also be preferably used.

  DHA-binding phospholipids can be produced as synthetic compounds, but can also be extracted from natural products. For example, it can be produced by extracting from a large fish egg that can be obtained on an industrial scale, a salted salmon or a salmon egg, a salmon roe, etc.

The DHA content of the DHA-binding phospholipid in the present invention is preferably 20% or more, and more preferably 25% or more. For example, DHA-binding phospholipids obtained by extracting ethanol from these fish egg raw materials as a solvent contain 20 to 35% of DHA, and 70% or more of phospholipids are phosphatidylcholine.
<Adiponectin>
Adiponectin is a secreted protein that is specifically synthesized and secreted by adipocytes, and is abundant in serum, but becomes unregulated in various obese forms of humans and mice. Research is progressing as an important molecule that affects it. Adiponectin is also known as Acrp30 (adipocyte complement-related protein of 30kDa), apM-1, Glatin-binding protein, Adipose most abundunt gene transcript 1, etc., and the gene names are also known as APM1, ACRP30, GBP28. ing. Adiponectin suppresses diabetes and arteriosclerosis by improving the decrease in insulin sensitivity, which is the cause of type 2 diabetes. In addition to suppressing arteriosclerosis by direct action on the arterial wall, it also protects liver function To do.

  As described above, adiponectin is specifically expressed in adipose tissue, but its concentration in the blood is decreased when fat is accumulated, although it is secreted specifically by adipocytes, it increases when fat is reduced. To do. In particular, the increase in visceral fat results in a decrease in blood concentration, which is also low in coronary artery disease patients and diabetic patients. That is, it can be said that the accumulation of visceral fat is suppressed if the adiponectin concentration in the blood increases. In the present invention, DHA or a derivative thereof is preferable because it can increase the concentration of adiponectin in blood without hindering absorption of lipid as an energy source.

  The adiponectin concentration in blood can be measured by ELISA using a commercially available adiponectin ELISA kit.

<Adiponectin elevating agent>
The adiponectin elevating agent of the present invention contains DHA or a derivative thereof as an active ingredient as described above, and can be used to suppress the accumulation of built-in fat.

  The adiponectin elevating agent of the present invention is applied to a built-in fat, for example, fat attached to the mesentery, the greater omentum, around the kidney, or around the epididymis.

  Although DHA or a derivative thereof contained in the adiponectin-elevating agent of the present invention can be used alone, it may form a salt, and as a salt of a candidate substance, a physiologically acceptable acid (for example, organic acid or inorganic acid) Acid) and bases (for example, alkali metals) are used, and physiologically acceptable acid addition salts are particularly preferable. Examples of such salts include salts with inorganic acids (eg, hydrochloric acid, phosphoric acid, hydrobromic acid, sulfuric acid, etc.), or organic acids (eg, acetic acid, formic acid, propionic acid, fumaric acid, maleic acid, And salts with succinic acid, tartaric acid, citric acid, malic acid, succinic acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid, and the like.

  The adiponectin elevating agent of the present invention can be formulated according to a conventional method, and the formulation may be solid or liquid. For example, tablet, pill, granule, dragee, capsule, emulsion, liquid, gel, syrup , Slurries and suspensions, and may contain a pharmaceutically acceptable carrier. Such carriers may be additives, water, pharmaceutically acceptable organic solvents, collagen, polyvinyl alcohol, polyvinyl pyrrolidone, carboxyvinyl polymer, sodium carboxymethylcellulose, sodium polyacrylate, sodium alginate, water soluble Dextran, sodium carboxymethyl starch, pectin, methylcellulose, ethylcellulose, xanthan gum, gum arabic, casein, agar, polyethylene glycol, diglycerin, glycerin, propylene glycol, petrolatum, paraffin, stearyl alcohol, stearic acid, human serum albumin, mannitol, sorbitol , Lactose, surfactants acceptable as pharmaceutical additives, and the like.

  In the formulation, an excipient can be added. As the excipient, fillers, binders, coagulants, slipping agents, disintegrating agents, pigments, sweeteners, fragrances, coating agents and the like can be used alone or in combination depending on the purpose. Furthermore, DHA or a derivative thereof can be emulsified with an emulsifier and used.

  The adiponectin elevating agent of the present invention contains DHA or a derivative thereof as an active ingredient, but other ingredients may be used in combination. Components used in combination include EPA, linolenic acid and fats and oils containing them because DHA is an n-3 fatty acid, and phospholipids, fat-soluble vitamins, fat-soluble physiologically active substances because DHA is a lipid. Etc. Since DHA is susceptible to oxidation in vivo after ingestion, fat-soluble antioxidants, tocopherol, α-lipoic acid, CoQ10, carotenoids, lutein, astaxanthin, plasmalogen, sesamin and other water-soluble antioxidants, Examples include ascorbic acid, anthocyanidin, catechin, glutathione, and gallic acid. In this case, the DHA concentration is preferably 5.0% to 98% by weight.

  These components can be used alone or in appropriate combination from the above depending on the form of the adiponectin elevating agent of the present invention.

  The administration form of the adiponectin elevating agent of the present invention can be either oral or parenteral. In the case of oral administration, administration by capsules having an appropriate dosage form as described above is possible. In the case of parenteral administration, pulmonary dosage forms (for example, those using a nephriser etc.), nasal dosage forms, transdermal dosage forms (for example, ointments, creams), injection dosage forms and the like can be mentioned. In the case of the injection form, it can be administered directly or indirectly systemically or locally, for example, by intravenous injection such as infusion, intramuscular injection, intraperitoneal injection, subcutaneous injection or the like. The dose of the adiponectin elevating agent of the present invention varies depending on age, sex, symptom, administration route, administration frequency, and dosage form. The administration method is appropriately selected from the above methods depending on the age and symptoms of the patient.

  The content of DHA or a derivative thereof contained in the adiponectin elevating agent of the present invention can be appropriately selected according to the dosage form as described above, for example, using the value obtained in this example, It can also be set based on the appropriate human intake calculated from the following two general rules.

  General rule (1) corresponds to 1g per kg of animal body weight in the case of 1% dietary confirmation in an animal experiment, and the amount corresponds to 1g of the appropriate intake for humans. Here, the application amount of DHA or the like that can be included in the adiponectin elevating agent of the present invention is 1.6 g in humans.

  According to general rule (2), about 1/50 of the effective amount of 1 kg body weight in rats and mice is an appropriate dose in humans. When a 200g rat is fed with 1.6% (calculated as 20g / day of food intake), the equivalent dose is 1.86g when converted to a 60kg human. According to this general rule, the daily intake of DHA or the like contained in the adiponectin elevating agent of the present invention is 0.5 to 25 g, preferably 0.5 to 5 g, more preferably 1 to 2 g. Can be set to

  In the present invention, the adiponectin increasing effect of DHA or a derivative thereof can be confirmed using an obese model animal. For example, administration of DHA or the like to an obese model animal and comparison with an animal that received corn oil not containing DHA as a control group show that the DHA administration group of the present invention has an effect of increasing adiponectin. As the obesity model animal, for example, OLETF (Otsuka Long-Evans Tokushima Fatty) rat, which is a model animal that causes overeating by an appetite regulating hormone receptor mutation and develops obesity, hyperlipidemia, and diabetes, can be used.

  Ingestion of docosahexaenoic acid (DHA) and its derivatives can increase adiponectin in the blood without interfering with lipid absorption, so that the growth of animals ingesting the adiponectin elevating agent of the present invention is It is not inferior to animals that did not take the adiponectin elevating agent of the invention. Animal growth can be confirmed by measuring the initial body weight, final body weight, weight gain, food intake, food intake efficiency, and the like of the animal.

  In addition, the adiponectin elevating agent of the present invention includes insulin, PAI-1 (Plasminogen activator inhibitor type-1), and CRP (CRP), which are molecular biological plasma parameters that are indicators of deterioration of obesity and arteriosclerosis. -Reactive protein) and liver profile, specifically triacylglycerol and cholesterol. Insulin, PAI-1, and CRP are obtained by ELISA, liver triacylglycerol is obtained by Fletcher method, and liver cholesterol is obtained by cholesterol oxidase / DAOS method using commercially available kits. Can be confirmed. In particular, even when the adiponectin elevating agent of the present invention is ingested, no influence is observed on free fatty acid and blood glucose level in plasma.

Furthermore, the adiponectin elevating agent of the present invention is safe without burden on the liver. This is clear from the fact that GOT and LDH significantly decreased due to DHA intake and GPT and ALP tended to decrease when the liver disorder index enzyme activity in blood was measured.
<Food for adiponectin elevation>
The adiponectin elevating agent containing the DHA of the present invention or a derivative thereof as an active ingredient is not limited to a pharmaceutical product, and may be processed as a food for elevating adiponectin. There is no particular limitation on the form of the food, and in addition to general processed foods, health foods, functional foods, nutritional supplements, foods and drinks including beverages and foods, or additives thereof can be used. . Specifically, although it can mix | blend with a supplement and a soft drink, it is not specifically limited.

  In addition, as an aspect of the food of the present invention, the dry powder, extract or purified product of the DHA of the present invention or a derivative thereof is directly mixed with food, or a liquid, gel, powder or solid food, for example, , Beverages, Tea, Soup, Jelly, Yogurt, Ice Cream, Sorbet, Frozen Yogurt, Pudding, Dressing, Mayonnaise, Sprinkle, Miso, Soy Sauce, Grilled Meat Sauce, Noodles, Ham and Sausage, etc. , Jam, milk, cream, butter, cheese and other powdered, solid or liquid dairy products, margarine, bread, and confectionery raw materials. The food of the present invention covers a wide variety of forms and is not limited to the above examples, but from the standpoint of suppressing visceral fat accumulation, the form added to foods containing a large amount of fats and sugars, the nutrition Supplemental and health food forms are preferred. Further, since DHA is vulnerable to oxidation, a capsule form or the like is also preferable.

  In addition to DHA, the food of the present invention may contain other additives depending on the form of the food. Examples of such additives include excipients, extenders, binders, moisture-proofing agents, preservatives, strengthening agents, thickeners, emulsifiers, sweeteners, acidulants, food additives, seasonings and the like. . Examples of food additives include vitamins, minerals, chitin, chitosan, lecithin, royal jelly and the like. Seasonings include sweeteners such as granulated sugar, honey, and sorbit, acidulants such as alcohol, citric acid, malic acid, and aragonic acid, flavorings, and pigments. Can be used to adjust. Moreover, you may use together the well-known raw material used for slimming and a diet relevant to suppression of visceral fat accumulation. Specific examples include Garcinia cambodia skin extract, hydroxycitric acid and salts thereof, grape seed extract, fruit polyphenols such as apples, mountain extract fruit extract, guava leaf extract, gymnema sylvesta leaf extract, ginkgo biloba extract, lipase inhibitor , Α- and β-amylase inhibitors, L-carnitine and animal meat peptides containing the same, green algae extracts such as Aosa and Aonori, brown algae extracts such as kombu, chili powder and its extract, garlic extract, purslane, puer tea leaves Examples thereof include powders and extracts thereof, chuchu leaf powder and extracts thereof, oolong tea leaf powder and extracts thereof, psyllium seed coat, xanthine derivatives, citrus aurantium extract, senna leaf or stem extract, and peel. Moreover, although the foodstuff of this invention contains DHA or its derivative (s), other oil-based components may be contained. Components used in combination include EPA, linolenic acid and oils and fats containing them, phospholipids, fat-soluble vitamins, fat-soluble physiologically active substances, and the like. Examples include fat-soluble antioxidants, tocopherol, α-lipoic acid, CoQ10, carotenoids, lutein, astaxanthin, plasmalogen, sesamin, and other water-soluble antioxidants, ascorbic acid, anthocyanidins, catechins, glutathione, gallic acid, etc. . These additives and the like can be used alone or in combination as appropriate depending on the form of the food of the present invention.

  The food of the present invention can be produced by a method usually performed by those skilled in the art. For example, in order to obtain a powdered food, an excipient such as dextrin, cyclodextrin, starch, maltose, etc. is added to DHA as necessary, and powdered by a drying method such as freeze drying or spray drying. Can be obtained. In addition, dextrin, lactose, starch or processed materials thereof, excipients such as cellulose powder, vitamins, minerals, fats and oils of animals and plants and seafood, proteins, sugars, pigments, flavors, other edible additives as necessary At the same time, it is processed into powders, granules, pellets, pills, tablets, etc. by methods usually performed by those skilled in the art, coated with gelatin, etc., formed into capsules such as hard capsules, soft capsules, or drinks, It can be used as a dietary supplement or health food. A beverage in which DHA is encapsulated in a water-insoluble capsule can be mixed with the beverage. Such a form is also preferable in terms of preventing air oxidation of DHA.

  In the food of the present invention, the amount of DHA or a derivative thereof can be appropriately set according to the type, form, purpose of use of the food, the type of food of the present invention, and the intake mode.

  You may set based on the human proper intake estimated from the present Example calculated from the above two general rules.

Hereinafter, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to these examples. The data obtained by the following experiment was subjected to a significant difference test using Student's t-Test.

Effects of DHA ethyl ester on body weight, food intake, food efficiency and organ weight The diet composition and breeding method were as follows. OLETF rats are obese model rats with chronic illness and were obtained from Hokudo Co., Ltd. 4-week-old male OLETF rats were divided into 2 groups (6 animals per group), and the control group (Con group) was fed with food according to the AIN-76 composition. The other group was DHA group, DHA ethyl ester. A diet supplemented with 1.6% (purity 97%) was fed. The diet composition is shown in Table 1.

  The above two groups were raised for 2 weeks, fasted for 9 hours, and sacrificed by abdominal aorta blood collection under ether anesthesia, and liver and adipose tissue were removed to obtain plasma from blood. The growth state is shown in Table 2.

  As a result, there was no difference between the groups in the initial body weight, the final body weight, the weight gain, the amount of food intake, and the food intake efficiency in the 2-week rearing (Table 2).

Regarding organ weights, there was a decrease in adipose tissue weight due to DHA feeding.

Measurement of plasma parameters Plasma lipid concentrations were measured by enzymatic methods. Plasma triglyceride concentration is triglyceride E-test Wako (GPO / DAOS method, Wako Pure Chemical), plasma phospholipid concentration is phospholipid E-test Wako (choline oxidase / DAOS method, Wako Pure Chemical), plasma cholesterol concentration Cholesterol CE-Test Wako (cholesterol oxidase / DAOS method, Wako Pure Chemical Industries), plasma free fatty acid concentration was measured using NEFA C-Test Wako (ACS / ACOD method, Wako Pure Chemical Industries).

  The blood glucose level was measured by the mutarotase / GOD method using glucose CII-Test Wako (Wako Pure Chemical Industries).

  Liver dysfunction markers include glutamate oxaloacetate aminotransferase (GOT) and glutamate pyruvate aminotransferase (GPT) (transaminase CII-Test Wako, POP / TOOS method, Wako Pure Chemical Industries), lactate dehydrogenase ( Lactate dehydrogenase CII-Test Wako, lactate substrate / tetrazolium salt method, Wako Pure Chemical), alkaline phosphatase (Alkaline Pospha K-Test Wako, phenyl phosphate substrate method, Wako Pure Chemical) were measured.

  As a result, the plasma triglyceride, cholesterol and phospholipid concentrations were significantly reduced by DHA intake (Table 3).

  No effects of DHA intake were observed on plasma free fatty acids and blood glucose levels. Significant decreases in liver injury index enzyme activity in blood were observed with GOT and LDH due to DHA intake (FIG. 1), and GPT and ALP also tended to decrease with DHA intake.

  Moreover, it was shown that the plasma adiponectin concentration increases significantly by DHA intake (FIG. 2).

  Next, plasma insulin (Levis / Rat Insulin Kit, Shiba Goat), leptin (Rat Leptin ELISA kit, Yauchihara Laboratory), adiponectin (mouse / rat adiponectin ELISA kit), PAI-1 (IMUCLONE RatPAI-1 ELISA kit, American Diagnostica Inc.) and CRP (Rat CRP ELISA kit, Alpha Diagnostic Inc.) were measured by ELISA.

  The results are shown in Table 4.

Insulin, PAI-1 and CRP concentrations also showed a downward trend due to DHA intake. There was no difference in leptin concentration between groups. Further, in the composition of the present invention, the adiponectin elevating agent decreases molecular biological plasma parameters that are indicators of deterioration of obesity and arteriosclerosis, specifically, insulin, PAI-1, DRP, It can be seen that it lowers the liver lipid profile, specifically triacylglycerol, cholesterol.

Measurement of liver lipid concentration Liver total lipid was extracted and concentrated by the method of Folch et al., Liver TG concentration was quantified by the Fletcher method, and liver phospholipid concentration was quantified by the method of Bartlett et al. Liver total cholesterol concentration was measured by cholesterol oxidase DAOS method using cholesterol E-test Wako (Wako Pure Chemical Industries).

  As a result, hepatic triglyceride concentration was significantly reduced by DHA intake (Table 5).

In addition, there was no effect of DHA intake on cholesterol and phospholipid concentrations.

Analysis of Fatty Acid Composition Plasma lipids were extracted by Bligh and Dyer method and then fractionated by thin layer chromatography. Analysis of the fatty acid composition was performed by gas chromatography.

  As a result, it was shown that fatty acid Δ6 desaturation in plasma and liver was significantly suppressed by DHA intake (Tables 6 and 7). Therefore, it has been shown that plasma and liver fatty acid Δ6 desaturation can suppress, for example, excessive production of arachidonic acid, which is a problem in diabetic patients.

Effects of DHA-bound phospholipid on body weight, food intake, food efficiency and organ weight The diet composition and breeding method were as follows.
4-week-old male OLETF rats are preliminarily raised for 1 week, divided into 2 groups (6 animals per group), and as a control group (Con group), a diet according to the AIN-76 composition is taken from 5 weeks of age. One group was taken as a DHA group and fed with 2% DHA-binding phospholipid. The diet composition is shown in Table 8.

  The DHA-binding phospholipid used this time was added dropwise to 1 L of acetone cooled at −80 ° C. by adding 100 g of DHA-binding phospholipid (Nihon Yushi Co., Ltd. Sanomega PC-DHA-N) extracted from salmon roe. The phospholipid fraction precipitated as a component was filtered and the solvent was distilled off to re-extract. The composition is shown in Table 9 and Table 10.

  For the lipid composition in Table 9, lipids were spotted on thin layer chromatography, and the developing solvent was separated with chloroform-methanol-water (65: 25: 4 (v / v)). After completion, detection was performed with 50% sulfuric acid, and quantification was performed using a densitometer signal ratio. In addition, the fatty acid composition analysis of Table 10 was performed by gas chromatography.

  The above two groups were bred for 4 weeks. After fasting for 9 hours, they were sacrificed by blood sampling of the abdominal aorta under ether anesthesia, and the liver and adipose tissue were removed, and serum was obtained from the blood. Table 11 shows the growth state of the rats.

As a result, there was no difference between the groups in the initial body weight, the final body weight, the weight gain, the amount of food intake, and the food intake efficiency in the 4-week rearing (Table 11).

Measurement of serum parameters Serum lipid concentration was measured by enzymatic method. That is, serum triglyceride concentration is triglyceride E-test Wako (GPO / DAOS method, Wako Pure Chemical), serum phospholipid concentration is phospholipid E-test Wako (choline oxidase / DAOS method, Wako Pure Chemical), serum cholesterol concentration Cholesterol CE-Test Wako (cholesterol oxidase / DAOS method, Wako Pure Chemical Industries), serum free fatty acid concentration was measured using NEFA C-Test Wako (ACS / ACOD method, Wako Pure Chemical Industries).

  The blood glucose level was measured by the mutarotase / GOD method using glucose CII-Test Wako (Wako Pure Chemical Industries).

  As liver dysfunction markers, glutamate oxaloacetate aminotransferase (GOT) and glutamate pyruvate aminotransferase (GPT) (transaminase CII-Test Wako, POP / TOOS method, Wako Pure Chemical Industries) were measured.

  As a result, serum cholesterol and phospholipid concentrations were significantly decreased by DHA-bound phospholipid intake (Table 12).

  Next, serum insulin (Levis / Rat Insulin Kit, Shibayagi), leptin (Rat Leptin ELISA kit, Yauchihara Laboratory), and adiponectin (mouse / rat adiponectin ELISA kit) were measured by ELISA.

  The results are shown in Table 13.

It was also shown that the serum adiponectin concentration increased significantly with DHA intake (Table 13, FIG. 3).

Measurement of liver lipid concentration Liver total lipid was extracted and concentrated by the method of Folch et al., Liver TG concentration was quantified by the Fletcher method, and liver phospholipid concentration was quantified by the method of Bartlett et al. Liver total cholesterol concentration was measured by cholesterol oxidase DAOS method using cholesterol E-test Wako (Wako Pure Chemical Industries).

  As a result, the liver triglyceride concentration was significantly reduced by DHA-linked phospholipid (Table 14).

It is a figure which shows that the raise of the liver damage marker enzyme activity in the blood was improved by ingestion of the DHA containing diet for 2 weeks. It is a figure which shows that the blood adiponectin density | concentration rises by ingesting a DHA containing meal for 2 weeks. It is a figure which shows that the blood adiponectin density | concentration raises by ingesting the DHA binding type phospholipid containing diet for 4 weeks.

Claims (4)

  An adiponectin elevating agent comprising docosahexaenoic acid or a derivative thereof as an active ingredient.   A food for increasing adiponectin, comprising docosahexaenoic acid or a derivative thereof as an active ingredient.   The adiponectin elevating agent according to claim 1, wherein the derivative of docosahexaenoic acid is docosahexaenoic acid ethyl ester or docosahexaenoic acid-binding phospholipid. The food according to claim 2, wherein the derivative of docosahexaenoic acid is docosahexaenoic acid ethyl ester or docosahexaenoic acid-binding phospholipid.

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