KR20110053141A - Composition for anti-diabetes comprising germinated brown rice extracts - Google Patents

Abstract

PURPOSE: A composition containing germinated brown rice extract for controlling serum glucose level is provided to suppress glucose absorption in the small intestine and to prevent or treat diabetes. CONSTITUTION: A composition for controlling serum glucose level contains solvent of germinated brown rice extract as an active ingredient. A method for preparing the germinated brown rice extract comprises: a step of drying germinated brown rice; a step of extracting the dried germinated brown rice with aqueous, organic solvent or mixture solvent thereof; a step of fractioning from non-polar solvent to polar solvent; and a step of drying the fraction.

Description

Composition for anti-diabetes comprising germinated brown rice extracts

The present invention relates to a composition for controlling serum glucose level comprising germinated brown rice extract as an active ingredient, and more particularly to a composition for preventing or treating diabetes, in particular insulin-independent diabetes.

Diabetes is a disease in which glucose in the blood is released through urine and is one of the chronic degenerative diseases that does not cure fundamentally. As life expectancy changes due to rapid economic development, life expectancy is increasing, whereas chronic degenerative diseases such as diabetes are increasing. The prevalence of diabetes in Korea is estimated to be about 5 to 10%. In 2007, the number of diabetics exceeded 4 million, 8.3 out of every 100 people with diabetes, but the number of diabetics is expected to increase to about 6.8 million by 2025. have. Diabetes is the third most serious disease in the world and the life span of patients is shortened by 5 to 10 years due to various complications, including complications.

Diabetes treatment is roughly divided into diet, exercise therapy and drug therapy and is determined by the patient's condition. Diabetes is more important to prevent and treat complications than the disease itself, so management can be said to maintain normal blood sugar and prevent complications. Meanwhile, diabetic drugs can be divided into three groups. ① Glycolytic inhibitors that inhibit sugar from being absorbed into the small intestine, ② Insulin resistance improving agent to help the action of insulin, ③ insulin secretagogues. Among these, glycolytic inhibitor oral hypoglycemic agents are mainly administered to patients with type 2 diabetes who do not have glycemic control by diet and exercise therapy. Many drugs have been developed in the meantime, but most of them have side effects, but only a few drugs are currently being used in the clinic because of their effectiveness or safety.

Currently, glycemic hypoglycemic agents, glymepiride, a sulfone urea agent, are mainly used, but hypoglycemia-induced loss of insulin secretion function is known as a side effect. Glucophage (glucophage), a biguanide-based drug, is a drug that is widely used in Korea, mainly to help insulin function in the liver to lower blood sugar, but side effects include gastrointestinal disorders, diarrhea, and lactic acidosis.

Alpha-glucosidase inhibitors reduce glucose uptake by inhibiting the breakdown of carbohydrates in the small intestine, and the drugs include acarbose, avoglibose, and miglitol. Boaz (acarbose) is currently used in Korea, boglibose (voglibose), miglitol (miglitol), etc. have recently been introduced in Korea, but side effects include diarrhea and upset stomach.

As a result of the side effects of the drug therapy, there is a growing trend to increase the anti-diabetic natural substances using food materials. Herb plants, such as mulberry leaves, coriander, ginseng, lotus flesh, evening primrose, gwanjaja, gojija, etc., soybeans, seaweeds, burdock, buckwheat and other foods are reported to have an effect on diabetes, or is being used as a folk remedy.

Brown rice ( Oryza sativa ) has less nutrients due to milling and is less preferred because of its low digestibility and poor texture, despite its richer vitamins, minerals and fiber than white rice. Recently, germinated brown rice products have been released to improve this. Germinated brown rice is a germination of brown rice to 1 ~ 5 mm by supplying proper temperature, moisture and oxygen, while maximizing the nutrition and function of brown rice, the taste is much softer than brown rice. Germinated brown rice is known to improve the cohesiveness as the brown rice shell is decomposed by the action of cell wall degrading enzymes during the germination process of brown rice to facilitate the absorption of moisture. Germinated brown rice contains GABA, vitamin E, squalene, sitosterol, campesterol, and stigmasterol as major constituents. Research on the utilization of various functional foods such as removing or removing them is being actively conducted. On the other hand, physiological and pharmacological activities have been reported to prevent hypertension and neurostable. In addition, research on the use of various functional food materials such as qualitative changes of stored protein during the germination, an increase in micronutrients, an increase in digestibility of constituent carbohydrates, and a reduction or elimination of semi-quantitative factors have been actively conducted. It has not been reported that there is an excellent effect of inhibiting the absorption of glucose in.

Under these backgrounds, the present inventors studied micromaterials having various physiological and pharmacological activities from germinated brown rice, and the number, extract, or organic solvent extract of germinated brown rice showed hypoglycemic action by inhibiting glucose absorption in the small intestine. By confirming that the present invention was completed.

It is an object of the present invention to provide a composition for regulating serum glucose levels comprising germinated brown rice water, an organic solvent or a mixed solvent extract thereof as an active ingredient. Here, the water used for the extraction may be cold or hot water.

Another object of the present invention is to provide a composition for the prevention or treatment of diabetes, in particular, insulin-independent diabetes, which has germinated brown rice extract as an active ingredient.

It is another object of the present invention to provide a composition for inhibiting glucose absorption in the small intestine having a germinated brown rice extract as an active ingredient.

Another object of the present invention is to provide a health functional food for diabetic disease improvement comprising the germinated brown rice extract as an active ingredient.

The present invention relates to a composition for controlling serum glucose level comprising the germinated brown rice extract as an active ingredient, specifically preventing the hypoglycemic agent diabetic comprising a germinated brown rice water, an organic solvent or a mixed solvent extract thereof as an active ingredient A therapeutic composition.

Germinated brown rice extract of the present invention is a food material that does not have toxicity, side effects, and the like, in particular, by inhibiting the absorption of glucose in the small intestine, not only can effectively prevent or treat diabetes, but also improve the health of various symptoms due to diabetes diseases. It can also be usefully used for nutraceuticals.

The present invention relates to a composition for regulating serum glucose level comprising germinated brown rice water, organic solvent or mixed solvent extract thereof as an active ingredient. The water used for the extraction may be cold or hot water.

Germinated brown rice extract is specifically an organic solvent of the present invention, more specifically C _{1 -10} can be extracted with an organic solvent, methanol, ethanol, isopropyl alcohol, butanol, pentanol, hexanol, cyclohexanol, diethyl ether , Tetrahydrofuran, ethyl acetate, ethyl acetoacetate, ethyl benzoate, dichloromethane, hexane, chloroform acetone or a mixed solvent thereof may be used.

Germinated brown rice extract of the present invention can regulate elevated serum glucose levels due to diabetes, specifically, the diabetes is insulin-independent diabetes.

Germinated brown rice extract of the present invention can regulate serum glucose levels through alpha-glucosidase inhibitory activity or glucose uptake inhibitory activity in the small intestine.

The present invention also relates to a powder for improving serum glucose level, food, health functional food or beverage comprising germinated brown rice water, organic solvent or mixed solvent extract thereof as an active ingredient. Here, the water used for the extraction may be cold or hot water.

The present invention also relates to a method for reducing serum glucose levels in mammals other than humans by ingesting a composition comprising germinated brown rice water, an organic solvent or a mixed solvent extract thereof as an active ingredient. Here, the water used for the extraction may be cold or hot water.

The present invention,

(a) drying the germinated brown rice;

(b) extracting the dried germinated brown rice with water, an organic solvent or a mixed solvent thereof;

(c) fractionating from nonpolar solvents to polar solvents in order; And

(d) drying the remaining fraction after fractionation.

A method for preparing germinated brown rice extracts. Here, the water used for the extraction may be cold or hot water.

Germinated brown rice extract of the present invention can be used without any conventional extraction method, but preferably the present invention is extracted at room temperature after grinding the germinated brown rice in a dry state, the extract is water or organic solvent, organic solvent when a particular fraction may be C _{1 -10} organic solvent, specifically, methanol, ethanol, isopropyl alcohol, butanol, hexane, chloroform, ethyl acetate, can be fractionated in an organic solvent such as acetone, and polar solvent from the non-polar solvent in order . More specifically, the extract of the present invention, after methanol extraction, the residue is fractionated in the order of hexane, chloroform, methyl acetate. The remaining can be obtained by filtration with methanol fraction (see Example 1 and FIG. 1).

Diabetes is divided into insulin-dependent diabetes (type 1 diabetes) and insulin-independent diabetes (type 2 diabetes). In developed countries, type 2 diabetes is increasing significantly. Type 2 diabetes is influenced by external environmental factors such as excessive intake of energy and lack of exercise, and high blood sugar symptoms due to increased insulin resistance and decreased beta cell function, accompanied by various complications.

In the present invention, the hypoglycemic effect has been found to be highly effective for insulin-independent diabetes while lowering the blood glucose level in blood by inhibiting glucose absorption in the small intestine.

One hypoglycemic action is the inhibition of alpha-glucosidase in the gastrointestinal tract. The hypoglycemic mechanism of alpha-glucosidase inhibits glucose uptake into the blood from the small intestine by inhibiting the breakdown of carbohydrates in the small intestine and delaying the absorption of glucose. Prior to the inhibition of glucose absorption in the small intestine, which is the core of the present invention, the germinated brown rice extract was primarily confirmed to have an effect on hypoglycemia by inhibiting alpha-glucosidase. Experimental Example 1 and FIG. 4

Ingested dietary carbohydrates are digested by amylase and glucosidase, broken down into glucose, and then absorbed and transported by small intestinal epithelial cells. Carriers involved in this transport are classified into the GLUT (sodium-independent glucose transporters) family and the SGLT (sodium-dependent glucose transporters) family, and glucose is absorbed in the small intestine by the action of SGLT1. SGLT1 is one of the carriers with the highest content of glucose in the small intestine and has been attracting attention in developing a material for preventing diabetes due to excessive absorption of glucose because it is present in BBMV (brush border membrane vesicle) of epithelial cells.

Therefore, the strategy of preventing and treating diabetes by inhibiting or inhibiting biosynthesis of SGLT1 can be directly controlled in terms of glucose uptake regardless of food intake and type, compared to conventional starch hydrolase inhibitors. Highly effective. Of these, GLUT is involved in glucose uptake studies and insulin action, and GLUT1 and 3, in particular, are present in the basal and are involved in glucose uptake into the blood from the small intestine. Studies on the substances that inhibit these transporters (transporters) are being conducted mainly on flavonoid-based substances, but only the inhibitory effect has been confirmed, but the mechanism of inhibition is still unknown. Recently, as the inhibitory effect of glucose absorption into the blood from the small intestine of the flavonoids quercetin has been revealed, it is expected that edible and medicinal biological resources will contain inhibitory active ingredients. If you can, it is the best way to prevent diabetes.

In the present invention, in order to confirm the inhibitory effect of glucose uptake into the blood from the small intestine extract from the germinated brown rice, Caco-2 cells were cultured, and the extracts of the present invention were treated according to the concentration. It was confirmed that it has a high blood sugar lowering effect compared to (Experimental Example 2).

As another aspect, the present invention relates to a composition for inhibiting SGLT1 using the germinated brown rice extract as an active ingredient.

By measuring the expression of sodium-dependent glucose transporters1 (SGLT1), hepatocyte nuclear factor-1α (HNF-1α), and hepatocyte nuclear factor-1β (HNF-1β), which are major factors for glucose uptake into the blood in the small intestine, Germinated brown rice extract of the present invention, specifically, germinated brown rice methanol extract was confirmed to have the activity of inhibiting the absorption of glucose into the blood in the small intestine (see Experimental Example 3 and Figures 6 to 8 results).

The term “hepatocyte nuclear factor-1α” in the present invention is called a transcription facter1 (TCF1), which is a homeodomain-contatining transcription facter and is an important substance for transcription factors. It is involved in metabolism in the liver, kidneys, pancreas and small intestine. It has been reported to interact with homeobox protein 2 (CDX2) and GATA-binding protein groups to regulate SGLT1 expression in the small intestine. In addition, the term "hepatocyte nuclear factor-1β" in the present invention is referred to as transcription facter2 (TCF2) and plays an upper role of homeodomain-contatining of HNF-1α. It also plays a role in helping metabolism in the liver, kidney, pancreas and small intestine, which has been reported to interact with HNF-1α to regulate SGLT1 expression in the small intestine.

As such, the transcription factors regulate SGLT1 expression, a transport protein for glucose uptake in the small intestine, thereby regulating a series of gene expressions involved in glucose uptake in the small intestine, particularly in the expression of HNF-1α and HNF-1β. It is an important transcription factor that regulates SGLT1 expression, and it is responsible for regulating glucose uptake in the small intestine.

In order to determine whether the germinated brown rice extract of the present invention inhibits glucose absorption in the small intestine, hepatocyte nuclear factor-1 (HNF-1α) and HNF-1β (hepatocyte nuclear factor-), which are transcription factors related to SGLT1 and SGLT1 expression As a result of confirming the expression of 1β), it was confirmed that the expression of all of the genes was reduced, and from this it was confirmed that the extract of the present invention has the inhibitory activity of glucose absorption in the small intestine (see Experimental Example 3 and Figures 6 to 8 results).

Hereinafter, the present invention will be described in more detail through examples according to the present invention and comparative examples not according to the present invention, but the scope of the present invention is not limited to the examples given below.

Example  1: Preparation of Search Samples from Germinated Brown Rice

Germinated brown rice extract showed high glucose uptake inhibitory activity during the anti-diabetic activity of extracts from various grains, spices, spices and edible algae. Thus, 40 kg of dried germinated brown rice was pulverized to separate the glucose absorption inhibiting active material contained in the germinated brown rice, extracted with 40 L of 100% methanol at room temperature, and the extract was concentrated and dried 44.5 g of hexane and chloroform. After fractionation in the order of ethyl acetate, the remainder was methanol fraction, and each was filtered through a filter cloth, and the filtrate was concentrated under reduced pressure and dried to prepare a sample (FIG. 1). In the case of extraction with water, 100% methanol was extracted with hot or cold water instead, and the extract was fractionated in the same manner as in the above method to prepare a sample.

< Antidiabetic  Measurement of Activity

Experimental Example  1: α- Glucosidase  Inhibitory Activity Test Method

The experimental method for measuring α-glucosidase inhibitory activity was carried out using a substrate 400 mM PNP-glucoside (glycoside) and 1 unit α-glucosidase (α-glucosidase, from Sacchromyces cerevisiae ). For experiments on each fraction, the reaction solution consisting of 750 ul of distilled water, 50 ul of 1 M KPB (pH 7.5), 50 ul of α-glucosidase (from sacchromyces cerevisiae ) and 100 ul of sample (1 mg / ml) at room temperature The reaction was carried out for 1 minute. 50 ul of PNP-glucoside was added to the reaction solution, and the absorbance after 4 minutes was measured with a spectrophotometer (405 nm).

<Equation 2>

Inhibitory effect (%) = [(Cc-Cs) / Cc] × 100

Cs: absorbance of the experimental zone treated with the sample

Cc: absorbance of the test zone without sample

Compared to the control group without the sample, it was confirmed that the germinated brown rice methanol extract treated by concentration significantly inhibited α-glucosidase present in the small intestine. Table 1 and FIG. 4

Table 1 Results of α-glucosidase inhibitory activity

sample Control (0 mg / ml) 1 mg / ml 0.1 g / ml 0.01 mg / ml Germinated brown rice 100 81.21 90.43 100.60 Brown rice 100 92.23 101.32 120.22

Experimental Example  2: Experimental Method of Glucose Absorption into Blood from Small Intestine

Experimental methods for measuring the inhibitory activity of glucose uptake into the blood in the small intestine were measured using Caco-2 cells and 2-NBDG (2- (N- (7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino-2 -deoxygluocose), ie, human colonic cancer cells (CaCO-2) at a concentration of 6 × 10 ⁴ cells / well, 24 well-plates, 37 ° C., CO ₂ for glucose uptake experiments for each fraction. After incubation for 13-14 days in the incubator and washed twice with PBS, 400 μl of PBS, 500 μl of 100 μM 2-NBDG, 100 μl of sample (1 mg / ml) were added and reacted at 37 ° C. for 2 hours. After removing the reaction solution and washing the cells three times with ice-cold PBS, the fluorescence intensity of 2-NBDG was measured with a spectrophotofluorometer (excitation: 485 nm, emission: 535 nm). In order to correct the effect of cell concentration on both measurements, the MTT assay was performed as follows: MTT on the well plate after fluorescence intensity measurement 50 μl of solution (5 mg / ml) was added and reacted for 3 hours at 37 ° C. and then absorbance was measured at 570 nm The glucose absorption inhibitory activity by the sample was calculated according to the following equation.

&Quot; (1) &quot;

Inhibitory effect (%) = [(Fc-Fs) / Fc] × 100

Fs: Fluorescence intensity of the experimental group treated with the sample

Fc: Fluorescence intensity of the experimental group without sample treatment

As a result of treatment of the germinated brown rice methanol extract by concentration, it was confirmed that the high glucose absorption inhibitory effect appeared when compared with the control group 100% untreated. Table 2 and FIG. 5

Table 2 Measurement Results of Glucose Absorption Inhibition

sample Control (0 mg / ml) 1 mg / ml 0.1 mg / ml 0.01 mg / ml Germinated brown rice 100 51.39 78.43 91.60 Brown rice 100 82.23 90.32 98.22

Experimental Example  3: germinated brown rice SGLT1 , HNF α-1, HNF Inhibition of β-1 Expression

The methanol extract of the present invention was added to the culture medium of the Caco-2 cell line on day 6, and after completion of the culture on day 14, RNA was extracted and key transcription factors SGLT1, HNFα-1 and HNFβ-1 primers involved in intestinal absorption. RT-PCR (reverse transcraptase-PCR) was performed using primers, followed by electrophoresis on agarose to compare the expression levels. As a result, it was confirmed that SGLT1, HNFα-1, HNFβ-1 gene expression was significantly suppressed when compared to the control group was not treated as a reference 100%. (FIGS. 6-8)

Figure 1 shows the process of obtaining a sample from germinated brown rice.

2 is a diagram showing α-glucosidase activity inhibitory activity according to the germination brown rice extraction method.

Figure 3 is a diagram showing the glucose uptake activity (Glucose uptake activity) inhibitory activity according to the germination brown rice extraction method.

Figure 4 shows the inhibitory activity of α-glucosidase activity by the concentration of germinated brown rice methanol extract.

5 is a diagram showing the glucose uptake inhibitory activity by the germinated brown rice methanol extract concentration.

6 is a diagram showing the SGLT1 mRNA expression inhibitory activity of the germinated brown rice methanol extract.

Figure 7 shows the inhibitory activity of HNFα-1 mRNA expression of germinated brown rice methanol extract.

8 is a diagram showing the inhibitory activity of HNFβ-1 mRNA expression of methanol extract.

Claims (10)

Composition for regulating serum glucose level comprising germinated brown rice water, organic solvent or mixed solvent extract thereof as an active ingredient. The method of claim 1 The composition for controlling the serum glucose levels, characterized in that the organic solvent is a C _{1 -10} organic solvent. The method of claim 1 The organic solvent is methanol, ethanol, isopropyl alcohol, butanol, pentanol, hexanol, cyclohexanol, diethyl ether, tetrahydrofuran, ethyl acetate, ethyl acetoacetate, ethyl benzoate, dichloromethane, hexane, chloroform, acetone And a solvent selected from the group consisting of mixed solvents thereof. The method of claim 1 Wherein the serum glucose level is elevated due to diabetes. The method of claim 4 Diabetes is an insulin independent diabetes. The method of claim 1 It is characterized by regulating serum glucose levels through the alpha-glucosidase inhibitory activity of germinated brown rice organic solvent extracts. Composition comprising a germinated brown rice extract as an active ingredient. The method of claim 1 It is characterized by regulating serum glucose levels through the inhibition of glucose uptake in the small intestine of germinated brown rice extract, Composition comprising a germinated brown rice extract as an active ingredient. The method of claim 1 Powder, food, dietary supplement or beverage for improving serum glucose level including germinated brown rice water, organic solvent or mixed solvent extract thereof as an active ingredient. A method for reducing serum glucose levels in mammals other than humans by ingesting a composition comprising germinated brown rice water, an organic solvent or a mixed solvent extract thereof as an active ingredient. (a) drying the germinated brown rice; (b) extracting the dried germinated brown rice with water, an organic solvent or a mixed solvent thereof; (c) fractionating from nonpolar solvents to polar solvents in order; And (d) drying the remaining fraction after fractionation. Process for preparing germinated brown rice extract.

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