JP2003009665A - Method for increasing yield of polyphenol in plant body and physiologically active substance thereof - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for effectively obtaining polyphenols as a physiologically active substance found only in minute amounts in a plant body, that is, a method for obtaining a plant of increased polyphenol content by raising the plant under specific conditions, and to obtain the polyphenols as the physiologically active substance having hepatopathy prophylactic and/or mitigating effect so as to use the substance in the pharmaceutical or functional food field. SOLUTION: The objective method for increasing the yield of polyphenols in plant body involves raising a polyphenol-generating plant under the conditions of loading a stress such as light stress or water stress on the plant. The other objective use of the polyphenols as a physiologically active substance is provided based on the finding that the polyphenol component obtained from the plant body of safflower has the effect of preventing and/or mitigating onset of hepatopathy, particularly hepatopathy due to toxic substance.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for increasing the yield of plant polyphenols or the use of plant polyphenols as physiologically active substances, particularly as a substance for preventing and / or reducing liver damage.

[0002]

2. Description of the Related Art Plants contain many polyphenol compounds. Plant polyphenols have been studied for a long time as the main factors of discoloration of foods derived from fruits and plants, and their components, content, and chemical characteristics ("Chemistry of Discoloration of Foods" Susumu Kimura, Toshiro Nakamura, Hiromichi Kato) , (1995) Korin Co., Ltd.). In recent years, the effect of the plant polyphenol component in food has begun to be noticed, starting from an epidemiological survey report in Europe that "a red wine drinker is less likely to suffer myocardial infarction even if the intake of oil and fat is large." Catechin, a polyphenol contained in green tea, has been shown to have many physiological actions such as antioxidative action, antibacterial action, and blood pressure elevation inhibitory action. In recent years, various types of tea beverages and tea products reflecting health consciousness have been revealed. Is becoming commercially available ("Antioxidant properties of tea ingredients" Satoshi Shiragi, Yasuhiko Hara FRAGRANCE JOURNAL, (1
990-11) 24-30). Rutin, which is abundant in buckwheat, has been shown to enhance the resistance of capillaries, and is used as a functional food material and antioxidant. ("Flavonoids as Antioxidants (2)" Foods &
Food Ingred. J. Jpn. 189 (2000) 47-53) ("Rutin")
Bio. Ind., 14 (1997) 57-58). Anthocyanins are widely present as a pigment that gives flowers, vegetables, fruits, cereals, etc. a red to purple color tone, and in the food-related field, not only the coloring function of food, but also the antioxidant function and antimutagenic activity. First, the usefulness of the health function was clarified (“Physiological function of anthocyanins as ingredients of food materials” Koji Igarashi Foods & Food Ingred. Jp.
n., 187 (2000) 17-28).

In addition, some physiological effects of plant polyphenols have been reported. For example, polyphenols obtained from fruits of roses such as apples are used as a lipid metabolism inhibitor in vivo (Japanese Patent Laid-Open No. 10-330278), teas, cacao, rosemary, etc. and extracted from plant materials. Those using polyphenols as active oxygen scavengers and the like (JP-A-2000-256)
No. 345), using polyphenols extracted from hops and the like as antioxidants (Japanese Patent Laid-Open No. 9-2).
No. 917), a method using a lentil seed coat extract as an antioxidant composition (Japanese Patent Publication No. 2001-500546), and the like. However, it cannot be said that the data on the individual food materials containing these polyphenols and the functionality of each component are sufficiently presented, and from the viewpoint of use as a bioactive agent or the viewpoint of use as a functional food. Therefore, the search for effective materials and components for disease prevention and treatment is awaited.

On the other hand, various living things on the earth maintain their lives while being constantly exposed to risk factors (environmental factors) caused by the environment. Mammals, including humans in particular, live while suffering from the influence of various environmental factors due to the wide range of movement and the amount of energy required within the individual in the form of latent diseases. In the recent human living environment, air pollution due to exhaust gas and food pollution due to mutagen chemical substances are one of the major causes of disease induction. In order to survive in such an environment, it is of course important to avoid inhalation and ingestion, but removal of these environmental factors that entered the body, detoxification, and environmental factors It is also important to control the harmful reactions that occur. From the viewpoint of disease prevention, research and research into food materials and ingredients having these effects (functionality) are being actively conducted (“active oxygen and medical food source” edited by Masayasu Inoue (1996), Kyoritsu Shuppan). . The plant polyphenol component has attracted particular attention as a promising candidate for a component having a disease-preventing function, due to the presentation of the above situation.

On the other hand, attempts to utilize plant components for prevention and treatment of diseases have been widely made. For example, some attempts have been reported to utilize plant components for the prevention and treatment of specific diseases such as liver dysfunction. For example,
What uses an extract of licorice for the prevention and treatment of liver damage (JP-A-7-135923), gluten, glutelin,
Use of cereal plant proteins such as prolamin as an alcoholic liver injury relieving agent (Japanese Patent Laid-Open No. 6-247865), extracts of Amaranthaceae, a plant of the Amaranthaceae family, intoxication caused by alcohol and drugs What is used as a prophylactic / therapeutic agent for liver damage and immune liver damage caused by viruses (WO96 / 36346), and one that uses an extract of Alphabacca as a liver damage inhibitory agent (JP-A-9-
No. 59168), using the essence of a plant of the genus Hippophae genus as a therapeutic or preventive agent for liver damage (JP-A-9-208484), and an extract of a plant of Kevlar-Pedra for inhibiting liver damage. Used as an agent (Japanese Patent Application Laid-Open No. 9-241176), and one using a plant extract of Mattico as a therapeutic agent for hepatitis (Japanese Patent Application Laid-Open No. 2000-
297043), and the like.

As described above, various attempts have been made to utilize physiologically active ingredients contained in plants in fields such as medicines and functional foods. In order to utilize a plant component having a physiological activity, it is first necessary to search for a plant containing the physiologically active component. However, even if a plant having physiological activity is found, it is necessary to efficiently obtain the active ingredient for practical use. Therefore, it is important to search for a plant body containing a large amount of active ingredients, but such a search itself is not easy. In general, a physiologically active ingredient is contained in a plant in a trace amount, so that it is necessary to concentrate the active ingredient, and the acquisition of the active ingredient requires a lot of labor under the present circumstances. This also applies to plant polyphenols obtained from plants. For that purpose, it is necessary to consider how to obtain a plant containing a large amount of polyphenols. As many reports have been made so far,
The composition and content of polyphenols vary greatly depending on the plant species and variety. Therefore, it is an effective method to search for a plant containing a large amount of polyphenol, but it is a laborious and difficult task. Even in one variety, it is not easy because the polyphenol component / content greatly varies depending on the growth stage. In addition, it is an effective means in the future to produce a plant body containing a large amount of effective polyphenol components by the genetic modification technology of plants which is becoming possible in the present age, but at the present time, it requires a great deal of research cost and is effective. is not. Therefore, it is desired to provide effective means for solving these problems.

[0007]

An object of the present invention is to effectively obtain polyphenols which are physiologically active substances, that is, to grow polyphenol-producing plants under specific conditions to remove polyphenols contained in the plants. It is intended to provide a method for increasing and effectively obtaining polyphenol, and to obtain a polyphenol physiologically active substance having an action of preventing and / or alleviating liver damage and aiming at its use in the field of medicine or functional food. .

[0008]

[Means for Solving the Problems] The present inventors have found that a polyphenol-producing plant can increase the polyphenol component in a plant by growing it under a stress-loaded condition. It came to completion. Furthermore, the present inventors have found that a polyphenol component obtained from a safflower plant and a substance that is one of its effective components have an action of preventing and / or reducing the occurrence of liver disorders, particularly liver disorders caused by toxic substances. Find out,
It has come to provide the physiologically active substance and its utilization.

That is, the present invention provides a method for increasing the yield of polyphenols in a plant (claim 1), which comprises growing a polyphenol-producing plant under conditions under stress to increase the polyphenol component in the plant. The stress is light stress and / or water stress, and the method for increasing the yield of plant polyphenols according to claim 1 (claim 2) or the polyphenol-producing plant is a plant seedling. The method for increasing the yield of plant polyphenols according to claim 1 or 2 (claim 3), or the plant producing polyphenols is safflower or buckwheat, and the plant polyphenols according to any one of claims 1 to 3. (Claim 4) and the increasing polyphenol component are luteolin-7-O-glucoside in safflower, rutin and buckwheat in buckwheat. It is made of sales of a plant polyphenol according to claim 4, wherein (claim 5) that the anthocyanin.

The present invention also relates to a method for producing a plant polyphenol (claim 6), which comprises growing a polyphenol-producing plant under stress, and extracting the polyphenol from the plant. A method for producing a plant polyphenol bioactive substance having a liver injury preventing and / or reducing action, which comprises subjecting a plant to a solvent extraction treatment (claim 7), or a plant of safflower, which is a young plant of safflower 7. A method for producing a plant polyphenol physiologically active substance according to claim 7 (claim 8), and a liver injury prevention characterized by using a polyphenol extract of a safflower plant as an active ingredient, and And / or a mitigating agent (claim 9) or a polyphenol extract of a safflower plant, which is added to a food material or food. Or functional food (claim 10)
Or a plant of safflower is a young plant of safflower, wherein the agent for preventing and / or reducing liver damage according to claim 9 or 10, or a functional food material or a functional food (claim 11). Or a hepatic disorder preventive and / or alleviating agent containing luteolin-7-O-glucoside, which is one of the plant polyphenols, as an active ingredient (claim 12).

[0011]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention comprises growing a polyphenol-producing plant under stress and increasing the polyphenol component in the plant. As the polyphenol-producing plant, various plants known to produce polyphenol can be used, but safflower and buckwheat can be advantageously used from the viewpoint of ease of cultivation and effects. Furthermore, the use of plant seedlings provides a particularly advantageous method for increasing the yield of polyphenols. As a method for growing the polyphenol-producing plant, an ordinary method is adopted except for stress load conditions, and there is no particular limitation. The stress to be applied includes various stresses such as light stress, water (moisture) stress, temperature stress, nutritional stress, drug stress, etc. However, considering the ease of adoption and the effect under growing conditions, light stress and Water and / or water stress can be used to advantage. To obtain plant polyphenols, extraction treatment with a solvent such as an organic solvent or water (hot water) from a plant grown under stress conditions is performed, but the solvent to be used should be appropriately selected from known ones. You can Solvents such as hot water and ethanol can be used particularly advantageously, provided that the use of other solvents is not excluded. The extract can be used as an extract, a concentrate thereof or a dried product thereof. The plant to be extracted can be used at any growth stage of the plant that grows under stress conditions and whose content is increased, but in the case of safflower or buckwheat, for example, those seedlings are advantageous. Can be used for. When the method of the present invention is applied to, for example, safflower seedlings, the total polyphenol content in the safflower seedlings and the specific polyphenol (luteolin-7-O-glucoside: 3 ', 4', 5,7-tetrahydroxyflavone- 7-O-glucoside) content increases, and when applied to buckwheat seedlings, it increases total polyphenol content and specific polyphenols (rutin and anthocyanin) content in buckwheat seedlings.

The plant polyphenol physiologically active substance having the preventive and / or alleviating effect on liver damage according to the present invention is obtained from safflower plants, especially safflower seedlings. The plant polyphenol physiologically active substance of the present invention has a preventive and / or alleviating action on liver damage, especially a preventive and / or alleviative action on liver damage due to toxic substances. The plant physiologically active substance of the present invention can be used as a liver damage preventive and / or alleviating agent, or by adding the substance to a food material or a food, as a functional food material or a functional food. Furthermore, the present invention provides a hepatic disorder and / or alleviation agent containing luteolin-7-O-glucoside, which is one of plant polyphenols, as an active ingredient.

[0013]

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

Example 1 (Method of increasing yield of plant polyphenol by growing polyphenol-producing plant under stress load condition) Light stress and water stress are used as stress load condition on the plant. As a condition for applying light stress, as a method for growing a plant, the light irradiation intensity during growth is increased to 5 times or more the light intensity under normal growth conditions (light stress is applied) for growth. As a result, the amount of polyphenol contained in the plant can be doubled or more. However, since the appropriate light stress condition for increasing the polyphenol content varies depending on the type and variety of plant, "it causes stress to plants but does not significantly inhibit growth."
It is necessary to set the conditions. Specifically, growth inhibition does not occur (elongation rate or wet weight increase rate does not decrease remarkably compared to the control), no serious photodamage occurs (chlorophyll a / chlorophyll b ratio is compared to the control). In comparison, it does not increase significantly). further,
The polyphenol content is further increased by growing under water stress in addition to light stress. Also at this time,
It is necessary to set growth conditions that do not cause excessive stress by monitoring the elongation rate or the wet weight increase rate and the chlorophyll a / chlorophyll b ratio described above. Specific examples will be shown below.

Example 2 (Method for increasing the yield of polyphenols in plants using safflower) By growing safflower seedlings under growth conditions under light stress and water stress, cotyledons and true leaves Increase the polyphenol content more than 2-fold.
At this time, in the true leaf, prevention against liver damage in mammals
The content of luteolin-7-O-glucoside, which is a polyphenol component having a reducing effect, is also doubled or more.
The details of the increase of polyphenol content in safflower seedlings due to light stress and water stress load are shown below.

A. Growth conditions Safflower seeds were sown on an equal amount of mixed soil of vermiculite and horticultural soil, and left in the dark for 48 hours until sprouting. After growing this for 7 days in an artificial weather chamber [bright (5000 lux, 12 hours), dark (12 hours), 25 ° C], it was divided into the following 3 experimental groups with different light conditions and water supply conditions in the light period. It was further grown for 7 days and subjected to analysis. Control group: illuminance, 5000 lux; water supply, total volume 170 ml
Was given 2-3 times a day. Light stress group: illuminance, 45000lux; water supply is the same as the control group. Light / water stress group: illuminance, 45000lux; water supply, 100 ml in total was given twice a day. When the wet weight / dry weight ratio of the leaves was determined in the light stress group, it was reduced by about 30% as compared with the control group. This indicates that under this experimental condition, water stress is also applied to the light stress group. In order to further clarify the effect of water stress, we prepared a light / water stress group with reduced water supply.

B. Extraction and Analysis In each experimental group, cotyledons and true leaves were collected, weighed wet and then lyophilized. The extraction was performed by the following operations. 1) 25 mg of freeze-dried powder was rubbed with quartz sand in a mortar. 2) 5 ml of dimethyl sulfoxide
After adding (DMSO) and stirring well, the mixture was centrifuged at 1500 × g and the supernatant was collected. 3) 5 ml DMS for precipitation
After adding O and stirring well, it was centrifuged again and the supernatant was collected. 4) Supernatants obtained by centrifugation twice were combined and used as an extract, which was stored at -80 ° C until analysis. The total amount of polyphenols in the extract was measured by the Folin-Denis method. Separation and analysis of each component of polyphenol
The separation was carried out by the high performance liquid chromatography method (HPLC) under the following separation conditions. Separation conditions: Column, Develosil
C-30-UG-5 (4.6 × 250 mm, Nomura Chemical); Mobile bed, (A) 5%
(v / v) acetonitrile / 1% (v / v) acetic acid, (B) 40%
(v / v) Acetonitrile (linear gradient at which solvent B becomes 100% in 180 minutes); column temperature, 25 ° C; detection,
330 nm; flow rate, 0.8 ml / min

C. Results First, in order to determine whether the stress treatment was excessive, the chlorophyll a / chlorophyll b ratio and the above-ground wet weight were compared with the control group. In the light stress group and the light / water stress group, the increase in the chlorophyll a / chlorophyll b ratio was about 15%, and it was confirmed that no serious photodamage occurred. The above-ground wet weight increased about twice in the light stress group, and hardly changed in the light / water stress group. Therefore, it was also confirmed that the stress treatment applied did not cause significant growth inhibition. FIG. 1 shows the total polyphenol content in the cotyledons and true leaves of each experimental group. The total polyphenol content in the cotyledons was hardly changed by light stress, but it was increased about 1.5 times in the true leaves. In the experimental group in which water stress was applied in addition to light stress, the total polyphenol content in cotyledons and true leaves increased more than 2-fold. Under these growth conditions, no remarkable growth inhibition or damage due to stress load such as light damage was observed. These results indicate that the total amount of polyphenols in seedlings can be increased by controlling the light / water (drying) stress. The DMSO extracts from the true leaves of the seedlings of each experimental group were individually identified by HPLC for the polyphenol components, and quantitative analysis was performed. From the results, luteolin-7-O in the true leaves of each experimental group was selected. -The glucoside content is shown in Figure 2. It was confirmed that the stress load condition increased more than 2-fold compared with the control group. Regarding the increase in the content of luteolin-7-O-glucoside in the plant body, under the experimental conditions shown here, light stress (water stress is also applied simultaneously) is sufficient, and light stress There was no further increase in the water stress group.

Example 3 (Method for increasing yield of polyphenols in plants using buckwheat) Polyphenol content in true leaves was obtained by growing buckwheat seedlings under growth conditions under light stress and water stress. Is more than doubled. At this time, rutin "having a capillary-enhancing effect" in the true leaf and anthocyanin "having an antioxidative function" in the stem also increase more than twice. The details of increase of polyphenol content in buckwheat seedlings due to light stress / water stress load are shown below. a. Growth conditions Buckwheat seeds were sown in an equal amount of mixed soil of vermiculite and horticultural soil, and left in the dark for 48 hours until sprouting. After growing this for 7 days in an artificial weather chamber [light (7500lux, 12 hours), dark (12 hours), 25 ° C], the light intensity in the light period was increased to 55000lux and grown for 7 days (light stress). And a control group in which the irradiation light intensity was not changed. In the light stress group, the wet weight / dry weight ratio of the leaves was reduced by about 30% as compared with the control group. As described in the growth conditions of the safflower seedlings in the previous section, under the experimental conditions, water stress was applied even in the light stress group. From the comparison of the chlorophyll a / chlorophyll b ratio and the wet weight, it was confirmed that photo-damage and growth inhibition due to stress treatment hardly occurred.

B. Extraction and analysis Same as in the previous section regarding the safflower seedlings, except for a part. For buckwheat, the true leaves and stems were analyzed. c. Results FIG. 3 shows the total polyphenol content of the true leaves and stems of the control group and the light stress group. The total amount of polyphenols in the true leaves increased about 3 times by the light stress. At this time, the total polyphenol content of the stem remained almost unchanged. In FIG. 4, the rutin content (a) in the true leaf and the anthocyanin content (b) in the stem were compared between the control group and the light stress group, respectively. In the light stress group, rutin in the true leaf increased about 7 times, and anthocyanin in the stem increased about 2.5 times. From these results, it was confirmed that under the growth conditions of the photo-stress group, a polyphenol component (rutin anthocyanin) effective for preventing diseases in mammals was effectively concentrated in plants.

Example 4 (Polyphenol extract of safflower plant and luteolin-7-O-glucoside preventive and alleviating action on liver damage) Polyphenol extract of safflower seedlings, or luteolin, which is one of its main components Pre-ingestion of -7-O-glucoside prevents or reduces the occurrence of liver damage caused by drugs and the like. Safflower seedling polyphenol extract and luteolin-7-O- against galactosamine-induced liver injury in rats
The protective effect of glucoside is shown below.

A. Preparation of safflower seedling polyphenol extract (PPE) As a plant sample, sowing safflower seeds on a planter containing commercially available horticultural soil, and use the above-ground parts (leaves and stems) of seedlings cultivated outdoors for 10 days. I was there. Demineralized water was added to the previously frozen sample, followed by boiling extraction for 30 minutes. From this hot water extract, a polyphenol-rich fraction was prepared as follows. After the extract was filtered, it was passed through an Amberlite XAD-7 column that had been equilibrated with 0.5% acetic acid in advance, and then the column was washed with 0.5% acetic acid. Then 5
Elution was performed sequentially with 0% methyl alcohol and 100% methyl alcohol, and the eluates were combined, demineralized water was added, and the mixture was concentrated under reduced pressure to remove methyl alcohol. After freeze-drying this concentrated liquid, safflower seedling polyphenol extract (Polyphene
ol extract: PPE) was used in the following experiments.

B. Experimental animals Wistar male rats aged 5 weeks (average initial weight: 76 g) were used. Breeding is done with individual gauges, room temperature is 22 ± 2 ° C, humidity is 40-60%, and light-dark cycle is 12 hours (06: 00-1).
8:00). Induction of liver damage by galactosamine (GalN) was performed according to a standard method (Reference 7: “Liver Injury-p
reventive Effect of Tea Theanine in Rats ”(2000)
He, P., Watanabe, N., Sugiyama, K. Journal of Food
Science, 65, 30-33); 200m on the 8th day after the start of breeding
It was caused by intraperitoneal administration of 40 mg / kg body weight of a g / dl GalN aqueous solution (pH adjusted to near neutral with a NaOH aqueous solution) intraperitoneally. The experiment groups were the following 4 groups, and the experiment was conducted using 6 animals in each group. ○ Control group (Con): On the 8th day of feeding with a basic diet, the same amount of physiological saline was administered instead of GalN, and the basic diet was subsequently fed. ○ Hepatopathy rat group (GalN): 8 days after feeding with basic diet
GalN was administered, followed by a basic diet. O Liver-damaged rats / PPE feeding group (GalN + PPE): After feeding a basic diet supplemented with 0.4% of safflower seedlings PPE for 8 days, GalN was administered, and subsequently the same feed was fed. ○ Hepatopathy rats / Luteolin-7-O-glucoside feeding group (GalN + LuG): Luteolin-7-O-glucoside (LuG)
Was fed for 8 days, then galactosamine was administered, and subsequently the same feed was fed. Lu
As G, a standard product isolated and purified in the laboratory was used, but the results described below were the same even if it was a commercial product having an equivalent purity.

The feed is 20.0% casein, 5.0% corn oil, 5.0% cellulose, 3.5% mineral mixture (AIN-93G manufactured by Oriental Yeast Co.), vitamin mixture (AIN- manufactured by Oriental Yeast Co., Ltd.). 93-VX) 1.0%, α-cornstarch: sucrose = 2: 165.5% was used as the basic diet. Safflower seedlings PPE (0.4%),
And, with LuG (0.1%) supplemented feed, α-corn starch: sucrose = 2: 1 amount was adjusted, and 100% in total.
I tried to become. Water was freely available during the experiment. Ga
lN was administered at 15:00 on the 8th day and 13: 0 on the 9th day
At 0 (22 hours after GalN administration), blood was collected from the heart under Nembutal anesthesia. GOT and G after separation of plasma
It was used for PT activity measurement.

C. Measurement of GOT activity and GPT activity in plasma Blood was directly collected from the heart using a syringe to which 20 μl of heparin was added in advance. Immediately, gently mix in a syringe and transfer to a test tube. Centrifuge (3000r
GOT and GPT activities were measured immediately after plasma was separated by pm, 15 ° C., 15 min). GOT activity and GPT activity were measured by using 100 μl of plasma obtained by the above-mentioned method as a measurement sample, and a commercially available kit (GOT-UV Test Wako (UV-r purchased from Wako Pure Chemical Industries, Ltd.)
ate method)). Activity was expressed in International Units (IU).

D. Results After breeding for 9 days, the liver weight was reduced by 10-15% in all the GalN-administered groups, and the addition of PPE and LuG to the feed did not improve the liver weight. GOT activity and GP in plasma, which are indicators of liver function (the lower the liver function, the more normal)
The T activity showed a 10-fold or more increase in the GalN group, confirming that liver injury was induced (FIG. 5). These increases in GOT activity and GPT activity were suppressed by feeding PPE (GalN + PPE). This result indicates that PPE is effective in preventing liver damage. With regard to LuG, which is one of the main polyphenol components contained in PPE, the inhibitory effect on liver damage was examined using GOT activity and GPT activity in plasma as indexes. The GOT activity of rats fed the diet supplemented with 0.1% of LuG (GalN + LuG) did not show a clear improvement as compared with the GalN group as compared with the PPE fed group (GalN + PPE), but tended to be lower. (Fig. 5a). G of GalN + LuG group
PT activity was suppressed lower by feeding LuG (Fig. 5b). These results indicate that LuG contributes to the liver damage prevention effect as one of the components of PPE, and a food material containing a large amount of LuG can be expected to be useful in preventing liver damage.

[0027]

INDUSTRIAL APPLICABILITY The method for increasing the yield of plant polyphenols according to the present invention can be applied to a wide range of polyphenol-producing plants, and polyphenols which are useful physiologically active components contained in the plant only in trace amounts Increasing the yield allows these practical uses. Therefore,
It is also important for practical use in order to use polyphenols having various useful physiological activities in medicines, functional foods and the like. In addition, the physiologically active substance having a liver injury prevention and / or alleviation effect, which is the polyphenol extract of safflower presented by the present invention, is a safe physiologically active substance obtained from a natural product, as a liver injury preventive agent or alleviation agent, Alternatively, it can be used as a functional food material. In particular, for liver damage caused by ingestion as a harmful substance to the human body from food and environment, active intake of these substances has the effect of preventing or reducing diseases caused by the damage, so In modern society, which is exposed to the risk factors caused by the environment, it is very useful for maintaining health.

[Brief description of drawings]

FIG. 1 is a diagram showing the total polyphenol content in cotyledons and true leaves of safflower seedlings when the plant polyphenol yield increasing method of the present invention is applied to safflower seedlings.

FIG. 2 shows luteolin-7-in the true leaves of safflower seedlings when the plant polyphenol yield increasing method of the present invention is applied.
It is a figure which shows O-glucoside content.

FIG. 3 is a diagram showing the total polyphenol content of true leaves and stems of buckwheat seedlings when the plant polyphenol yield-increasing method of the present invention is applied to buckwheat seedlings.

FIG. 4 is a diagram showing the contents of rutin in true leaves and anthocyanins in stems of buckwheat seedlings when the plant polyphenol harvesting method of the present invention is applied to buckwheat seedlings.

FIG. 5 is a diagram showing GOT activity (FIG. A) and GPT activity (FIG. B) when a safflower seedling polyphenol, which is a physiologically active substance of the present invention, was administered to hepatopathy rats.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) // C07H 17/07 C07H 17/07 (72) Inventor Hiroaki Oya 4-3-4 Tokamachi, Yamagata City, Yamagata Prefecture F-term (reference) 2B022 AA01 AB15 DA01 4B018 MD08 ME14 MF01 4C057 BB02 DD01 KK02 4C086 AA01 EA11 MA04 MA52 NA14 ZA75 4C088 AB26 AC01 BA10 CA06 MA52 NA14 ZA75

Claims (12)

[Claims] 1. A method for increasing the yield of polyphenols in a plant, which comprises growing a polyphenol-producing plant under conditions under stress to increase the polyphenol component in the plant. 2. The method for increasing the yield of plant polyphenols according to claim 1, wherein the stress is light stress and / or water stress. 3. The method for increasing the yield of plant polyphenols according to claim 1, wherein the polyphenol-producing plant is a plant seedling. 4. The method for increasing the yield of plant polyphenols according to claim 1, wherein the polyphenol-producing plant is safflower or buckwheat. 5. The increasing polyphenol component is luteolin-7-O-glucoside in safflower and rutin and anthocyanin in buckwheat.
A method for increasing the yield of plant polyphenols as described. 6. A method for producing a plant polyphenol, which comprises growing a polyphenol-producing plant under conditions under stress and extracting the polyphenol from the plant. 7. A process for producing a plant polyphenol physiologically active substance having a liver damage preventive and / or alleviating action, which comprises subjecting a safflower plant to a solvent extraction treatment. 8. The method for producing a plant polyphenol physiologically active substance according to claim 7, wherein the safflower plant is a safflower seedling. 9. Prevention of liver damage and / or the use of a polyphenol extract of a safflower plant as an active ingredient.
Or a mitigating agent. 10. A functional food material or a functional food, which is obtained by adding a polyphenol extract of a safflower plant to a food material or a food. 11. The liver damage preventive and / or alleviating agent, or functional food material or functional food according to claim 9, wherein the safflower plant is a safflower seedling. 12. An agent for preventing and / or reducing liver damage, which comprises luteolin-7-O-glucoside, which is one of plant polyphenols, as an active ingredient.