KR100826560B1 - Composition for improving diabetes - Google Patents

Abstract

An extract of Scytosiphonaceae seaweed is provided to lower blood sugar, differentiate 3T3-L1 preadipocytes into adipocytes and promote accumulation of neutral fats, thereby being used for treating or preventing diabetes without side effects. A composition for improving diabetes or preventing diabetic complications comprises an extract of Scytosiphonaceae seaweed as an effective ingredient, wherein the Scytosiphonaceae seaweed is selected from the group consisting of Endarachne binghamiae Agardh, Scytosiphon lomentaria Link, Petalonia fascia Kuntze, Colpomenia bullosa Yamada, Hydroclathrus clathratus Howe, and Colpomenia sinuosa DERBES et SOLIER. Further, the extract of Scytosiphonaceae seaweed is extracted by polar or non-polar solvent.

Description

Composition for Improving Diabetes {Composition for Improving Diabetes}

1a to 1k respectively, ethanol extract of brown seaweed, methylene chloride extract of brown seaweed, chloroform extract of brown seaweed, hexane extract of brown seaweed, hexane extract of brown seaweed, ethyl acetate extract of brown seaweed, Of blood sugar after administration of methanol extract, seaweed ether ether, seaweed iron butanol extract, seaweed iron ethanol extract, hexane fraction of seaweed iron ethanol extract, ethyl acetate fraction of seaweed iron ethanol extract, and butanol fraction of seaweed iron ethanol extract over 8 weeks The change is observed.

Figures 2a and 2b is a photograph taken by each extract solvent, the cells stained with Oil Red O dye, to determine whether the seaweed extract has the activity of differentiating 3T3-L1 adipocytes into adipocytes, and accumulates neutral fat It is a graph showing each extract by analyzing the degree quantitatively.

Figures 3a and 3b is to determine whether the ring hawk extract, anthill extract, long bulrush extract, net basket extract and bulrush extract have the activity of differentiating 3T3-L1 adipocytes into adipocytes, respectively Cells stained with Red O dye were taken by extractant and graphs of quantitative analysis of triglyceride accumulation.

4 is a graph showing the results of glucose uptake of adipocytes when ethanol extracts of insulin and / or seaweed are treated to adipocytes.

Figure 5 graphically shows whether the ethanol extract of Wakame iron interacts with troglitazone or rosiglitazone in the transcriptional activity of PPAR-γ.

The present invention relates to a diabetic improver composition comprising a ring extract and seaweed extract.

Diabetes refers to a condition in which insulin, a glucose-regulating hormone secreted from the beta cells of the pancreas, is insufficient or does not function properly, causing blood glucose in the blood not to be used as energy, accumulating in the blood, causing high blood sugar, and detecting urine glucose.

Diabetes is generally divided into insulin dependent diabetes mellitus (type I diabetes mellitus) and insulin independent diabetes mellitus (type II diabetes mellitus), depending on whether insulin is essential for the treatment.

Insulin-dependent diabetes is caused by the destruction of pancreatic beta cells due to autoimmune mechanisms, viral infections, drugs, and other diseases of the pancreas. It is also called pediatric diabetes because it occurs in younger generations. It is named because it is difficult to maintain life if insulin is not supplied from the outside.

Insulin-independent diabetes is caused by the lack of insulin in the pancreatic beta cells, but the amount is reduced and its action is caused, mainly after 30s, it is also called adult diabetes. It is called insulin-independent diabetes because it does not necessarily need to supply insulin from the outside to maintain life, but this does not mean that insulin therapy is not necessary to treat hyperglycemia.

Diabetes is further classified by the above two types of diabetes in addition to diabetes, which occurs in young people with a history of malnutrition, and gestational diabetes, in which insulin is inhibited by the release of various hormones during pregnancy. Sometimes.

On the other hand, if diabetes persists for a long time, glucose is not properly absorbed into the body, and various metabolic disorders occur due to an imbalance in nutrient supply, thereby causing various diabetes complications.

Diabetes complications include nerve complications caused by damage to nerve fibers and nerve membranes, diabetic retinopathy (non-proliferative retinopathy, proliferative retinopathy, diabetic cataracts), renal failure, sexual dysfunction, skin diseases (allergic), hypertension, atherosclerosis, stroke ( Stroke, heart disease (myocardial infarction, angina pectoris, heart attack), and gangrene.

The antidiabetic agents are mainly obtained by oral hypoglycemic agents such as sulfonylureas, biguanides, and alpha-glucodiase inhibitors, and insulin or DNA recombination from cows and pigs. Insulins such as insulin are used, but these drugs can cause hypoglycemia (sulfonylurea-based drugs), kidney toxicity (biguanide-based drugs), lactic acidosis (biguanide-based drugs), diarrhea and upset stomach (alpha-glucosidase inhibitors). And many adverse side effects, including immunorejection (insulin), the World Health Organization (WHO) strongly recommends the use of natural products with fewer side effects for diabetes (Grover JK, Vats. V., Rathi. SS, Journal of Ethnopharmacology, 73, pp 461-470, 2000).

Natural products reported to be effective in the treatment of diabetes include Ajuga multiflora Bunge extract (Korean Patent No. 0242283), Pasqueflower root extract (Korean Patent No. 0213667), and Apolana acid derivative compounds isolated from Jangnabi hungry mushrooms (Korea) Patent No. 457690), polysaccharides isolated from Korean Angelica gigas (International Publication No. 2001-60386), bile of ruminant animals (US Pat. No. 6645355), and the like.

The present invention has also been completed in view of using natural products for the treatment of diabetes.

An object of the present invention is to provide a diabetic improver composition using a natural extract of ringberry and seaweed.

Other objects or specific aspects of the present invention will be presented below.

In one aspect, the present invention provides a diabetic improver composition comprising a ring extract and seaweed extract.

Plants belonging to the ring hawk and algae include Endarachne binghamiae Agardh, Scytosiphon lomentaria Link, Antling ( Petalonia fascia Kuntze), Colpomenia bullosa Yamada, and Basket ( Hydroclathrus clathratus Howe) A total of 17 species are known, including Colpomenia sinuosa DERBES et SOLIER. Among them, the species inhabited in Korea have been reported (Ga Youn Cho, Hwan Su Yoon, Han-Gu Choi, Kazuhiro). Kogame and Sung Min Boo., Phylogeny of the Famil Scytosiphonaceae (Phaeophyta) from Korea based on Sequences of plastid-encoded RuBisCo Spacer Region.Algae. 16 (2): 145-150, 2001., Giljeon Chungsaeng. , Marine Algae of Japan, National Civil War Happiness 1998., Kang-Won Won.

The inventor (s) extracts or further fractionates the ring and seaweeds of all the species reported to inhabit in Korea as identified in the following Examples and Experimental Examples, or further fractions thereof, and the blood sugar of the extract, fractions, etc. The effects of dropping and inducing differentiation of 3T3-L1 adipocytes into adipocytes were examined.

As a result, it was confirmed that the extracts of all ring hawk and algae had hypoglycemic effect, induction effect of differentiation of 3T3-L1 adipocytes into adipocytes.

 3T3-L1 adipocytes are well known biological properties that are widely used in diabetes research. They differentiate into adipocytes when insulin is treated with DEX (Dexamethasone) and IBMX (1-methy-3-isobutylxanthine). It is a cell used for the search for insulin substances because it has (Nobuya, M., Gen, I., Motozumi, O., Yasunao, Y., Shigeo, K., Tatsuya, H., Katsuaki, K., Hideshi, K., Kazuwa, N. Antihyperglycemic mechanism of M16209, an antidiabetic agent, in 3T3-Ll adipocytes., Life Sciences 60: 1821-1831, 1997; Sherrie, RT, Troglitazone Enhances Differentiation, Basal Glucose Uptake, and Glut1 protein Levels in 3T3-L1 Adipocytes., Endocrinology 137: 4706-4712, 1996), Substances that induce differentiation of 3T3-L1 adipocytes into adipocytes are considered to be effective in improving diabetes (Kletzien, RF, Clarke, SD). , Vlrich, RG, Enhancement of adipocyte differentiation by an insulin-sensitizing agent, Mo lecular Pharmacology 41 (2): 393-398).

The present invention is provided based on the experimental results as described above.

In the present specification, including the claims, “ring hawk and seaweed” should be understood to refer to all seaweeds belonging to the ring family, regardless of their species.

As mentioned above, there are about 17 species of plants belonging to Korimae and Algae, but the species inhabited in Korea has been reported by seaweed iron, hooked hawk, anthill, long-tailed horse, net basket, and red-tailed horse. The inventor (s) extracted these six ring hawks and seaweeds with ethanol (80%) and examined whether the ethanol extracts had the activity of differentiating 3T3-L1 adipocytes into adipocytes. It was confirmed that it has activity. As a result of these experiments, as long as any seaweed is classified as a ring hawk and algae in taxonomic terms, the extracts will have a differentiation-inducing activity of adipocytes into adipocytes of 3T3-L1 adipocytes. It is expected that one of ordinary skill in the art, based on the disclosure of the present specification, will be able to ascertain and select whether the extract of such algae has such activity within its usual capacity.

Therefore, the ring hawk and seaweed in the present specification should be taken as a meaning including all seaweeds belonging to the other ring hawk and seaweed in addition to the six kinds of seaweed. Nevertheless, in the following Examples and Experimental Examples, 3T3-L1 adipocytes may be understood as one of the above 6 types of seaweeds, in which the activity of inducing differentiation into adipocytes is directly confirmed, and more preferably, the following Examples and Experimental Examples It can be understood as the seaweed that appeared to have the highest activity in.

In the present specification, including the claims, " ringberry seaweed extract " should be understood to mean any form of extract obtained by any extraction method, regardless of the extraction method.

Generally, extraction methods for plants can be roughly classified into methods using an extraction solvent, a method of compressing an extract to obtain an extract in the form of a essence, and a method of obtaining an extract in an aqueous state after removing solids by hydrolysis and crushing the extract. have.

Although obtained in the following examples of the present invention is a method using an extraction solvent, even in the case of the extract of the aqueous solution obtained by crushing the extract of the extract form or the extraction object and the solid content can be seen in a more purified form than that As long as it is confirmed that the extracts (particularly fractions) of the following examples have a diabetic improvement effect, it should be understood that the extracts are included in the range of the hawk and algae extracts.

In addition, it should be understood that any method such as cold soaking, refluxing, warming, and ultrasonic wave may be applied to the extracting solvent as long as it is extracted through the step of dipping the ring solvent and the algae to be extracted.

In the following examples, but extracted by cold soaking, for example, in order to save the extraction time, it can be extracted by refluxing or warming, and can also be extracted by radiating ultrasonic waves in order to preserve the activity of the useful component to the maximum.

The inventor (s) is not shown in the following Examples and Experimental Examples, but the hypoglycemic effect even when refluxed or warmed seaweeds in sea bream and seaweed or extracted by releasing ultrasound, the effect of 3T3-L1 adipocytes into adipocytes In the differentiation-inducing activity and the like it was confirmed that it does not bring a special difference with the extract of the following examples.

In addition, when using an extraction solvent, both a polar solvent and a nonpolar solvent can be used. The present inventor (s) first prepared ethanol extracts of 6 kinds of ring plums and seaweeds to examine the effect of differentiation-inducing activity of adipocytes into adipocytes of 3T3-L1 adipocytes. . As a result, it was confirmed that the extracts of all species had this activity, and in particular, seaweed fermentation among Korimae and algae had the highest activity. In addition, the inventor (s) selected seaweeds from six kinds of ring and seaweeds based on the experimental results and extracted them using a non-polar solvent such as hexane in addition to extracting them using a polar solvent such as ethanol (in the following example). Although methylene chloride, chloroform, hexane, ethyl acetate, methanol, ether, butanol, etc. were used in addition to these extracting solvents, all of the seaweed iron extracts lowered blood sugar and differentiated 3T3-L1 adipocyte cells into adipocytes. It was confirmed that there is induction activity. These experimental results show that both polar and nonpolar solvents can be used as extraction solvents. Therefore, when using an extraction solvent to obtain a ring solvent and seaweed extract, it should be understood whether the extraction solvent is polar or nonpolar. However, for reasons of convenience and economy, for example, alcohol (preferably lower alcohol having 5 or less carbon atoms) mixed with water or distilled water, pure alcohol, ethyl acetate, acetone, chloroform, methylene chloride, benzene, It is preferable to use alkanes having 5 to 10 carbon atoms, toluene, kerosene and the like, such as n-pentane, cyclopentane, n-hexane, cyclohexane, n-heptane, n-octane, n-nonane and n-decane, It is more preferable to use ethanol or a mixed solvent of ethanol with water or distilled water when considering the toxicity to humans.

On the other hand, even when obtaining a fraction of a more purified form in the crude extract is understood whether the solvent used to obtain the fraction is a polar solvent or a non-polar solvent. As shown in the following examples and experimental examples, when the ethanol extract of Wakame iron was partitioned into a hexane layer, an ethyl acetate layer, and a butanol layer, all the fractions had a hypoglycemic effect, and the fat tax of 3T3-L1 adipocytes. It is because it is confirmed that they have a differentiation-inducing activity of captive and triglyceride accumulation promoting activity. Preferred embodiments of the solvent used to obtain the fractions herein are valid as mentioned above in connection with the extraction solvent.

On the other hand, in the present specification, including the claims, "diabetes" is understood to include both insulin-dependent type 1 diabetes and insulin-independent type 2 diabetes according to the general classification, furthermore diabetes and gestational diabetes due to malnutrition It is understood to include. Treatment of all diabetes, including insulin dependent type 1 diabetes and insulin independent type 2 diabetes, usually includes hypoglycemic agents and / or insulin (refers to insulin or insulin-like agents). In the sense that it is used, the ring hawk and seaweed extracts of the present invention which have a hypoglycemic effect (particularly seaweed iron extract) and have been found to act as an insulin-like substance are insulin-dependent type 1 diabetes mellitus and insulin-independent type 2 diabetes mellitus. This is because it can be usefully used for all other diabetes.

Also in this specification, including the claims, “improvement” is meant to include treatment, prevention, and alleviation of symptoms.

Meanwhile, in the diabetic improver composition of the present invention, the extract of Korimae and seaweed, which are the active ingredients, may be included in any amount within the range that does not have toxicity to the human body depending on the type of diabetes mellitus, the degree of diabetes symptoms, and the like. Based on the total weight of the composition may be included from 0.001% to 99.99% by weight, preferably from 0.001% to 20% by weight.

The diabetic improver composition of the present invention may include, in addition to the ring extracts and seaweed extracts, which are the active ingredients, compounds or natural products that are known to have an antidiabetic effect so as to enhance or enhance diabetes improvement effects. One sulfonylureas drug, biguanides, alpha-glucodiase inhibitor, insulin, Ajuga multiflora Bunge extract (Korean Patent No. 0242283) , Pasqueflower Root Extract (Korean Patent No. 0213667), Apanaic Acid Derivative Compound Isolated from Janna Beetle Mushroom (Korean Patent No. 457690), Polysaccharides Isolated from Angelica gigas (International Publication No. 2001-60386), Ruminant Animals Bile (US Pat. No. 6645355) and the like, but are not limited thereto.

In particular, the diabetic improvement composition of the present invention further comprises an insulin agent, such as insulin obtained from cows or pigs, insulin obtained by genetic recombination, insulin-like substances, etc., to enhance and elevate its diabetic improvement effect, or PPAR-gamma antagonist. It is preferable to further include a thiazolidinediones (TZDs) -based drugs such as introglitazone, rosiglitazone, pioglitazone, and the like. It is because in the following Examples and Experimental Examples that insulin has the effect of increasing the glucose absorption effect of fat cells of wakame iron extract and troglitazone, rosiglitazone, etc. have the effect of increasing the PPAR-γ activity of wakame iron extract to be. In general, substances that promote the glucose uptake effect of adipocytes differentiated from 3T3-L1 adipocytes or promote the activity of peroxisome proliferator-activated receptor-γ (PPAR-γ) are likely to have diabetes improvement effects. In addition, the synergistic effect in the glucose uptake effect of adipocytes and the synergistic effect in promoting PPAR- [gamma] activity may be synergistic in diabetic improvement effect. (Willson, TM, Cobb, JE, Cowan, DJ, Correa, ZD, Prakash, SR, Beck, KD, Moore, LB, Kliewer, SA, Lehmann, JM, The structure-activity relationship between peroxisome proliferator-activated receptor gammar agonism and the anti-hyperglycemic activity of thiazolidinediones.J. Med Chem. 39: 665-668, 1996., Berger, J., Bailey, P., Biswas, C., Cullinan, CA, Doebber, TW, Hayes, NS, Saperstein, R., Smith, RG, Leibowitz, MD, Thiazolidinediones produce a conformational change in peroxisomal proliferator-activated receptor-gamma: binding and activation correlate with antidibetic actions in db / db mice., Endocrinology 137: 4189-4195, 1996)

In PPAR-γ, 3T3-L1 adipocytes are converted into adipocytes by insulin added together with DEX and IBMX, which prevents the growth of the cells and initiates the conversion into adipocytes as well as the differentiation of adipocytes. It is a transcription factor known to play an important role in regulation (Lowell, BB, PPAR-gammar: adipogenic regulator and thiazolidinedione receptor. Cell 99: 239-242, 1999., Spiegelman, BM, PPAR-gamma: an essential regulator of adipogenesis and modulator of fat cell function.Diabetes 47: 507-514, 1998.).

The diabetic improver composition of the present invention can be understood as a pharmaceutical composition in specific embodiments.

The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier in addition to the ring medium and seaweed extract as an active ingredient. Such pharmaceutically acceptable carriers are commonly used in pharmaceutical preparations, such as lactose, dextrose, Sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propyl Hydroxybenzoate, talc, magnesium stearate, mineral oil, and the like.

The pharmaceutical compositions of the present invention may also further comprise lubricants, wetting agents, sweetening agents, flavoring agents, emulsifying agents, suspending agents, preservatives and the like as additives.

The carrier may be included in the pharmaceutical composition of the present invention from about 1% to about 99.99% by weight, preferably from about 90% to about 99.99% by weight, based on its total weight, the additive being about 0.01% by weight To about 20% by weight.

On the other hand, the pharmaceutical composition of the present invention can be administered by various routes, for example, orally or rectally, or by injection into veins, muscles, subcutaneous, intrauterine dura, and the like.

The pharmaceutical compositions of the present invention may be prepared in unit dose form or formulated into pharmaceutically acceptable carriers and / or excipients or incorporated into multi-dose containers. In this case, the formulation may be in the form of a solution, suspension, or emulsion, or may be in the form of an excipient, an excipient, a powder, a granule, a tablet, or the like.

The daily dosage of the pharmaceutical composition of the present invention is usually 0.001 ~ 150 mg / kg body weight range, it can be administered once or divided into several times. However, since the dosage of the pharmaceutical composition of the present invention is determined in view of various related factors such as the route of administration, the age, sex, weight of the patient, and the severity of the patient, the dosage may limit the scope of the present invention in any aspect. It should not be understood as.

The diabetic improver composition of the present invention may be regarded as a food composition in a specific embodiment.

The food composition of the present invention may be prepared from gums, vitamin complexes, dietary supplements, special nutritional supplements, functional drinks, and the like.

The food composition of the present invention, in addition to the extract of the ring hawk and seaweeds, glucose, monosaccharides such as fructose, disaccharides such as maltose, sucrose, or polysaccharides such as dextrin, cyclodextrin can be added, and xylitol, sorbitol, erythrate Sugar alcohols such as itol may also be added, sweeteners such as taurine, stevia extract and the like may also be added, and other food additives may also be added.

In another aspect, the present invention provides a composition for preventing diabetic complications comprising the extract of Korimae and seaweed as an active ingredient.

Considering that it is generally necessary to reduce hyperglycemia to normal levels for the prevention of diabetic complications (Kim, Sang-Sang, Chang-Soo Na: Effect of Phenollic Compounds Extracted from Pear on Streptozotocin-induced Hyperglycemic Mice, Korean Journal of Food and Nutrition, 31 ( 6), 1107-1111, 2002), the composition for preventing diabetic complications of the present invention, which comprises an extract of an algae and an algae extract having an effect of lowering blood sugar, as an active ingredient, alone or for the prevention of diabetic complications, Can be used in combination.

In the present specification, including the claims, the "diabetic complication" is meant to include all diseases secondary to the onset of diabetes, such as neurological complications, diabetic retinopathy, renal failure, Sexual dysfunction, skin diseases, high blood pressure, atherosclerosis, stroke, heart disease, gangrene and the like.

Meanwhile, in the composition for treating or preventing diabetic complications of the present invention, the aforementioned meanings are effective as they are related to the meaning of Korimae and seaweed extract, the use as a pharmaceutical composition, and the use as a food composition. Do.

Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these examples and experimental examples.

<Example> Preparation Example of Kori-mae and Seaweed Extract

Example 1 Preparation of Seaweed Soup Extract

<Example 1-1> Preparation Example 1 of seaweed iron extract

6 kg of wakame iron was washed with tap water to remove salt, lyophilized and ground. Crushed wakame iron was extracted twice with room temperature of 80% ethanol, concentrated under reduced pressure and lyophilized to remove the extraction solvent to prepare a powder of wakame iron powder.

<Example 1-2> Preparation Example 2 of seaweed iron extract

In the same manner as in <Example 1> to prepare a seaweed iron extract, methylene chloride was used as the extraction solvent instead of ethanol.

<Example 1-3> Preparation Example 3 of Wakame Iron Extract

In the same manner as in <Example 1> to prepare a seaweed iron extract, methanol was used as the extraction solvent instead of ethanol.

<Example 1-4> Preparation Example 4 of Wakame Iron Extract

In the same manner as in <Example 1> to prepare a seaweed iron extract, chloroform was used as the extraction solvent instead of ethanol.

<Example 1-5> Preparation Example 5 of Wakame Iron Extract

To prepare a seaweed iron extract in the same manner as in <Example 1>, butanol was used as the extraction solvent instead of ethanol.

<Example 1-6> Preparation Example 6 of Wakame Iron Extract

In the same manner as in <Example 1> to prepare a seaweed iron extract, hexane was used as the extraction solvent instead of ethanol.

Example 1-7 Preparation Example 7 of Seaweed Iron Extract

In the same manner as in <Example 1> to prepare a seaweed iron extract, ethyl acetate was used as the extraction solvent instead of ethanol.

<Example 1-8> Preparation Example 8 of the seaweed iron extract

In the same manner as in <Example 1> to prepare a seaweed iron extract, an extraction solvent was used instead of ether ethanol.

<Example 1-9> Preparation Example 9 of Wakame Iron Extract

The extract obtained in <Example 1-1> was suspended in distilled water, hexane (Hexane) was added to the solvent fractionation and the hexane fraction obtained by dark concentration was removed to remove the extraction solvent to obtain a hexane fraction in powder form.

<Example 1-10> Preparation Example 10 of Wakame Iron Extract

Solvent fractions other than the hexane fraction obtained in <Example 1-9> were solvent fractionated with ethyl acetate (EtOAc), and the obtained ethyl acetate fraction was concentrated under reduced pressure to remove the extraction solvent to obtain an ethyl acetate fraction in powder form.

Example 1-11 Preparation Example 11 of Seaweed Iron Extract

Solvent fractions other than the ethyl acetate fraction obtained in <Example 1-10> were solvent fractionated with butanol (BuOH), and the obtained butanol fraction was concentrated under reduced pressure to remove the extraction solvent to obtain a butanol fraction in powder form.

Example 2 Preparation of Ringberry Extract

6 kg of ring medium was washed with tap water to remove salt, and lyophilized and then ground. The pulverized ring medium was extracted with 80% ethanol twice at room temperature, concentrated under reduced pressure and lyophilized to remove the extraction solvent, thereby preparing a ring type extract in powder form.

Example 3 Preparation of Anthill Extract

6 kg of anthill iron was washed with tap water to remove salt, and lyophilized and then ground. The pulverized ants were extracted with 80% ethanol twice at room temperature, concentrated under reduced pressure, lyophilized to remove the extraction solvent, and ant extracts were prepared in powder form.

Example 4 Preparation of Long Bulrush Horse Extract

6 kg of a long fire end was washed with tap water to remove salts, and lyophilized and then ground. The pulverized long bulrush horsetail was extracted with 80% ethanol twice at room temperature, concentrated under reduced pressure, and lyophilized to remove the extraction solvent, thereby preparing a long bulgarae extract in powder form.

Example 5 Preparation of Basket Basket Extract

6 kg of the basket was washed with tap water to remove salt, lyophilized and ground. The pulverized net basket was extracted with 80% ethanol twice at room temperature, concentrated under reduced pressure and lyophilized to remove the extraction solvent to prepare a net basket extract in powder form.

Example 6 Preparation of Fireweed Extract

6 kg of end of fire was washed with tap water to remove salts, and lyophilized and then ground. The pulverized bulge horse was extracted with 80% ethanol twice at room temperature, concentrated under reduced pressure and lyophilized to remove the extraction solvent to prepare a bulge horse extract in powder form.

Experimental Example Hypoglycemic and Insulin-like Effects of Seaweed Iron Extract

<Experiment 1> Blood sugar drop test of Wakame iron extract

Four-week-old ICR male mice purchased from Orient Co., Ltd. were adapted for three weeks with solid feed and water, and then randomly selected 10 animals were divided into two groups and reared at random. In addition, the rest of the mice induced diabetes by intraperitoneally administering 40 mg / kg once a day for 5 days by dissolving streptozotocin, known to cause pancreatic beta cells to cause diabetes, in 0.01M citric acid (pH 4.5). Diabetes was confirmed 5 days after the intraperitoneal administration of streptozotocin for 5 days, then fasted for 4 hours, blood was collected from tail vein blood, and blood glucose was measured using a glucodoctor blood glucose meter (Allmedicus, Korea). Mice with blood sugar of 150 mg / dl or more were considered to be diabetic and used as a diabetic group.

The diabetic group was randomly divided into 10 groups of diabetic control group (normal feed) and each seaweed extract of the above example for 8 weeks by feeding and freely feeding with water, taking into account blood sugar and weight similarly. 1-1> to <Example 1-8> of the extract was supplied by mixing with feed at 1% by weight and the extract of <Example 1-9> to <Example 1-11> was mixed with feed at 0.5% by weight. Supply). The breeding environment was maintained at a temperature of 23 ± 2 ° C., a humidity of 50 ± 5%, and a night and day cycle for 12 hours.

Blood glucose in the blood was fasted for 4 hours while freely ingesting water, and blood was collected from the tail vein of the mice at the same time interval every week and then measured after 8 weeks using a glucodoctor blood glucose meter (Allmedicus, Korea).

Experimental results are shown graphically in FIGS. 1A-1J.

Referring to Figures 1a to 1j it can be seen that each of the seaweed extract of the embodiment has a hypoglycemic effect.

Experimental Example 2 Experiments Inducing Differentiation of Adipocytes

Experimental Example 2-1 Induction of Adipocyte Differentiation and Triglyceride Accumulation Experiment

The 3T3-L1 progenitor cell line was purchased from the American Type Culture Collection (ATCC) and cultured in Dulbecco's modified Eagle's medium (DMEM) medium with 10% bovine calf serum and 1% penicillin / streptomycin in a 37 ° C, 5% CO ² incubator. It was. Differentiation into adipocytes was carried out in 6 well plates at 2 × 10 ⁵ cells / well concentration in DMEM medium containing 10% fetal bovine serum (FBS) and 1% penicillin / stereptomysin. Differentiation induction medium (10% FBS, 1% penicillin / streptomycin, 1μM dexamethasone (DEX), 0.5 mM 3-isobutyl-1-methylxanthine (IBMX)) was replaced by induction for 2 days. Induced cells were incubated for 10 days with 10% FBS-DMEM medium, changing medium once every two days. The experimental group was treated with 25 ㎍ / ㎖, 50 ㎍ / ㎖, 100 ㎍ / ㎖ of seaweed extract of <Example 1-1> to <Example 1-8> and <Example 1-11> in the medium , The brown seaweed extract of <Example 1-9> was treated with 12.5 μg / ml, 25 μg / ml, 50 μg / ml and the brown seaweed extract of <Example 1-10> was 5 μg / ml and 10 μg /. Induction of differentiation into adipocytes by treatment with ㎖, 20 ㎍ / ㎖ and the degree of triglyceride accumulation by the Oil Red O staining method to observe whether the Wakae extract extract has the same properties as insulin. For Oil Red O staining, cells were washed twice with phosphate-buffered saline (PBS), fixed with 10% formalin for 1 hour, and washed three times with distilled water. And dilute water 0.6% Oil Red O (Sigma, USA) diluted with isopropanol and distilled water to 6: 4, filtered and treated the medium and stained for about 1 hour. Here, the -Control group was incubated in 10% FBS-DMEM medium for 8 days in confluent cells, and the + Control group was incubated for 2 days in adipose cell differentiation induction medium that was not treated with Wakame extract of Example 6 days. The medium was changed every 10 days with 10% FBS-DMEM medium, and Rosi was treated with 5 μM of thiazolidinedione-based rosiglitazone, a diabetes treatment, to be cultured in the same manner as the experimental group.

Quantitative analysis of the accumulation of triglycerides was performed by washing the cells treated with Oil Red O three times with distilled water, eluting the dye with isoprophanol containing 4% NP-40, and then using ELISA (Bio-Tek, USA) at 520 nm. The absorbance was measured using

The results are shown in FIG.

Figure 2a is a photograph of the plate stained with the Oil Red O dye by the extraction solvent, and Figure 2b is a graph showing the extraction solvent by quantitatively analyzing the degree of triglyceride accumulation.

In FIG. 2A, -C is a picture of -Control, + C is a picture of + Control, and Rosi is a picture of Rosiglitazone treated.

In FIG. 2B, con. Is the case of the + Control group in FIG. 2A.

Referring to FIGS. 2A and 2B, it can be seen that almost all seaweed extract extracts 3T3-L1 precursor adipocytes differentiating adipocytes and promoting accumulation of triglycerides in a concentration-dependent manner.

Experimental Example 2-2 Induction of Adipocyte Differentiation and Triglyceride Accumulation Experiment 2

The experiment was conducted in the same manner as in <Experimental Example 2-1>, but the extracts of <Examples 2 to 6> were used, and the concentrations of the extracts were 50 µg / ml, 100 µg / ml and 200 µg / ml. .

The results are shown in FIG.

Figure 3a is a photograph taken by the extraction target plate stained with Oil Red O dye, Figure 3b is a graph showing the extraction by quantitative analysis of the accumulation of triglycerides.

In FIG. 3A, -C is a picture of -Control, + C is a picture of + Control, and Rosi is a picture of Rosiglitazone treated.

In FIG. 3B, con. Is the case of + Control group in FIG. 3A and Rosi is the case of rosiglitazone treatment group.

Referring to Figures 3a and 3b, almost all the extracts can be seen that the concentration-dependent 3T3-L1 precursor adipocytes differentiate the adipocytes and promote the accumulation of triglycerides.

Experimental Example 2-3 Glucose Uptake of Adipose Cells

In the glucose absorption experiment, 3T3-L1 adipocytes differentiated from more than 90% obtained in <Experimental Example 2-1> were washed with serum free DMEM medium, and then cultured in the same medium for 2 hours, followed by Krebs-Ringer-Hepes (KRP). ) Was washed with buffer and incubated for 30 minutes in the same buffer. 37 MBq / L 2-deoxy-D with or without any 1 μM / L insulin (Sigma) and / or 50 mg / ml, 100 mg / ml, 200 mg / ml seaweed extract of Example 1-1 -KRP buffer containing [ ³ H] glucose and 1mM glucose was reacted at 37 ℃ for 10-20 minutes. Thereafter, the cells were washed with PBS buffer to stop the glucose uptake reaction, and the cells were homogenized with 1% Triton X-100 solution. Homogenized cells were measured for glucose uptake with a scintillation counter.

In this experiment, only the seaweed iron extract of <Example 1-1> was used as a representative.

The results are shown in FIG.

Referring to Figure 4 it can be seen that the wakame iron extract of <Example 1-1> promotes the absorption of glucose of adipocytes in a concentration-dependent manner. In addition, it can be seen that the Wakame iron extract of <Example 1-1> synergistically interacts with insulin in the absorption of glucose.

Experimental Example 2-4 Activity Test for PPAR-γ

The activity against PPAR-γ of Wakame Iron Ethanol Extract of Example 1-1 was evaluated in COS-1 cells using a luciferase assay. Dispense 96 × wells into 2 × 10 ⁴ cells per well at 37 ° C, 5% CO ₂ Incubated for 24 hours under conditions. PM-hPPARγ (plasmid expressing a fusion protein of 0.2 μg / well, GAL4 DNA-binding domain and human PPAR-γ ligand-binding domain), p4xUASg according to the proposed protocol using FuGENE 6 transfection reagent (Roche, USA) -tk-luc (0.2 μg / well, reporter plasmin containing four repeating upstream activating sequences for the GAL4 DNA-binding domain and thymidine kinase gene promoter before luciferase cDNA), pRL-CMV (0.2 ng / well) and pSK A reporter vector mixture consisting of -CBP (0.2 μg / well) was introduced into the cells. After 24 hours, the seaweed iron ethanol extract of <Example 1-1> was added to the medium at a concentration of 1 μg / ml, 5 μg / ml and 10 μg / ml with troglitazone or rosiglitazone 10 μM, which is one of the antidiabetic agents, for 24 hours. It was further cultured. Luciferase activity was measured with a luminometer (Berthold, Germany) using the Dual-Glo Luciferase Activity kit (Promega, USA).

In this experiment, only the ethanol extract of <Example 1-1> was used as a representative.

The results are shown in FIG.

Referring to FIG. 5, it can be seen that the Wakame iron extract of <Example 1-1> increased PPAR-γ transcriptional activity in a concentration-dependent manner even in a luciferous reporter assay analyzed in COS-1 cells. In addition, it can be confirmed that the Wakame iron extract of <Example 1> synergistically interacts with troglitazone or rosiglitazone, which is a concentration-dependent drug of thiazolidinedione.

<Statistical processing>

All experimental results were expressed as mean ± standard deviation, and statistical significance was analyzed by student's t-test. The in vivo test results were expressed as mean ± standard error using the SPSS statistical program, and Duncan's multiple range test was performed with one-way ANOVA test to test the significant difference between the experimental groups at p <0.05 level.

As described above, the present invention can provide a diabetic improver composition. Diabetes improver composition of the present invention is expected to be a treatment or prevention of diabetes with little side effects by including a natural seaweed extract as an active ingredient.

Claims (16)

Diabetes improver composition comprising an extract of Korimae and seaweed as an active ingredient. The method of claim 1, Said hookberry and seaweed extract is diabetic improver composition, characterized in that the extract selected from the group consisting of seaweed extract, hookberry extract, anthill extract, long bulrush horse extract, net basket extract, and bulgarae extract. The method of claim 1, The ringberry and algae extract is a diabetic improver composition, characterized in that the seaweed extract. The method of claim 1, The ring berry and seaweed extract is an diabetes improving composition, characterized in that the extract obtained by extraction with a polar solvent or a non-polar solvent. The method of claim 3, The seaweed iron extract is a diabetic improver composition, characterized in that the extract obtained by extraction with a polar solvent or a non-polar solvent. The method of claim 1, The extract of algae and algae is an extract obtained by extracting with alcohol or mixed solvent of water or distilled water, alcohol, ethyl acetate, acetone, chloroform, methylene chloride, benzene, alkanes having 5 to 10 carbon atoms, toluene, or kerosene. Diabetes improver composition. The method of claim 3, The seaweed iron extract is an extract obtained by extraction with alcohol, mixed solvent with water or distilled water, alcohol, ethyl acetate, acetone, chloroform, methylene chloride, benzene, alkanes having 5 to 10 carbon atoms, toluene, or kerosene. Diabetes improver composition. The method of claim 1, The ring mae and seaweed extract is diabetic improver composition, characterized in that obtained by extraction with a mixed solvent of ethanol or water or distilled water of ethanol. The method of claim 3, The Wakame iron extract is diabetic improver composition, characterized in that obtained by extraction with a mixed solvent of ethanol or water or distilled water of ethanol. The method according to claim 1 or 3, The composition is an diabetes improving composition, characterized in that it further comprises an insulin agent or thiazolidinediones-based drugs. The method of claim 1, The composition is a diabetic improver composition, characterized in that the pharmaceutical composition. The method of claim 1, The composition is a diabetic improver composition, characterized in that the food composition. A composition for preventing diabetic complications comprising Korimae and seaweed extract as an active ingredient. The method of claim 13, The composition is a diabetic complication prevention composition, characterized in that the pharmaceutical composition. The method of claim 13, The composition is a diabetic complication preventive composition, characterized in that the food composition. The method of claim 13, Diabetic complications are neurological complications, diabetic retinopathy, renal failure, sexual dysfunction, skin diseases, hypertension, atherosclerosis, stroke, heart disease or necrotic composition, characterized in that the necrosis.

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