JP2005075749A - Cell differentiation promoter - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a cell differentiation promoter which contains an ingredient having an action for promoting the differentiation of fat cells and extracted from the rhizomes and/or leaves of Zingiber officinale having been eaten for long years and can be taken as a safe food everyday. <P>SOLUTION: This cell differentiation promoter from precursor fat cells to fat cells, containing an ingredient extracted from the rhizomes and/or leaves of Zingiber officinale; the cell differentiation promoter from enlarged fat cells to small normal fat cells; a visceral fat-increasing inhibitor; an agent for preventing and improving life-style related diseases originated from fat cells. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a cell differentiation promoting agent, and in particular, an active ingredient is a component extracted from rhizome and / or leaves of ginger that has been eaten for many years, and the function of promoting differentiation from a preadipocyte to an adipocyte, The present invention relates to a cell differentiation promoting agent having the property of reducing the number of cells and increasing the number of small fat cells. To prevent and improve a wide range of lifestyle-related diseases such as diabetes, hyperlipidemia, hypertension, arteriosclerosis, cancer, etc. It relates to health food ingredients and medicines.

In Japan, eating habits have become richer, and is now called the age of satiety, and due to excessive caloric intake and lack of exercise, obesity or diabetes is rapidly increasing. Currently, one in three men in their 30s is overweight or obese, and nearly 40% of those in their 40s and 50s are obese. Obesity is known to cause hyperlipidemia, arteriosclerosis and diabetes as complications.
Obesity is a condition in which fat is abnormally accumulated in fat cells and the cells are enlarged. Until now, fat cells have been considered to be a tissue for storing surplus energy.
Recently, however, adipocytes secrete 10 or more hormones and cytokines including leptin, adiponectin, TNF-α, resistin, and free fatty acids, and have been found to be active endocrine organs. In particular, it is known that secretion is vigorous in visceral fat, and hypertrophic adipocytes excessively secrete TNF-α, resistin, free fatty acid and the like, which are called insulin resistance-inducing molecules. This is the cause of insulin resistance and is thought to lead to lifestyle-related diseases such as diabetes, hypertension, hyperlipidemia, and arteriosclerosis. That is, visceral fat accumulation is closely related to lifestyle-related diseases.

Therefore, in order to prevent lifestyle-related diseases, it is considered important to convert the enlarged fat cells into small fat cells. Adipocyte differentiation is mainly controlled by PPARγ called a nuclear receptor transcription factor.
In order to increase the number of small adipocytes, a method of enhancing the action of PPARγ to promote differentiation of adipocytes from preadipocytes can be considered. Thiazolidine drugs used as antidiabetic drugs are potent PPARγ agonists (agonists) and are said to have the effect of reducing obese adipocytes by apoptosis as they differentiate into small adipocytes. Actually, it is known that insulin sensitivity is increased clinically and has a hypoglycemic effect (see, for example, Patent Document 1).

By the way, the main cause of lifestyle-related diseases is that normal small fat cells are enlarged into large fat cells, and accordingly, insulin resistance or a pathogenic substance is excessively secreted.
Therefore, in order to prevent and ameliorate various lifestyle-related diseases, it is necessary to stop the progress from small adipocytes to hypertrophic fat cells, or to reduce hypertrophic fat cells. For that purpose, it is considered important to take a material that reduces such large fat cells and promotes differentiation into small fat cells as food safer than meals every day.
However, there are no examples of food materials that have been studied from such a viewpoint, and materials having such functionality are not known.

Japanese Patent No. 3176694

  The present invention eliminates such conventional problems and uses as an active ingredient a component extracted from ginger rhizomes and / or leaves that have been eaten for many years while having the effect of promoting adipocyte differentiation. An object of the present invention is to provide a cell differentiation promoting agent that can be taken as a safe food every day.

Thus, the present inventors have screened plants having a PPARγ agonist-like action with the aim of promoting differentiation of adipocytes as an index for the purpose of reducing hypertrophic adipocytes and increasing small adipocytes. As a result, it was found that the extract of ginger promotes differentiation from preadipocytes to adipocytes, and the glucose uptake capacity of adipocytes was also increased. Based on such knowledge, the present invention has been completed.
The present invention according to claim 1 provides an agent for promoting cell differentiation from preadipocytes to adipocytes, comprising as an active ingredient a component extracted from the rhizome and / or leaves of ginger.
The present invention according to claim 2 provides an agent for promoting cell differentiation from hypertrophic fat cells to small normal fat cells, comprising as an active ingredient components extracted from rhizomes and / or leaves of ginger.
This invention which concerns on Claim 3 provides the visceral fat increase inhibitor which uses the component extracted from the rhizome and / or leaf of ginger as an active ingredient.
The present invention according to claim 4 provides a preventive or ameliorating agent for an adipocyte-derived lifestyle-related disease comprising as an active ingredient a component extracted from rhizomes and / or leaves of ginger.
The present invention according to claim 5 provides the cell differentiation promoting agent according to claim 1, wherein the component extracted from the rhizome and / or leaves of ginger is ginger phenylpropanoid.
The present invention according to claim 6 provides a food containing the cell differentiation promoting agent according to claim 1.

The adipocyte differentiation-promoting agent of the present invention has the effect of increasing normal small fat cells and activating sugar and lipid metabolism by promoting differentiation of preadipocytes into adipocytes. In addition, by reducing hypertrophic fat cells, it has preventive and therapeutic effects on lifestyle-related diseases such as diabetes, hyperlipidemia, hypertension, obesity, cancer caused by the increase of hypertrophic fat cells Conceivable.
Since the adipocyte differentiation promoter of the present invention is derived from ginger that has been eaten for many years, it is safe and very excellent for daily ingestion and use. In addition, since it has strong activity even at low concentrations, it can be used by adding to various foods, processed foods, pharmaceuticals, etc. Can be used.

The present invention according to claim 1 is an agent for promoting cell differentiation from preadipocytes to adipocytes, comprising as an active ingredient components extracted from rhizomes and / or leaves of ginger.
The agent for promoting cell differentiation of the present invention according to claim 1 comprises an ingredient extracted from rhizome and / or leaves of ginger as an active ingredient, and is extracted from rhizome or leaves of ginger or from both rhizomes and leaves. Ingredients are active ingredients. A dried product can be used as the rhizome and / or leaf of ginger, and the dried product can be used after being pulverized.
The extraction can be performed using an organic solvent such as methanol, ethanol, butanol, diethyl ether, etc., and it is particularly preferable to perform the extraction using ethanol, methanol, especially ethanol. As shown in the examples described later, in particular, the butanol layer in water / butanol separation of methanol or ethanol extract is active, and the ether-insoluble fraction is stronger than glycerol-3- Phosphate dehydrogenase activity (hereinafter abbreviated as GPDH activity) and a high amount of triglyceride (TG) are observed.

The cell differentiation promoting agent of the present invention according to claim 1 can be used as an extract (extract extract) as it is or after concentrating and drying it, and as a functional food preparation, pharmaceutical, etc., tablets, powders, It can be in various forms such as granules, capsules, liquids, etc., and can be taken orally or taken into the body by intravenous injection. In addition, a nutritional supplement can be added to the preparation.
The use amount of the cell differentiation promoting agent of the present invention according to claim 1 is basically an effective amount for promoting differentiation into adipocytes, but the extract amount is preferably 0.01 to 10 g, preferably Is preferably used once or several times a day so that about 0.05 to 1 g is ingested.

The present invention according to claim 2 is the same as that of the present invention according to claim 1, except that the active ingredient is a component extracted from the rhizome and / or leaves of ginger. It is different in that it is not an agent for promoting cell differentiation from fat cells to fat cells but an agent for promoting cell differentiation from enlarged fat cells to small normal fat cells.
According to the second aspect of the present invention, hypertrophic fat cells can be reduced and small normal fat cells can be increased by the action of promoting cell differentiation from preadipocytes to fat cells.

Next, the present invention according to claim 3 is a visceral fat increase inhibitor, although the component extracted from the rhizome and / or leaves of ginger is the same as that of the present invention according to claim 1. .
According to the third aspect of the present invention, increase in visceral fat and hypertrophy can be suppressed.

Moreover, although this invention which concerns on Claim 4 uses as an active ingredient the component extracted from the rhizome and / or leaf of ginger similarly to this invention which concerns on Claim 1, prevention of a fat cell origin lifestyle-related disease It is an improving agent.
According to the present invention according to claim 4, diabetes, hyperlipidemia, hypertension, arteries derived from adipocytes are suppressed by suppressing hypertrophy of visceral fat that causes excessive secretion of cytokines particularly associated with lifestyle-related diseases. Can broadly prevent and improve lifestyle diseases such as sclerosis and cancer.

The present invention according to claim 5 is the cell differentiation promoting agent according to claim 1, wherein the component extracted from the rhizome and / or leaves of ginger is a phenylpropanoid of ginger.
Specific examples of ginger phenylpropanoids include gingerol and gingerol.
As shown in Examples below, ginger phenylpropanoids have been shown to have a strong fat differentiation promoting action.

Furthermore, the present invention according to claim 6 is a food containing the cell differentiation promoting agent according to claim 1.
That is, the cell differentiation promoting agent according to claim 1 is used as it is or by appropriately adding a nutritional supplement or the like, thereby widely preventing and improving lifestyle-related diseases such as diabetes, hyperlipidemia, hypertension, arteriosclerosis and cancer. Can be used as a functional food.

  Note that the present invention according to claims 2 to 4 can be limited as in claims 5 and 6 as in the case of claim 1.

  EXAMPLES Next, although an Example demonstrates this invention, this invention is not restrict | limited by these.

Production Example 1
3 kg of methanol was added to 1 kg of raw ginger (rhizome), and extraction under heating was performed. The resulting extract was concentrated and dried by a rotary evaporator. Next, 500 ml each of water and n-butanol were added to the concentrated and dried solid, shaken and stirred, and then allowed to stand to separate into an aqueous layer and a butanol layer. The aqueous layer and butanol layer were dried under reduced pressure, and diethyl ether was added to the butanol-soluble material and shaken and stirred. Thereafter, the mixture was centrifuged (3,000 rpm, 10 minutes), and the supernatant was separated into an ether-soluble substance and a precipitate. Further, ether was further added to the precipitate to separate a soluble substance and an insoluble substance several times. By such an operation, 1.8 g of ether soluble fraction, 0.3 g of ether insoluble fraction, and 17.8 g of water soluble fraction were obtained.

Production Example 2
Add 3 liters of ethanol to 1 kg of raw ginger (rhizome), perform heating under reflux extraction as in Production Example 1, concentrate the resulting extract with a rotary evaporator and dry to solid, and extract an ethanol extract (ethanol extract) 20 g was obtained.

Example 1
Mouse-derived preadipocytes 3T3-L1 cells were cultured at 37 ° C. in DME medium (Dulbecco's modified Eagle medium) containing 10% fetal calf serum using a 6-well multiplate. The medium was changed every 2-3 days. After the cells reached a dense state, differentiation was induced with the following representative reagents. That is, differentiation induction treatment was performed by adding three types of mixed solutions of dexamethasone, methylisobutylxanthine and insulin or insulin to the medium. At this time, the test object obtained in Production Example 1 was added. The differentiation induction treatment was carried out for 2 days, and then the culture was continued in the original medium, but insulin and the test substance were continuously added when the medium was changed. About 1 week later, GPDH activity in the medium, which is an index of differentiation into adipocytes, was quantified. This enzyme is an important enzyme in sugar and lipid metabolism, and its activity increases when preadipocytes differentiate into adipocytes. Therefore, if this enzyme activity increases, differentiation has occurred. In addition, when differentiation into adipocytes occurs, the amount of TG in the cell increases. At the same time, the amount of TG in the cell was measured and used as an indicator of differentiation.

The results are shown in FIGS. FIG. 1 is a graph showing GPDH activity and TG amount [relative values based on the case where differentiation induction treatment with insulin is performed (control: 100)] for the ginger ether-soluble fraction in Production Example 1. . FIG. 2 is a graph showing GPDH activity and TG amount [relative values based on the case where differentiation induction treatment with insulin is performed (control: 100)] for the ginger ether insoluble fraction in Production Example 1. FIG. 3 is a graph showing GPDH activity and TG amount [relative values based on the case where differentiation induction treatment with insulin is performed (control: 100)] for the ginger aqueous fraction in Production Example 1.
The numerical values in the figure show the GPDH activity produced by various concentrations of the test substance based on the GPDH activity when the differentiation induction treatment with insulin was performed on the ginger ether-soluble fraction in Production Example 1 (control: 100). Relative values compared, and relative values comparing TG amounts produced by various concentrations of the test substance with reference to the amount of TG when differentiation induction treatment with insulin is performed (control: 100).
As is apparent from the figure, both GPDH activity and TG amount increased by the addition of 100 g / ml ginger ether insoluble fraction or ginger ether soluble fraction. In the ginger ether insoluble fraction, high values were shown even at concentrations of 10 μg / ml and 30 μg / ml.

Example 2
Using the ethanol extract obtained in Production Example 2, the effect of promoting differentiation into mouse-derived preadipocytes 3T3-L1 cells was examined in the same manner as in Example 1. By adding 100 μg / ml of ethanol extract, both GPDH activity and TG amount showed high values, GPDH activity increased 2.1 times and TG amount increased 1.8 times.

Example 3
As for gingerol, a typical spicy component of ginger, the fat differentiation promoting action was examined in the same manner as in Example 1. The results are shown in FIG. FIG. 4 is a graph showing the GPDH activity and TG amount of gingerol [both relative values based on the case where differentiation induction treatment with insulin is performed (control: 100)].
As a result, the GPDH activity and the amount of TG increased in a concentration-dependent manner, showing 4.2 times the activity at a concentration of 100 μM compared to the control. Therefore, it was shown that gingerol has a fat differentiation promoting action.

Example 4
Using the ginger ether insoluble fraction of Production Example 1 which showed strong activity in Example 1, the glucose uptake ability into mouse-derived preadipocytes 3T3-L1 cells was measured. FIG. 5 is a graph showing the glucose uptake amount of the ginger ether insoluble fraction.
As a result, at a concentration of the ginger ether insoluble fraction of 100 μg / ml, a glucose uptake activity of 3.5 times higher than that without addition was shown.

Example 5
Using gingerol, which showed a strong differentiation promoting action in Example 3, the ability of glucose uptake into mouse-derived preadipocytes 3T3-L1 cells was measured. FIG. 6 is a graph showing the glucose uptake amount of gingerol.
As a result, the glucose uptake ability increased in a dose-dependent manner at a concentration of 0.03 mM to 1.0 mM of gingerol.

It is a graph which shows the GPDH activity and the amount of TG [both relative values based on the case where differentiation induction treatment with insulin is performed (control: 100)] for the ginger ether soluble fraction in Example 1. It is a graph which shows the GPDH activity and TG amount about the ginger ether insoluble fraction in Example 1 [both are relative values when the differentiation induction treatment with insulin is performed as a reference (control: 100)]. It is a graph which shows the GPDH activity and the amount of TG [the relative values based on the case where differentiation induction treatment with insulin is performed (control: 100)] for the ginger aqueous layer fraction in Example 1. It is a graph which shows the GPDH activity and the amount of TG of gingerol in Example 3 [both are relative values when the differentiation induction treatment with insulin is performed as a reference (control: 100)]. 4 is a graph showing the glucose uptake amount of a ginger ether insoluble fraction in Example 4. 6 is a graph showing the glucose uptake amount of gingerol in Example 5.

Claims (6)

  An agent for promoting cell differentiation from preadipocytes to adipocytes, comprising as an active ingredient a component extracted from rhizomes and / or leaves of ginger.   An agent for promoting cell differentiation from hypertrophic fat cells to small normal fat cells, comprising as an active ingredient components extracted from rhizomes and / or leaves of ginger.   A visceral fat increase inhibitor comprising as an active ingredient a component extracted from rhizomes and / or leaves of ginger.   A preventive or ameliorating agent for adipocyte-derived lifestyle-related diseases, comprising as an active ingredient a component extracted from rhizomes and / or leaves of ginger.   The cell differentiation promoting agent according to claim 1, wherein the component extracted from the rhizome and / or leaves of ginger is ginger phenylpropanoid. A food containing the cell differentiation promoting agent according to claim 1.

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