KR101467407B1 - Antidiabetic food composition with the extract of Dendropanax morbifera - Google Patents

Abstract

The present invention relates to a glycemic control food composition. More specifically, the present invention relates to a food composition, wherein a Dendropanax morbifera extract is contained as an active ingredient so as to prevent and alleviate diabetes by increasing insulin secretion and decreasing glucagon secretion in order to reduce blood glucose levels in vivo.

Description

[0001] The present invention relates to an antidiabetic food composition containing an extract of Dillardia chinensis as an active ingredient,

The present invention relates to a food composition for controlling blood sugar, and more particularly, to a food composition containing an extract of Euphorbiaceae which is capable of reducing blood sugar levels in the body and preventing and improving diabetes as an active ingredient.

Diabetes mellitus, not only itself, but also various complications can lead to death, and the danger is emphasized, and the prevalence is high around the world. In particular, the incidence of diabetes and complications in Korea is higher than the OECD average, and the incidence rate is increasing in Korea due to the change to the pattern of living in advanced countries.

Diabetes is a type of metabolic disease that is caused by insufficient secretion of insulin in the pancreatic cells or failure to function normally. It is characterized by hyperglycemia, which increases the blood glucose level. Hyperglycemia has been reported to cause hyperglycemia by abnormal hyperglycemia of glucagon, accompanied by overproduction of glucose, body fat breakdown and protein waste. Diabetes also causes metabolic control abnormalities in vivo, which can lead to complications such as the circulatory system if proper treatment and management is not performed.

In diabetes, there is no problem with insulin secretion, type 1 diabetes and insulin secretion that do not regulate insulin secretion because of insufficient secretion of insulin, and insulin secretion is insufficient. There is type 2 diabetes. Type 1 diabetes requires insulin therapy. Type 2 diabetes is based on lifestyle modification and can be supplemented with additional medications.

The agents used for the treatment of diabetes are divided into oral preparations and injections. Oral preparations are divided into insulin secretagogues and insulin sensitizers. In the case of an oral preparation, there is a problem that the time to eat varies depending on the action time of the medicine, and the side effects appear slightly different. In the case of injections, they are administered by subcutaneous injection. However, the administration method differs depending on the time of administration. The effect of blood glucose lowering is more rapid than that of eating medicine, and it is safe to use even in an environment where the medicine to be consumed is not available. However, there is a difficulty in rejecting the needle and the method of administering it is not easy.

Although many prescriptions have been developed for the treatment of diabetes, including oral and injectable drugs, a method for treating diabetes mellitus has not yet been developed. Because pharmacotherapy is based on chemicals such as insulin or chemical hypoglycemic agents, side effects and resistance to drug use are constantly occurring. Therefore, it is necessary to develop a therapeutic agent for diabetes using natural products.

On the other hand, Dendropanax morbifera is a dicotyledonous plant native to southern coast of Korea and Jeju Island, and is an evergreen tree of Araliaceae. Recently, its efficacy has been actively researched and its physiological activities such as anticancer activity and antioxidant activity of Hokutogi extract have been reported to be limited and used in pharmaceuticals and food compositions.

Korean Patent Laid-Open No. 10-2012-0101743 (a title of the invention: a composition for preventing or treating diabetes comprising dendron-panoxide as an active ingredient) is a dendropanoxide DP ) As an active ingredient for preventing or treating diabetes mellitus. Korean Patent No. 10-1333007 (the name of the invention: a method of extracting yellow marigold extract) is a method for purifying a mixture of a mixture of vinegar, a sugar ingredient and water added to a ground raw material after washing and cutting the same, And the fermentation and extraction processes are carried out.

It is an object of the present invention to develop and provide a food composition having blood glucose controlling effect by increasing insulin secretion in blood and decreasing glucagon secretion.

In order to achieve the above object, the present invention hwangchil tree (Dendropanax morbifera ) as an active ingredient. The ethyl acetate extract used in the present invention is preferably one in which ethyl acetate as an extraction solvent has been removed.

It is also called ginseng tree or ginseng tree because it contains a large amount of saponin. It grows in Jeju Island, Wando Island, Dae Huksan Island, Geomundo, Jeolla Province, Eocheongdo, The major ingredients of Huangchil are sesquiterpene, saponin, benzyl, beta-Elemene, alpha-Muurolene, germarcrene D and beta Sitosterol, and the like. It is known that Hwangchilchae contains liver function improvement, whitening, wrinkle improvement and hard tissue improvement.

In the present invention, the term "comprising as an active ingredient" means that the extract of ethyl gallate ethyl acetate of the present invention is added to the composition to such an extent that it can exhibit a blood glucose controlling effect.

Meanwhile, in the food composition for controlling blood sugar level of the present invention, the ethyl acetate extract may be obtained by, for example, (a) adding ethanol to woody spruce tree to obtain an ethanol extract; (B) suspending the obtained ethanol extract in water, adding n-hexane and fractionating it; Separating the water layer after fractionation of step (b) and separating by adding ethyl acetate; And (d) separating the ethyl acetate layer after fractionation of step (c).

Meanwhile, in the food composition for controlling blood sugar of the present invention, the ethanol extract of step (b) may be ethanol-free.

Meanwhile, the food composition for controlling blood sugar of the present invention may further include step (e) of removing ethyl acetate as an extraction solvent after the step (d).

On the other hand, in the food composition for controlling blood glucose of the present invention, it is preferable that 0.1 to 50% by weight of the ethyl acetate extract of Hwanyeolmum is added to the food composition. When the content of the extract is less than 0.1% by weight, the blood glucose controlling effect is not exhibited, and when it exceeds 50% by weight, the extract is not economical.

On the other hand, if the form of the composition is a food composition, the present invention can add a formulation base and excipients to make various formulations. Examples of formulations are cereals, caffeinated beverages, regular beverages, dairy products, chocolate, bread, snacks, confectionery, tea, pizza, jelly, noodles, gums, ice creams, alcoholic beverages, alcoholic drinks, vitamin complexes, But it is not necessarily limited thereto.

The food composition of the present invention containing as an active ingredient the extract of U. chrysanthemum extract of the present invention has an excellent effect in controlling blood glucose by increasing insulin secretion and decreasing glucagon secretion.

FIG. 1 shows a process for preparing the extract of Fusarium oxyspora or fraction of the present invention.
FIG. 2 is a graph showing blood glycated hemoglobin concentration (HbA1c) of the extract of U. In Figure 2, the results were expressed as mean ± standard error (n = 6), and statistical analysis was performed using the ANOVA test (p <0.05).
FIG. 3 is a graph showing the activity of glucose-6-phosphatase (G6Pase) of the extract of U. In Fig. 3, the results were expressed as mean ± standard error (n = 6), and statistical analysis was performed using the ANOVA test (p <0.05).

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the scope of the present invention is not limited to the following embodiments, and includes modifications of equivalent technical ideas.

[Example 1: Preparation of ethyl acetate fraction extract of Fusarium oxysporum]

2 ㎏ of dried Dendropanax morbifera stem was cut into 0.5 cm size, ground and put into an extraction container, 20 L of ethanol was added, and the resultant was heated to 50 캜 by a distillation circulation apparatus and filtered with 3M paper To obtain an extract liquid phase. Extraction was carried out once for 6 hours, after which the solvent was concentrated under reduced pressure to obtain 96 g of an ethanol extract.

A total of 1.6 L of distilled water was suspended in 80 g of the obtained Hwanyeobu tree ethanol extract, and the suspension was added to a fractionating funnel, followed by extraction with 1.6 L of n-hexane. After about 1 hour, the n-hexane fraction-extracted supernatant was taken to obtain an n-hexane fraction extract. The fractionation procedure was repeated three times, after which the supernatant was concentrated under reduced pressure to obtain 6.8 g of n-hexane fraction extract.

The distilled water fraction extract obtained in the step of obtaining the above yellowish tree n-hexane fraction extract was concentrated under reduced pressure. Thereafter, a total of 1.5 L of distilled water was added to suspend the suspension, and the suspension was put into a fractionation funnel, and 1.5 L of ethyl acetate was added to extract the fraction. After about 1 hour, the ethyl acetate fraction-extracted supernatant was taken to obtain an ethyl acetate fraction extract. The fractionation procedure was repeated three times, after which the supernatant was concentrated under reduced pressure to obtain 17.3 g of an ethyl acetate fraction extract (see Fig. 1).

[Comparative Example 1: Preparation of ethanol extract of Huchulchinda tree]

2 ㎏ of dried Dendropanax morbifera stem was cut into 0.5 cm size and pulverized and placed in an extraction container. 20 L of ethanol was added thereto, and the mixture was heated at 50 캜 in a distillation-circulating apparatus and filtered with 3M ™ paper to obtain an extract liquid &Lt; / RTI &gt; Extraction was carried out once for 6 hours, after which the solvent was concentrated under reduced pressure to obtain 96 g of an ethanol extract (see Fig. 1).

[Comparative Example 2: Preparation of water extract of Huaculi tree]

2 kg of the dried Dendropanax morbifera stem was cut into 0.5 cm size and pulverized. Then, 20 L of water was added to the extraction vessel, and the mixture was heated at 50 캜 in a distillation circulating apparatus and filtered with 3M ™ paper to obtain an extract liquid &Lt; / RTI &gt; Extraction was carried out once for 6 hours, after which the solvent was concentrated under reduced pressure to obtain 138 g of water extract (see Fig. 1).

[Comparative Example 3: Preparation of extract of Huchulchu n-hexane fraction]

A total of 1.6 L of distilled water was added to 80 g of the extract of Yellow Hut tree obtained in the above Comparative Example 1, suspended in the fraction funnel, and 1.6 L of n-hexane was added thereto. After about 1 hour, the n-hexane fraction-extracted supernatant was taken to obtain an n-hexane fraction extract. The fractionation procedure was repeated three times, after which the supernatant was concentrated under reduced pressure to obtain 6.8 g of n-hexane fraction extract (see Fig. 1).

[Comparative Example 4: Preparation of n-butanol fraction extract from Hwangcholgak extract]

The distilled water fraction obtained in the step of obtaining the ethyl acetate fraction extract obtained from Example 1 was concentrated under reduced pressure. Thereafter, a total of 1.2 L of distilled water was added to suspend the mixture, and the mixture was added to a fractionation funnel, and 1.2 L of n-butanol saturated with water was added thereto. About one hour later, the supernatant fractionated with n-butanol was taken to obtain an n-butanol fraction extract. The fractionation procedure was repeated three times, after which the supernatant was concentrated under reduced pressure to obtain 41.2 g of n-butanol fraction extract (see Fig. 1).

[Comparative Example 5: Preparation of a water fraction extract from Hwangcholgak extract]

The distilled water fraction obtained in the step of obtaining the yellowtail n-butanol fraction extract was concentrated under reduced pressure to obtain 14.7 g of a water fraction extract (see Fig. 1).

[Experimental Example 1: Fasting blood glucose measurement]

In this experiment, we used C57BL / Ks01aHsd-db / db mouse, which shows the clinical characteristics of type 2 diabetes due to leptin receptor mutation, including obesity, obesity, insulin resistance, hyperglycemia and hyperinsulinemia. A 5-week-old C57BL / Ks01aHsd-db / db male mouse of 30 g received from the Central Animal Research and Development Center was adapted from the feeder room during the week, and the normal state was observed during the adaptation period. . The breeding environment was maintained at a temperature of 23 ± 3 ℃ and a humidity of 50 ± 5% for 12 hours. Weights were measured for the animals that were judged to be healthy during the refinement period, and the individuals close to the average weight were selected and grouping was performed using the random method.

In this experiment, 50 ㎎ / ㎏ group of extract of Huangchu tree extract, 50 ㎎ / ㎏ of Huangchu tree ethanol extract, 50 ㎎ / ㎏ of Huchulchia n - hexane fraction extract and 50 ㎎ / , 50 ㎎ / ㎏ group of extract of yellowtail n - butanol fraction and 50 ㎎ / ㎏ of extract of yellowtail wood fraction. The experiment was carried out with 6 groups per group.

Experimental materials were tested at a purity of 100%, and were orally administered with 0.5% carboxymethylcellulose (CMC) just prior to administration. The control and experimental groups were orally administered with 0.5% carboxymethylcellulose (CMC) once a day for a total of 6 weeks. Water extract, ethanol extract, n-hexane fraction extract, ethyl acetate fraction extract group Example 1), n-butanol fraction extract, and water fraction extract administration group were orally administered once a day for a total of 6 weeks.

The animals were fasted for 12 hours at 1, 2, 4, and 6 weeks and then blood was collected from the tail vein and blood glucose was measured at fasting using a glucose meter (GlucoDr.supersensor, Allmedicus, Korea). The results are shown in the following Table 1 and are expressed as mean ± standard error (Mean ± S.E) (n = 6).

Experimental group 1 week Blood sugar (mg / dL) 2-week blood sugar (mg / dL) 4 weeks Blood sugar (mg / dL) 6 weeks Blood sugar (mg / dL) Diabetic control 254.7 ± 33.1 447.1 + - 24.3 504.5 ± 21.7 651.9 ± 31.6 Water extract 276.3 ± 20.1 473.9 ± 29.4 484.1 ± 22.5 625.2 ± 14.9 Ethanol extract 238.1 ± 30.7 488.2 ± 32.4 462.3 ± 28.9 545.7 ± 34.9 n-hexane fraction extract 263.3 ± 15.3 431.8 ± 19.7 487.4 ± 27.5 612.9 + - 31.4 Ethyl acetate fraction extract 244.8 ± 27.4 404.9 ± 23.0 431.4 + - 14.7 488.4 ± 22.1 n-butanol
Fraction extract 241.7 ± 19.7 433.9 ± 28.9 487.1 ± 30.5 544.3 ± 18.9 Water fraction extract 260.5 + - 24.0 464.5 ± 31.8 523.3 ± 34.9 679.1 ± 40.1

As a result of the experiment, the fasting blood glucose level was lower in all of the other groups than the control group except the water fraction extract. Especially, the ethyl acetate fraction extract showed the lowest fasting blood glucose level when compared to the diabetic control group, even when it was compared to the 6th week (Table 1).

[Experimental Example 2: Oral glucose tolerance test]

This experiment was conducted to determine whether glucose metabolism is smooth by measuring oral glucose tolerance. Glucose tolerance refers to the glucose metabolism ability of glucose absorbed in the body to be lowered by insulin secretion and burned in peripheral tissues to regulate blood sugar.

The animals were fasted for 12 hours, and blood was collected from the tail vein. Fasting glucose was measured, and then 50% glucose solution (0.1 g glucose / 100 g BW) was orally administered. After 30 minutes, 60 minutes and 120 minutes , And blood glucose were measured. The results are shown in Table 2 below and are expressed as mean ± standard error (Mean ± S.E) (n = 6).

Experimental group Fasting Blood Sugar (mg / dL) 30 min Blood glucose (mg / dL) 60 min Blood glucose (mg / dL) 120 min Blood glucose (mg / dL) Diabetic control 447.1 ± 17.3 621.7 + - 22.8 565.8 ± 14.7 521.3 ± 13.8 Water extract 423.9 ± 19.3 618.3 ± 14.6 576.1 ± 10.9 533.9 ± 12.9 Ethanol extract 441.7 ± 9.8 577.0 + - 13.7 524.3 ± 16.4 483.5 ± 20.3 n-hexane fraction extract 429.9 ± 16.4 614.9 ± 13.3 544.1 ± 20.1 505.3 ± 11.9 Ethyl acetate
Fraction extract 434.3 ± 15.4 581.2 + - 22.1 483.5 ± 17.4 429.2 + - 16.8 n-butanol
Fraction extract 429.4 ± 23.4 599.1 + - 16.9 522.4 ± 21.2 484.5 ± 8.9 Water fraction extract 461.0 + - 14.1 611.7 ± 15.4 570.5 ± 17.6 531.6 ± 20.7

As a result, the fasting blood glucose level was 447.1 ± 17.3 mg / dL in the diabetic control group, but increased to 621.7 ± 22.8 mg / dL after 30 minutes of glucose administration and 521.3 ± 13.8 mg / dL after 120 minutes. Among the groups administered orally, the ethyl acetate fraction extract was 581.2 ± 22.1 mg / dL 30 minutes after oral glucose administration and dropped to 429.2 ± 16.8 mg / dL after 120 minutes to the lowest blood glucose level. In the water fraction extract, the fasting blood glucose level was found to be higher than that of the diabetic control group to the blood glucose level measured 120 minutes later. These results show that the ethyl acetate fraction extract has superior blood glucose control ability than other samples (Table 2).

[Experimental Example 3: Measurement of glycated hemoglobin concentration in blood]

In this experiment, the concentration of HbA1c, a part of hemoglobin in red blood cells, was measured. Since HbA1c is glycosylated by non-enzymatic binding with glucose during the blood cell's survival period, the degree of glycation of HbA1c is an indicator of blood glucose concentration.

The glycated hemoglobin concentration (HbA1c) was determined by using a Nycocard reader (Axis-Shield Poc. AS, Norway) using reflectometry after blood sampling from the tail vein after fasting for 12 hours in the experimental animals for 6 weeks Respectively.

As a result, the glycated hemoglobin of the diabetic control group was measured as 8.1 ± 0.3%. The ethanol extract, ethyl acetate fraction, and n - butanol fraction of the experimental group were significantly lower than those of the diabetic control group in the oral administration group, and the ethyl acetate fraction and the n - butanol fraction were respectively 6.5 ± 0.2 and 7.1 ± 0.2% There was no significant difference in the experimental group. The ethyl acetate fraction extract showed the lowest glycated hemoglobin significantly. Therefore, it was confirmed that the extract of the ethyl acetate fraction of sulfuric acid was more effective in reducing blood glucose than the other samples (FIG. 2).

[Experimental Example 4: Measurement of insulin and glucagon secretion]

In this experiment, the plasma insulin concentration (AKRIN-010T, Shibayagi Co. Ltd, Japan) and the glucagon concentration (PG090, Yanaihara institute Inc, Japan) were measured using an ELISA kit. The results are shown in Table 3 below and the mean ± standard error (Mean ± SE) (n = 6) was described.

Insulin is a hormone that reduces blood sugar and improves glucose uptake and promotes glycogen, protein and fatty acid synthesis. Glucagon, on the other hand, is secreted when glucose is lowered, promoting glucose synthesis to increase blood sugar and maintain blood sugar. Therefore, when the secretion of insulin is reduced, the ability to reduce blood sugar is lowered.

Experimental group insulin
(ng / mL) Glucagon
(ng / mL) Insulin / glucagon Diabetic control 1.19 ± 0.09 2.08 ± 0.09 0.57 ± 0.09 Water extract 1.19 ± 0.11 2.07 ± 0.03 0.58 + 0.07 Ethanol extract 1.21 ± 0.11 2.04 ± 0.08 0.59 + - 0.10 n-hexane fraction extract 1.21 ± 0.13 2.08 ± 0.08 0.58 0.11 Ethyl acetate
Fraction extract 1.31 + 0.09 1.98 ± 0.05 0.66 + 0.07 n-butanol
Fraction extract 1.24 ± 0.07 2.04 + 0.03 0.61 ± 0.05 Water fraction extract 1.17 + 0.07 2.07 ± 0.03 0.57 ± 0.05

1.21 ± 0.11 ng / mL of ethanol extract, 1.21 ± 0.13 ng / mL of n-hexane fraction extract, 1.31 ± 0.09 ng / mL of ethylacetate fraction extract and n-hexane fraction of n-hexane fraction were compared with 1.19 ± 0.09 ng / - butanol fraction extract, 1.24 ± 0.07 ng / mL. Among them, the highest concentration of insulin was observed in the experimental group treated with ethyl acetate fraction.

The ratio of insulin to glucagon was 0.58 ± 0.07 for water extract, 0.58 ± 0.07 for ethanol extract, 0.58 ± 0.11 for n-hexane fraction, 0.66 ± 0.07 for ethyl acetate fraction and 0.61 ± 0.05 for n-butanol fraction, And increased in the ethyl acetate fraction extract (Table 3).

[ Experimental Example  5: liver glucose-6-phosphatase ( G6Pase ) Activity measurement]

In this experiment, the activity of glucose-6-phosphatase (G6Pase), which is an enzyme for increasing blood glucose, was measured by hydrolyzing glucose-6-phosphate in the liver to increase glucose synthesis.

After 0.2 M glucose-6-phosphate and enzyme source were added to a 0.1 M Tris-maleate buffer (pH 6.5) solution at a concentration of 0.1 M, the mixture was reacted at 30 ° C for 20 minutes, The supernatant was collected by centrifugation. Acetate buffer (pH 4.0), ammonium molybdate solution and reducing reagent were added to the supernatant, and the absorbance was measured at a wavelength of 680 nm. The activity of the enzyme was expressed as the amount of phosphate produced per minute of 1 mg of microsomal protein.

 The results were as follows: 54.3 ± 2.1 nmol / min / mg protein, 55.7 ± 2.7 nmol / min / mg protein, 52.9 ± 1.9 nmol / min / mg protein, and 53.1 ± 2.9 nmol / min protein in the diabetic control group min / mg protein, ethyl acetate fraction, 48.7 ± 1.3 nmol / min / mg protein, n - butanol fraction and 52.8 ± 2.7 nmol / min / mg protein and 55.1 ± 3.4 nmol / min / mg protein respectively. The experimental group with the lowest activity compared to the diabetic control group was the ethyl acetate fraction extract (Fig. 3).

From these results, it was confirmed that the sulfuric acid ethyl acetate fraction extract of the present invention is more effective in controlling blood glucose than the extracts and fraction extracts of Comparative Examples 1 to 5.

Claims (4)

delete It contains ethyl acetate extract of Dendropanax morbifera as an active ingredient,
The ethyl acetate extract,
Adding ethanol to the seedlings to obtain an ethanol extract (a);
(B) suspending the obtained ethanol extract in water, adding n-hexane and fractionating it;
Separating the water layer after fractionation of step (b) and separating by adding ethyl acetate; And
(D) separating the ethyl acetate layer after fractionation of the step (c), and extracting the ethyl acetate fraction from the fraction.
3. The method of claim 2,
The ethanol extract of step (b)
Wherein the ethanol is removed.
3. The method of claim 2,
The food composition for controlling blood glucose,
(E) removing the ethyl acetate as an extraction solvent after the step (d).

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