JPH10298094A - Lipid metabolism improver - Google Patents

Abstract

(57) Abstract: A lipid metabolism-improving agent, an antioxidant and a food containing one or more extracts selected from tamarillo, pomegranate and feijoa. [Effect] Safe and effective.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an agent for improving lipid metabolism derived from natural products, an antioxidant and a food containing the same.

[0002]

2. Description of the Related Art In recent years, in Japan, the number of patients with arteriosclerosis causing myocardial infarction, cerebral infarction and the like has been increasing. Although various causes are considered for the progression of arteriosclerosis, blood lipids, hypertension and smoking are among the three major risk factors.

[0003] Among them, the blood lipid concentration is increased by excessively ingesting a food containing lipid and cholesterol.
That is, the ingested cholesterol is solubilized in the body and then absorbed into the blood. Therefore, if cholesterol is insolubilized, an increase in blood lipid concentration can be suppressed. From such findings, cholestyramine and chitosan have been found as substances that insolubilize cholesterol.

[0004] On the other hand, it is also known that oxidative denaturation of low density lipoprotein (LDL) in blood is mainly involved in the progression of arteriosclerosis. In other words, the denatured LDL oxidized in the body is taken up through the macrophage scavenger receptor, whereby the infiltration of the macrophages progresses, and these fibers fibrillate and form atherosclerotic foci having calcification foci. Because

[0005] Antioxidants such as vitamin A, vitamin E, and polyphenol have been found as substances that prevent oxidative denaturation of LDL. Furthermore, since LDL contains a large amount of cholesterol, administration of a substance that suppresses cholesterol absorption as described above can also be an effective means of preventing LDL oxidative degeneration.

[0006]

Accordingly, an object of the present invention is to suppress the absorption of cholesterol in the body,
It is an object of the present invention to provide a novel substance that can improve lipid metabolism in blood by preventing DL oxidative degeneration and prevent various circulatory diseases such as arteriosclerosis.

[0007]

Means for Solving the Problems In view of such circumstances, the present inventor has conducted intensive studies and as a result, found that Tamarillo, pomegranate and / or pomegranate.
Or that the Feijoa extract insolubilizes cholesterol and suppresses absorption in the body, and that the tamarillo, pomegranate and / or feijoa extracts have antioxidant activity, and that the tamarillo extract has antioxidant activity against LDL Thus, the present invention was found to be useful as a drug or food for improving lipid metabolism, and completed the present invention.

That is, the present invention provides a lipid metabolism improving agent containing one or more extracts selected from tamarillo, pomegranate and feijoa.

The present invention also provides an antioxidant containing one or more extracts selected from tamarillo, pomegranate and feijoa.

The present invention also provides a low-density lipoprotein antioxidant containing one or more extracts selected from tamarillo, pomegranate and feijoa.

Further, the present invention provides a food for improving lipid metabolism or LDL antioxidant, comprising one or more extracts selected from tamarillo, pomegranate and feijoa.

Further, the present invention provides a low-density lipoprotein antioxidant food containing one or more extracts selected from tamarillo, pomegranate and feijoa.

[0013]

DETAILED DESCRIPTION OF THE INVENTION Tamarillo (English name) used in the present invention.
Tree tomato) is an evergreen fruit having a height of 2 to 3 m cultivated in tropical and subtropical highlands. Fruits are 5 long
７7 cm, slightly sweet, slightly sour and full of magenta or black seeds. Red tamarillos are generally available on the market, and there are also other varieties such as gold tamarillos.

[0014] Pomegranate (garnet, pomegranate) is an evergreen fruit that is widely cultivated in the subtropical region and has a height of 10 m. The fruit is spherical with a diameter of 7-8 cm, and its interior is divided into 6 chambers. Among the pomegranates, the typical ones are paper shell and road wonderful, and there are various other types such as crystal pomegranate and large red pomegranate.

[0015] Feijoa is grown at a height of 3 in subtropical and temperate regions, such as the Mediterranean coast.
~ 5m of evergreen fruit. The fruit is about 5 cm egg-shaped and its outer skin is pale green. The main varieties include Coolidge, Mammoth, Gracie and the like.

When the above three kinds of fruit extracts are used as lipid metabolism improving agents or antioxidants, their varieties are not particularly limited, and any varieties may be used.
Since these have been eaten by humans for some time, there is no problem in their safety, etc., even if the extract is used as an agent or food. Therefore, a variety may be selected in consideration of taste, color, and the like according to the application.

Methods for obtaining an extract from tamaro, pomegranate and feijoa include water extraction, hot water extraction, and organic solvent extraction. Preferred examples of the organic solvent used here include alcohols such as ethanol and methanol.

However, since the extraction effect differs depending on the combination of the fruit and the solvent, it is preferable to select a preferable combination. For example, tamarillo and pomegranate are water,
The active ingredient for insolubilizing micelle cholesterol can be satisfactorily extracted using either hot water or an organic solvent.However, for Feijoa, the extraction efficiency of the active ingredient is not favorable in water, and extraction with hot water or an organic solvent is difficult. preferable.

The organic solvent extract such as an ethanol extract showed high activity from any fruit, but overall activity was higher than that of water (excluding feijoa) or hot water extract. Was low. However, depending on the application, an organic solvent extract such as alcohol may be more easily used, so that it is preferable to select an extraction solvent according to the purpose.

The extraction can be performed by a conventional method, for example, a method in which the above-mentioned fruit is shredded and immersed in a solvent.

The extract thus obtained has a micelle cholesterol insolubilizing activity and is useful as an agent for improving lipid metabolism and the like. This activity was not changed by the pH of the extract. Further, all of these extracts show antioxidant activity, the antioxidant activity of the tamarillo extract is slightly stronger than quercetin or BHT at a concentration of 8% by weight, and the antioxidant activity of Feijoa extract is 8% by weight of α- It was comparable to tocopherol and vitamin C.

The present inventors further note that these extracts are L
It was also confirmed to have DL antioxidant activity. In particular, the tamarillo extract prevented the oxidative denaturation of both the lipid and protein parts of the low density lipoprotein.

In the present invention, when an extract of tamarillo, pomegranate or feijoa is used as a medicine or food for improving lipid metabolism or for antioxidation, the compounding amount is preferably 1 to 50% in terms of weight of solids, 5 to 30% is more preferable, and 10 to 25% is particularly preferable.

[0024]

The present invention will be described below in more detail with reference to examples, but the present invention is not limited to these examples.

Example 1 Production of Various Fruit Extracts and Measurement of Their Micellar Cholesterol Insolubilizing Activity (1) Preparation of Various Extracts The edible portion of each commercially available food material was cut into pieces (wet weight: 30 g). Add water or ethanol (10 ml) and add 4
C. overnight. After that, the extract obtained by centrifugation or filtration and adjusted to pH 7.0 with 1N-NaOH was used as a sample for measuring micelle cholesterol insolubilizing activity.
In addition, hot water extraction was performed by adding water (10 ml) to the above shredded (30 g) and treating at 100 ° C. for 20 minutes.

(2) Preparation of micelle cholesterol In 75 ml of phosphate buffer (150 mM, pH 7.0), 2 g of oxygel (oxgall (DIFCO)), 921 mg of cholesterol (manufactured by Wako Pure Chemical Industries, Ltd.), 135 mg of lysophosphatidylcholine (SIGMA) Were added and dissolved, and then 702.2 mg of monooleic acid (manufactured by Tokyo Chemical Industry Co., Ltd.) was added and mixed, and a phosphoric acid buffer was added to make a total volume of 100 ml. Sonicate at room temperature while stirring the solution (SONI
FR, small chip). After stirring this emulsion and micelle solution for a while, 100,000 × g (RP50
-2, 40,000rpm), 16-18 ultrasonic separation at 25 ° C
Time went. After ultracentrifugation, only the transparent micelle layer was collected to obtain artificial lipid micelles.

(3) Measurement of micellar cholesterol insolubilizing activity Each sample extracted with water, hot water and ethanol was mixed with cholesterol-dissolved artificial micelles (cholesterol concentration: 10 mg / dl) at a ratio of 1: 1 to 37 ° C. For 1 hour in the culture room. Centrifuge the mixture (10,0
(00 rpm, 15 min), and determine the concentration of cholesterol remaining in the supernatant with Detamina TC555 (manufactured by Wako Pure Chemical Industries, Ltd.).
It measured using. Using only each extraction solvent as cholesterol, the cholesterol insolubilization rate was calculated from the following equation.

[0028]

(Equation 1)

Tables 1 and 2 show the cholesterol insolubilization rate of each sample, and Table 3 shows the top 10 species having the highest activity. In the samples obtained by any of the extraction methods, the activities of tamarillo and pomegranate were strong, and the activity of the feijoa extract was also strong except for the water extraction.

[0030]

[Table 1]

[0031]

[Table 2]

[0032]

[Table 3]

Example 2 Investigation of Addition Concentration of Tamarillo Extract As a result of measuring tamarillo squeezed juice, the solid content was 12.8.
%Met. Therefore, the solid content was changed by dilution, and the micelle insolubilization rate was examined. Tamarillo squeezed juice and micelle cholesterol were mixed at a weight ratio of 1: 1 and the insolubilization rate was measured (FIG. 1).

Example 3 Examination of LDL Antioxidant Activity of Tamarillo Extract The presence or absence of inhibition of the production of oxidized LDL, which is a risk factor for arteriosclerosis, was examined for a tamarillo extract having a strong micellar cholesterol insolubilizing activity. At this time, the antioxidant activity against apoprotein was evaluated by the inhibitory activity against LDL oxidation using Cu ²⁺ as an oxidizing agent.

(1) Preparation of Sample The edible portion of tamarillo was squeezed with a mixer (wet weight: 0.2 g), and water (400 microliters) was added to the mixture.
After dilution twice, a water-extracted sample was obtained. Similarly, methanol (400 microliters) was added and left overnight at 4 ° C. to obtain a methanol sample. The methanol sample was then centrifuged (8000 rpm for 5 min) to obtain a supernatant. These samples were further diluted 5-fold at the time of measurement and used for measurement of antioxidant activity. The final concentration of the sample was 6.7% by wet weight / liter.

(2) Preparation of LDL LDL was collected from Syrian hamster. Syrian hamsters (male, 10 to 14 weeks old) were preliminarily reared on an MF diet for 5 days, and then reared on an MF diet supplemented with 0.5% cholesterol and 5% lard for 2 weeks. 24 days before dissection
After fasting for an hour, blood was collected from the abdominal aorta, and EDTA plasma was prepared by a conventional method. LD from this plasma by ultracentrifugation
After collecting the L fraction, phosphate buffered saline (P
(BS) at 4 ° C. for 24 hours. After measuring the protein concentration by the Lowry-Folin method, it was appropriately diluted to obtain LDL for oxidation reaction.

(3) Measurement of LDL Oxidation Inhibitory Activity The activities of apoprotein denaturation inhibition and LDL lipid oxidation inhibition were measured using two systems. Apoprotein denaturation inhibition (agarose electrophoresis: hereinafter referred to as AGE method) L prepared in (2) in 20 μl of 250 mM phosphate buffer
DL (final concentration 250 μg / ml) 60 μg and sample 1
0 μl was added, and 10 μl of 100 μM CuSO ₄ was added thereto.
After the addition of 1 liter, the mixture was incubated at 37 ° C. for 5 hours to oxidize, 20 μl of 2.5 mM EDTA was added, and the reaction was terminated by cooling. A sample using methanol instead of a sample was used as a control, and a sample to which EDTA was added without incubation was used as a blank.

2 μl of the reaction solution was applied to a 1% agarose gel (multi-track LDH isozyme gel), and
After electrophoresis at 0 V for 45 minutes, cholesterol staining was performed with Corcorest A (Nippon Chemifa Co., Ltd.). 3 gels
After drying at 7 ° C., the moving distance of each lane was measured. The blank was assumed to be in a state in which LDL was not oxidized at all, and the rate at which the increase in mobility was suppressed by the addition of the sample as compared to the control was defined as the rate of inhibition of oxidation denaturation. The calculation formula is shown below. The results are shown in FIG.

(BC) ÷ (BA) × 100 = oxidation inhibition rate (%) (A: moving distance of blank LDL, B: moving distance of control, C: moving distance at sample addition)

LDL lipid oxidation inhibition (TBARS method) L prepared in (2) was added to 20 μl of a 250 mM phosphate buffer.
DL (final concentration 250 μg / ml) 60 μg and sample 1
Add 0 μl. 1 ml of 0.05N hydrochloric acid and T
After adding 500 μl of BA (thiobarbital) reagent, the mixture is reacted in boiling water for 45 minutes. After the completion of the reaction,
2 ml of butanol containing 5% methanol are added. Then, after shaking extraction for 10 minutes, centrifugation (3000 rpm, 5
, The room temperature) to separate the butanol layer. The absorbance at 535 nm of the thiobarbital reactant (TBARS) was measured, and the rate at which the production of TBARS was suppressed was defined as the oxidation inhibition rate. The results are shown in FIG.

In the figure, R stands for Red Tamarillo and G stands for Gold Tamarillo. In the agarose electrophoresis, the red of the water extraction was slightly higher, and the activity of about 50% was observed in other samples. In TBARS, the activity of red in the water extraction was weak, but about 30 to 50% of other activities were observed. From the above results, activity was confirmed in any of the red, gold, and fractionation methods.

Example 4 Evaluation of antioxidant activity of Tamarillo extract and Feijoa extract The antioxidant activity of the Tamarillo extract and Feijoa extract was examined by comparing with a specified concentration of a known substance. (1) Preparation of sample Antioxidants are classified into water-soluble and fat-soluble ones. Water-soluble substances (vitamin C, quercetin) are dissolved in 80% methanol, fat-soluble substances (α-tocopherol, B
HT) used was dissolved in 100% ethanol.
The concentration of these samples was 1 mM. For the tamarillo extract and the feijoa extract, an 8% solution obtained by diluting the juice with ion-exchanged water was used.

(2) Measurement of Antioxidant Power DPPH (2,2-diphenyl-1-picrylhydrazyl), which is a stable free radical, comes into contact with an antioxidant to form a blue-violet color (an absorption wavelength of 517 nm). ) To be colorless. That is, D
Reduction of PPH was confirmed by a decrease in absorbance at an absorption wavelength of 517 nm. Actually, by diluting the antioxidant (1 mM) and the tamarillo extract and the Feijoa extract stepwise, and comparing the changes in reactivity according to the dilution,
The antioxidant power was compared.

(3) Evaluation of antioxidant power The 8% tamarillo solution had a slightly stronger antioxidant power than 1 mM BHT and quercetin. The 8% Feijore solution had an antioxidant power equivalent to 1 mM vitamin C and 1 mM α-tocopherol. (FIG. 4)

[0045]

Since the lipid metabolism improving agent of the present invention is extracted from edible fruits, it is safe and has good palatability, and can be used as it is or added to various foods. Can be. Further, it can be manufactured simply and at low cost. Furthermore, the tamarillo, pomegranate and feijoa extracts also have antioxidant activity and are therefore useful as antioxidants.

[Brief description of the drawings]

FIG. 1 is a graph showing the concentration and activity of a tamarillo extract.

FIG. 2. LDL of Tamarillo by agarose electrophoresis
It is a figure which shows denaturation inhibitory activity.

FIG. 3. L of Tamarillo using TBARS generation as an index
It is a figure which shows DL denaturation inhibitory activity.

FIG. 4 is a diagram showing a comparison of the antioxidant power of the Tamarillo extract and Feijoa extract with other substances.

Claims (6)

[Claims] 1. An agent for improving lipid metabolism comprising one or more extracts selected from tamarillo, pomegranate and feijoa. 2. An antioxidant containing one or more extracts selected from tamarillo, pomegranate and feijoa. 3. A low-density lipoprotein antioxidant containing one or more extracts selected from tamarillo, pomegranate and feijoa. 4. A lipid metabolism-improved food containing one or more extracts selected from tamarillo, pomegranate and feijoa. 5. An antioxidant food containing one or more extracts selected from tamarillo, pomegranate and feijoa. 6. A low-density lipoprotein antioxidant food containing one or more extracts selected from tamarillo, pomegranate and feijoa.