KR101047796B1 - Cold grass extract having an effect of improving insulin resistance and a product containing the same as an active ingredient - Google Patents

Abstract

The present invention relates to a cold grass ethanol extract and a product containing the same as an active ingredient, the cold grass ethanol extract of the present invention inhibits the expression of SOCS-3 protein of adipocytes to improve insulin resistance, lower blood sugar, body fat and Since it has the effect of inhibiting weight gain, it can be usefully used as a medicine or a functional food for preventing and treating metabolic syndrome.

Insulin resistance, cold grass (Veronicastrum sibiricum), diabetes, obesity, metabolic syndrome, SOCS-3 protein

Description

{Veronicastrum sibiricum extracts for improving insulin resistance and products containing them as effective ingredient}

The present invention relates to a cold grass ethanol extract and a product containing the same as an active ingredient, and more particularly, to inhibit the expression of SOCS-3 protein of adipocytes to improve insulin resistance, lower blood sugar, increase body fat and weight gain. It relates to a cold ethanol extract having an inhibitory effect and a pharmaceutical composition for preventing and treating metabolic syndrome containing the same as an active ingredient, fermented milk, beverages and functional foods.

Insulin resistance is a condition in which insulin moves through the blood to target tissues, but no reaction occurs in the target tissues, and the main role of insulin is to lower blood sugar and lipolysis. Therefore, those who show insulin resistance have high blood sugar and high insulin levels in the blood. If these symptoms persist, lipolysis continues in fat cells, glucose uptake does not occur in muscles, and glucose production occurs in the liver. Eventually, insulin resistance continues and progresses to obesity, fatty liver and diabetes. Causes of insulin resistance include excessive intake, lack of exercise, aging, smoking, or drug side effects. In recent years, many studies have been conducted to improve such insulin resistance.

SOCS protein (suppressor of cytokine signaling protein) is a cytokine signaling inhibitor and is expressed by various signaling agents including insulin and plays an appropriate role in stopping their action. When insulin is delivered to cells through the blood, insulin binds to the insulin receptor on the cell membrane, and there is an insulin receptor subunit-1 (IRS-1) inside the cell that carries signals into the cell. In particular, in adipocytes, SOCS-3 protein inhibits the activation of insulin receptors and induces the breakdown of insulin receptor sub-types (IRS-1), which disrupts insulin signal transduction (K. Egawa et al., Endocrinology 141 (2000). 1930-1935, R. Zhande et al., Mol Cell Biol 24 (2004) 8929-8937). SOCS-3 protein is overexpressed in adipose tissues of obese and diabetic model animals such as ob / ob or db / db mice, demonstrating that SOCS-3 protein is closely related to insulin resistance (Emanuelli B et al., J Biol Chem 276 (2001) 47944-47949). In addition, the rats inducing obesity by feeding high-fat diet and not by genetic manipulation showed that SOCS-3 protein is overexpressed and is a major cause of insulin resistance in obesity (Ueki K et al., Proc Natl Acad Sci USA (2004) 10422-10427).

On the other hand, cold grass (冷 草, Veronicastrum sibiricum ) is a perennial herb of Hyunsamaceae, eaten as young sprouts and herbs, and dried roots are used as a herbal medicine. Hypothalamus, each, rheumatism, arthritis, dry stomach, expectorant, migraine, edema, diuresis, pain, paralysis, constipation. It also has antipyretic effects on the common cold and has been used to control pain, gynecology and circulatory diseases. The roots are known to contain saponins, and outposts have coumarin, ascorbic acid and alkaloids. They are registered in the Food Code (August 2, 2002) for legal use as food ingredients. At present, in Korea, patents for cold water extracts containing cold grass extracts in Korea Patent Registration No. 0512322 and patents on the efficacy of constipation improvement effect of cold grass extracts in Korea Patent Registration No. 0512322 have been analyzed and analyzed components of cold grass etc. Although there is a paper (Sook-Yeon Lee et al., 19 (1988) 34-38), the fact that cold tea extract is effective in improving insulin resistance or lowering blood sugar has not been disclosed or taught in any literature.

In this regard, the present inventors confirmed the effect of ethanol extract of cold grass ( Veronicastrum sibiricum ) among 200 kinds of natural extracts to inhibit the expression of SOCS-3 protein in adipocytes to maintain insulin signaling and improve insulin resistance. Through animal experiments, the hypoglycemic effect and the anti-obesity effect were verified and the present invention was completed.

Accordingly, the present invention improves insulin resistance by inhibiting the expression of SOCS-3 protein in adipocytes, lowers blood sugar, inhibits body fat and weight gain, and cold ethanol extract and metabolic syndrome containing the same as an active ingredient. An object of the present invention is to provide a prophylactic and therapeutic pharmaceutical composition, fermented milk, a beverage and a dietary supplement.

In order to achieve the above object, the present invention is to inhibit the expression of SOCS-3 protein of adipocytes to facilitate the signaling of insulin, lowering blood sugar and inhibiting obesity as cold ethanol extract and an effective ingredient It is characterized by providing a pharmaceutical composition for the prevention and treatment of metabolic syndrome containing, fermented milk, beverages and health functional food.

Hereinafter, the present invention will be described in detail.

The ethanol extract of the cold grass of the present invention is collected by drying the roots of the cold grass, crushed and added to 1L 50% ethanol per unit mass (kg) to react at room temperature for 7 days to obtain a cold grass extract, then filtered through a filter paper After lyophilization to prepare a powder.

In addition, the composition for the prevention and treatment of metabolic syndrome comprising the cold ethanol extract of the present invention as an active ingredient can be administered in various oral dosage forms at the time of actual clinical administration, when formulated, commonly used fillers, extenders , Diluents or excipients such as binders, wetting agents, disintegrants, surfactants and the like. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient such as starch, dextrin, and calcium carbonate in the cold ethanol extract. (Calcium carbonate), gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium, stiarate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. have.

A typical daily dosage of the composition for preventing and treating metabolic syndrome is in the range of 100 to 1,000 mg / kg body weight, preferably 200 to 500 mg / kg body weight, and may be administered once or in several divided doses. However, the actual dosage of the active ingredient should be determined in view of several related factors such as the route of administration, the age, sex, weight of the patient and the severity of the patient, and the dosage does not limit the scope of the present invention in any aspect.

In addition, cold ethanol extract according to the present invention can be used with confidence even when taking for the purpose of prevention because there is little toxicity and side effects.

On the other hand, the fermented milk containing the cold ethanol extract of the present invention as an active ingredient is a lactic acid bacteria culture medium, cold grass ethanol extract lyophilized powder and mixed fruit syrup combined at a constant rate at 150 bar and then cooled to 10 ℃ or less and packed in a container To prepare fermented milk.

On the other hand, the beverage containing the cold ethanol extract of the present invention as an active ingredient is a mixture of fruit juice syrup, cold ethanol extract lyophilized powder and water in a uniform ratio and homogenized at 150 bar and then cooled to below 10 ℃ glass bottles, plastic bottles Functional beverages are prepared by packaging in small packaging containers.

On the other hand, the health functional food containing the cold ethanol extract of the present invention as an active ingredient, in addition to the cold ethanol extract freeze-dried powder, vitamin B1, B2, B5, B6, E and acetate esters, nicotinic acid amide as a nutritional supplement Oligosaccharides may be added, and other food additives may be added.

As described above, the present invention improves insulin resistance by inhibiting the expression of SOCS-3 protein in adipocytes, pharmaceutical compositions containing cold grass ethanol extracts that have a hypoglycemic and anti-obesity effect, fermented milk, beverages, functional foods, etc. It can be applied to a product having the effect of preventing and improving metabolic syndrome.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are not intended to limit the scope of the present invention, and ordinary changes by those skilled in the art are possible within the scope of the technical idea of the present invention.

≪ Example 1 >

Preparation of cold ethanol extract

Cold grass was collected by drying the roots and stems, and then crushed and added to 1 L of 50% ethanol per unit mass (kg) and reacted at room temperature for 7 days to obtain an extract. The extract was filtered on a filter paper and then lyophilized to powder to prepare a cold ethanol extract.

<Example 2>

Preparation of pharmaceutical composition using cold ethanol extract

Hereinafter, an example of the preparation of a pharmaceutical composition for preventing and treating metabolic syndrome containing cold ethanol extract of the present invention as an active ingredient will be described, but the present invention is not intended to be limited thereto.

Manufacture of powder

According to the preparation method of powder in the Pharmacopoeia General Formulation, it is prepared in the following component content per packet.

Cold ethanol extract (dry powder) ...................... 300 mg

Lactose 100 mg

Talc ........................................ 10mg

Manufacture of tablets

According to the preparation method of tablets in the Pharmacopoeia General Formulation, it is prepared in the following component contents per tablet.

Cold ethanol extract (dry powder) ....... 300 mg

Corn starch ........................... 100 mg

Lactose ......................................... 100 mg

Magnesium Stearate ................................................... 2mg

Manufacture of capsules

According to the preparation method of capsules in the pharmacopeia formulation, it is prepared in the following component content per capsule.

Cold ethanol extract (dry powder) ........ 300 mg

Corn starch ........................... 100 mg

Lactose ... Mg

Magnesium Stearate ......................................... 2 mg

Preparation of liquid

According to the method for preparing a liquid formulation in the Pharmacopoeia General Formulation, it is prepared in the following component content per 100 ml of liquid formulation.

Cold ethanol extract (dry powder) ....................... 1g

Isomerized sugar ......................................... 10g

Mannitol ......................................... 5 g

Purified water ......................................

<Example 3>

Preparation of Fermented Milk Using Cold Ethanol Extract

Method for producing a fermented milk consisting of a freeze-dried powder of the lactic acid bacteria culture medium and the cold ethanol extract of Example 1 and mixed fruit juice syrup is as follows.

The lactic acid bacteria culture medium was stirred at 95.36% of crude milk and 4.6% by weight of skim milk powder (or mixed milk powder), and the specific gravity at 15 ° C was 1.0473 to 1.0475, the titratable acidity was 0.200 to 0.220%, the pH was 6.65 to 6.70, and the brix at 20 ° C. (Brix ^o ) was mixed to 16.3 ~ 16.5%. Then, UHT heat treatment (sterilized at 135 ℃ for 2 seconds), cooled to 40 ℃, Streptococcus thermophilus bacteria and lactose (Valley laboratory, USA) each added 0.02% by weight Incubated for 6 hours, the total lactic acid bacteria in BCP medium was prepared by 1.0 × 10 ⁹ cfu / ㎖ or more, the titratable acidity is 0.89 ~ 0.91%, pH is 4.55 ~ 4.65.

Mixed fruit syrup consists of 10% by weight liquid fructose, 3% by weight white sugar, 3% by weight brown sugar, 10% by weight mixed juice concentrate (56 Brix ^o ), 0.1% by weight pectin, 0.05% by weight fresh fruit mix essence and 73.85% by weight purified water After mixing by stirring at 30 ~ 35 ℃ UHT heat treatment (sterilized for 2 seconds at 135 ℃) was prepared by cooling.

75% by weight of the lactic acid bacteria culture medium prepared by the above method, 0.1% by weight of the freeze-dried powder of the cold ethanol extract of Example 1 and 24.9% by weight of the mixed fruit juice syrup prepared by the above method was homogeneous at 150 bar and then 10 ° C. or less. Cooled to prepare a fermented milk containing cold grass ethanol extract as an active ingredient having the effect of preventing and treating metabolic syndrome.

<Example 4>

Preparation of Beverage Using Cold Ethanol Extract

Method for producing a functional beverage consisting of a freeze-dried powder and mixed juice syrup of the cold ethanol extract of Example 1 is as follows.

Mixed fruit syrup consists of 10% by weight liquid fructose, 3% by weight white sugar, 3% by weight brown sugar, 10% by weight mixed juice concentrate (56 Brix ^o ), 0.1% by weight pectin, 0.05% by weight fresh fruit mix essence and 73.85% by weight purified water After mixing by stirring at 30 ~ 35 ℃ UHT heat treatment (sterilized for 2 seconds at 135 ℃) was prepared by cooling.

29.9% by weight of the mixed juice syrup prepared by the above method, 0.1% by weight of the lyophilized powder of the cold ethanol extract of Example 1 and 70% by weight of sterile purified water were homogenized at 150 bar, and then cooled to 10 ° C. or lower. Functional drinks containing cold ethanol extracts as effective ingredients were prepared by packaging them in small packaging containers such as glass bottles and plastic bottles to prevent and treat metabolic syndrome.

Example 5

Preparation of health functional food using cold ethanol extract

0.1 parts by weight of the freeze-dried powder of the cold ethanol extract of Example 1 to the nutritional supplements (vitamins B1, B2, B5, B6, E and acetate esters, nicotinic acid amide) and oligosaccharides of the cold ethanol extract of Example 1 10 parts by weight based on 100 parts by weight of the lyophilized powder was added and mixed in a high speed rotary mixer. 10% by weight of sterile purified water was added to the mixture, mixed, and molded into granules having a diameter of 1 to 2 mm. The molded granules were dried in a vacuum dryer at 40 to 50 ° C., and then passed through 12 to 14 meshes to uniformly prepare granules. The granules prepared as described above were extruded by appropriate amounts into tablets or powders or filled into hard capsules to produce hard capsule products.

<Test Example 1>

Inhibitory Effects of Ethanol Extracts from Cold Water on Adipose Cell SOCS-3 Protein Expression

1-1 Adipose Cell SOCS-3 Protein Expression

Adipocytes were pre-differentiated into mouse progenitor cells (3T3-L1 preadipocytes) in sterile 6 well plates. On day 10 of differentiation, fully differentiated adipocytes were treated with insulin at concentrations of 1, 3, and 5 μg / mL, and then cultured in a 37 ° C. CO2 incubator for 5 hours, and proteins were extracted to obtain SOCS through western blot. It was confirmed that expression of -3 protein was induced.

As can be seen in Figure 1 showing the results, it was confirmed that the expression of SOCS-3 protein when 5μg / mL of insulin was added.

Inhibition of Adipose Cell SOCS-3 Protein Expression by 1-2 Cold Ethanol Extracts

Induction of SOCS-3 protein by treating insulin with 5μg / mL of the precursor adipocytes as in Test Example 1-1, followed by 50, 100, 200μg / mL of cold ethanol extract, respectively, by concentration of SOCS It was confirmed whether the expression of the -3 protein is inhibited.

On the other hand, induction of the expression of SOCS-3 protein as in Test Example 1-1 and treated with only insulin as a control.

The results are shown in FIG.

As can be seen in Figure 2, it can be seen that the expression of SOCS-3 protein is inhibited when the cold ethanol ethanol extract 100μg / mL or more treatment, according to the insulin receptor subtype that is degraded by SOCS-3 protein ( Insulin receptor subunint-1 (IRS-1) expression was improved in adipocytes treated with cold ethanol extract.

<Test Example 2>

Effect of Cold Ethanol Extract on Insulin Resistance

2-1 Fasting Blood Glucose Measurement

In order to confirm the effect of improving the insulin resistance of the cold ethanol extract according to the present invention was carried out animal experiments using a high-fat diet. The experimental animals were C57BL / 6 mice, which were used after adapting a 3 week old male with a high fat diet for 1 week. Seven rats of all test groups were completely randomized and the test groups consisted of three groups. The high-fat diet was D12451 (45% fat, 5.252 cal / g) from Research diets. The experimental group (high-fat diet group) and the normal diet-only test group (normal diet group) were used as controls. Cold ethanol extract added to the feed was lyophilized and powdered, and then mixed with a high fat diet at a concentration of 1% (w / w) to make a 1% cold grass administration group.

The experimental diet was administered for a total of four weeks, and body weight and feed intake were measured at two intervals per week. After 4 weeks of dietary administration, the experimental animals were fasted for 6 hours and fasting glucose was measured.

The results are shown in Table 1.

As can be seen in Table 1, the 1% cold grass administration group was confirmed that the fasting blood sugar was significantly lower than the high-fat diet group.

2-2 Insulin Sensitivity Measurement

In addition, the glucose-load test was performed in a fasted state to measure insulin sensitivity of rats (high-fat diet, normal diet, 1% cold grass administration group) administered the experimental diet for 4 weeks of Test Example 2-1. 6 hours of fasting to the test animals were administered orally administered glucose (glucose) concentration of 2g / kg concentration for 30 minutes each 2 hours to measure the change in blood glucose.

The results are shown in Fig.

As can be seen in Figure 3, rats fed a high-fat diet, high blood sugar and 120 minutes after fasting blood sugar, such as fasting blood sugar control can be seen later than normal diet group, while cold ethanol extract high-fat diet The experimental group fed (1% cold grass administration group) was mixed with low fasting blood sugar as in the normal diet group, and after a certain period of time after the glucose load was confirmed to be controlled to fasting blood glucose.

As such, when the cold ethanol extract is taken for a certain period of time, the blood sugar is lowered, and even when there is a sugar load such as food intake, the blood sugar is stabilized rapidly. Thus, the use of the cold ethanol extract of the present invention may be considered as a hypoglycemic agent. It means that there is.

<Test Example 3>

Anti-obesity Effect of Cold Ethanol Extracts

In order to confirm the anti-obesity effect of the cold ethanol extract according to the present invention, the weight change and the amount of fat tissue formed around the epididymis after 4 weeks in the animal experiment of Test Example 2 every two weeks for 4 weeks Was measured.

The results are shown in Table 2.

As can be seen in Table 2, the high-fat diet group gained twice as much weight as the normal diet group, whereas the 1% cold grass group showed similar weight gain as the normal diet group. In addition, the fat diet of the high-fat diet group increased 2.5 times compared to the normal diet group, but the 1% cold grass group fed the high-fat diet, but the increase of fat mass decreased compared to the high-fat diet group.

Through these facts, the cold ethanol extract was found to have an effect of inhibiting body fat increase and weight gain and finally having an excellent anti-obesity effect.

Figure 1 is a protein analysis picture to determine whether the expression of SOCS-3 protein is induced by treating the concentration of insulin in rat fat cells (3T3-L1 adipocyte).

2 is a protein analysis picture showing that the expression of SOCS-3 protein is inhibited and the expression of IRS-1 is improved by treating cold grass ethanol extract with insulin in mouse fat cells.

3 is a graph showing the change in blood glucose with time of the experimental animal after oral administration of glucose.

Claims (8)

delete delete delete A pharmaceutical composition for preventing and treating diabetes, characterized in that it contains cold ethanol extract having an effect of improving insulin resistance by inhibiting the expression of SOCS-3 protein in adipocytes as an active ingredient. delete delete delete A pharmaceutical composition for the prevention and treatment of obesity, characterized in that it contains a cold grass ethanol extract having an effect of inhibiting body fat and weight gain as an active ingredient.

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