KR20140084928A - Composition for treating diabete and diabete-induced complication containing an extract from Agrimonia pilosa - Google Patents

Abstract

The present invention relates to a composition including an agrimonia pilosa extract as an effective element for treating or preventing diabetes and diabetic complication and, more particularly, to a composition including an agrimonia pilosa extract or a fraction thereof as an effective element for preventing or treating diabetes and diabetic complication.

Description

[0001] The present invention relates to a composition for preventing or ameliorating diabetes and diabetic complications comprising an extract of strawberry and herb as an active ingredient.

The present invention relates to a method for producing the < RTI ID = The present invention relates to a composition for preventing and / or ameliorating diabetic and diabetic complications comprising, as an active ingredient, an extract of pilosa or a fraction of the above extract.

Diabetes mellitus is a chronic disease that has been attracting attention worldwide because of the recent increase in the level of living standards of modern people and the increase in the longevity of human life.

Diabetes mellitus is a metabolic syndrome characterized by a deficiency of insulin hormone or insulin resistance in the beta cells of the pancreas, as well as hyperglycemia resulting from both of these defects. Such diabetes can be divided into insulin-dependent diabetes mellitus (IDDM) and insulin-dependent diabetes mellitus (NIDDM) (Type II) caused by insulin resistance and insulin secretory damage.

Diabetes mellitus, chronic hyperglycemia, as well as lipid metabolism, lipid and protein metabolic disorders are also caused. The condition is diverse and is directly caused by hyperglycemia, and includes diabetic peripheral neuropathy, diabetic retinopathy, diabetic nephropathy, diabetic cataract, keratosis, and diabetic atherosclerosis in the retina, the kidney, the nervous system, and the cardiovascular system. The number of diabetic patients is steadily increasing due to changes in lifestyle such as overeating, lack of exercise, westernized eating habits, stress, drinking and smoking.

In Type I and Type II diabetes, diverse complications such as heart disease, bowel disease, ophthalmic disease, neurological disease, and stroke occur, which can lead to chronic neuropathy and cardiovascular disease due to elevated blood glucose and insulin levels over a long period of time And short-term hypoglycemia and hyperglycemia reaction cause acute complications.

Diabetes mellitus is medication, exercise and diet, and insulin medicines and various blood glucose drugs are used depending on the patient's symptoms. However, diabetes is a complex disease characterized by overgrowth of glucose in the liver, insulin resistance, and decreased glucose tolerance in muscle and adipocytes. Therefore, specific treatment alone can not prevent the occurrence of various side effects. Because of the use of insulins and chemicals, drug therapy is a constant problem for drug side effects and patient resistance. Recently, diabetes treatment has been used to treat diabetes, And to further prevent and treat diabetic complications.

Accordingly, the inventors of the present invention discovered that the antioxidative effect of the strawberry herb has excellent effects in the treatment of diabetic and diabetic complications, thus completing the present invention.

Accordingly, one example of the present invention provides a composition for preventing and / or ameliorating diabetic and diabetic complications, comprising at least one member selected from the group consisting of extracts and fractions of strawberry extract as an active ingredient.

Another example provides a composition for preventing and / or ameliorating diabetic and diabetic complications comprising at least one selected from the group consisting of extracts and fractions of strawberry seeds.

Another example provides an antioxidant composition containing at least one selected from the group consisting of extracts and fractions of a strawberry herb as an active ingredient.

Hereinafter, the present invention will be described in more detail.

There is provided a composition for preventing and / or ameliorating diabetic and diabetic complications comprising, as an active ingredient, a strawberry herb extract and / or a fraction of the above extract.

It is a perennial herbaceous plant belonging to the rose family, and it is known that 10 species are distributed in the temperate zone of the northern hemisphere and Brazil and South America. In Korea, it grows frequently in the roadside of the mountains and fields, and it is also called Seonghakcho, Yonga, Yongaeko, Hwanghaeko, Hwangyongjang, Gijunseok. Its height grows about 15 ~ 60 cm. It has white hairs all over and has a leaflet shape or long elliptical leaflet staggered. Small flower of yellow gathering on flower stand in June ~ August. Traditionally, it has been widely used for folk medicine, green juice, etc. and has been widely used for medicines such as lung cancer, liver cancer, esophageal cancer, tumor, pain relief, topography, branching, blood, hematoma, uterine bleeding and fever. In recent years, many substances from strawberry herb extracts have been used as pharmaceutical raw materials, health foods, wild plants and vegetable nutrients. Vitamin K, tannins, essential oils, luteolin-7-glucosidase, apigeni-7- glucosidase, phenolic glycosides, agrimoniin, agrimonolide, etc. are known, and the anticancer effect of agrimoniin is well known.

In the present invention, Agrimonia pilosa (Agrimonia pilosa ) was used as a short - term screening method for the prevention of diabetic complications. The inhibitory activity of alpha - glucosidase was confirmed as a short - term screening method for inhibiting the formation of advanced glycation endproducts and preventing diabetes.

There is no particular limitation on the form of the strawberry herb used in the present invention, and the composition of the present invention is characterized by containing an extract and / or a fraction obtained from the strawberry herb as an active ingredient. The strawberry extract may be obtained by a conventional method of extracting straws and herbs using at least one solvent selected from the group consisting of water and C ₁ to C ₄ linear or branched alcohols or mixtures thereof.

       In one embodiment of the present invention, in order to achieve better antidiabetic and antidiabetic effects, the extract may be an ethanol extract obtained by extracting 60-80% (v / v) ethanol of a strawberry herb, The fraction may be obtained by fractionating the above extract into at least one selected from the group consisting of hexane, methylene chloride, ethyl acetate and butanol. More preferably, the fraction may be selected from the group consisting of fractions obtained by applying at least one selected from the group consisting of hexane, methylene chloride, ethyl acetate and butanol in the order of polarity, . For example, the hexane fraction obtained by fractionating the ethanol extract with hexane, the methylene chloride fraction obtained by fractionating the hexane fraction, and the remaining aqueous layer fractionated with methylene chloride. The methylene chloride fraction is separated and the remaining aqueous layer is fractionated with ethyl acetate to obtain the ethyl acetate fraction The butanol fraction obtained by separating the ethyl acetate fraction and the remaining aqueous layer with butanol, and the water fraction remaining after separating the butanol fraction.

        The strawberry herb extract may be produced according to a conventional method for producing plant extracts, and may be preferably a pressure extraction method, a reflux extraction method, a warm-up extraction method, an ultrasonic pulverization extraction method and the like, , But is not limited thereto.

In addition, the extract prepared as described above can be concentrated or removed by concentration filtration and / or lyophilization in a subsequent vacuum filtration step. Therefore, the strawberry herb extract in the present invention is used to mean a dried product dried by a conventional method and a concentrate concentrated in a conventional manner.

The present invention provides a therapeutic effect of diabetic and diabetic complications of the extract of strawberry and herbal compositions and the fraction composition.

Diabetic complications are diabetic complications that lead to hyperglycemia and protein reactions that irreversibly progress to produce advanced glycation endproducts (AGEs) that cross-link with other proteins in the blood or tissue. Specifically, the diabetic complications in which the extracts and / or fractions of the present invention have therapeutic effects may be diabetic vascular disorders, diabetic neuropathy or diabetic infections, and preferably diabetic retinopathy, diabetic retinopathy, May be diabetic cataract, diabetic nephropathy, diabetic neuropathy, diabetic vascular complication, and the like, more preferably diabetic retinopathy, diabetic cataract or diabetic nephropathy .

Alpha-glucosidase is the most important carbohydrase and facilitates the degradation of the glucose linkage at the end of the carbohydrate digestion process. The alpha-glucosidase inhibitor (AGI) has an effect of lowering digestion and absorption of carbohydrates in the duodenum and the like, and thus can be used for the prevention and treatment of adult diseases such as diabetes, obesity, and hyperglycemia.

The therapeutic activity of the present invention for diabetic complications was tested by inhibiting the formation of Advanced Glycation Endproducts (AGEs) and inhibiting alpha-glucosidase activity.

The final glycation products are formed by non - enzymatic glycation through sugar - protein binding and schiff base and amadori product. However, in a chronic hyperglycemic state, perhaps a lipid product is reversibly accelerated by a irreversible reaction that produces a final glycation product and is mass-produced and accumulated in tissues and serum. Thus, the production of the final glycated product is one of the major causes of severe diabetic complications.

It is known that the concentration of the final glycated product in the blood of patients with type 2 diabetes is highly correlated with the presence or severity of cardiovascular disease and also correlates with various animal model studies. The end glycation products produced by non-enzymatic glycation of chronic hyperglycemia and biologic proteins are one of the major causes of diabetic complications. Diabetic complications often occur despite the normal recovery of blood sugar. The final glycation product is a protein glycosylation product accumulated in tissues in proportion to the concentration of hyperglycemia, and exhibits a specific absorbance and fluorescence. Especially, amino groups of proteins cause cross-link reaction with each other. Once the final glycation products are produced, they are not degraded even when blood glucose is normally regulated, and accumulate in the tissues, abnormally changing the structure and function of the tissue, and are known to be involved in atherosclerosis, DNA mutation, senescence, Alzheimer's disease and diabetic complications have.

The toxicity of aminoguanidine (aminoguanidine), which is known as an inhibitor of the final glycation end product, has been shown to increase. Therefore, it is necessary to develop a safe natural product free of toxicity.

As a result of testing the inhibitory activity against the final glycation products of the extracts and fractions of the strawberry herb extract according to the present invention, they were found to have an effect of inhibiting the final glycation end product formation (see Table 1 and Fig. 1).

The extracts and / or fractions of the strawberry herb according to the present invention showed excellent antidiabetic effect.

Alpha-glucosidase is a digestive enzyme present in the villi of the small intestine and serves to hydrolyze disaccharides and small sugars to monosaccharides which are digestion and absorption state of the monohydrate. Therefore, the alpha-glucosidase inhibiting substance inhibits the hydrolysis of carbohydrates (lowering digestion absorption rate) at the upper part of the plant including the duodenum, thereby suppressing the increase of blood glucose level. Therefore, . ≪ / RTI >

The anti-diabetic effect was evaluated by testing the alpha-glucosidase inhibitory effect of the strawberry herb extract and / or fractions. As a result, the strawberry herb extract and / or fraction showed excellent antidiabetic effect (see Table 2 and FIG. 2).

The extracts and / or fractions of the present invention have excellent antioxidant activity. DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging effect was tested to test the antioxidant effect of the composition of the present invention. DPPH is a chemically stable compound with free radicals and is widely used to confirm antioxidant ability when it is contacted with a substance having antioxidant activity. The electron donating action using DPPH is used for the purpose of inhibiting lipid oxidation in foods by donating electrons to free radicals and is being used in the body to suppress aging by free radicals.

The DPPH radical scavenging activity of the extracts and / or fractions of the strawberry herb extracts was evaluated and the antioxidant activity was evaluated. As a result, the extracts and / or fractions of the strawberry herb extracts showed excellent antioxidant ability (see Table 3 and FIG. 3).

       The ABTS radical scavenging activity method developed on the basis that the absorbance of the cation radical of ABTS is inhibited by the antioxidant can be compared with the value of the standard substance Trolox and it is possible to measure not only the antioxidant activity in vivo but also the antioxidant activity And is widely used as a method for measuring performance.

When ABTS and potassium persulfate are left in cows and ABTS + is produced, ABTS + is cleaved by the antioxidant power of the extract, and decolorization of the specific cyan color of the radical is discontinued. The ABTS + of ABTS + can be measured by the absorbance value.

The antioxidant activity of the extracts and / or fractions of the strawberry herb extracts was evaluated by ABTS radical scavenging ability. As a result, the extracts and / or fractions thereof showed excellent antioxidant ability (see Table 4 and FIG. 4).

The composition of the present invention can be used as a nutritive agent, a vitamin, an electrolyte, a flavoring agent, a coloring agent, a thickening agent, a pectic acid and its salt, an alginic acid and its salt, an organic acid, a protective colloid thickener, a pH Stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. The components can be added independently or in combination. The content of the additional ingredient is preferably in the range of 0.1 to 20 parts by weight per 100 parts by weight of the extract or fraction of the sandalwood extract or in the range of 100 to 10,000,000 parts by weight per 100 parts by weight of lycoricidine, no.

The content of the extracts and / or fractions of the present invention may be appropriately adjusted according to the symptoms of the disease, the progress of the symptoms, the condition of the patient and the like, for example, 0.0001 to 99.9% by weight, Preferably 0.001 to 50% by weight, but is not limited thereto. The content ratio is a value based on the dried amount from which the solvent is removed.

The composition may further contain an appropriate carrier, excipient and diluent conventionally used in the production of a pharmaceutical composition. The composition may be administered orally or parenterally in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, An external preparation, a suppository or a sterilized injection solution, and the like.

When the composition is formulated, it is prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations may contain at least one excipient and / or lubricant, etc. Liquid formulations for oral administration include, Suspensions, solutions, emulsions, syrups, and the like. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, suppositories, and the like.

The preferred dosage of the composition varies depending on the condition and the weight of the patient, the degree of the disease, the drug form, the administration route and the period, but can be appropriately selected by those skilled in the art. For the more preferable effect, the dose of the composition of the present invention is preferably from 0.1 mg / kg to 20 mg / kg per day on the basis of the active ingredient, but it is not limited thereto. Several doses can be administered. The compositions of the present invention may be administered to a mammal, including a human, in various ways. All modes of administration may be expected, for example, by oral, intravenous, intramuscular, subcutaneous injection, and the like. The pharmaceutical dosage form of the composition of the present invention may be used in the form of a pharmaceutically acceptable salt of the active ingredient, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable combination.

In another embodiment, there is provided a composition for preventing and / or ameliorating diabetic and diabetic complications comprising the extracts and / or fractions of strawberry as described above. In the present invention, the term " food " means a natural product or a processed product containing one or more nutrients, preferably a state of being able to be directly eaten through a certain processing step, , Health functional foods, beverages, food additives and beverage additives. Examples of the food include various foods, beverages, gums, tea, vitamin complex, and functional foods. In addition, in the present invention, the food may contain special nutritional foods (e.g., crude oil, spirits, infant food, etc.), meat products, fish products, tofu, jelly, noodles (Such as soy sauce, soybean paste, hot pepper paste, mixed sauce), sauces, confectionery (eg snacks), dairy products (eg fermented milk, cheese), other processed foods, kimchi, pickled foods But are not limited to, natural flavors (eg, ramen soup, etc.), vitamin complexes, alcoholic beverages, alcoholic beverages and other health supplement foods. The health functional food, beverage, food additive or beverage additive can be produced by a usual production method.

The food composition may comprise a pharmaceutically acceptable food-aid additive and may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of a functional food.

In addition, the amount of the strawberry extract and / or fraction may be in the range of 0.00001% to 50% by weight of the total food, and when the food is a beverage, the amount of the extract is preferably 0.001 g to 50 g, preferably 0.01 g to 10 g, but is not limited thereto.

The composition comprising the extract of strawberry and herb according to the present invention is a composition derived from a natural product which is excellent in the therapeutic effect of diabetic and diabetic complications and has no toxicity and side effects, and thus can be safely applied to the treatment of diabetic and diabetic complications.

FIG. 1 is a straw herbs (Agrimonia pilosa) (a graph showing the AGEs) Inhibitory effect (Each bar represents the meanSD (n = 3) extract and advanced glycation end products of the fractions; Values with different letters are significantly different (p <0.05); AG: aminoguanidine; 70 % EtOH ex .: 70% ethanol extract n- Hex fr .: n -hexane fraction MC fr .: methylene chloride fraction EtOAc fr .: ethyl acetate fraction water n- BuOH fr .: n -butanol fraction DW fr .: water fraction).
FIG. 2 is a graph showing the effect of inhibiting alpha-glucosidase activity on the extract of strawberry and herb extracts compared with the extract of Nobleman's stem extract. (Each bar represents the mean SD (n = 3); Values with different letters are significantly different (p <0.05); N- BuOH fr .: n -hexane fractions; MC fr .: methylene chloride fractions; EtOAc fr .: ethyl acetate fractions; water n- BuOH fr .: n -Butanol fraction; DW fr .: water fraction).
Figure 3 is a graph depicting the DPPH radical inhibition of the extracts and fractions of strawberry seeds. 70% EtOH ex .: 70% Ethanol (n = 3); Values with different letters are significantly different Extract: n- Hex fr .: n -hexane fraction, MC fr .: methylene chloride fraction, EtOAc fr .: ethyl acetate fraction, water n- BuOH fr .: n -butanol fraction, DW fr .: water fraction).
FIG. 4 is a graph showing the ABTS radical inhibition rates of the extracts and fractions of strawberry herbs. 70% EtOH ex .: 70% Ethanol (n = 3); Values with different letters are significantly different Extract: n- Hex fr .: n -hexane fraction, MC fr .: methylene chloride fraction, EtOAc fr .: ethyl acetate fraction, water n- BuOH fr .: n -butanol fraction, DW fr .: water fraction).

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these examples are merely illustrative of the present invention, and the scope of the present invention is not limited by these examples.

Material preparation

Agrimonia pilosa was purchased from Gangwon Province and Yanggu Kuk, and was purchased from DPGH (1,1-diphenyl-2-picrylhydrazyl), DMSO, BSA (Albumin from bovin serum), Methylglyoxal, Aminoguanidine was purchased from Sigma (USA), -glucosidase was purchased from Wako (Japan), and all the reagents used for the analysis were Express.

Sample Preparation Example 1

The strawberry herb extracts and fractions of the present invention are prepared by adding 10 times 70% ethanol of 1000 g of the dried strawberry herb, and extracting it in an aqueous solution at 65 for 3 hours, followed by filtration to obtain an extract. The extraction process is repeated three times. The obtained extract is concentrated under reduced pressure at 40 or less and then lyophilized to obtain a powdery extract.

Sample Preparation Example 2

The sandwich herb obtained in the above-mentioned Sample Preparation Example 1 To 100 g of the extract, 500 L of distilled water was added, suspended, 500 L of hexane was added thereto, and the mixture was separated into a hexane-soluble fraction and a water-soluble fraction. The same procedure was repeated three times to collect only the hexane soluble fraction, followed by drying under reduced pressure to obtain hexane soluble extract powder. The methylene chloride, ethyl acetate, butanol soluble fraction and water soluble fraction were collected and dried under reduced pressure Powder was obtained.

       Example 1. Inhibitory effect test of the final glycation products (AGEs)

In order to test the therapeutic effect of diabetic complications in this example, the effect of inhibiting the final glycation products (AGEs) of the extracts and fractions of straw sandal herb extracts by the 24-hour short-term screening method using methylglyoxal was evaluated by increasing the fluorescence Were measured.

A sample (11 mg / mL) obtained by dissolving DMSO (dimethylsulfoxide) in BSA (5 mg / mL) and methylglyoxal (2 mg / mL) (the extract and fraction of the strawberry herb obtained in Example 1 above) was added and dissolved in 50 mM phosphate buffer (pH 7.4) was used to adjust the total volume to 5 mL. (355 nm, emission: 460 nm) were measured before and after the reaction using a spectrofluorometer (Fluoroskan Ascent FL, Thermo Scientific, USA). The inhibition rate of the absorbance was calculated by comparing the absorbance increase value of the sample based on the absorbance increase value of the control group.

Equation 1 = (S ₁ -S ₀ ) / (C ₁ -C ₀ ) * 100

(S ₀ : sample 0hr absorbance response value, S ₁ : sample 24hr absorbance response value, C ₀ : control 0hr absorbance response value, and C ₁ : control 24hr absorbance response value). In addition, fluorescence inhibition rates were compared using aminoguanidine (Sigma Chemical Co., St. Louis, Mo., USA) as a standard substance known to have an inhibitory effect on the final glycation end product.

The obtained results are shown in Table 1 and Fig. In the case of strawberry herb extract, the 70% ethanol extract showed a 50.59% inhibition effect at the concentration of 11 mg / mL and the EtOAc fraction in the fraction was 11 mg / mL And 85.42%, respectively.

Table 1. Inhibitory effect of extracts and fractions of strawberry herb extracts on the formation of AGEs Sample Concentration
(mg / mL) Inhibition (%) 70% EtOH ex. 11 50.590.90 n- Hex fr. 11 17.063.12 M.C fr. 11 71.872.79 EtOAc fr. 11 85.4233.77 n- BuOH fr. 11 66.062.71 D.W fr. 11 1.160.04 Aminoguanidine 1) 11 85.020.43

¹⁾ Positive control substances (conventional inhibitors of AGE formation)

Example  2. Alpha- Glucosidase Inhibition  exam

The active principle of alpha-glucosidase is the conversion of p- nitrophenyl-Alpha-D-glucoside into alpha-D-glucose and p- nitrophenol by alpha-glucosidase. Yellow can be obtained by measuring the final absorbance using a spectrophotometer at a wavelength of 405 nm

The samples used in this experiment were dissolved at a concentration of 1 mg / mL. -Glucosidase was dissolved in phosphate buffer (pH 7.0) to prepare enzyme solution. 10 L of sample was added to 60 L of enzyme solution, pre-incubated at 37 for 15 min, and p- nitrophenyl-D-glucopyranoside was dissolved in phosphate buffer ), 30 L of a substrate solution was added and the absorbance was measured at 405 nm. Acarbose was used as a positive control, and each sample was repeated three times and averaged.

The obtained results are shown in Table 2 and Fig. The results showed that 70% ethanol extract showed the inhibition of 82.79% at the concentration of 1 mg / mL, and that the EtOAc fraction in the fractions was 1 mg / mL And 98.25%, respectively.

       Table 2. Alpha-glucosidase inhibition of strawberry herb extracts and fractions Sample Concentration
(mg / mL) Inhibition (%) 70% EtOH ex. One 82.790.90 n- Hex fr. One 12.821.11 M.C fr. One 37.653.30 EtOAc fr. One 98.251.31 n- BuOH fr. One 75.821.01 D.W fr. One 69.241.11 Acarbose ¹⁾ One 37.871.21

¹⁾ Positive control substances (conventional alpha-glucosidase inhibitors)

Example  3. DPPH Antioxidant Effect Test

DPPH is a chemically stable compound with free radicals and is widely used to confirm antioxidant ability when it is contacted with a substance having antioxidant activity. Thus, in this example, the antioxidative effect of DPPH was evaluated by evaluating the free radical scavenging effect (hydrogen donating ability against DPPH) of the extracts and fractions of strawberry herb extracts and fractions.

The strawberry herb extract and each fraction were diluted with DMSO (dimethyl sulfoxide). To a 96-well microplate, 270 L of 5 10 ^-4 M DPPH solution (dissolved in 99% ethanol) was added to 30 L of the sample prepared at a constant concentration and left to stand at room temperature for 30 minutes. The resulting reaction solution was measured by electron donating ability at an absorbance of 570 nm. The inhibitory activity of each fraction was calculated from the reduced absorbance versus the positive control.

Formula = 1- (Control / Sample) * 100

At this time, ascorbic acid (Showa Chemicals Inc., Japan), which is known to have an antioxidative effect, was used as a positive control for the activity comparison, and DPPH radical scavenging activity was measured by the same method as described above.

Table 2 shows the results of DPPH radical scavenging ability of the extracts and fractions of straw sandal herb extracts. 3 times repeatedly.

The obtained results are shown in Table 3 and FIG. The results showed that the 70% ethanol extract showed a DPPH radical scavenging ability of 31.26% at the concentration of 1 mg / mL, and the EtOAc fraction in the fractions showed 1 mg / mL, the highest DPPH radical scavenging activity of the fractions was 54.02%.

 Table 3. DPPH radical scavenging effect of extracts and fractions of strawberry herb extract Sample Concentration
(mg / mL) Inhibition (%) 70% EtOH ex. One 31.265.17 n- Hex fr. One 33.742.31 M.C fr. One 40.330.08 EtOAc fr. One 54.021.21 n- BuOH fr. One 32.140.78 D.W fr. One 35.841.89

Example  4. ABTS Antioxidant Effect Test

The ABTS radical scavenging experiment is based on the formation of ABTS ^{O +} (ABTS radical cation), which is characterized by the colorless, reddish ABTS is oxidized. This cyan ABTS ^{O +} can be assayed for its antioxidant capacity by reacting with oxidizable substances and reducing the absorbance of the reacted material to the original colorless ABTS

        The ABTS radical scavenging activity of the sample was measured as follows. ABTS radicals were generated by reacting 7.4 mM ABTS and 2.6 mM potassium persulfate in a dark place for 12 hours. A sample of 10 L extract was added to each well of a 96-well plate, and 290 L of ABTS solution was added. The reaction was allowed to proceed for 30 minutes and the absorbance at 750 nm was measured. The degree of decrease of the absorbance by the amount of radical scavenging was measured, and the radical scavenging ability was calculated by the following equation.

         Formula = 1- (Control / Sample) * 100

For activity comparison, trolox (Calbiochem, Canada), which is known to have an antioxidative effect, was used as a positive control, and ABTS radical scavenging activity was measured by the same method as described above.

The obtained results are shown in Table 4 and Fig. As a result of the measurement of the fluorescence increase degree based on the blank and the control group, the 70% ethanol extract showed a ABTS radical scavenging ability of 67.48% at a concentration of 1 mg / mL in the case of the strawberry herb extract, and the EtOAc fraction layer of 1 mg / mL, the highest ABTS radical scavenging activity of the fractions was shown to be 92.54%.

Table 4. ABTS radical scavenging effects of extracts and fractions of strawberry herb extracts Sample Concentration
(mg / mL) Inhibition (%) 70% EtOH ex. One 67.481.85 n- Hex fr. One 72.350.49 M.C fr. One 82.602.11 EtOAc fr. One 92.540.12 n- BuOH fr. One 73.580.80 D.W fr. One 61.031.24

Statistical processing

Statistical analysis of all data was performed using the SAS statistical program (SAS institute, 1987). The results were expressed as means and standard errors per experiment group. The significance of the treatment groups was tested at the p <0.05 level by the Duncans multiple range test.

Claims (7)

An extract obtained by extracting a strawberry herb with at least one selected from the group consisting of water and a straight chain or branched alcohol or a mixture of C ₁ to C ₄ and an extract obtained by extracting the extract from water, hexane, methylene chloride, ethyl acetate and butanol And fractions obtained by fractionating at least one selected from the above-mentioned fractions as an active ingredient, a composition for preventing or ameliorating diabetic and diabetic complications The extract of claim 1, wherein the extract is obtained by extracting 60 to 80% (v / v) ethanol of a strawberry herb and the extract is fractionated into at least one selected from the group consisting of hexane, methylene chloride, ethyl acetate and butanol A composition for preventing or improving diabetic and diabetic complications, containing at least one selected from the group consisting of the obtained fractions as an active ingredient An extract obtained by extracting strawberry seeds with 60 to 80% (v / v) ethanol The hexane fraction obtained by fractionating the above extract with hexane and the hexane fraction are separated and the aqueous layer remaining is fractionated with methylene chloride Methylene chloride fraction and the methylene chloride fraction are separated and the remaining aqueous layer is separated into ethyl acetate and ethyl acetate fraction and the remaining water fraction is separated from the ethyl acetate fraction, Composition for preventing or improving diabetic complications 4. The composition according to any one of claims 1 to 3, wherein the strawberry herb is a composition for preventing or improving diabetic complications due to diabetes The composition according to any one of claims 1 to 3, wherein the diabetic complication is a composition for preventing or improving diabetic complications characterized by diabetic vascular disorder, diabetic neuropathy or diabetic infectious disease 6. The method of claim 5, wherein the diabetic complication is characterized by diabetic retinopathy, diabetic cataract, diabetic nephropathy, diabetic neuropathy, and diabetic vascular complications Composition for the prevention or amelioration of diabetic complications A composition for preventing or ameliorating diabetic and diabetic complications comprising the composition according to any one of claims 1 to 3

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