JP6829550B2 - How to use anti-obesity foods and carob powder - Google Patents

Description

The present invention relates to anti-obesity foods and methods of using carob powder.

In recent years, the increase in the obese population has become a serious social problem, especially in developed countries. Obesity is a lifestyle-related disease and can be a factor that causes lifestyle-related diseases such as hyperlipidemia and diabetes. Therefore, the number of patients with lifestyle-related diseases tends to increase as the obese population increases. In addition, there is a strong demand for the elimination of obesity not only for health reasons but also for cosmetic reasons.

Obesity is a condition in which body fat is excessively accumulated in the body. Examples of the method for reducing body fat in the body include diet therapy and exercise therapy. However, diet therapy must be strictly implemented, and excessive dietary restrictions may adversely affect health. In addition, exercise therapy needs to be performed continuously over a long period of time. For this reason, it is often difficult to carry out continuously in a busy modern society, and it is not always an easy method.

Therefore, in recent years, there has been a demand for finding a natural substance that can be easily and safely ingested and that can suppress obesity. For example, anti-obesity foods using powdered shells having nacre and extracts of plants belonging to the genus Maple have been proposed (Patent Documents 1 and 2).

By the way, carob powder is known as a natural food material having functionality. Carob powder is an ingredient obtained by drying the pods (flesh) of carob beans (carob beans) cultivated mainly in the vicinity of the Mediterranean Sea and processing them into powder, and is used as a substitute for cocoa powder. The functions of carob powder as a food material include anti-inflammatory properties that can reduce inflammatory diseases such as the gastrointestinal tract (Patent Document 3) and antioxidant properties that can suppress the oxidation of tissues that cause aging and the like. (Non-Patent Document 1) and the like are known.

Japanese Unexamined Patent Publication No. 2006-143614 JP-A-2009-268452 Japanese Unexamined Patent Publication No. 2003-339352

Kumazawa et al., "Antioxidant Activity of Polyphenols in Carob Pods" Journal of Agricultural and Food Chemistry, 2002, Vol.50, No.2, p.373-377

However, the anti-obesity foods proposed in Patent Documents 1 and 2 are not always sufficiently high in anti-obesity property, and there is room for further improvement. Further, in Patent Document 3 and Non-Patent Document 1, there is no report that carob powder is a food material having anti-obesity property.

The present invention has been made in view of the problems of the prior art, and the subject thereof is to provide an anti-obesity food having high versatility as a food material and having excellent anti-obesity. There is. Another object of the present invention is to provide a method for using carob powder, which can provide an anti-obesity food having excellent anti-obesity.

As a result of diligent studies to solve the above problems, the present inventors have found that the above problems can be solved by adopting the following configuration, and have completed the present invention. That is, according to the present invention, the following anti-obesity products are provided .
[1] contains roasted carob powder as an active ingredient, which pre Kiabu roast carob powder, after roasting less than 45 minutes 30 minutes or more at 120 to 180 ° C. The portion of the sheath of the carob, was prepared by grinding der is, SomosomoSae the enlargement of the fat cells by reducing the fat droplets are accumulated in the fat cells, an anti-obesity food suppress obesity.

Further, according to the present invention, the following methods of using carob powder are provided .
[2 ] Roasted carob powder is added to a food to impart an anti-obesity effect to the food by reducing fat droplets accumulated in adipocytes to suppress adipocyte enlargement and suppressing obesity. a method, before Kiabu decoction carob powder, after partial roasting below 120 to 180 ° C. 45 minutes 30 minutes or more at the sheath of the carob, using the der Luke Rob powder that prepared by grinding.

According to the present invention, it is possible to provide an anti-obesity food having high versatility as a food material and having excellent anti-obesity. Further, according to the present invention, it is possible to provide a method of using carob powder which can provide an anti-obesity food having excellent anti-obesity.

It is a graph which shows the result of the adipose progenitor cell differentiation induction test. It is a graph which shows the result of the adipose progenitor cell differentiation induction test. It is a graph which shows the result of the radical scavenging test. It is a graph which shows the result of the nitric oxide production test.

Hereinafter, embodiments of the present invention will be described, but the present invention is not limited to the following embodiments. The anti-obesity food of the present invention contains carob powder as an active ingredient. The details will be described below.

Carob powder is an ingredient obtained by drying the pod (flesh) of carob bean (carob bean) and then pulverizing it. Carob powder is known as a food material containing a large amount of dietary fiber and calcium, a low content of lipid, and no stimulants such as caffeine and theobromine. Further, since carob powder is a food material having a peculiar sweetness and aroma and slightly brownish, it is a material for producing chocolate-flavored foods and the like as a substitute for cocoa (cacao) powder. It is known to be used as.

As a result of diligent studies, the present inventors have found that carob powder has anti-obesity properties. Here, the term "anti-obesity" as used herein means a property of suppressing obesity by reducing fat droplets accumulated in adipocytes and suppressing hypertrophy of adipocytes. Such anti-obesity property of carob powder is a previously unknown function discovered by the present inventors for the first time. By using carob powder having such a function, it is possible to impart anti-obesity to the food and obtain the anti-obesity food of the present invention having anti-obesity.

In addition, as described above, carob powder is also a food material having anti-inflammatory and antioxidant properties. Therefore, the anti-obesity food containing carob powder as an active ingredient is a highly functional food having anti-obesity as well as anti-inflammatory and antioxidant properties.

As the carob powder, roasted carob powder obtained by roasting raw carob powder can be used in addition to so-called raw (unroasted) carob powder, which is powdered after drying the pod of carob bean. .. Depending on the roasting conditions (difference in the degree of roasting), the roasted carob powder can be used in, for example, carob powder Light (light roasting), carob powder Dark (medium roasting), carob powder Extra Dark (deep roasting), etc. Can be categorized. It is presumed that the composition of the functional components contained in the obtained roasted carob powder differs due to the difference in roasting conditions (difference in the degree of roasting), but the composition of each such component is specified. It is practically difficult to do.

The carob powder contained as an active ingredient in the anti-obesity food of the present invention is preferably roasted carob powder. Further, as the roasted carob powder, carob powder Light (light roasting) is preferable. By using roasted carob powder, especially lightly roasted roasted carob powder (carob powder Light), the anti-obesity property of the obtained anti-obesity food can be further enhanced.

The amount of carob powder contained in the anti-obesity food of the present invention can be appropriately set according to the type and purpose of the food. Specifically, the amount of carob powder in the anti-obesity food may be about 5 to 30% by mass based on the entire anti-obesity food. If the content of carob powder is less than 5% by mass, anti-obesity may be insufficient. On the other hand, if the content of carob powder exceeds 30% by mass, the taste of food may be substantially affected.

Since the anti-obesity food of the present invention has excellent anti-obesity properties, it is a food for specified health use and functionality aimed at obtaining effects such as anti-obesity, diet, anti-hypertriglyceridemia, and anti-arteriosclerosis. It is useful as food, nutritional supplement, health food, etc. It is expected that the effect will be exhibited by ingesting such foods on a daily basis.

Specific embodiments (food examples) of the anti-obesity food of the present invention include flour products such as bread and cake and their dough; rice and rice; snacks, biscuits, cookies, chocolate, sweets, ice cream, manju and the like. Confectionery; processed meat products such as ham and sausage; marine products such as kamaboko and chikuwa; beverages such as milk powder, tea, water, liquor, fruit juice, milk, and other soft drinks; and the like.

Various additives can be added to the anti-obesity food of the present invention, if necessary, as long as the effects of the present invention are not impaired. Specific examples of additives include excipients, pH adjusters, refreshing agents, suspending agents, defoaming agents, thickeners, solubilizing agents, disintegrants, binders, lubricants, coating agents, etc. Colorants, flavoring agents, surfactants, plasticizers, minerals, vitamins, fragrances and the like can be mentioned.

Hereinafter, the present invention will be specifically described based on examples, but the present invention is not limited to these examples. In addition, "part" and "%" in Examples and Comparative Examples are based on mass unless otherwise specified.

<Preparation of carob powder>
The pods (flesh) of Ceratonia siliqua native to Spain were placed in an oven set at 120-180 ° C and roasted for 30 minutes. Then, it was pulverized to prepare roasted carob powder (carob powder Light). Further, the carob powder Dark and the carob powder Extra Dark, which are roasted carob powders, were prepared in the same manner as in the case of the carob powder Light described above, except that the roasting time was set to 45 minutes and 60 minutes, respectively. Further, the dried carob pod (flesh) was crushed without roasting to prepare unroasted carob powder. Hereinafter, unroasted carob powder will be referred to as "carob R". Further, the carob powder Light, the carob powder Dark, and the carob powder Extra Dark obtained by roasting are referred to as "carob L", "carob D", and "carob E", respectively.

<Preparation of sample for activity evaluation>
To 500 mg of the prepared four types of carob powder (carob R, L, D, and E) and 500 mg of commercially available cocoa powder (cocoa B and H), 10 mL of methanol was added, and the mixture was extracted with ultrasonic waves for 30 minutes. The supernatant was filtered after standing. 10 mL of 80% methanol was added to the residue, and the mixture was extracted with ultrasonic waves for 30 minutes. After standing, the supernatant was filtered and mixed with the supernatant obtained by filtering first to prepare an extract. The solvent was removed from the extract obtained using an evaporator to obtain each methanol extract. 0.5 mL of water was added to each of the obtained methanol extracts to suspend the mixture, and the mixture was subjected to a conditioned and equilibrated column (trade name “Sep-Pak C18”, manufactured by Japan Waters Corp.). After washing with 0.5 mL of water (twice), the mixture was eluted with 0.5 mL of methanol to obtain an eluate. The solvent was removed from the eluate obtained by using an evaporator to obtain a dry matter (extract). Dimethyl sulfoxide (DMSO) was added to each of the obtained extracts to prepare a sample for activity evaluation having a concentration of the extract of 100 mg / mL.

<Adipose progenitor cell differentiation induction test>
Mouse-derived preadipocytes (3T3-L1) were placed in 100 μL wells of 96-well plates at 1 × 10 ⁴ cells / well and cultured at 37 ° C. in a 5% CO ₂ incubator for 2 days. Differentiation-inducing medium was prepared by adding 3-isobutylmethylxanthine (IBMX) and dexamethasone (Dex) to DMEM medium at 0.5 mM and 0.1 μM, respectively. Samples for activity evaluation were added to the prepared differentiation-inducing medium, and dilution series of 25, 50, and 100 μg / mL were prepared for each sample for activity evaluation. The medium in the wells of the 96-well plate was removed, 100 μL of the differentiation-inducing medium containing the sample for activity evaluation was added, and the cells were cultured in an incubator at 37 ° C. and 5% CO ₂ for 2 days. In addition, only the differentiation-inducing medium containing no sample for activity evaluation was added in an amount of 100 μL each, and the cells were cultured as untreated wells.

Insulin was added to DMEM medium at a concentration of 0.1 μg / mL to prepare a differentiation maintenance medium. Samples for activity evaluation were added to the prepared differentiation maintenance medium, and dilution series of 25, 50, and 100 μg / mL were prepared for each sample for activity evaluation. The medium in the wells of the 96-well plate was removed, 100 μL of the differentiation-inducing medium containing the sample for activity evaluation was added, and the cells were cultured in an incubator at 37 ° C. and 5% CO ₂ for 3 days. In the untreated wells, only 100 μL of the differentiation maintenance medium containing no sample for activity evaluation was added and cultured. After culturing for 3 days, the same operation was performed, and after culturing in an incubator at 37 ° C. and 5% CO ₂ for 3 days, the cell viability and fat accumulation rate were calculated by the WST-8 method and the Oil Red O method shown below, respectively. ..

(Cell viability)
The WST-8 / PBS solution prepared by adding WST-8 to PBS at 2% was added in 100 μL each into the wells of the 96-well plate from which the medium had been removed. After culturing in an incubator at 37 ° C. and 5% CO ₂ for 3 hours, the absorbance at 450 nm was measured using an absorption plate reader. The "cell viability (%)" of the cells treated with each activity evaluation sample was calculated from the measured absorbance values, assuming that the absorbance value of the untreated well was 100%. The test was performed with n = 3. The results are shown in FIGS. 1 and 2.

(Fat accumulation rate)
The WST-8 / PBS solution was removed from each well of the 96-well plate, 100 μL of formalin was added, and the mixture was allowed to stand overnight at 4 ° C. Formalin was removed, 100 μL of a 60% isopropanol aqueous solution was added, the cells were washed, and then the isopropanol aqueous solution was removed. 100 μL of Oil Red O solution (5 mg / mL Oil Red O in isopropanol: water = 6: 4) was added to each well, allowed to stand at room temperature for 10 minutes, and then the Oil Red O solution was removed. After performing the washing operation of adding 100 μL of ultrapure water three times, the ultrapure water was completely removed. After adding 100 μL of isopropanol to each well and shaking for 3 minutes, the absorbance at 520 nm was measured using an absorption plate reader. Assuming that the absorbance value of the untreated well was 100%, the "fat accumulation rate (%)" of the cells treated with each activity evaluation sample was calculated from the measured absorbance value. The test was performed with n = 3. The results are shown in FIGS. 1 and 2.

(Results of adipose progenitor cell differentiation induction test)
As shown in FIGS. 1 and 2, when the sample for evaluating the activity of carob R, carob L, and carob D was used, when the target hesperetin was used (12.5, 25, and 50 μg / mL). ), A decrease in the effective cell accumulation rate was observed. When the cells using the carob L activity evaluation sample at each concentration were observed under a microscope, a decrease in intracellular lipid droplets could be confirmed. In addition, no cytotoxicity to adipose progenitor cells was observed in any of the samples (cell viability> 80%). On the other hand, it was found that no decrease in the effective cell accumulation rate was observed when the samples for evaluating the activity of cocoa B and cocoa H were used (Fig. 2).

<Radical scavenging test>
2,2-diphenyl-1-picrylydrazyl (DPPH) was dissolved in ethanol to prepare a 0.1 mM DPPH solution. Meanwhile, ethanol was used to prepare dilution series of 50, 100, and 200 μg / mL activity evaluation samples (final concentrations 25, 50, and 100 μg / mL). A dilution series of 100 μL of each prepared sample for activity evaluation was put into a well of a 96-well plate, and another 100 μL of DPPH solution was put into the well, and the mixture was stirred using a vortex mixer. The 96-well plate was shielded from light with aluminum foil and allowed to stand at room temperature for 30 minutes, and then the absorbance at 517 nm was measured using an absorption plate reader. The "DPPH radical scavenging rate (%)" of each activity evaluation sample was calculated from the measured absorbance value, assuming that the absorbance value of the untreated well was 100%. The test was performed with n = 3. The results are shown in FIG.

(Result of radical scavenging test)
As shown in FIG. 3, concentration-dependent DPPH radical scavenging activity was observed in all activity evaluation samples.

<Nitric oxide production test>
Mouse macrophage-like cells (J774.1) were placed in wells of 96-well plates at 1 × 10 ⁵ cells / well in 100 μL portions and cultured overnight in an incubator at 37 ° C. and 5% CO ₂ . The medium was prepared by adding lipopolysaccharide (LPS) to RPMI-1640 medium at a concentration of 20 μg / mL. Activity evaluation samples were added to the prepared medium, and dilution series of 50, 100, and 200 μg / mL were prepared for each activity evaluation sample (final concentrations 25, 50, and 100 μg / mL). After 100 μL of the medium containing the sample for activity evaluation was added to the wells of the 96-well plate after culturing (total amount 200 μL), the cells were cultured in an incubator at 37 ° C. and 5% CO ₂ for 1 day. In addition, only the medium containing no sample for activity evaluation was added by 100 μL each, and the mixture was cultured as an untreated well. After culturing for 1 day, the nitric oxide production rate and the cell viability were calculated by the grease reagent method and the MTT method shown below, respectively.

(Nitric oxide production rate)
The wells of the new 96-well plate were filled with 100 μL of grease reagents (1% sulfanilamide, 0.1% N-1-naphthylethylenediamine, 2.5% phosphoric acid, and ultrapure water). After adding 100 μL of the cultured medium supernatant and allowing it to stand at room temperature for 20 minutes, the absorbance at 550 nm was measured using an absorption plate reader. Assuming that the absorbance value of the untreated well was 100%, the "nitric oxide production rate (%)" of the cells treated with each activity evaluation sample was calculated from the measured absorbance value. The test was performed with n = 3. The results are shown in FIG.

(Cell viability)
The medium supernatant was removed from each well of the cultured cells, 100 μL of MTT / PBS solution prepared to contain 5 μL of 5 mg / mL MTT solution was added, and the cells were cultured in an incubator at 37 ° C. and 5% CO ₂ for 3 hours. .. After culturing for 3 hours, the supernatant was removed, and 100 μL of dimethylsulfoxide was added to dissolve intracellular formazan. Absorbance at 450 nm was measured using an absorption plate reader. The "cell viability (%)" of the cells treated with each activity evaluation sample was calculated from the measured absorbance values, assuming that the absorbance value of the untreated well was 100%. The test was performed with n = 3. The results are shown in FIG.

(Results of nitric oxide production test)
As shown in FIG. 4, an effective decrease in nitric oxide production was observed when the activity evaluation samples of carob D and carob E were used, especially when the activity evaluation sample of carob E was used. .. In addition, no cytotoxicity to mouse macrophage-like cells was observed in any of the samples (cell viability> 86%).

The anti-obesity food of the present invention is useful as a functional food for the purpose of obtaining effects such as anti-obesity.

Claims (2)

Contains roasted carob powder as an active ingredient,
Before Kiabu decoction carob powder, after roasting less than 45 minutes 30 minutes or more at 120 to 180 ° C. The portion of the sheath of the carob state, and are not prepared by grinding,
An anti- obesity food that suppresses obesity by shrinking fat droplets accumulated in fat cells and suppressing hypertrophy of fat cells . It is a method of blending roasted carob powder into a food to give the food an anti-obesity effect that suppresses obesity by reducing the fat droplets accumulated in the fat cells to suppress the hypertrophy of the fat cells. ,
Before Kiabu decoction carob powder, after partial roasting below 120 to 180 ° C. 45 minutes 30 minutes or more at the sheath of the carob, using the Der Luke Rob powder that prepared by grinding.

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