JP2006328056A - Obesity-preventing agent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an obesity-preventing agent in which in vivo lipid metabolism is improved and accumulated fat is actively decreased and body weight is healthfully decreased. <P>SOLUTION: The obesity-preventing agent comprises a plant body of a cruciferous plant, e.g. Japanese horseradish, preferably a leaf of Japanese horseradish or an extract obtained by carrying out pulverizing treatment and grinding-down treatment of the cruciferous plant, extraction treatment of the cruciferous plant with a solvent and drying treatment of the cruciferous plant alone or in combination thereof as an active ingredient. The obesity-preventing agent improves lipid metabolism and healthfully decreases the fat amount in the body and body weight. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to an anti-obesity agent that prevents or ameliorates obesity by ingesting a cruciferous plant and its components, and in particular, prevents obesity by improving in vivo lipid metabolism to reduce in vivo fat and reduce body weight. It relates to the agent.

  Japan currently has 7.4 million diabetics, 15 million hypertension and 23 million hyperlipidemia patients, and the estimated number of patients in all lifestyle-related diseases is several thousand. As a cause of these lifestyle-related diseases, it has been pointed out that dietary habits are high calorie intake such as animal fat and chronic lack of exercise. By continuing these lifestyle habits, it is thought that the amount of body fat increases, leading to weight gain and obesity.

  If such a state is left unattended, it may cause diabetes, arteriosclerosis, etc., and may be fatal due to ischemic heart disease or cerebrovascular disorder. It is very important to prevent such diseases.

  In general, moderate exercise and control of calorie intake are encouraged for the prevention and improvement of obesity, but the current situation is that it is difficult to exercise as expected in modern busy lifestyles. Therefore, it has been desired to develop a material that can prevent or improve obesity by easy oral intake.

  A dietary fiber is mentioned as what is generally used as a diet material. Dietary fiber shortens the intestinal residence time of food, further suppresses carbohydrate absorption and moderates the increase in blood sugar level. As a result, insulin secretion that promotes conversion of carbohydrates to neutral fat is reduced, and fat accumulation is suppressed. Glucomannan (Japanese Patent Laid-Open No. 2002-186469), Prickly Pair Cactus (Japanese Patent Laid-Open No. 2003-70444) and the like have been devised as materials that expect these effects.

  In addition, citrus fruits (Japanese Patent Laid-Open No. 2005-40107), Katuava (Japanese Patent Laid-Open No. 2004-292368) and the like have been devised as materials that inhibit the activity of lipase, an enzyme essential for digestion and absorption of fat.

  However, all of these materials suppress digestion and absorption of carbohydrates and lipids and prevent accumulation in the body, and do not actively burn or excrete absorbed and accumulated fat.

Therefore, there has been a demand for a material that can be continuously consumed without difficulty in daily eating habits, actively reduces fat in the body, and reduces body weight healthily.

  In general, when fat cells in the body become enlarged, a peptide hormone, leptin, is secreted and acts on the feeding center in the hypothalamus of the brain to suppress appetite, while also acting on brown adipocytes and energy metabolism It is thought to promote the increase of Control of the function of leptin, other lipid-metabolizing hormones and brown adipocytes is important for the prevention and improvement of obesity.

As a result of diligent research, the present inventors have found that cruciferous plants or extracts thereof have an effect of preventing and improving obesity, in particular, improving lipid metabolism in vivo and reducing body weight. The invention has been completed.
JP 2002-186469 A JP 2003-70444 A Japanese Patent Laid-Open No. 2004-159637 JP-A-2005-40107 JP 2004-292368 A JP 2005-8572 A

  It is an object of the present invention to provide an anti-obesity agent that improves lipid metabolism in vivo, actively reduces accumulated fat to reduce weight healthyly, and improves the above-mentioned drawbacks of the known techniques. It is in.

  In order to achieve the above-described object, the antiobesity agent of the present invention contains a cruciferous plant or an extract thereof as an active ingredient to improve lipid metabolism in vivo or inhibit fat degradation. It is characterized by actively reducing the accumulated fat by suppressing absorption and healthy weight loss.

  The present invention contains a cruciferous plant or an extract thereof as an active ingredient, and improves the lipid metabolism in the living body by ingesting it as a food or as a pharmaceutical product or quasi-drug, so that the body weight is healthy. There is an effect of reducing.

  Hereinafter, the present invention will be described in detail.

  Examples of the cruciferous plant used in the present invention include horseradish, horseradish, cabbage, watercress, Brussels sprouts, cauliflower, radish, leach radish, rapeseed, broccoli, Takana, mustard, turnip, Chinese cabbage. Among these, this wasabi is preferable, and this wasabi leaf is particularly preferable. In the present invention, these cruciferous plants are used singly or in combination.

The active ingredients of the above cruciferous plants are obtained by physical treatment means such as pulverization or grated plant bodies, or by solvent extraction treatment means such as water, methanol, ethanol, acetone, ethyl acetate, diethyl ether, dichloromethane, dichloroethane and the like. Further, it can be obtained by a drying treatment means such as hot air drying, freeze drying or a combination of these treatments.

  The active ingredients of the cruciferous plants used in the present invention are flavonoids such as apigenin, luteolin, kaempferol and their glycosides / multimers, phenylpropanoids such as ferulic acid and sinapinic acid, and their glycosides / multiples. Body, including other polyphenols.

  Active ingredients other than polyphenols include isothiocyanates, specifically allyl isothiocyanate, secondary butyl isothiocyanate, 3-butenyl isothiocyanate, 4-pentenyl isothiocyanate, 5-hexenyl isothiocyanate, 5- Methylthiopentyl isothiocyanate, 6-methylthiohexyl isothiocyanate, 7-methylthioheptyl isothiocyanate, 8-methylthiooctyl isothiocyanate, 4-methylsulfinyl butyl isothiocyanate, 5-methylsulfinyl pentyl isothiocyanate, 6-methylsulfinyl hexyl isothiocyanate, 7-methylsulfinyl heptyl isothiocyanate, 8-methylsulfinyl octyl isothiocyanate.

  Further, as active ingredients, it contains indoles such as 5-methoxy-indole-3-acetonitrile, chlorophyll and the like.

  The anti-obesity agent of the present invention can be taken alone, but can also be used by adding it to various foods and beverages or medicines. Specifically, liquid foods such as soft drinks, tea beverages, drinks, alcoholic beverages, solid foods such as confectionery, breads, noodles and seasonings, pharmaceuticals such as powders, granules, capsules, tablets, etc. It can be added to quasi drugs.

Extraction of polyphenols contained in this wasabi leaf This wasabi leaf extract was prepared. The wasabi was cut into 4 kg, 16 kg of 50% ethanol was added to 4 kg, and extracted for 1 hour with stirring. The extract was filtered, concentrated with an evaporator and freeze-dried, and pulverized with a food mill to obtain 70 g of a green powder. This was subjected to the test as this wasabi leaf extract.

  The results of subjecting this wasabi leaf extract to high performance liquid chromatography (HPLC) are shown in FIG. As shown in FIG. 1, plural kinds of flavonoids and phenylpropanoids were detected from this wasabi leaf extract.

Extraction of isothiocyanates contained in this wasabi rhizome This wasabi rhizome extract was prepared. 1 kg of grated wasabi rhizome was put in a stainless steel sealed container and kept at 25 ° C. for 3 hours to generate isothiocyanates. Thereafter, the pungent component was removed for 1 hour while sucking the air in the container with a vacuum pump. 3 liters of 50% ethanol was added to this wasabi rhizome from which the pungent taste was removed, followed by stirring and extraction at room temperature for 1 hour, followed by filtration with a pressure filter to obtain an extract. The extract was concentrated with an evaporator, and then freeze-dried and pulverized with a food mill to obtain 55 g of a brown powder. This was subjected to the test as the wasabi rhizome extract.

  The results of subjecting this wasabi leaf extract to mass spectral gas chromatography (GCMS) are shown in FIG. As shown in FIG. 2, multiple types of isothiocyanates were detected from this wasabi leaf extract.

Examination of anti-obesity effect using diet-induced obese rats This wasabi leaf extract was obtained by the method described in Example 1, and this wasabi rhizome extract was obtained by the method described in Example 2.

  Thirty Std Wister rats (male, age 6 weeks) were raised for 28 days in individual cages made of stainless steel. The temperature of the animal room was maintained at 24 ± 1 ° C., and the animals were reared at a humidity of 50%, a light period of 08: 00-20: 00, and a dark period of 20: 00-08: 00.

  Rats had a high fat diet group (control group) so that the average body weight was uniform, a high fat diet group (Example 1 group) containing 5% of this wasabi leaf extract of Example 1, and this wasabi of Example 2. The feed composition was shown in Table 1 by dividing into a high fat diet group (Example 2 group) containing 5% rhizome extract. Feed intake was limited so that the feed intake was the same in all groups.

  Body weight and feed intake were measured three times a week and water was ad libitum. On the 28th day after breeding, the animals were fasted overnight and then sacrificed by decapitation under ether anesthesia.

  Rats subjected to the test were dissected. The pancreas, kidney, liver, gastrocnemius muscle, inguinal adipose tissue, perirenal adipose tissue, accessory epididymal adipose tissue, and brown adipose tissue were removed and weighed. A part of the liver was placed in a neutral buffered formalin solution to prepare a pathological specimen. Other organs were quickly frozen in liquid nitrogen and stored at −80 ° C. with serum.

  Neutral lipids of the extracted liver were measured. About 15 m of Folch's solution was added to 2.5 g of liver and homogenized. A well-turbid homogen solution was extracted in a 50 ml volumetric flask for one day. After extraction, the volume was made up with a Folch solution, mixed, and then filtered through a No. 2 filter paper to prepare a sample. 150 μl of the subject group and 120 μl of the other group were placed in a test tube and left at room temperature to evaporate the solvent.

  Cholesterol cholesterol, triacylglycerol, and phospholipid concentrations were measured with cholesterol CII kit Wako, triglyceride G-test Wako, and phospholipid test Wako kits, respectively.

  Cholesterol CII Kit Wako, Triglyceride G-Test Wako, Phospholipid Test Wako, Urea Nitrogen B Test Wako, each concentration of serum cholesterol, triacylglycerol, phospholipid, urea nitrogen, creatinine, free fatty acid, GOT, GPT , Creatinine-test Wako, NEFA-C test Wako, GOT-UV test Wako, GPT-UV test Wako.

  The concentrations of adiponectin, leptin, and TNF-α in the serum were measured using the Mouse / rat adiponectine ELISA kit, Rat Leptin ELISA kit, Yanaihara, Rat TNF-α ELISA kit, and TFB, respectively.

  Brown adipose tissue β3-adrenergic receptor (hereinafter referred to as β3-AR), peroxisome growth promoting receptor γ (hereinafter referred to as PPARγ), CAAT / enhancer binding protein α (hereinafter referred to as C / EBPα), PPARγ Coactivator 1 (PGC-1), medium chain acyl CoA dehydrogenase (hereinafter referred to as “MCAD”) mRNA expression level and white adipose tissue MCAD mRNA expression level were measured using real-time PCR (ABI PRISM® 7000 Sequence Detection). System).

  The results are shown below. Due to intake restrictions, there was no difference in the amount of food consumed during the breeding period among the three groups. The final body weight of the Example 1 group and the Example 2 group was significantly less than that of the control group. (Table 2)

  The pancreas, kidney, liver and gastrocnemius muscle weights were not significantly different among the three groups. There was no significant difference between the three groups in the weight of the white adipose tissue around the kidney and the white adipose tissue around the accessory testis. there were. The brown adipose tissue of Example 1 group and Example 2 group increased significantly compared with the control group. (Table 3).

  Serum total cholesterol, triacylglycerol, phospholipid, free fatty acid, creatinine, urea nitrogen, GPT, and GOT concentrations were not significantly different among the three groups. The liver TBARS and kidney TBARS concentrations in the Example 1 group tended to decrease compared to the control group. (Table 4)

  The concentrations of TNF-α and adiponectin in the serum were not significantly different among the three groups, but the leptin in the Example 1 group and the Example 2 group tended to increase. (Table 5)

  Brown adipose tissue β3-AR (FIG. 3), PPARγ (FIG. 4), PGC-1 (FIG. 5), MCAD (FIG. 6) increased significantly, and C / EBPα (FIG. 7) tended to increase. . The white adipose tissue MCAD (FIG. 8) tended to increase.

  The diet-induced obese rats used in this study are similar to human visceral obesity induced by lifestyle habits, and this wasabi leaf is effective as an obesity-preventing food that prevents and ameliorates obesity caused by lifestyle-related diseases confirmed.

  In vitro lipase inhibitory activity was measured. 2 g each of this wasabi leaf, horseradish leaf, and wasabi rhizome were lyophilized and ground.

  10 ml of hexane was added to the above sample and homogenized with a glass / polytron homogenizer. This was left at room temperature for 24 hours. This was filtered and concentrated, and 10 ml of tetrahydrofuran was added to obtain a fat-soluble fraction.

  To the residue after filtration, 10 ml of tetrahydrofuran was added and stirred, followed by extraction for 24 hours, and this supernatant was used as a water-soluble fraction.

Tetrahydrofuran 85 in 100 μl tetrahydrofuran containing 0.01 mg of the above sample.
400 μl of 0.1 μl Macilvan buffer PH 7.0 was added and stirred in a small test tube.

  After adding 0.15 mM 4-methylumbelliferon oleate (4-MU oleate) dissolved in tetrahydrofuran and stirring, pre-incubated 0.05 ml (0.05 units) Polysine pancreatic lipase was added and heated at 37 ° C. for 20 minutes. The reaction was stopped by adding 0.1N hydrochloric acid. To this, 2 ml of citrate buffer was added to prepare PH 4.3.

  The decomposition rate to 4-MU by lipase was measured with a fluorometer (wavelength 450 nm, excitation wavelength 320 nm). For the inhibition rate, the control was 100%, and IC50 was calculated. Inhibitory activity was calculated by the following formula.

  Inhibition rate% = (control 4-MU fluorescence-sample addition 4-MU fluorescence) / control 4-MU fluorescence × 100

  The results are shown below. None of the extracted components had a weak lipase inhibitory activity in vitro. (Fig. 9)

  These results suggest that the anti-obesity effect in rats is mainly due to increased energy metabolism accompanied by an increase in brown adipose tissue, and that lipase inhibitory effects in the gastrointestinal tract are also involved. .

The effect of this wasabi leaf on obesity prevention / amelioration was confirmed in humans. A hard capsule enclosing 150 mg of this wasabi leaf extract extracted by the method described in Example 1 was prepared.

  Six healthy men were ingested hard capsules with this wasabi leaf extract three times a day for three months.

  The results are shown below. At 3 months after ingesting this wasabi leaf, the body weight was significantly reduced compared to before ingestion. (FIG. 10) From this, it was confirmed that ingestion of this wasabi leaf is effective in suppressing body weight even in humans.

It is a HPLC chromatograph of this wasabi leaf 50% ethanol extract. It is a GCMS chromatograph of this wasabi leaf 50% ethanol extract. It is a comparison graph with the control | contrast of brown adipose tissue (beta) 3-AR. It is a comparison graph with the brown adipose tissue PPARγ control. It is a comparison graph with the control | contrast of the brown adipose tissue PGC-1. It is a comparison graph with the control of white adipose tissue MCAD. It is a comparison graph with a brown adipose tissue C / EBPα control. It is a comparison graph with the brown adipose tissue MCAD control. It is the graph showing the lipase inhibitory activity of this wasabi leaf and a horseradish leaf. It is the graph showing transition of the body weight when this human wasabi leaf was ingested by the human.

Claims (10)

  It contains cruciferous plants or their components as a main component, improves lipid metabolism in the living body, or inhibits fat breakdown and suppresses absorption, thereby reducing fat in the body and reducing body weight. Anti-obesity agent.   The obesity preventing agent according to claim 1, wherein the cruciferous plant is wasabi and / or wasabi leaf.   The cruciferous plant according to claim 1, wherein the cruciferous plant is one or more selected from the group of horseradish, cabbage, watercress, Brussels sprouts, cauliflower, radish, leach radish, rapeseed, broccoli, Takana, mustard, turnip, and Chinese cabbage Item 2. The anti-obesity agent according to Item 1.   The obesity-preventing agent according to claim 1, wherein the component contained in the cruciferous plant is one or more of the group of flavonoids, phenylpropanoids, and isothiocyanates. 5. The isothiocyanate of claim 4, wherein the isothiocyanates are 4-methylsulfinyl butyl isothiocyanate, 5-methylsulfinyl pentyl isothiocyanate, 6-methylsulfinyl hexyl isothiocyanate, 7-methylsulfinyl heptyl isothiocyanate and 8-methylsulfinyl octyl isothiocyanate. Claim 4 selected from the group
The antiobesity agent described in 1. In Claim 4, the isothiocyanates are allyl isothiocyanate, secondary butyl isothiocyanate, 3-butenyl isothiocyanate, 4-pentenyl isothiocyanate, 5-hexenyl isothiocyanate, 5-methylthiopentyl isothiocyanate, 6-methylthio. The antiobesity agent according to claim 4, selected from the group of hexyl isothiocyanate, 7-methylthioheptyl isothiocyanate and 8-methylthiooctyl isothiocyanate.   The obesity-preventing agent according to claim 1, wherein the component of the cruciferous plant is obtained by pulverizing or grateing the cruciferous plant, performing extraction treatment with a solvent, or drying treatment alone or in combination. .   The cruciferous plant or a component thereof according to claim 1, wherein the cruciferous plant or its component suppresses an increase in white adipose tissue weight, promotes an increase in brown adipose tissue weight, or promotes an increase in leptin in vivo. The antiobesity agent according to claim 1, which improves lipid metabolism in the body.   The cruciferous plant or a component thereof according to claim 1, wherein leptin, β3-adrenergic receptor, peroxisome growth promoting receptor γ, CAAT / enhancer binding protein α, PPARγ coactivator 1, medium chain acyl CoA dehydrogenase in vivo The obesity-preventing agent according to claim 1, which improves lipid metabolism in a living body by promoting an increase in one or a plurality of types of indices. A food or drink or a pharmaceutical comprising the obesity-preventing agent according to claim 1 containing a cruciferous plant or a component thereof as a main component.

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