KR20120016960A - Composition comprising laminaria japonica for preventing and treating obesity or hyperlipidemia and atherosclerotic-vascular diseases - Google Patents

Abstract

PURPOSE: A composition containing Laminaria japonica is provided to reduce neutral lipid in feces and neutral fat, cholesterol, phospholipid, and hydroxyl radical in serum. CONSTITUTION: A pharmaceutical compsotion for preventing and treating obesity or hyperlipidemia or arteriosclerotic vascular diseases contains Laminaria japonica as an active ingredient. The Laminaria japonica is used in the form of dry powder, polar solvent soluble extract, or polar solvent insoluble extract.

Description

Composition comprising Laminaria Japonica for preventing and treating obesity or hyperlipidemia and atherosclerotic-vascular diseases}

The present invention relates to a composition for the prevention and treatment of obesity or hyperlipidemia and atherosclerotic vascular disease containing kelp as an active ingredient.

[Reference 1] Hill JO, Lin D, Yakubu F and Peter JC. (1992): Development of dietary obesity in rats, influence of amount and composition dietary fat. Int J obes realt Metab Disore. 16: 321-333

Lee H. K., Recent Progress in Obesity Research (1990): Diseases associated with obesity, Koran J. Nutr. 23 (5), 341-346

Document 3 Bidlack WR and Tappel AL. (1973): Damage to microsomal membrane by lipid peroxidation. Lipids. 8: 177-178

[4] Valko M, Leibfritz D, Monocol J, Cronin MT, Mazur M and Telser J. (2007): Free radicals and antioxiidants in normal physiological functions and human disease. Int. J biochem Ceoo biol. 39: 44-84

[Reference 5] National Statistical Office Republic of Korea: Annual Report on the Cause of Death Statistics 2008 (2010)

[Ref. 6] Ministry of Health & Welfare (2010): 2008 National Health and Nutrition Survey Results Press Release, December 8, 2009 (Morning)

[Reference 7] Kim EM, Na MY, Park MS and Baik HJ (2000): Manual of medical nutrition therapy Vol. 2, Korean Dietetic Ass. pp.159

Connor WE, Stone DB, Hodges RE (1994): The interrelated effects of dietary cholesterol and fat upon human serum lipid level. J of Clin Invest 43: 1691-1696

Lee KJ (2002): Effects of Diet in Sea Tangle from East Coast on Serum Composition and Calcium Absorption in Rats, MS thesis, Graduate School of Kangnung National University

10. Ministry of Maritime Affairs and Fisheries (1999): Statistical year book of maritime affairs and fisheries. pp.1092-1093

[11] Lahaye M. (1991): Marine algae as sources of fibers: Determination of soluble and insoluble dietary fiber concentrates in some sea vegetable. J Sci Food Agric 54: 587-594

Nishino T, Aizu Y, Nagumo T. (1991): The relationship between the molecular weight and the anticoagulant and activity of two types of fucan sulfates from the brown scawced. Ecklonia kurom, Agric Biol Chem., 55: 791-797

13 Hsueh WA. and Anderson PW. (1992): Hypertension, the endothelial cell, and the vascular complications of diabetic mellitus. Hypertension, 20: 253-263

Anderson JW, Gustafson NJ (1988): Hypocholesterolemic effects of oat and bean products, Am J Clin Nutr 48: 749-753

[15] Hopewell R, Yeater R (1993): Ullrich I, Soluble fiber: Effect on carbohydrate and lipid metabolism, Prog Food Nutr Sci 17 (2), 159-182

Nishide E, Anzai H, Uchida N (1993): Effect of alginates on the ingestion and excretion of cholesterol in the rat, J Applied Phycology, 5, 207-211.

Schneeman BO, Tietyen J (1994): Dietary Fiber, In: Shills ME, Olson JA, Shike M, eds, Modern Nutrition in Health and Disease, Lea & Febiger, 1, 89-100.

Kang MS and Kang JS (2001): Hypocholesterolemic Effect of Tangerine Pulp, Sea Tangle of Prickly Pear Cactus on Lipid Level, Intestinal Cholesterol Absorption, Platelet Aggregation and Liver Tissue in Hypercholesterolemic Rats, Korean J Nutrition, 34 (2) , 141-149

[19] Yim MJ (2007): Antiatherosclerosis Animal study and the ket Funtional Compound Identification of the Alginate extracted Residue of Sea Tangle, MS thesis, Graduate School of Kangnung National University

20, Choi JH, Kim DI, Park SH, Kim DW and Koo JG (1999): Effects of Sea Tangle (Laminaria japonica) and Fucoidan Components on the Attack of Oxygen Radicals in Kidney, J. Korean Fish. Soc, 32 (6): 758-763

[21] Choi JH, Kim DI, Park SH, Kim DW and Lee JS (1999): Effects of Sea Tangle (Laminaria japonica) and Fucoidan Components on Chronic Degenerative Diseases, Korean J. Life Science, 9 (4), 430 -438

22. Kang JY (2008): Anti-inflammatory Activities of Undaria pinnatifida and Laminaria japonica (Phaeophyta), MS thesis, Graduate School of Pukyong National University

23. Tiidus PM. et al. (1995): Sports Med, 20, p. 12-13, Britton, C., et al. (1979): Hydroperoxide metablism in mammalian organ, Physiol. Rev., 59, pp. 527-605, Ishinaga M, et al. (1994): daily intake of fatty acids, sterols, and phospholipids by Japanese women and serum cholesterol, J. Nutr. Sci. Vitaminol, 40, pp. 557-567

24. Kumar CT., Et al. (1992): Mol. Cell Biochem., 111, pp. 109-115

25 Branen., AL, J. Am. (1975): Oil. Chem. Soc., 55, pp. 123-126 Ito N., et al. (1983): J. Natl. Cancer Inst., 70, pp. 343-349

[26] Lim DK., Et al. (1996): Korean J. Food Sci. Technol., 28, pp. 83-89, Kim HK., Et al. (1995): Korean J. Food Sci. Technol., 27, pp. 80-85 Cha JY. (1999), J. Korean Soc, Food Nutr., 28, pp. 1131-1136, Kim MH., Et al. (1999): Korean J. Food Sci. Technol., 31, pp. 273-279

[27] Korea Fisheries Association (1996): Korean Fisheries Yearbook, 540-541

[Reference 28] Kim, Jong-Gun (1993): A Study on the Extraction Method of Seaweed for Development of Seaweed Natural Seasoning, 韓國 食 文化 硏 究 論, 621-651

29 W Kim and Choi HS (1994): Development of Combined Methods for Effective Extraction of Sea Mustard, Korean J. Food Sci. Technol., 26 (1), 44-50

30. Kim WJ, Lee JK and Chang YS (1994): Development of Combined Method for Extraction of Sea Tangle, Korean J. Food Sci. Technol, 26 (1), 51-56

Lee JK, Lee SR and Kim WJ (1994): Effect of Removal of Viscous Materials on Physicochemical Properties of Sea Tangle Extract, Korean J. Food Sci. Technol, 26 (2), 127-132

32 Folch J., Lees M., Sloane Stanley GH. (1957): A simple method for the isolation and purification of total lipids from animal tissues, J. Biol. Chem., 226, 497-509

33 Fringe CS and Dunn RM (1980). : The Colormetric method for determination of serum total lipids based on the sulfo-phosphanilli reaction an J.clin pathol 52, 82-92

34. W. Richmond (1976). : Use of cholesterol oxidase for assay of total and free cholesterol in serum by continuous flow analysis. Clin. Chem, 22, 1579

35. M.W. Mc Gowan, J.D. Artiss, D. R. Stramdbergh. (1983): Aperoxidase-coupled method for the colorimetric determination of serum triglycerides. Clin. Chem. 29, 538

36. P. S. Chen, T. Y. Toribara, H. Warnerm (1956): Microdetermination of phosphorus. Anal. Chem. 1756-1760

37. A. Noma, S. Matsushita, T. Komori (1986): High-density lipoprotein cholesterol levels of very old people in the diagnosis of dementia. Oxford Journals. 15. 267-270

38. Friedwald WT, Levy RL, Fedreicson DS (1972): Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without was of the preparative ultracentrifuge, Clin. Chem., 18, 499-506

39. K. Yagi (1987): Lipid peroxides and human diseases. Chemistry and Physics of Lipids. 45. 337-342

[40] Kobatake Y, Saito M, Kuroda K, Kobayashi S, Innami S. (1987): Influence of fish consumption on serum lipid and lipid peroxide concentrations in middle aged subjects, J Japan Soc Nutr & Food Sci 40, 103- 110

Oyanagui, Y. (1984): Reevaluation of assay methods and establishment of kit for superoxide dismutase activity. Anal, Biochem, 42, 290-296

42 Lowry O. H., Rosebrough. N. J., Farr, A. L., and Rardall, R. J. (1951): J. Biol. Chem., 193, 265

43. Valery M. Dembitsky, Olga A. Rozentsvet, Elena E. Pechenkina. (1990): Glycolipids, phospholipids and fatty acids of brown algae species. Phytochemistry, Volume 29, Issue 11, Pages 3417-3421

Modern people are consuming excessive amounts of saturated fat and cholesterol due to the demands of the western diet, lifestyle changes and simplicity, which lead to cardiovascular diseases such as obesity, hyperlipidemia, arteriosclerosis and myocardial infarction, and lipid peroxides. It is known to cause degenerative processes in vivo, promote cancer and aging, and change or destroy biological membranes (1-3). Blood cholesterol has been shown to be a risk factor for ischemic cerebrovascular disease and coronary artery disease, and free radicals have been shown to be a cause of chronic diseases such as aging, cerebrovascular disease, cardiovascular disease and cancer (4).

The three major causes of death in Korea in 2008 were malignant neoplasms (ceramic), cerebrovascular disease, and heart disease (including ischemic heart disease and other heart diseases), accounting for 48.1% of all deaths. Although the decrease in heart disease (including ischemic heart disease and other heart diseases) was increased, the statistical results show that (5). In addition, the Ministry of Health and Welfare announced the Comprehensive National Health Promotion Plan of the Health Plan 2010 in the first plan ('02 ~ '05) in 2002, and presented the overall goal of extending the life expectancy and securing health equity as the overall goal in 2010. The second plan ('06 ~ '10) has been established and implemented with the goal of Diseases are closely related to dietary changes.The results of the 2008 National Health and Nutrition Survey indicate that chronic diseases are obesity (over 19 years old), 30.7%, high blood pressure (over 30 years), 26.8%, diabetes (over 30 years), 9.7%, high cholesterol The prevalence rate was 10.9% (over 30 years). Hypertension and diabetes mellitus have decreased in recent decades, but obesity and hypercholesterolemia have increased. The percentage of energy and fat-intake consumers (1 year old and over) was 4.9%, down from 2005 but increasing from 2007. Hypertriglyceridemia was 17.3% ('07), an increase of 6.3% over 10 years. In 2005, animal foods accounted for 21.6% and energy intakes exceeded 20%, the highest since the National Nutrition Survey began in 1969 (6).

Hyperlipidemia refers to an abnormally increased state of blood lipids (triglycerides and cholesterol), and an increase in blood cholesterol or triglycerides is a risk factor for atherosclerosis (7). Many risk factors such as age increase, sex, obesity, poor eating habits, blood pressure, blood sugar, stress, smoking, and the environment have been reported as causes of hyperlipidemia, and are known as the three major risk factors for coronary artery disease along with hypertension and smoking. 8).

Seaweed has long been used as a source of nutrients, seasonings, minerals and dietary fiber for pregnant women in Korea and Japan, but in recent years, Korea's seaweed industry has suffered from slow exports to Japan, domestic consumption limitations, changes in dietary habits, and various competitions. The company is facing limitations due to the expansion of counterpart food supply. The main components of seaweed are carbohydrates and minerals. Especially, most of carbohydrates have been recognized for their value as non-digestible dietary fiber. Brown seaweed kelp is a perennial seaweed, and its general lifespan is 2 ~ 3 years (9), and it is the second most produced seaweed (우), seaweed and seaweed (10). Kelp contains about 32-75% of dietary fiber, 51-85% of which is water-soluble dietary fiber. Since water-soluble dietary fiber has a higher viscosity than insoluble dietary fiber, it forms a gel with high viscosity. Therefore, water-soluble dietary fiber can reduce glucose absorption and increase glucose tolerance of diabetics by prolonging the residence time of gastric contents and inhibiting absorption. There is a report (11). Especially Brown algae are rich in fiber, minerals, and proteins, and are rich in water-soluble polysaccharides such as alginic acid, fucan, and laminaran (12). These water-soluble polysaccharides are not toxic even after high intake, and fucan and alginic acid derivatives are known to have antidiabetic, anticoagulant, anti-inflammatory, antiviral and anti-tumor effects (13). Kelp is a water-soluble, viscous dietary fiber, such as alginic acid, pectin, absorbs or binds cholesterol, and is reported to lower LDL-cholesterol (14-17). A common mechanism of action is the inhibition of absorption of dietary cholesterol or bile acids (18). Kelp, which is rich in alginic acid, has higher iodine content and higher magnesium and iron content than other seaweeds. Alginate is a major component of kelp viscous, and it is known to show lowering cholesterol in blood, extracorporeal discharge of heavy metals, lowering blood pressure, and anticancer function in addition to general functions of dietary fiber such as intestinal action. (19). Alginate has been mainly studied as a biologically active ingredient such as seaweed and kelp, but the viscosity is very large and has various problems in beverage development (20). Recently, fucoidan, which has been spotlighted as a new bioactive ingredient of brown algae, has been found to have a very high potential (21). Fucoidan is an acidic polysaccharide containing a sugar called fucose in its molecular structure. It is known that Fucoidan has excellent blood flow improvement functions such as anti-cancer, anti-cholesterol, blood coagulation inhibition, blood pressure control, and lipid metabolism improvement functions. Is said to contain more than 5 g of fucoidan. In addition, there is a small amount of the amino acid laminine in kelp has a blood pressure lowering function (19).

In addition, according to the research results (22) that seaweed has an antioxidant effect, we are trying to find a drug that has an antioxidant effect. Antioxidation refers to the prevention of various oxidation reactions in the body. Lipids present as biofilms or lipoproteins are attacked by free radicals in the body to form various kinds of peroxides. Decomposition products are highly reactive and convert and increase the structure and function of surrounding biomolecules, leading to aging and various chronic diseases. In vivo, there are several antioxidant defenses that can neutralize these free radicals and protect the body. Human diseases and aging occur during metabolism in the body: O2- (superoxide), NO (nitric oxide), NO2 (nitrogen dioxide), OH (hydroxyl), ROO (proxyl), RO (alkoxyl), HO2 (hydroperoxyl) radicals It is due to oxidation such as radical and there are free radicals which are such harmful radicals in vivo. As a means of defending the living body, superoxide dismutase (SOD), an antioxidant enzyme in liver and red blood cells, is converted to H ₂ O ₂ by reducing superoxide radicals. H ₂ O ₂ is an antioxidant enzyme such as catalase (CAT; catalase), glutathione peroxidase (GSH-px; glutathione-peroxidase), and antioxidants such as vitamin E and glutathione to remove free radicals. Protect against damage (23). However, when these antioxidant defensive free radicals are too low to control production, oxidative stress and damage of tissues are promoted. Excess free radicals and lipid peroxides produced in the body are injured by oxidative stress such as protein oxidation and DNA damage. Increase (24). Therefore, many synthetic or natural antioxidants have been developed for the reduction of oxidative stress, but synthetic antioxidants have a problem in that they are vulnerable to safety (25). Using carotenoids, flavonoids, and polyphenols, which are antioxidant substances present in plant extracts, efforts have been made to develop safer and more natural antioxidants (26).

Currently, the processing methods using these seaweeds were mainly salted products and dried products (27). Alginate in seaweeds was extracted and beverages were made, or seaweed extracts were processed and distributed in extract form, and effective methods for seaweed extraction Much has been studied (28). However, when industrially processed in the form of extract using kelp, the extraction yield of the solid content concentration is about 7-8%, which is rather low, and the extraction residue containing a large amount of soluble components is discarded as it is (29-31). The health function of kelp is known to be fucosterol extracted from ethanol (19). The purpose of this study was to verify the health functionalities such as anti-arteriosclerosis, anti-hyperlipidemia, and antioxidant effects.

Therefore, the present inventors have studied the composition of the prevention and treatment of anti-obesity, hyperlipidemia and atherosclerotic vascular disease of kelp, and caused obesity and dietary hyperlipidemia by raising a sample prepared with the addition of an obesity inducing ingredient (beef tallow) It was confirmed that the triglyceride and cholesterol content in the kelp group of the animal model was decreased, and the lipid content in the liver tissue and feces was reduced, and triglyceride, cholesterol, and phospholipid in the animal model of hyperlipidemia induced by high cholesterol diet. To reduce the lipid peroxide and Hydroxyl radical content, and enhance the SOD activity in the serum, the present invention was confirmed to be useful for the prevention and treatment of obesity or hyperlipidemia and atherosclerotic vascular disease.

In order to accomplish the above object, the present invention provides a pharmaceutical composition for the prevention and treatment of obesity or hyperlipidemia and atherosclerotic vascular disease containing kelp as an active ingredient.

In addition, the present invention provides a health functional food for the prevention and improvement of obesity or hyperlipidemia and atherosclerotic vascular disease containing kelp as an active ingredient.

The kelp as defined herein includes a dry powder, a polar solvent soluble extract, or a polar solvent insoluble extract form.

The (1) kelp dry powder as defined herein includes a kelp dry powder (hereinafter referred to as “LJ-B”) dried and powdered by conventional drying methods such as sunlight drying, natural drying, forced drying, and the like; (2) The kelp polar solvent soluble extract is a kelp extracted from the dry powder with a lower alcohol such as water, methanol, ethanol, butanol and the like, preferably a polar solvent such as a mixed solvent of water and ethanol. Polar solvent soluble extract (hereinafter referred to as “LJ-1”); And (3) kelp polar solvent insoluble extract (hereinafter referred to as “LJ-P”), which is an insoluble residue in the polar solvent, and these extracts are characterized by containing a large amount of kelp polysaccharides such as alginic acid.

Hyperlipidemia and atherosclerotic vascular diseases as defined herein are specifically hyperlipidemia, arteriosclerosis, heart failure, hypertensive heart disease, arrhythmia, congenital heart disease, myocardial infarction, angina pectoris, stroke or peripheral vascular disease, preferably hyperlipidemia or arteriosclerosis to be.

Hereinafter, a method for obtaining a kelp polar solvent soluble extract and a kelp polar solvent insoluble extract of the present invention will be described in detail.

For example, after finely powdering dried kelp with a grinder, the kelp powder is placed in an Erlenmeyer flask, and then a lower alcohol such as water, methanol, ethanol, butanol and the like, and a mixture of water and ethanol, preferably More preferably, 80 to 100% ethanol is added at a ratio of 1: 15 (w / v), preferably 1: 10 (w / v) and mixed, and then 1 to 50 ° C, preferably 5 ° C. A first step of extracting by ultrasonic extraction method, reflux extraction method, hot water extraction method, cooling extraction method or supercritical extraction method, preferably ultrasonic extraction method for 10 minutes to 50 minutes, preferably 20 minutes to 40 minutes at 45 ℃; The extract was filtered with a filter paper, and concentrated and lyophilized at 5 ° C. to 45 ° C. with a rotary vacuum concentrator, followed by preparation of a second step of storing at −5 ° C. to −60 ° C., preferably −10 ° C. to −50 ° C. The kelp polar solvent soluble extract of the present invention is obtained through the process, and the remaining residue is dried to obtain a kelp polar solvent insoluble extract.

In the first step, the kelp is natural or cultured acid, preferably Laminaria Japonica.

The pharmaceutical composition of the present invention comprises 0.1 to 50% by weight of the kelp relative to the total weight of the composition.

However, the composition is not limited thereto, and may vary depending on the condition of the patient, the type of disease, and the progress of the disease.

Since the kelp itself of the present invention has little toxicity and side effects, it is a drug that can be used safely even when taken for long periods of time.

The compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions.

The composition of the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions, Examples of carriers, excipients and diluents that can be included in the composition containing the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid preparations contain at least one excipient such as starch, calcium carbonate and sucrose in the powder. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The preferred dosage of the composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. However, for the desired effect, the kelp of the present invention is preferably administered at 0.5 g / kg to 5 g / kg, preferably at 1 g / kg to 3 g / kg. The administration may be carried out once a day or divided into several doses. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.

The composition of the present invention can be administered to mammals such as rats, mice, livestock, humans, etc. by various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.

In addition, the present invention provides a health functional food for the prevention and improvement of obesity or hyperlipidemia and atherosclerotic vascular disease containing kelp as an active ingredient.

The composition comprising the kelp of the present invention can be used in various ways, such as drugs, foods and beverages for the prevention and improvement of obesity or hyperlipidemia and atherosclerotic vascular disease. Foods to which the kelp of the present invention may be added include, for example, various foods, beverages, gums, teas, vitamin complexes, health supplements, and the like, and may be used in the form of powders, granules, tablets, capsules, or beverages. have.

Kelp of the present invention may be added to food or beverage for the purpose of preventing and improving obesity or hyperlipidemia and atherosclerotic vascular disease. At this time, the amount of the kelp in the food or beverage is generally added to the health food composition of the present invention 1 to 5% by weight of the total food weight, the health beverage composition is 0.02 to 10 g, preferably based on 100 ml Can be added in a ratio of 0.3 to 1 g.

In addition to containing the kelp as an essential ingredient in the indicated ratio, the health beverage composition of the present invention is not particularly limited in the liquid component and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of the above-mentioned natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose such as maltose, sucrose and the like and polysaccharides such as dextrin, cyclodextrin and the like Sugar, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of said natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 mL of the composition of the present invention.

In addition to the above-mentioned composition, the composition of the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and intermediates (cheese, chocolate etc.), pectic acid and its salts, Salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the compositions of the present invention may contain flesh for the production of natural fruit juices and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

The kelp of the present invention reduces lipid content in liver tissues and feces in animal models causing hyperlipidemia, decreases triglycerides, cholesterol, phospholipids, lipid peroxide and hydroxyl radicals in serum, and enhances SOD activity in serum. It can be used in pharmaceutical compositions and health functional foods useful for the prevention and treatment of obesity or hyperlipidemia and atherosclerotic vascular disease.

Hereinafter will be described in detail by the reference examples, examples and experimental examples of the present invention.

However, the following Reference Examples, Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following Reference Examples, Examples and Experimental Examples.

Example  1. Extracts Extraction residue  Produce

The dried kelp (Laminaria Japonica) purchased on the market is finely powdered with a general domestic grinding machine (Hanil, Korea) to obtain a kelp powder (hereinafter referred to as “LJ-B”). 100 g of the obtained kelp powder was placed in a 1000 mL Erlenmeyer flask, followed by adding 95% ethanol at a ratio of 1: 5 (w / v) and mixing, followed by ultrasonic extraction at 30 ° C. for 30 minutes (sonication; power sonic 520, Hwashin, Korea). Whatman No. 2, filtered through filter paper, concentrated and freeze-dried at 40 ℃ with a rotary depressurizer (Rotavator; R-200, Buchi, switzerland) and stored at -40 ℃ kelp ethanol extract of the present invention (hereinafter referred to as "LJ-1" 3.1 g of the obtained residue was concentrated under reduced pressure and dried to obtain 95 g of ethanol extract residue (hereinafter referred to as “LJ-P”) under the reduced pressure, and was used as the following animal test sample.

Reference Example  1. Preparation for experiment

1-1. Experimental Animal and Breeding Method

After purchasing from SPF male 140 ± 10g Sprague Dawley (SD) rats (Hyochang Science (Daegu) Co., Ltd.) used for this test, only healthy animals were used for the experiment through quarantine, purification and breeding for 1 week. The breeding environment of this study was conducted in the laboratory animal breeding room (TECNIPLAST, Italy) of Kyungsung University pharmacy set at a temperature of 22 ± 3 ℃, a relative humidity of 50 ± 10% and an illumination time of 12 hours (07: 00 ~ 19: 00). The diet was fed a solid feed for experimental animals (Central Experimental Animal, Seoul), and drinking water was freely ingested. Only water was fasted for 24 hours before the experiment time to verify the animal dietary effect. At this time, the experimental animals were treated within a certain time (10:00 AM ~ 12:00 AM) in consideration of the daily variation of enzyme activity.

1-2. Kelp Experiment Feed Animal diet  Dosing method

As animal testing material Raw materials of kelp grind, kelp ethanol extract and kelp ethanol extract residue were used, and 4% tween 80 diluted with physiological saline was administered to the experimental animals. The dose and duration of administration were determined once a day using oral needle zonde in each experimental group for 6 weeks after each sample was set to 200 mg / kg body weight, which is the dose that is considered to be effective for this experiment. Administered at time.

1-3. Statistical analysis

The results obtained in this experiment were expressed as mean ± standard deviation, and statistical significance was verified by Duncan's multiple range test.

Experimental Example  1. Hyperlipidemia experiment

1-1. Induction of hyperlipidemia

The composition of the basic diet and hyperlipidemia of experimental animals is shown in Table 1. The hyperlipidemic diet caused hyperlipidemia by raising the sample prepared by reducing the amount of sucrose by 15.5% and corn oil 5% by adding 20.5% beef tallow to the basic diet for 4 weeks (see Table 1). .

Foundation and Hyperlipidemic  Composition (g / kg diet ) Configuration Basic diet Hyperlipidemia Casein 200 200 DL-Methionine 3 3 Corn starch 150 150 Sucrose 500 345 Fiber ^{1 )} 50 50 Corn oil 50 - AIN-mineral Mixture ^{2 )} 35 35 AIN-vitamin Mixture ^{3 )} 10 10 Choline bitartate 2 2 Beef tallow - 205 ¹⁾ Cellulose: Sigma Co. LTD., USA
^{2) The} mineral mixture according to the method of Rogers and Haper (1965) is as follows (g / kg diet): calcium phosphate divalent 500.0, sodium chloride 74.0, calcium citrate monohydrate 220.0, calcium sulfate 52.0, magnesium oxide 24.0, magnesium carbonate 3.5, ferrous citrate 6.0, zinc carbonate 1.6, cupuric carbonate 0.3, potassium iodide 0.01, chromium potassium sulfate 0.55, sucrose finely adjusted to 1,000
³⁾ Vitamin mixture (g / kg diet): Thiamine HCl 0.6, Biotin 0.02, Riboflavin 0.6, Cyanocobalamin 0.001, Pyridoxine HCl 0.7, Retinyl Acetate 0.8, Nicotinic Acid 3.0, DL-Tocopherol 3.8, Ca-Pantothenate 1.6, Finely powdered 7-dihydrocholesterol 0.0025, folic acid 0.2, methionine 0.005, sucrose to 1,000

1-2. Serum and Enzyme  pharmacy

After the sample was injected, the animal was lightly anesthetized with CO ₂ , and blood was collected from the abdominal aorta. The collected blood was left for 30 minutes and centrifuged at 3,000 rpm for 10 minutes to separate serum and lipid, lipid peroxide, hydroxyl radical. The content and superoxide dismutase activity were measured.

1-3. Body weight and fat tissue weight measurement

Body weight change was measured every week from the start of the experiment, measured 24 hours before the treatment of the experimental animals to calculate the weight change for the initial weight. The weight of adipose tissue was calculated by extracting fat around the abdominal cavity and testicles.

1-4. Liver tissue  And Feces  Heavy lipid content measurement

Liver tissue was weighed and stored frozen at -70 ℃, feces collected for the last 4 days. Triglycerides and cholesterol were measured according to the method of Folch et al. (32). In other words, the stools were lyophilized and ground to be powdered. The liver tissues were cut and then ground with a teflon homogenizer. 10 times the amount of solvent (chloroform: methanol = 2: 1) was added to the sample, and the lipids were repeatedly extracted. The filtrate was put in a water container and concentrated under reduced pressure to obtain lipids. Methanol was added thereto to dissolve well, and then quantified using a kit using an enzyme method.

1-5.Total Lipids Total lipid Content measurement

By the method of Fringe and Dunn (33) using the principle of the sulfo-phospho-vanillin reaction, the absorbance was measured at 540 nm for the pink color, which was heated with concentrated sulfuric acid and added with vanillin phosphate. The blood content was expressed in mg / dl according to the standard calibration curve.

1-6. Total cholesterol ( Total cholesterol Content measurement

Experiments were carried out using a kit (AM 202-K, Asan) prepared by the enzyme method of Richmond et al. (34). Enzyme reagents (containing 20.5 U / l cholesterol, oxidase 10.7 U / l, and sodium hydroxide 1.81 g / l) were dissolved in enzyme reagent solution (potassium phosphate monobasic 13.6 g / l, phenol 1.88 g / l). 3.0 ml of the enzyme solution prepared in 20 μl of the sample was added to the dissolved solution, followed by incubation at 37 ° C. for 5 minutes, and the absorbance was measured at a wavelength of 500 nm using the reagent blank as a control. The blood content was expressed in mg / dl according to the standard calibration curve.

1-7. Triglyceride ( Triglyceride Content measurement

Experiments were performed using a kit (AM 157S-K, Asan) prepared according to the method of McGowan et al. (35). Enzyme reagent (containing lipoprotein lipase 10800U, glycerol kinase 5.4U, peroxidase 135000U, and L-α-glycero phosphooxidase 160U) in ice cold phase was dissolved in enzyme reagent [N, N-bis (2-hydroxyethyl) -2-aminomethane sulfonic acid 0.427 g / dl], 3.0 ml of enzyme solution prepared in 20 µl of the sample was added to the solution, and incubated at 37 ° C. for 10 minutes to measure absorbance at a wavelength of 550 nm based on a reagent blank. The blood content was expressed in mg / dl according to the standard calibration curve.

1-8. Phospholipids ( Phospholipid Content measurement

Experiments were performed using a kit (Iatron Chem. Co.) prepared by the enzyme method of Chen et al. (36). In an ice cold solution, 20 µl of enzyme reagent (containing phospholipase 3.9U, choline oxidase 5.6U, peroxidase 3.6U, 4-aminoantipyrine 0.3252 mg) dissolved in enzyme reagent solution [containing tris (hydroxymethyl) -aminomethane 6.057 mg]. 3.0 ml of the prepared enzyme solution was added and incubated at 37 ° C. for 20 minutes to measure absorbance at a wavelength of 500 nm. The content was expressed in mg / dl according to the standard calibration curve.

1-9. High density lipoprotein - cholesterol ( HDL -C) content determination

The experiment was carried out using a kit (AM 203-K, Asan) prepared by the enzyme method of Noma et al. (37). 0.2 ml of sedimentation reagent (containing dextran sulfate 0.1% and magnesium chloride 0.1M) was added to 20 µl of serum and mixed well. The mixture was left at room temperature for 10 minutes and centrifuged at 3000 rpm for 10 minutes. 0.1 ml of the supernatant was mixed well with 3.0 ml of enzyme solution, incubated at 37 ° C. for 5 minutes, and the absorbance was measured at a wavelength of 500 nm using the reagent blank as a control. The content was expressed in mg / dl according to the standard calibration curve.

1-10. LDL - cholesterol  Content measurement

Low density lipoprotein-cholesterol (LDL-C) content was calculated by Equation 1 according to the method of Fridewald et al. (38).

1-11. Blood Lipid peroxide  Content measurement

According to the method of Yagi et al. (39), add 4.0 ml of 1 / 12N H ₂ SO ₄ to 20 µl of serum, mix 0.5 ml of 10% phosphotungstic acid, and leave at room temperature for 5 minutes. Then, 2.0 ml of 1 / 12N H ₂ SO ₄ and 0.3 ml of 10% phosphotungstic acid were added thereto, followed by centrifugation. 1.0 ml of a solution of 4.0 ml of distilled water, a mixture of 0.67% thiobarbituric acid and acetic acid in a 1: 1 manner was added thereto, and the mixture was reacted at 95 ° C. for 60 minutes, cooled at room temperature, and 5.0 ml of n-BuOH was added. After centrifugation for a minute, the resulting red n-BuOH was taken and absorbance was measured using a spectrofluorometer (Ex: 515 nm, Em: 553 nm). The absorbance was measured by reacting tetraethoxypropane 0.5 nmole with the standard solution in the same manner, and the serum lipid peroxide content was calculated by Equation 2 below.

1-12. Blood Hydroxyl radical  Content measurement

According to the method of Kobatake et al. (40), 34.8 μl of serum was added to 333.3 μl as 0.54M NaCl, 0.1M potassium phosphate buffer (pH 7.4), 10mM NaN ₃ , 7mM deoxyribose, 5mM ferrous ammonium sulfate and distilled water, and vortex well. The mixture was allowed to stand at 37 ° C. for 15 minutes. 67 μl of serum was taken and mixed with 75 μl of 8.1% sodium dodecyl sulfate, 500 μl of 20% acetic acid and 25 μl of distilled water. 333 μl of 1.2% thiobarbituric acid was added thereto, followed by heating in a water bath (100 ° C.) for 30 minutes, cooling at room temperature, and centrifugation at 700 × g for 5 minutes to measure absorbance at a wavelength of 532 nm. The content of hydroxyl radical (nmole / mg protein) was quantified by the calibration curve.

1-13. Blood Superoxide dismutase ( SOD Activity measurement

Quantification was performed according to Oyanagui's method (41). The serum was diluted 100-fold with potassium phosphate buffer and 100 μl of this was added to the test tube, where 500 μl of re-distilled water, 200 μl of reagent A (3 mM hydroxylamine / 3 mM hypoxanthine) and reagent B (7.5 mU / ml xanthine oxidase (XO) with Add 200µl of 0.1mM EDTA-2Na), mix well, and let stand for 40 minutes in 37 ℃ water bath. 2.0 ml of Reagent C (300 of sulfanilic acid / 5.0 mg N-1-naphthyl-ethylenediamine in 500 ml of 16.7% acetic acid) was added to the reaction solution, which was allowed to stand at room temperature for 20 minutes, and then the absorbance was measured at 550 nm. The superoxide dismutase activity in serum was measured. Enzyme activity was expressed as U / mg protein of superoxide dismutase. One unit of SOD was calculated as 50% inhibition of adrenochrome production.

1-14. Protein Quantitation

Protein content was measured using bovine serum albumin (Sigma, Fr. V) as a standard according to the method of Lowry et al. (42).

2. Experimental results

2-1. Of kelp ethanol extract sample Extraction yield  And principal components

The 95% yield of ethanol extraction of the dry crushed kelp was 1.76% as shown in Table 2. As reported by Lim (19), this kelp ethanol extract contained almost no polysaccharides such as alginic acid, and was composed mainly of fat components such as pigments, chlorophyll and essential oils (see Table 2).

Kelp extraction yield
(Unit: g / 100g) Solvent (v / w) Extraction yield (%) 95% ethanol 1.76

2-2. Kelp In the diet  Weight change in animals caused by

High fat diet shown in Table 1 was administered to rats for 4 weeks in an excessive amount to induce hyperlipidemia, and the kelp powder, kelp ethanol extract, and kelp ethanol extract residues were orally administered 200mg / kg every day for 6 weeks, and 1 week Body weight change was measured every time (see Table 3). Kelp samples were killed 24 hours after the last dose. The rats fed the normal diet weighed 220.8g per 6 weeks, and the rats fed the high-fat diet were 331.7g, resulting in an obese rat with an increase of 50.2%. The kelp powder was effective for weight loss until 2 weeks, but after 3 weeks, the kelp ethanol extract residue showed a significant weight loss compared to the control group. The weight of hyperlipidemic rats administered with 200mg / kg kelp ethanol residue was 268.9g after 6 weeks, which is 18.9% less than the control group, indicating that kelp ethanol residue was effective in weight loss. Choi et al. (22) reported that a tendency to lose weight when ingesting kelp is indicative of the effect of obesity, and Lee (23) reports that the East Coastal kelp diet is effective in inhibiting weight gain. These results suggest that kelp ethanol extract residue is more effective in weight gain inhibition than kelp itself, and may be useful as an anti-obesity and diet-related functional agent.

These results suggest that fatty acids such as fucosterol do not have a dietary effect of kelp, but that polysaccharides such as ilgin acid are the main ingredients of dietary effect. However, the detailed mechanism and the like have been analyzed since it can be revealed only by analyzing the cholesterol, TG, antioxidant enzyme activity, oxidation promoting or inhibitory hormone in various tissues and blood after the animal diet.

Normal and 6 weeks High-fat diet  Ingested Rat  Effect of Kelp on Weight Aid Dose
(mg / kg) One 2 3 4 5 6 (Weeks) Weight (g) Normal 69.7 ± 8.26 b 117.9 ± 10.6 b 138.5 ± 19.4 e 181.7 ± 29.6 b 211.7 ± 27.2 f 220.8 ± 28.6 f Control group 88.5 ± 9.52 a 161.3 ± 21.3 a 221.7 ± 30.5 abc 251.2 ± 21.1 a 306.8 ± 20.5 ab 331.7 ± 31.7 a Kelp dry powder 200 85.6 ± 7.49 a 159.9 ± 17.3 a 201.4 ± 18.7 abcd 235.5 ± 13.5 a 271.4 ± 18.3 de 285.6 ± 14.9 cde Kelp Ethanol Extract 200 86.8 ± .857 a 163.3 ± 19.8 a 227.6 ± 16.2 a 241.1 ± 19.2 a 287.2 ± 17.2 abcd 288.7 ± 18.3 cde Kelp ethanol extract residue 200 86.9 ± 7.78 a 162.6 ± 18.6 a 192.3 ± 17.3 d 227.9 ± 18.6 a 262.4 ± 18.6 e 268.9 ± 15.9 e The figures are expressed as mean ± S.D of the six experiment results. The figures following the same letter are not significantly different (p <0.05).

2-3. Kelp In the diet  Weight variation of adipose tissue in animals by

The results of observing the effect of diet with kelp samples on the adipose tissue of rats in dietary hyperlipidemia induced rats are shown in Table 4 (see Table 4). After four weeks of oral administration of kelp samples to rats with dietary hyperlipidemia, the amount of adipose tissue was measured. Decreased. When 200mg / kg of kelp ethanol extract residue was administered, it was 9.37mg / g, 37% lower than the control group 14.9mg / g. The testicular fat was orally administered with 200mg / kg of kelp ethanol extract residue, which was 9.41mg / g, which was reduced by 24% compared with the control 12.4mg / g. This result is consistent with the results obtained by oral administration of kelp alginic acid extract residue in the dietary obesity-induced rats again by supercritical harmful extraction for 2 weeks, and then measured the weight of the abdominal cavity (24). It was confirmed that the object sample had the most fat inhibitory effect. The results clearly suggested that fatty acids such as fucosterol produced dietary effects of kelp but that polysaccharides such as ilginic acid were the main ingredients of dietary effects.

Add kelp sample for 4 weeks Dietary  Effect on Adipose Tissue in Rats Aid Dose
(mg / kg) Abdominal cavity Epididymis mg / g body weight Normal 6.253 ± .19 f 7.83 ± 1.97 f Control group 14.9 ± 2.13 a 12.4 ± 1.36 abc Kelp dry powder 200 12.0 ± .125 cd 11.2 ± 0.82 bcd Kelp Ethanol Extract 200 13.8 ± 1.46 abc 11.8 ± 0.91 abcd Kelp ethanol extract residue 200 9.37 ± 1.48 e 9.41 ± 0.87 e The figures are expressed as mean ± S.D of the six experiment results. The figures following the same letter are not significantly different (p <0.05).

2-4. Kelp In the diet  Changes in Serum Lipid Components in Animals

The oral administration of kelp samples in hyperlipidemia-induced rats shows the results of changes in serum lipid components (Table 5). Phospholipids were reduced to 120.5 mg / dl compared to the control 148.7 mg / dl when 200 mg / kg of kelp ethanol extract residue was administered. Triglycerides, however, were 200.3 mg / dl in the control group and were significantly decreased by 21% to 158.2 mg / dl when 200 mg / kg of kelp ethanol extract residue was administered. Administration of kelp raw material and kelp ethanol extract 200mg / dl decreased to 183.2mg / dl and 187.8mg / dl respectively by 8% and 6%, respectively. appear.

After oral administration of kelp sample for 4 weeks Serum Lipids  Concentration change result Aid Dose
(mg / kg)  Phospholipid Triglyceride mg / dl Normal 120.4 ± 14.6 d 68.7 ± 7.26 f Control group 148.7 ± 10.2 a 200.3 ± 12.4 a Kelp dry powder 200 136.9 ± 10.1 abc 183.2 ± 10.2 bc Kelp Ethanol Extract 200 141.9 ± 11.2 abc 187.8 ± 8.43 ab Kelp ethanol extract residue 200 130.5 ± 10.3 cd 158.2 ± 10.1 e The figures are expressed as mean ± S.D of the six experiment results. The figures following the same letter are not significantly different (p <0.05).

2-5. Kelp In the diet  Serum in Animals by cholesterol  Content change and arteriosclerosis index

Table 6 shows the effects of dietary hyperlipidemia-induced dietary hyperlipidemia and changes in serum cholesterol content and atherosclerosis in rats treated with oral kelp samples (see Table 6). Dietary hyperlipidemia induced blood cholesterol levels of 91.6 mg / dl and 200 mg / kg of kelp raw material and kelp ethanol extract residue, respectively, 81.7 mg / dl (decreased by 10%) and 74.3 mg / dl (18% reduction), the kelp ethanol extract residue is more effective in lowering the total cholesterol content in the blood. HDL-cholesterol induced dietary hyperlipidemia was 31.8 mg / dl, but 200 mg / kg of kelp raw material, kelp ethanol extract and kelp ethanol extract residue was increased by 35.2 mg / dl (about 10%). 34.3 mg / dl increased by 7% and 37.3 mg / dl increased by 17%. In the case of LDL-cholesterol-induced dietary hyperlipidemia, 7.59 mg / dl of rats and 200 mg / kg of kelp ethanol extract residue were 6.62 mg / dl, which was significantly decreased by 12% compared to the control group. The results of these two experiments demonstrate that kelp ethanol extract residues are effective for hyperlipidemia. In addition, AI (Atheroscrelosis index, arteriosclerosis index) was 1.88mg / dl in rats induced dietary hyperlipidemia, and 1.32mg was obtained by oral administration of kelp raw material, kelp ethanol extract and kelp ethanol extract residue, respectively. / dl, 1.58mg / dl and 0.99mg / dl significantly decreased to 2.97%, 15% and 47%, respectively. It can be seen that the kelp ethanol extract residue sample has an excellent effect of lowering the arteriosclerosis index. Hyperlipidemia is due to increased synthesis of triglycerides in the small intestine, increased synthesis of triglycerides in the liver, decreased synthesis of HDL-cholesterol, increased synthesis of LDL-cholesterol and secretion (11). Showed. These results indicate that dietary seaweed drinks in rats induced dietary hyperlipidemia and arteriosclerosis resulted in a 15-20% decrease in serum total lipid content compared to dietary hyperlipidemia-induced rats (12). It is similar to the study (13) that HDL-cholesterol was elevated by oral administration of 200mg / kg of seaweed kelp and alginic acid extracted seaweed kelp residue from sea coast. The kelp ethanol extract residue was considered to be an antihyperlipidemic or anti-arteriosclerosis formulation because it has an excellent effect on the increase of HDL-cholesterol, LDL-cholesterol reduction and arteriosclerosis index.

Effects of Serum Cholesterol Contents and Atherosclerosis Index on Rats Treated with Oral Tangle for 4 Weeks Aid Dose
(mg / kg) Cholesterol (mg / dl) AI Total HDL LDL Normal 56.8 ± 6.25 g 41.6 ± 2.43 a 2.62 ± 0.55 a 0.37 ± 0.09 h Control group 91.6 ± 5.17 a 31.8 ± 1.18 f 7.59 ± 0.93 b 1.88 ± 0.16 a Kelp dry powder 200 81.7 ± 4.19 de 35.2 ± 1.45 cd 7.03 ± 0.53 bc 1.32 ± 0.11 e Kelp Ethanol Extract 200 88.5 ± 3.16 abc 34.3 ± 1.46 cde 7.38 ± 0.63 bc 1.58 ± 0.08 c Kelp ethanol extract residue 200 74.3 ± 3.24 f 37.3 ± 1.18 b 6.62 ± 0.52 c 0.99 ± 0.06 g Values are expressed as mean ± SD of the results of experiment 6. The figures following the same letter are not significantly different (p <0.05).
AI (Atheroscrelosis Index) = (total cholesterol-HDL cholesterol) / HDL cholesterol

2-6. Kelp In the diet  Lipids in liver tissue in animals and cholesterol  Content change

Table 7 shows the results of experiments on the change of lipid and cholesterol content in hepatic tissues when oral administration of kelp samples to rats induced dietary hyperlipidemia (see Table 7). In the rats induced dietary hyperlipidemia, the total lipid content was 34.9 mg / g of tissue, and 200 mg / kg of kelp ethanol extract residue was 29.4 mg / g of tissue, which was significantly 15.7% of the control group. Decreased. In the case of triglyceride, oral administration of 200mg / kg of kelp ethanol extract residue to 26.4mg / g of control tissue, 23.9mg / g of tissue significantly decreased by 9.5% compared to the control. And the content of cholesterol was 6.25mg / g of control tissue, oral administration of 200mg / kg of kelp ethanol extract residue was reduced to 5.83mg / g of tissue, although there was no significant difference. This suggests that administration of kelp ethanol extract residues affects lipid metabolism of hepatic tissues, which indirectly proves effective in diseases such as hyperlipidemia (12). These results showed a similar result to Miyashita et al. (13) report that the administration of kelp to hyperlipidemic rats was able to suppress the increase of body weight and liver tissue to some extent.

Effect of Lipid and Cholesterol Contents in Hepatic Tissues after Oral Administration of Kelp Samples for 4 Weeks Aid Dose
(mg / kg)      Total lipid Triglyceride Cholesterol mg / g of tissue Normal 16.8 ± 1.98 f 9.37 ± 0.97 d 2.63 ± 0.49 b Control group 34.9 ± 2.17 a 26.4 ± 2.11 a 6.25 ± 0.37 a Kelp dry powder 200 30.9 ± 1.33 de 24.9 ± 1.16 abc 5.99 ± 0.31 a Kelp Ethanol Extract 200 33.8 ± 1.39 ab 25.8 ± 1.31 abc 6.13 ± 0.40 a Kelp ethanol extract residue 200 29.4 ± 1.21 e 23.9 ± 1.25 c 5.83 ± 0.36 a The figures are expressed as mean ± S.D of the six experiment results. The figures following the same letter are not significantly different (p <0.05).

2-7. Kelp In the diet  In animals by Feces  Formation change of neutral lipid concentration

Table 8 shows the results of experiments on the change in the production of triglyceride levels in feces when oral administration of kelp samples to rats induced dietary hyperlipidemia (see Table 8). Dietary hyperlipidemia induced fecal weight was 4.07 g / day and 200 mg / kg of kelp ethanol extract residue was orally administered at 4.58 g / day, which was significantly increased by 12.5% compared to the control group. After oral administration of 200mg / kg kelp ethanol extract, the fecal weight of rats was 4.26g / day, which was 4.7% lower than that of the control group. Fecal weight of rats orally administered with kelp samples was significantly different from those of normal-diet rats and feces, and the kelp ethanol extract residue increased by 27.9% and the kelp raw material increased by 23.7%. Cholesterol content in dietary hyperlipidemia induced fecal cholesterol was 5.13 mg / g, and when oral administration of kelp ethanol extract 200 mg / kg was 5.31 mg / g, but it was decreased by 3.5%. The triglyceride content in the feces of dietary hyperlipidemia was 12.7mg / g, and 200mg / kg of kelp ethanol extract showed no significant difference to 13.5mg / g but decreased by 6.3%.

In this experiment, the oral administration of 200mg / kg of kelp samples increased the amount of feces compared to normal diet and hyperlipidemia-induced rats. Increases viscosity, inhibits lipid absorption, inhibits bile acid resorption in the small intestine, promotes sterol excretion into feces, and short-chain fatty acids, a fermentation metabolite of dietary fiber in the large intestine, inhibits the synthesis of cholesterol. By promoting the movement of the colon to shorten the time to pass through the intestines and increase the bowel movements to prevent constipation (13) Oral administration of kelp samples to rats that induced hyperlipidemia resulted in dietary fiber of kelp Cholesterol content and triglyceride content increased. In addition, Lee was administered to rats in the East Coast of Korea, and fecal amount increased according to the amount of kelp added, which appears to be due to the effects of alginate and fucoidan, which are non-digestible polysaccharides contained in kelp. The amount was increased, and accordingly, the weight gain was estimated to be suppressed and it was similar to the results of the study confirming the effect of bowel movement due to the intake of kelp. Kim et al. Measured the effect of constipation by administering kelp extract to rats that caused constipation and reported that the intake of dietary fiber shortened the intestinal transit time while increasing defecation (14). Jang et al. (15) showed that the cholesterol and bile acids excreted in the stool significantly increased in diabetic rats fed with kelp methanol extract, thus contributing to the prophylactic effects of atherosclerosis, a complication that is likely to occur due to diabetes. There are reports that you can. Sandberg et al. (16) reported that sodium alginate increased fat excretion by 140% because alginate was excreted in combination with fatty acids. The results suggest that increased fecal cholesterol and triglyceride levels may be related to the action of polysaccharides such as alginate, a dietary fiber component in ethanol residues.

When oral administration of kelp sample for 4 weeks Feces  Effect of Formation of Neutral Neutral Lipid Concentration Aid Dose
(mg / kg)  Fecal dry weight Cholesterol Triglyceride g / day mg / g Normal 3.58 ± 0.39 d 1.43 ± 0.31 c 7.53 ± 1.25 f Control group 4.07 ± 0.41 c 5.13 ± 0.53 b 12.7 ± 2.36 cde Kelp dry powder 200 4.43 ± 0.26 abc 5.42 ± 0.37 ab 15.1 ± 1.37 b Kelp Ethanol Extract 200 4.26 ± 0.34 abc 5.31 ± 0.32 b 13.5 ± 1.25 bcde Kelp ethanol extract residue 200 4.58 ± 0.26 a 5.87 ± 0.43 a 17.6 ± 1.39 a The figures are expressed as mean ± S.D of the six experiment results. The figures following the same letter are not significantly different (p <0.05).

2-8.Tashima In the diet  In liver tissue in animals TBA Fall concentration

Table 9 shows the results of dietary hyperlipidemia and the effect of oral administration of kelp samples on blood lipid peroxidation (see Table 9). The TBA value of dietary hyperlipidemia-induced rats was 52.4 MDA nmol / mg protein, which was 1.95 times higher than that of rats fed normal diet, whereas the kelp powder samples, kelp ethanol extract, and kelp ethanol extract residues in the hyperlipidemic rats. After oral administration of 200mg / Kg body weight daily, 19.1%, 8.6%, and 30.0%, respectively, showed significant inhibition of lipid peroxidation. In particular, when kelp ethanol extract residue was administered orally at 200 mg / kg body weight, 36.7 MDA nmol / mg protein significantly inhibited lipid peroxidation by 30% compared to the control group, while kelp ethanol containing a large amount of fat such as fucosterol and chlorophyll Extract diet group inhibited lipid peroxidation only 8.6%. By the way, the kelp ethanol extract residue and algae polyalgae such as alginic acid and kelp itself reduced the lipid peroxidation significantly by 30% and 19.1%, respectively. This suggests that polysaccharides such as alginic acid, the main component of kelp, are mainly involved in the inhibition of peroxidation by animal diet in kelp.

Radicals have been shown to cause aging and chronic diseases such as cerebrovascular disease, cardiovascular disease, and cancer by damaging lipids, proteins and DNA (14). Among many radicals, TBARS is most commonly used as an indicator of lipid damage by radicals (15).

In general, lipid peroxides cause various diseases related to the induction of degenerative processes and changes in cancer, aging, biofilm change and destruction (16). The antioxidant activity of kelp ethanol extract residues may be effective in preventing such diseases. It has been reported that the amount of MDA in the tissues increased due to the increase in oxidative stress during diabetes, and Cho et al. Ingested the kelp powder into diabetic rats and reported the functional effect of the antioxidant effect of kelp as the kidney lipid peroxide was reduced.

Effect of kelp samples on oral blood lipid peroxidation after 4 weeks Aid Dose
(mg / kg) TBARS MDA nmol / mg protein Normal 26.9 ± 3.57 h Control group 52.4 ± 2.15 a Kelp dry powder 200 42.4 ± 1.87 f Kelp Ethanol Extract 200 47.9 ± 2.16 bcd Kelp ethanol extract residue 200 36.7 ± 1.42 g The figures are expressed as mean ± S.D of the six experiment results. The figures following the same letter are not significantly different (p <0.05).

2-9. Kelp In the diet  Blood in animals caused by hydroxy radical  Generated and superoxide  dismutase (SOD) activity

Table 10 shows the results of dietary hyperlipidemia induced by oral administration of kelp sample and its effect on free radicals in blood (see Table 10). The effects on SOD activity were 1.69 Unit / mg protein in hyperlipidemia-induced rats. Oral administration of 200mg / kg of kelp powder, kelp ethanol extract, and kelp ethanol extract daily resulted in 1.79, 1.74, 1.89 Unit / mg protein, respectively. 5.9%, 2.95%, and 11.8% more than the control group. On the other hand, the effects on the production of hydroxyl radicals were 5.87 nmol / mg protein in hyperlipidemic rats. 200 mg / kg of kelp powder, kelp ethanol extract, and kelp ethanol extract residue were administered orally at 5.18, 5.36, 4.93, respectively. nmol / mg protein decreased by 11.8%, 8.7%, and 16.0% than the control group. Blood lipid peroxidation and free radicals are known to cause cytotoxicity and cause aging and pathology of various diseases and detoxification by the action of detoxification mechanisms (17-18).

The results of animal experiments showed no significant difference in the amount of SOD enzyme that removes active oxygen after oral administration of kelp raw material, kelp ethanol extract and its extract residue, but it was the largest increase in oral administration of kelp ethanol extract residue. In the rats with kelp powder itself, the polysaccharide remained intact than the ethanol extract orally administered rats, whereas the amount of hydroxyl radical, the representative active radical in the body, was highest in the kelp ethanol extract residue group with the highest alginate. Was keeping low. It was confirmed by animal experiment that kelp ethanol extract residue has the greatest antioxidant effect in the body.

Effect of kelp sample on oral blood for 4 weeks after oral administration Aid Dose
(mg / kg)      Hydroxy radical SOD nmol / mg protein Unit / mg protein Normal 3.01 ± 0.37 e 3.53 ± 0.26 a Control group 5.87 ± 0.25 a 1.69 ± 0.17 bcd Kelp dry powder 200 5.18 ± 0.26 cd 1.79 ± 0.16 bcd Kelp Ethanol Extract 200 5.36 ± 0.30 bc 1.74 ± 0.12 bcd Kelp ethanol extract residue 200 4.93 ± 0.21 d 1.89 ± 0.14 b Values are expressed as mean ± SD of the results of experiment 6. The figures following the same letter are not significantly different (p <0.05).
Unit: The unit of SOD is defined as the inhibition of reduction of alkaline DMSO-mediated adrechrome for 50% in blood.

Hereinafter, the preparation examples of the composition including the kelp of the present invention will be described, but the present invention is not intended to limit the present invention but is only specifically described.

Formulation example  One. Powder  Produce

LJ-1 20 mg

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation example  2. Preparation of Tablets

LJ-1 10 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation example  3. Manufacture of capsule

LJ-P 10 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Formulation example  4. Preparation of injections

LJ-B 10 mL

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO ₄ , 12H ₂ O 26 mg

(2 ml) per 1 ampoule according to the usual injection preparation method.

Formulation example  5. Liquid  Produce

LJ-B 20 mL

10 g of isomerized sugar

5 g of mannitol

Purified water

Each component was added to purified water in accordance with the usual liquid preparation method and dissolved, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was adjusted to 100 ml with purified water, And sterilized to prepare a liquid preparation.

Formulation example  6. Manufacture of health food

LJ-P 1000 mL

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

0.2 μg of vitamin B12

Vitamin C 10 mg

10 μg biotin

Nicotinic acid amide 1.7 mg

50 μg folic acid

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

The composition ratio of the above-mentioned vitamin and mineral mixture is mixed with a composition suitable for a health food in a preferred embodiment, but the compounding ratio may be arbitrarily modified. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation example  7. Manufacture of health drinks

LJ-1 1000 mL

Citric acid 1000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Add 900 ml of purified water

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was heated at 85 DEG C for about 1 hour with stirring, and the solution thus prepared was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, &Lt; / RTI &gt;

Although the composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment, the blending ratio may be arbitrarily varied according to the regional and national preferences such as the demand level, the demanding country, and the intended use.

Claims (7)

A pharmaceutical composition for the prevention and treatment of obesity or hyperlipidemia and atherosclerotic vascular disease containing kelp as an active ingredient. The method of claim 1,
The kelp is a pharmaceutical composition comprising a dry powder, polar solvent soluble extract, or polar solvent insoluble extract form. The method of claim 2,
(1) kelp dried powder includes kelp dried powder dried and powdered by conventional drying methods such as sunlight drying, natural drying, forced drying, etc .; (2) The kelp polar solvent soluble extract is a kelp polar solvent soluble extract obtained by extracting the dried powder by a conventional extraction process with a polar solvent such as water, methanol, ethanol, butanol, lower alcohols and mixtures thereof; And (3) a pharmaceutical composition comprising a kelp polar solvent insoluble extract that is an insoluble residue in the polar solvent. The method of claim 1,
The hyperlipidemia and atherosclerotic vascular disease are hyperlipidemia, arteriosclerosis, heart failure, hypertensive heart disease, arrhythmia, congenital heart disease, myocardial infarction, angina pectoris, stroke or peripheral vascular disease. After finely powdering the dried kelp in a grinder, the kelp powder was put into an Erlenmeyer flask and 80 to 100% ethanol was added at a ratio of 1: 15 (w / v), mixed, and then mixed at 10 ° C. to 50 ° C. A first step of extracting by ultrasonic extraction, reflux extraction, hot water extraction, cooling extraction or supercritical extraction for minutes to 50 minutes; The kelp polar solvent soluble in claim 1 comprising a second step of manufacturing the extract by filtration with filter paper, concentrating it at 5 ° C. to 45 ° C. with a rotary vacuum condenser, and freeze drying and storing it at −5 ° C. to −60 ° C. Obtaining an extract, and drying the remaining residue to prepare a kelp polar solvent insoluble extract. Health functional food for the prevention and improvement of obesity or hyperlipidemia and atherosclerotic vascular disease containing kelp as an active ingredient. 7. The dietary supplement of claim 6 which is a powder, granule, tablet, capsule or beverage.