JP6613515B2 - Myoblast proliferation promoter and focus adjustment function improving agent - Google Patents

Description

  The present invention relates to a myoblast proliferation promoter containing an Asteraceae plant.

In the Asteraceae plants, chrysanthemums have traditionally been used as tea, liquor, herbal medicines as longevity flowers, and are still used for food.
These Asteraceae plants and their extracts have various functionalities. Examples of what has been reported so far include, for example, an odor generation suppressing action (Patent Document 1), an allonidase activity inhibiting action (Patent Document). 2) Improvement of skin damage caused by exposure to ultraviolet rays (Patent Document 3), antioxidant effect (Patent Document 4), promotion of collagen production (Patent Document 5), whitening effect (Patent Document 6), and the like.
However, almost no other effects are currently recognized.

JP 2002-114660 A Japanese Patent Laid-Open No. 2003-12489 Japanese Patent Laid-Open No. 2005-298481 JP 2008-247839 A JP 2013-35776 A JP 2013-173735 A

  In order to solve the above-mentioned conventional problems, the present invention finds that Asteraceae plants have a myoblast proliferation promoting action, and is excellent in safety as a new use using Asteraceae plants. In addition, the present invention provides a myoblast proliferation promoter and a focus adjustment function improving agent having excellent actions and effects such as enhancement of muscle strength and suppression of atrophy, acceleration of muscle strength metabolism, recovery of visual acuity and prevention of vision loss associated therewith. Objective.

  The inventors of the present invention have made extensive studies in view of the above problems, and as a result, have found that a plant belonging to the genus Asteraceae has a myoblast proliferation promoting action, and led to the present invention.

  That is, the myoblast proliferation promoter and the focus adjustment function improving agent of the present invention contain an Asteraceae plant.

  Moreover, it is preferable that the myoblast proliferation promoter of this invention uses the flower of the Asteraceae genus plant as a raw material, and it is more preferable that the Asteraceae genus plant is purple chrysanthemum.

Since the myoblast proliferation promoter of the present invention promotes muscle metabolism, it can help increase muscle strength and can help recover muscle strength that has been reduced without being used due to injury or disease.
The focus adjustment function improving agent of the present invention can promote the growth of myoblasts, so that the increase in ciliary muscle muscles promotes the reduction of visual acuity and the improvement of visual acuity, as well as prevention of presbyopia and recovery of presbyopia. It is considered possible.

The figure which showed the myoblast proliferation rate of Example 1 and Comparative Examples 1-3

  Hereinafter, the myoblast proliferation promoter in the present invention will be described. In addition, this invention is not limited to the following embodiment.

Here, the asteraceae plant used in the myoblast proliferation promoter in the present invention is a perennial plant, and among these, chrysantheum morifolium syn. Or its related plants are preferably used. .
Chrysanthemum grows naturally in Japan and is easily available, and its flower part has been used medicinally for a long time in China. In Japan, it is drunk as tea or liquor or used for cooking as food, and is said to be effective for dizziness, headaches, and eye strain.
Among these chrysanthemums, it is more preferable to use purple chrysanthemums which are edible chrysanthemums. This purple chrysanthemum is said to be "prolonged life" because it can live long when eaten, and is cultivated in Yamagata and Niigata prefectures. It is.

  Asteraceae plants can use stems, leaves, fruits, pericarps, seeds, seed coats, roots, flowers, etc., as well as crushed materials (including juices, powders, etc.) or extracts as raw materials. However, it is preferable to use flowers from the work surface.

As a method for pulverizing Asteraceae plants, either wet or dry methods may be used, and pulverization conditions are not particularly limited.
The pulverizer is not particularly limited, and a commercially available pulverizer can be used as appropriate. For example, a high-pressure homogenizer, an ultrasonic pulverizer, an airflow pulverizer, a high-speed rotary impact pulverizer, a ball mill, a bead mill, etc. Is mentioned.
The pulverization treatment may be repeated a plurality of times as necessary so that the particle size of the pulverized product is within a preferable range.

The extraction method of the extract from the Asteraceae plants is not particularly limited, and can be appropriately selected according to the purpose. For example, a method of extracting with water or an organic solvent is preferable, and examples thereof include a warm extraction method and a supercritical fluid extraction method. The extraction method may be used alone or a plurality of methods may be combined.
Examples of the extraction solvent include water, lower alcohols such as methanol, ethanol, propanol, and isopropanol, polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, dipropylene glycol, and glycerin, ethyl ether, and propyl. Solvents such as ethers such as ether, esters such as butyl acetate and ethyl acetate, and ketones such as acetone and ethyl methyl ketone can be used, and one or more of these are selected and used. Further, physiological saline, phosphate buffer, phosphate buffered saline, or the like may be used. Furthermore, you may use 1 type, or 2 or more types of supercritical fluids and subcritical fluids, such as water, a carbon dioxide, ethylene, propylene, ethanol, methanol, ammonia. It is also possible to extract under pressure using an autoclave or the like. When an organic solvent is used as the extraction solvent, it is preferable to use an organic solvent that is acceptable for the production of foods or drugs. For example, ethanol, edible fats and oils, etc. are mentioned.
The obtained extract can be used as it is, but the concentrated and dried solid can be dissolved again in water or a solvent, or decolorized, deodorized, desalted, etc. as long as these physiological functions are not impaired. You may use after performing the refinement | purification process of this, or performing the fractionation process by column chromatography. For storage, it can be freeze-dried after purification and dissolved in a solvent when used.

The extract of Asteraceae plants obtained by the extraction can be used as it is, but it can be used again as long as the concentrated and dried solids are not dissolved again or the myoblast proliferation promoting effect is not impaired. It may be used after performing purification treatment such as dilution or decolorization, deodorization, desalting, etc., or fractionation treatment by column chromatography.
The form is not particularly limited. For example, there are solutions, suspensions, semi-solids (eg, gels, pastes, etc.), solids (eg, powders, granules, tablets, etc.). These forms can be appropriately prepared by a known method.

The myoblast proliferation promoter in the present invention may contain other components.
Examples of other components include fats and oils, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, metal soaps, pH adjusters, preservatives, fragrances, powders, UV absorbers, Thickeners, dyes, antioxidants, stabilizers, chelating agents, excipients, extenders, cooling agents, dispersants, diluents, antifoaming agents, solubilizers, disintegrants, binders, lubricants , Wetting agents, coating agents, coloring agents, flavoring agents, reversible agents, capsule bases and the like. As other components, a functional component may be added.

In preparing the myoblast proliferation promoter of the present invention, the Asteraceae plants (including pulverized products and extracts) may be used as they are, but water (purified water, etc.), edible oil (corn oil) Etc.) may be used after being dissolved or dispersed.
In addition, the content of the Asteraceae plants (including pulverized products and extracts) contained in the myoblast proliferation promoter is not particularly limited, and depends on various conditions such as purpose, shape, and intended use. The content can be appropriately set in a wide range, and a range of 0.0001 to 100% by mass is preferably selected.
Even when a myoblast proliferation promoter is included as a component of the composition, the content is not particularly limited, and the content can be appropriately set in a wide range according to conditions, and the myoblast proliferation promoter is 0.0001. A range of ~ 90% by weight, preferably 0.001-50% by weight, is selected.

The myoblast proliferation promoter of the present invention is not particularly limited as an ingestion method, ingestion amount, ingestion frequency, ingestion time, and ingestion target, and can be appropriately selected according to the purpose.
There is no restriction | limiting in particular as said ingestion method, According to purposes, such as oral and dermal, it can select suitably. In the case of an oral composition, the myoblast proliferation promoter can be ingested easily and continuously. The oral composition may be taken as it is, or may be taken by dissolving or dispersing in a solution such as water. Oral composition is cream, tablet, capsule, caplet, soft capsule, stick, plate, block, gel, jelly, gummy, wafer, biscuit, bowl, chewable, You may shape | mold in syrup shape, stick shape, etc.
Moreover, when it is set as the composition for transdermal, you may make it apply locally.

  The intake of the myoblast proliferation promoter is not particularly limited, and can be appropriately selected in consideration of various factors such as the age, weight, and constitution of the individual to be ingested.

Hereinafter, the present invention will be specifically described by way of examples. The present invention is not limited to these examples.

<Activation of myoblasts>
Example 1
A 96-well plate of L6 (rat skeletal muscle-derived myoblasts) subcultured in a standard medium (DMEM medium containing fetal bovine serum inactivated at 10% by volume) to give 1 × 10 ^ 4 cells / well And precultured for 24 hours.
Next, a powder of purple chrysanthemums (flowers), which is an Asteraceae genus plant, was prepared as a sample. Note that MurasakiKiku used as a sample, cyanidin as anthocyanin 3-o- (6 '' -o- monomer B sulfonyl-beta-D-glucopyranoside) and cyanidin 3-o- (3 '', 6 '' - o- dimaleate b sulfonyl-beta-D-glucopyranoside), etc. are included.
A standard medium dissolved so that the concentration of the sample was 6.25, 12.5, 25, 50, 100 μg / mL and a standard medium containing no sample were prepared and replaced with the DMEM medium in which L6 was cultured. Thereafter, the cells were further cultured for 24 hours.
Next, the cultured medium is discarded, washed with serum-free DMEM, Cell Counting Kit-8 (manufactured by Dojindo Laboratories) diluted 1:30 with serum-free DMEM is added to each well, and 2 Time culture was performed.
After culturing, the absorbance at 450 nm was measured using a plate reader. The proliferation rate of myoblasts was calculated as proliferation rate = (absorbance of cells added with sample / absorbance of cells not added with sample) × 100.

(Comparative Example 1)
The procedure was the same as Example 1 except that black soybean powder was used as a sample instead of purple chrysanthemums. The black soybean of Comparative Example 1 contains cyanidin-3-glucoside as anthocyanin.

(Comparative Example 2)
Example 1 was repeated except that grape seed powder was used as a sample instead of purple chrysanthemums. The grape seed of Comparative Example 2 contains proanthocyanidins, which are related compounds of anthocyanins.

(Comparative Example 3)
Instead of purple chrysanthemum, the same procedure as in Example 1 was performed except that lutein powder extracted from marigold belonging to the genus Asteraceae was used as a sample.
The above results are shown in FIG.

  FIG. 1 and Table 1 indicate that in Example 1, the proliferation rate of myoblasts increased as the sample concentration increased, and there was a positive correlation between the sample concentration and the proliferation rate of myoblasts. In particular, when the sample concentration was 50 μg / mL, the proliferation rate of myoblasts was 153%, indicating that a high myoblast proliferation effect was obtained.

Compared to Example 1, in black soybean (Comparative Example 1) and grape seed (Comparative Example 2), which are known to contain anthocyanins, in Comparative Example 1, the growth rate of myoblasts increased in sample concentration. In Comparative Example 2, the growth rate of myoblasts is not more than 110% without changing the sample concentration. Conversely, when the sample concentration reaches 50 μg / mL, the growth rate is increased. However, the proliferation of myoblasts was suppressed by 28%.
Therefore, in plants such as black soybeans containing anthocyanins, which are considered to be good for the eyes, and grapes containing anthocyanin-related compounds, myoblast proliferation was not promoted. In purple chrysanthemum, an excellent myoblast proliferation promoting action was confirmed.

Further, in Comparative Example 3 using lutein extracted from Marigold belonging to the genus Asteraceae as a sample, the proliferation rate of myoblasts was 115 to 120%, but the value was lower than the result of purple chrysanthemums of Asteraceae. Met.
From this, it was confirmed that among the plants belonging to the genus Asteraceae, purple chrysanthemum flowers have a superior myoblast proliferation promoting effect compared to plants of other genera.

From the above, it can be expected that muscles increase by increasing muscle strength and recovering muscles by promoting muscle metabolism due to the myoblast proliferation promoting action by Asteraceae plants.
In addition, by increasing and recovering muscle strength around the eyes (mainly ciliary muscles), the reduction of vision and the improvement of visual acuity are promoted, and the eye focus adjustment function that is effective for presbyopia prevention and presbyopia recovery is improved. It can be used as an agent.

<Clinical trial on improvement of eye focus adjustment function>
(Example 2)
6 healthy adult males and 4 adult females (average age 31.5 ± 4.4 years old) as subjects, 2 supplements containing purple chrysanthemum flower powder (purple chrysanthemum contains 51 mg in total) for 14 days Ingested with water every morning, and then conducted a questionnaire on visibility.
Thereby, the presence or absence of the improvement of the focus adjustment function of eyes in the myoblast proliferation promoter of the present invention was confirmed.
During clinical trials, the use of health foods, supplements, and quasi-drugs that are effective for improving blood flow other than the test foods described below was prohibited.
In addition, during clinical trials, hunger and eating were avoided, sufficient sleep was taken, and physical condition was maintained. During clinical trials, food was consumed as much as possible.

Survey results,
(1) It became easier to focus on the eyes.
(2) The blur of eyes at the time of long desk work has been eliminated or the frequency of blurring of eyes has decreased.
(3) After a long time of desk work, the focus came straight away even when looking at the distance.
(4) The night vision (including when driving) became brighter.
Etc., visibility of all subjects was improved.

  From the above, it was confirmed that the myoblast proliferation promoter of the present invention has an effect of improving the focus adjustment function of the eyes.

  According to the present invention, as a new use using a plant belonging to the genus Asteraceae, it is excellent in safety, and is excellent in enhancing muscle strength and suppressing atrophy, promoting muscle metabolism, recovery of visual acuity associated therewith, prevention of visual loss, and the like. It is possible to provide a myoblast proliferation promoting agent and a focus adjusting function improving agent having actions and effects.

Claims (1)

  Purple chrysanthemum containing cyanidin 3-o- (6 ″ -o-monomalonyl-β-D-glucopyranoside) and cyanidin 3-o- (3 ″, 6 ″ -o-dimalonyl-β-D-glucopyranoside) Metabolism promoters that contain flowers of flowers (excluding agents for restoring visual acuity, preventing vision loss or improving focus adjustment function).

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