JP7352275B2 - Composition for improving menopausal symptoms - Google Patents

Description

Application of Article 30, Paragraph 2 of the Patent Act March 31, 2019, https://www. mdpi. com/1420-3049/24/7/1259/pdf

Article 30, Paragraph 2 of the Patent Act applies April 1, 2019, https://www. mdpi. com/1420-3049/24/7/1272/pdf

The present invention relates to a composition for improving menopausal symptoms.

Cassis is also called black currant, black currant, black currant, etc., and it grows in cool regions such as New Zealand, and in Japan, it is a fruit tree plant with the largest production volume in Aomori Prefecture.

In 2015, the Ministry of Agriculture, Forestry and Fisheries certified Aomori Cassis as the first product under the Geographical Indication Protection System. Blackcurrant is rich in vitamins, minerals, and anthocyanins, and is known to be effective in preventing dark circles and reducing eye strain (see Non-Patent Document 1).

Molecules with estrogenic activity exist in plants and are collectively called phytoestrogens (phytoestrogens). Phytoestrogens are so-called plant-based female hormones, and isoflavones contained in soybeans and resveratrol contained in grapes are known as a type of phytoestrogens (see Non-Patent Document 2 and Non-Patent Document 3). .

The present inventors have revealed that anthocyanins contained in blackcurrants have estrogenic activity via estrogen receptors (ER) α and ERβ, and are a type of phytoestrogens (Non-patent Document 4 and (See Non-Patent Document 5). In addition, the present inventors have reported that the amounts of collagen, elastin, and hyaluronic acid were increased by ingesting a blackcurrant extract to ovariectomized rats (see Non-Patent Document 6).

Gopalan, A. et al., Food Funct 2012, 3, 795-809. Limer, J. L., et al, Breast Cancer Research, 2004, 6(3), 119-127. Mahmoud, A. M., et al., Journal of Steroid Biochemistry and Molecular Biology, 2014, 140, 116-132. Nanashima, N. et al., Mol Nutr Food Res 2015, 59, 2419-2431. Nanashima, N. et al., Molecules 2018, 23, 74 Nanashima, N. et al., Nutrients, 2018, 10, 495

According to Non-Patent Documents 4 to 6, it is known that anthocyanins contained in blackcurrants function as phytoestrogens.

However, according to our investigation, the amount of collagen in the skin of ovariectomized rats that ingested blackcurrant extract increased more than that in sham-operated rats. This suggests that the collagen-increasing effect of the blackcurrant extract may occur not only in female individuals with reduced estrogen secretion, but also in healthy female individuals. Therefore, it cannot be denied that the collagen increasing effect of the blackcurrant extract may be caused by components other than phytoestrogens (anthocyanins) contained in the blackcurrant extract.

Furthermore, it has not been known until now that blackcurrant extract improves the symptoms peculiar to menopausal females to the same extent as those of non-menopausal females.

Therefore, the present invention aims to provide a composition containing as an active ingredient a substance that has the effect of improving the symptoms peculiar to female individuals in menopause to at most the same level as that of female individuals who are not in menopause. , the problem to be solved by the invention.

The inventors of the present invention have conducted extensive studies to solve the above problems, and have surprisingly found that ingestion of blackcurrant extract can reduce hair thinning, weight gain, and intra-abdominal fat observed in menopausal females. It has been found that it has the effect of improving symptoms such as increase in weight to the same level as in non-menopausal female individuals.

In addition, blackcurrant extract improved nitric oxide (NO) production in vascular endothelial cells to the same level as 17β-estradiol, and also increased endothelial nitric oxide synthase (eNOS) in vascular tissues of ovariectomized rats. The expression level was improved to the same level as that of sham-operated rats. Thus, the present inventors found that a blackcurrant extract has a vasodilatory effect and an ameliorating effect on blood vessel stiffness through NO and eNOS.

Furthermore, the present inventors found through metabolomic analysis that black currant extract contains not only anthocyanins but also β-estradiol and 17α-estradiol. Furthermore, it was surprisingly found that blueberry fruit powder also contained β-estradiol and 17α-estradiol at the same level as blackcurrant extract. This indicates that blueberry fruit powder has the same effect on improving menopausal symptoms as blackcurrant extract. All of these findings were discovered for the first time by the present inventors.

Based on such knowledge, the present inventors succeeded in creating a composition for improving menopausal symptoms that contains blackcurrant, blueberry, and/or an extract thereof as an active ingredient. The present invention was completed based on such knowledge and successful examples.

Therefore, according to one aspect of the present invention, the following compositions and methods [1] to [9] are provided.
[1] Contains at least one active ingredient selected from the group consisting of blackcurrants, blueberries, and their extracts, and has the effect of improving menopausal symptoms other than skin deterioration due to decreases in collagen, elastin, and hyaluronic acid. , a composition for improving menopausal symptoms.
[2] The composition according to [1], wherein the menopausal symptom is at least one menopausal symptom selected from the group consisting of hair thinning, vascular sclerosis, weight gain, and fat gain.
[3] The composition according to any one of [1] to [2], wherein the active ingredient is at least one active ingredient selected from the group consisting of blackcurrant and blackcurrant extract.
[4] The composition according to any one of [1] to [3], wherein the composition is an oral composition.
[5] A composition for hair growth, a composition for vasodilation, a composition for improving vascular stiffness, and a composition for suppressing weight gain, containing at least one active ingredient selected from the group consisting of blackcurrants, blueberries, and extracts thereof. composition, composition for suppressing fat gain, or composition for anti-hepatitis.
[6] The composition according to [5], wherein the composition is a composition to be ingested by an adult female.
[7] Improvement of menopausal symptoms, comprising the step of improving the menopausal symptoms of a female individual suffering from menopausal symptoms by administering the composition according to any one of [1] to [6] to the female individual suffering from menopausal symptoms. Method.
[8] A method of increasing hair growth, dilating blood vessels, and softening blood vessels in an adult woman by administering the composition according to any one of [1] to [6] to the adult woman. method, method to reduce weight, method to reduce fat or method to reduce hepatitis.
[9] Use of at least one active ingredient selected from the group consisting of blackcurrants, blueberries, and extracts thereof for producing the composition according to any one of [1] to [6].

According to the composition of one embodiment of the present invention, menopausal symptoms such as thinning hair, vascular hardening, weight gain, and fat gain can be alleviated by ingesting blackcurrant, blueberry, extracts thereof, and the like as active ingredients in female individuals who are not in menopause. can be expected to improve to the same extent.

FIG. 1 shows the results of metabolomic analysis of BCE and blueberry fruit powder as described in the Examples. FIG. 2 is a diagram showing the results of measuring the expression level of the KRT19 gene by real-time PCR as described in the Examples. Figure 3 shows the results of HE staining of hair follicles in rat skin tissues (A to C; scale bar is 20 μm) and the number of hairs per hair follicle as described in the Examples. It is a figure which shows the result (D) obtained by measurement. FIG. 4 is a diagram showing the results of immunostaining using anti-Ki67 antibodies (A to D; scale bar is 100 μm) and the measurement results of Ki67-positive cell percentage (E), as described in the Examples. . FIG. 5 shows the results of fluorescent immunostaining using an anti-KRT19 antibody (scale bar is 100 μm) as described in the Examples. FIG. 6 shows the results of fluorescent immunostaining (scale bar: 100 μm) using antibodies for KRT15 and CD34, as described in the Examples. FIG. 7 shows the change in body weight of each group of rats before and after rearing for 3 months as described in the Examples. FIG. 8 shows the change in intraperitoneal fat weight before and after 3 months of rearing of each group of rats as described in the Examples. FIG. 9 shows the changes in liver weight of each group of rats before and after 3 months of rearing as described in the Examples. FIG. 10 is a diagram showing a photograph obtained by HE staining intraperitoneal adipose tissue after 3 months of rearing as described in Examples. FIG. 11 is a diagram showing the measurement results of the diameter of adipocytes in intraperitoneal adipose tissue as described in the Examples. FIG. 12 is a diagram showing the results of real-time PCR analysis of the expression level of TNFα in liver cells as described in the Examples. FIG. 13 is a diagram showing the results of real-time PCR analysis of the expression level of IL-6 in liver cells as described in the Examples. FIG. 14 shows the results of RT-qPCR analysis as described in the Examples. A in HUVEC, and B in EA. The mRNA expression level of eNOS in hy926 cells is shown. C, EA in the presence of 100 nM fulvestrant. Results in hy926 cells are shown. Data are shown as the mean ± SEM of at least three independent experiments. *p<0.05, **p<0.01 versus untreated control cells. C3G represents cyanidin-3-glucoside, C3R represents cyanidin-3-rutinoside, D3G represents delphinidin-3-glucoside, and D3R represents delphinidin-3-rutinoside. FIG. 15 shows the results of NO measurements in the absence (A) and presence (B) of 100 nM fulvestrant, as described in the Examples. Data are presented as the mean ± SEM of at least three independent experiments. *p<0.05, **p<0.01 vs. untreated control cells. FIG. 16 is a diagram showing the results of immunohistochemical staining of vascular endothelial cells of a menopausal model rat as described in the Examples. A shows an image of eNOS protein expression at low magnification (×200). B shows an image 400 times larger than the image of the boxed area shown in A. C shows an image using tissue not treated with primary antibody as a negative control. D shows the results of scoring the staining intensity of eNOS as the mean score±SEM. *p<0.05.

The details of the present invention will be described below, but the technical scope of the present invention is not limited only by the items in this section, and the present invention can take various embodiments as long as the object is achieved.

Unless otherwise specified, each term herein is used in the sense commonly used by those skilled in the art and should not be construed as having an unduly limiting meaning. Further, since the speculations and inferences made in this specification were made based on the knowledge and experience of the present inventors, the technical scope of the present invention is not limited solely to such speculations and inferences. It is not something that will be done.

A "composition" is not particularly limited in its commonly used meaning, but is, for example, a combination of two or more substances, specifically a combination of an active ingredient and another substance. Examples include solids that are a combination of two or more active ingredients, and more specifically, solids that are a combination of one or more active ingredients and one or more solid carriers or solvents. Examples include compositions and liquid compositions.

The number of digits of the integer value and the number of significant figures match. For example, 1 has 1 significant digit, and 10 has 2 significant digits. Furthermore, in a decimal value, the number of digits after the decimal point and the number of significant figures match. For example, 0.1 has one significant digit, and 0.10 has two significant digits.

"And/or" means any one of the listed related items, any combination of two or more, or all combinations.

"Content" is synonymous with concentration, and means the ratio of the amount of the component to the total amount of the composition. However, the total content of the components does not exceed 100%.

"~" in the numerical range is a range that includes the numbers before and after it; for example, "0% by mass to 100% by mass" means a range of 0% by mass or more and 100% by mass or less. .

"Menopause" refers to the transition period from the reproductive period to the non-reproductive period as a woman ages, and specifically refers to a total of 10 years, five years before and after menopause. In women undergoing menopause, the female hormone estrogen decreases due to decline in ovarian function or aging, and physiological and sexual functions decrease or disappear.

"Menopausal symptoms" are symptoms seen in women who are undergoing menopause. Menopausal symptoms include vascular symptoms such as facial flushing, hot flashes, tachycardia, sweating, headache, and stiff shoulders; feeling of fatigue, palpitations, muscle pain, joint pain, lower back pain, cold feet and lower back, frequent urination, and urination. Physical symptoms such as incontinence, dizziness, decreased vision, skin deterioration, thinning hair, weight gain, increased fat, increased neutral fat, and hepatitis; decreased vision; psychological symptoms such as decreased memory, depressive symptoms, decreased concentration, etc. can be mentioned.

"Symptom-improving effect" is not particularly limited in its commonly used meaning, but includes, for example, the effect of moving a symptom-free state toward a symptom-free state; This includes actions that do not make symptoms worse, actions that maintain a symptom-free state, and actions that prevent a person from progressing from a symptom-free state to a symptom-free state. Specifically, "symptom-improving effects" include prevention and delay of the occurrence of symptoms and reduction of the risk of occurrence; improvement of symptoms; prevention and delay of worsening of symptoms; maintenance of functions that cause symptoms and prevention of worsening. and delays. "Improvement" of symptoms includes alleviation, suppression, inhibition, recovery, treatment, and prevention of symptoms.

The composition of one embodiment of the present invention contains at least an active ingredient that is blackcurrant, blackcurrant extract, blueberry, blueberry extract, or two, three, or four types thereof. The composition of one embodiment of the present invention has an effect of improving menopausal symptoms by containing these active ingredients.

The scientific name of blackcurrant is Ribes nigrum or Ribes nigrum L. It is the fruit of a temperate deciduous shrub fruit tree, also called black currant, black currant, black currant, etc. Blackcurrant fruit is used in jellies, jams, ice creams, cordials, and liqueurs. As long as the blackcurrant includes the fruit, other parts such as blackcurrant leaves, stems, roots, and flowers may be mixed in as well.

Blackcurrants may be harvested as they are (fresh fruits), but since blackcurrant juice residue contains a large amount of β-estradiol and 17α-estradiol in addition to anthocyanins, the juice must be squeezed after harvesting. Preferably, it is a processed product that has been subjected to processing such as processing or drying. In addition, if it takes time to process the blackcurrant after it is collected, it is preferable to store it by storage means commonly used by those skilled in the art, such as low-temperature storage, in order to prevent deterioration of the blackcurrant.

The method of squeezing is not particularly limited as long as it is a method that uses commonly used plant fruits to separate squeezed juice and juice residue, but for example, blackcurrants may be squeezed using a mechanical device such as a squeezer or juicer. Examples include a method of obtaining a juice residue by squeezing the juice using a squeezing means and collecting the crude solid content by means such as sieving or filtration as necessary. The juice residue is preferably subjected to drying treatment.

The drying method is not particularly limited as long as it is a commonly used method for drying plant fruits. For example, if the mass of blackcurrant after drying (dry mass) is equal to the mass before drying (wet mass). For example, drying may be carried out so that the amount of carbon dioxide is 80% or less, preferably 50% or less, more preferably 30% or less, even more preferably 10% or less. The drying treatment may be performed by any method known to those skilled in the art, such as hot air drying, high pressure steam drying, electromagnetic wave drying, freeze drying, and air drying, but in order to prevent the components contained in blackcurrant from deteriorating or decomposing. Freeze-drying is preferred.

A blackcurrant extract is obtained by subjecting commonly used blackcurrant or processed blackcurrant products (hereinafter referred to as blackcurrant etc.) to an extraction treatment. The extraction treatment method is not particularly limited as long as it is a commonly used method for extracting desired components contained in plant fruits. For example, blackcurrant etc., and β-estradiol and 17α-estradiol contained in blackcurrant. Examples include a method that includes obtaining an extract by contacting the target with a solvent that is soluble in the target.

As the extraction solvent, examples of solvents that are soluble in β-estradiol and 17α-estradiol include water, organic solvents, and mixed solvents of water and organic solvents. Examples of organic solvents include lower alcohols such as methanol, ethanol, propanol, isopropanol, and butanol; lower esters such as ethyl acetate and methyl acetate; dimethyl sulfoxide, acetonitrile, acetone, hexane, glycerin, and propylene glycol; preferably, A mixed solvent of water and an organic solvent, more preferably a mixed solvent of water and ethanol. Extraction conditions are not particularly limited as long as they are conditions suitable for dissolving β-estradiol and 17α-estradiol in blackcurrants, etc., and the type and use of extraction solvent may vary depending on the condition of blackcurrants etc. and the amount used. The amount can be set as appropriate.

By performing solid-liquid separation treatment after the extraction treatment, it is possible to separate the extract from the residue of the raw material, such as black currant. Although the solid-liquid separation means is not particularly limited, for example, known solid-liquid separation means such as filtration and centrifugation can be used.

The extract may be subjected to concentration treatment and/or drying treatment as necessary. The concentration treatment method is not particularly limited as long as it is a commonly known method for reducing the volume of a liquid. For example, the extract is reduced in pressure as necessary at room temperature or under heating to form a concentrated liquid. Examples include methods including obtaining. The degree of concentration is not particularly limited, but the volume may be reduced to 1/2 to 1/10 or less, for example. The concentrated liquid is preferably subjected to a drying process such as freeze drying, hot air drying, or spray drying, preferably freeze drying process. Furthermore, it is preferable that the dried concentrate is subjected to pulverization treatment. The method of pulverization treatment is not particularly limited, and examples thereof include methods using a crusher, blender, mixer, mill, and the like.

A specific method for producing a blackcurrant extract containing β-estradiol and 17α-estradiol using blackcurrant as a raw material will be described below, but the method for producing a blackcurrant extract is not limited to the following.

Freeze-dry the squeezed residue of blackcurrant fruit to obtain a dried squeezed residue, and then subject the obtained dried squeezed residue to an extraction treatment using aqueous ethanol to obtain an extract. The extracted liquid is sequentially subjected to a filtration process using a filter, a concentration process, a freeze-drying process, a pulverization process, and a mixing process to obtain a blackcurrant extract. By sieving the blackcurrant extract, a blackcurrant extract with a specific particle size, for example 80 mesh, is obtained.

Blueberries are not particularly limited as long as they are fruits of deciduous shrubs and fruit trees of the Ericaceae family, Vaccinium genus, which are commonly known as blueberries. As long as the blueberry includes the fruit, it may be mixed with other parts of the blueberry such as leaves, stems, roots, flowers, etc.

Blueberries may be either ones in the state they are collected (fresh fruits) or processed products that have been subjected to processing such as squeezing or drying after being collected. When blueberries are subjected to drying treatment, the method of drying treatment is preferably freeze-drying. If it takes time to process blueberries after they are harvested, it is preferable to store them by storage means commonly used by those skilled in the art, such as low-temperature storage, to prevent deterioration of the blueberries.

For the blueberry extract, the description of the blackcurrant extract described above can be referred to as appropriate.

The blackcurrant extract and the blueberry extract may be obtained by the above-described production method or commercially available. For example, blackcurrant extract is commercially available as "CaNZaC-35" (Koyo Sho Company).

The climacteric symptom-improving effect of the composition of one embodiment of the present invention is not particularly limited as long as it improves at least one type of menopausal symptoms. However, the effect of improving menopausal symptoms that the composition of one embodiment of the present invention has excludes the effect of improving skin deterioration caused by a decrease in collagen, elastin, and hyaluronic acid, but the cause is different from the decrease in collagen, elastin, and hyaluronic acid. This includes the effect of improving skin deterioration due to

The menopausal symptom-improving effect of the composition of one embodiment of the present invention is preferably an effect of improving hair thinning, vascular sclerosis, weight gain, fat gain, or two, three, or four of these symptoms. The degree of climacteric symptom-improving effect of the composition according to one embodiment of the present invention may be such that it improves the effect on female individuals undergoing menopause compared to when the composition according to one embodiment of the present invention is not used. , preferably to the same level as a non-menopausal female individual.

The effect of improving menopausal symptoms can be confirmed and evaluated by a method of comparison using menopausal model animals and sham-operated animals as described in the Examples below. For example, compared to sham-operated animals, menopausal model animals have a decrease in the number of hairs per hair follicle, a decrease in eNOS expression level and NO synthesis in skin tissue, an increase in body weight, and an increase in intraperitoneal fat. , and/or the diameter of adipocytes increases.

If the reduction in the number of hairs per hair follicle in a menopausal model animal is improved by ingesting the composition of one embodiment of the present invention, the number of hairs per hair follicle is preferably comparable to the number of hairs per hair follicle in a sham-operated animal. If this is the case, it can be said that the composition of one embodiment of the present invention has the effect of improving the menopausal symptom of thinning hair.

If the reduction in eNOS expression level and/or NO synthesis amount in vascular tissue in a menopausal model animal is improved by ingesting the composition of one aspect of the present invention, preferably the reduction in eNOS expression level and/or NO synthesis amount in vascular tissue in a sham-operated animal is improved. If the amount of NO synthesis is improved to the same level as the amount of NO synthesis, it can be said that the composition of one embodiment of the present invention has the effect of improving menopausal symptoms, which is blood vessel hardening.

If the weight gain in a menopausal model animal is improved by ingesting the composition according to one embodiment of the present invention, preferably to the same level as the weight gain in sham-operated animals, the composition according to one embodiment of the present invention It can be said that this product has the effect of improving menopausal symptoms, which is weight gain.

If intraperitoneal fat increases and/or fat cell diameter increases in menopausal model animals by ingesting the composition of one aspect of the present invention, preferably intraperitoneal fat in sham-operated animals is improved. If the increase and/or the diameter of adipocytes is improved to the same level as that of adipocytes, it can be said that the composition of one embodiment of the present invention has an effect of improving menopausal symptoms, which is an increase in fat.

If the increase in the expression level of hepatitis-related genes in menopausal model animals is improved by ingesting the composition of one embodiment of the present invention, it is preferably improved to the same level as the expression level of hepatitis-related genes in sham-operated animals. Therefore, it can be said that the composition of one embodiment of the present invention has an effect of improving hepatitis that may occur as a result of fat accumulation in the liver.

The composition of one embodiment of the present invention is expected to have various effects such as hair growth effect, anti-vascular disease effect, and anti-obesity effect by having an effect of improving menopausal symptoms. Compositions, compositions for anti-vascular diseases, compositions for anti-hypertension, compositions for reducing vascular resistance, compositions for anti-obesity, compositions for improving QOL (Quality of life), preferably the above-mentioned compositions for adult women. It can take various forms. For example, the composition of one aspect of the invention helps reduce body weight, abdominal fat (visceral fat and/or subcutaneous fat), BMI (Body Mass Index), and/or waist circumference in obese adult women. It can be a composition for

Blackcurrant and blueberry, which are active ingredients, have been ingested by humans for a long time and are highly safe. Therefore, the composition of one embodiment of the present invention is highly practical and can be used as is or by processing. , applicable to various uses.

The composition of one aspect of the present invention is not particularly limited in its intake method, and can be applied orally or parenterally, for example. Parenteral applications include, but are not limited to, injections and infusions such as intradermal, subcutaneous, intravenous, and intramuscular administration; transdermal; and inhalation through mucous membranes such as the nose and pharynx. The composition of one embodiment of the present invention is preferably an oral composition for easy and long-term ingestion.

The subject that receives the composition of one embodiment of the present invention is not particularly limited, and includes, for example, animals, especially mammals, such as humans, dogs, cats, cows, horses, pigs, and sheep. Among these, humans are preferred. The ingesting individual may be a healthy female individual in the climacteric period, but is preferably a female individual exhibiting menopausal symptoms.

The composition of one aspect of the present invention is not particularly limited as long as it is in a commonly used form, and examples include solid, liquid, gel, suspension, cream, sheet, stick, powder, granule, It can take various forms such as granules, tablets, rods, plates, blocks, pastes, capsules, and caplets.

The composition of one embodiment of the present invention may contain the active ingredient in combination with other ingredients as long as the composition has an effect of improving menopausal symptoms. That is, the composition of one embodiment of the present invention may contain various other ingredients in addition to the active ingredient as long as the object of the present invention can be achieved.

Other ingredients include, for example, carbohydrate sweeteners, stabilizers, emulsifiers, starch, starch processed products, starch decomposition products, salt, flavoring agents, coloring agents, acidulants, flavor materials, nutrients, fruit juice, eggs, etc. Examples include additives used in normal food processing and pharmaceutical processing, such as animal and plant foods, excipients, fillers, binders, thickeners, perfume oils, preservatives, buffers, and brighteners. The amounts of other components to be used are not particularly limited as long as they do not interfere with solving the problems of the present invention, and can be set as appropriate.

Examples of excipients include lactose, glucose, sucrose, mannitol, potato starch, dextrin, corn starch, calcium carbonate, calcium phosphate, calcium sulfate, crystalline cellulose, licorice powder, gentian powder, and the like. Examples of the binder include starch, gum tragacanth, gelatin, syrup, polyvinyl alcohol, polyvinyl ether, polyvinylpyrrolidone, hydroxypropylcellulose, methylcellulose, ethylcellulose, carboxymethylcellulose, and the like. Examples of the disintegrant include starch, agar, gelatin powder, sodium carboxymethylcellulose, calcium carboxymethylcellulose, crystalline cellulose, calcium carbonate, methylcellulose, ethylcellulose, and carboxymethylcellulose. Examples of the lubricant include talc and magnesium stearate. The coloring agent is not particularly limited and may be one that is allowed to be added to foods and drinks and oral pharmaceuticals.

When preparing tablets or granules, if desired, white sugar, gelatin, purified shellac, glycerin, sorbitol, ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyvinyl pyrrolidone, cellulose phthalate acetate, hydroxypropyl methyl cellulose phthalate, methyl methacrylate, It may be coated using a methacrylic acid polymer or the like, or it may be coated in multiple layers. Furthermore, granules or powders can be packed into capsules made of ethyl cellulose or gelatin to form capsules.

The content of the active ingredient in the composition of one embodiment of the present invention is not particularly limited as long as it has an effect of improving menopausal symptoms, but as an oral composition, for example, the amount per intake is 0. The amount is from 1 mg to 10,000 mg, preferably from 1 mg to 1,000 mg, more preferably from 10 mg to 500 mg, and even more preferably from 20 mg to 100 mg; based on the total amount of the composition, from 0.1% by mass. 99.9% by weight, preferably 0.5% to 99% by weight, more preferably 1% to 90% by weight, even more preferably 5% to 90% by weight, even more preferably 10% to 90% by weight %.

The amount ingested of the composition of one aspect of the present invention is not particularly limited, and can be appropriately set according to the characteristics of the ingesting individual such as the age, physique, and symptoms of the ingesting individual; the method of ingestion; and the site of application. The intake amount of the composition according to one aspect of the present invention is, for example, about 0.1 g to about 10 g/day, preferably 1 g to about 10 g/day, for a general adult female (body weight: 50 kg) when converted from a human equivalent dose. 5g/day.

The number of times the composition of one embodiment of the present invention is ingested is not particularly limited, but is, for example, once to three times a day, and the number of times can be increased or decreased as appropriate depending on the degree of menopausal symptoms of the individual ingesting it and the amount of intake per time. can do. The intake period of the composition of one aspect of the present invention continues for a certain period of time, that is, 3 days or more, preferably 1 week or more, more preferably 1 month or more, still more preferably 3 months or more, and even more preferably 6 months or more. Administer directly. The composition of one embodiment of the present invention is preferably ingested every day, but does not have to be ingested every day as long as it is continuously ingested during the period.

The composition of one aspect of the present invention may take the form of a food or drink composition, a pharmaceutical composition, or the like containing the composition alone or containing the composition. Therefore, one specific embodiment of the present invention is a composition for food or drink, a composition for pharmaceuticals, a composition for quasi-drugs, which contains an active ingredient or contains the composition of one embodiment of the present invention. These include cosmetic compositions and animal feed compositions. The composition of one embodiment of the present invention is also useful as a composition for internal and external cosmetics (inner cosmetics) that can be taken orally.

When the composition of one embodiment of the present invention is a composition for food or drink, it can take the form of any food or drink; for example, pharmaceutical food such as capsules, tablets, granules, jelly, and paste; fruit juice drinks; Beverages such as , vegetable juices, fruit and vegetable juices, tea drinks, coffee drinks, sports drinks, soft drinks, and health drinks; Dairy products such as yogurt and cheese; Processed food and drinks such as soups, noodles, puddings, and jams; Chewing gum and candy Confectionery such as tablets, gummy jelly, chocolate, biscuits, and snacks; Frozen desserts such as ice cream, sherbet, and frozen desserts; Rice crackers such as rice crackers; Added to yokan and other soybean-based confections for general food and drink products. It can be in the form of

A specific embodiment of the food/beverage composition is, for example, a functional food/beverage product that has certain functionality for living bodies. Functional foods and beverages include, for example, foods and beverages for specified health uses, foods and beverages with functional claims, foods and beverages with nutritional functions, foods and beverages with health functions, foods and beverages for special purposes, dietary supplements, health supplements, supplements, and beauty beverages. In addition to so-called health foods and beverages such as these, it includes foods and beverages for specific persons such as foods and beverages for infants, foods and beverages for pregnant women, and foods and beverages for the elderly. Furthermore, functional food and drink products include health food and drink products to which health claims based on the food standards of Codex (FAO/WHO Joint Food Standards Committee) are applied.

The packaging form of the composition for food and beverages is not particularly limited and can be selected as appropriate depending on the form to be applied. For example, it may be made of coated paper, plastic such as PET or PTP, metal such as aluminum, glass, etc. Examples include packs, bottles, cans, pouches, and single-layer or laminated film bags.

When the composition of one embodiment of the present invention is a pharmaceutical composition, it can take the form of any pharmaceutical, but examples include oral preparations, injections (muscular, subcutaneous, intradermal), nasal spray preparations, etc. Ru. In addition to the active ingredient, the pharmaceutical composition may contain other physiologically active substances having an effect of improving menopausal symptoms, as long as the object of the present invention can be achieved. Furthermore, the pharmaceutical composition of one embodiment of the present invention may be used in combination with other pharmaceuticals containing other physiologically active substances as active ingredients.

The method for producing the composition of one embodiment of the present invention is not particularly limited, and includes, for example, a method of mixing the active ingredient and optionally other ingredients and molding the mixture into a desired dosage form.

The method of using the composition of one embodiment of the present invention is not particularly limited, but for example, when the composition of one embodiment of the present invention is an oral composition, the oral composition may be used as is, or with water or the like, or It can be taken orally by diluting it with water etc. and eating or drinking it. Depending on the preference of the individual using the composition, the composition of one embodiment of the present invention may be mixed with other solids or liquids and taken orally. When the composition of one embodiment of the present invention is made into an orally disintegrating dosage form, it can be orally ingested without water.

Another aspect of the invention is a method of utilizing the composition of one aspect of the invention or the active ingredient in the composition. A method of one embodiment of the present invention is a method for improving menopausal symptoms, which comprises the step of improving the menopausal symptoms of a female individual suffering from menopausal symptoms by administering the composition of one embodiment of the present invention to the female individual suffering from menopausal symptoms. . The above items can be referred to as appropriate for administration methods and methods for confirming the effect of improving menopausal symptoms in female individuals.

A method according to another embodiment of the present invention provides a method for increasing hair, dilating blood vessels, and softening blood vessels in an adult female by administering the composition according to one embodiment of the present invention to the adult female. method, method of suppressing weight gain, method of suppressing fat gain, or method of reducing the risk of hepatitis. Another aspect of the method of the present invention is the use of at least one active ingredient selected from the group consisting of black currants and blueberries and extracts thereof to produce the composition of one aspect of the present invention.

In the method of one embodiment of the present invention, various steps and operations can be added before or after the above-described steps, or during the steps, as long as the problems of the present invention can be solved.

Hereinafter, the present invention will be explained in more detail with reference to Examples, but the present invention is not limited to these Examples, and the present invention can take various embodiments as long as the problems of the present invention can be solved. .

[Example 1 Evaluation of thinning hair improving effect of blackcurrant extract and blueberry fruit powder on menopausal model]
(1-1) Blackcurrant extract and blueberry fruit powder As the blackcurrant extract (BCE), "CaNZac-35" (Koyo Sho Co., Ltd.), a commercially available BCE powder, was used. BCE contained polyphenols at 37.6g/100g-BCE and anthocyanins at 38.0g/100g-BCE. BCE also contained anthocyanins C3G (5.6% by mass), C3R (32.0% by mass), D3G (16.8% by mass), and D3R (45.3% by mass).

As the blueberry fruit powder, one obtained by subjecting blueberry fruit to freeze-drying treatment and then subjecting it to pulverization treatment was used.

The results of metabolome analysis of BCE and blueberry fruit powder are shown in FIG. As shown in FIG. 1, it was found that BCE and blueberry fruit powder contained β-estradiol and 17α-estradiol.

(1-2) Human dermal papilla cells Human dermal papilla cells (HFDPC) were purchased from Promocell. HFDPCs were maintained using a dermal papilla cell growth medium (Promocell). Cell culture was performed at 37°C in a humidified incubator containing 5% _CO2 .

(1-3) Gene analysis by microarray and IPA Dermal papilla cell basic medium (phenol red free; PromoCell) was used as the medium. After culturing HFDPC for 24 hours using a test medium containing 1 μg/ml BCE and a control medium consisting only of dermal papilla cell basal medium, cells were collected, and then the collected cells were washed twice with PBS. Total RNA was extracted from the washed cells using "RNeasy mini kit" (Qiagen).

RNA labeling and hybridization were performed according to the procedure of Agilent one-color microarray-based gene expression analysis. Specifically, 100 ng of total RNA was linearly amplified and then labeled using Cy3-dCTP. The obtained labeled cRNA was purified using "RNeasy mini kit". The labeled and fragmented cRNA was hybridized to "SurePrint G3 Human GE Microarray" (8x60K Ver. 3.0; Agilent Technologies). Microarray datasets are available at http://www. ncbi. nlm. nih. It is available under gov/geo accession number GSE121730.

Furthermore, for genes whose expression increased by more than 2.0 times by using the test medium containing BCE, pathway analysis was performed using IPA (Ingenuity Pathway Analysis) software (Ver. 36601845). A z-score algorithm was utilized to reduce the possibility of false positive results. A z-score of 2.0 or higher indicated that the expression of the transcript was significantly increased, and a z-score of −2.0 or lower indicated that the expression was significantly decreased.

As described above, changes in gene expression in human dermal papilla cells due to BCE were comprehensively explored.

(1-4) Gene analysis using real-time PCR In order to investigate the expression level of keratin 19 (KRT19), a dermal papilla stem cell marker that showed large fluctuations in microarrays, we used RNA extracted from human dermal papilla cells to perform ``TB Green''. Real-time PCR was performed using "Premix Ex Taq II (registered trademark) Tli RNaseH Plus" (TaKaRa).

(1-5) Evaluation of hair thinning improvement effect using ovariectomized rats. 12-week-old ovariectomized rats (“OVX female Sprague-Dawley”; Claire Japan) were used as menopausal model animals, and 12-week-old sham-operated animals were used as sham-operated animals. Sham rats ("Sprague-Dawley"; Claire Japan) were used. Six ovariectomized rats were divided into two groups, one of the three rats was fed a normal diet consisting only of "AIN-93M" (Claire Japan) (the "OVX control" group), and the other three were fed a normal diet consisting of only "AIN-93M" (Claire Japan). A test feed consisting of "AIN-93M" containing % BCE by mass was ingested ("OVX BCE" group). In addition, in order to eliminate the influence of the surgical procedure, three sham-operated rats that underwent only anesthesia and skin incision and whose ovaries were not removed were allowed to ingest normal food ("Sham" group). Rats in each group had free access to food and water.

After raising the rats in each of the above groups for 3 months, skin tissues were collected according to a conventional method, fixed in 10% formalin neutral buffer, and paraffin-embedded tissues were created into tissue blocks by embedding in paraffin. was cut into continuous sections with a thickness of 3 μm, placed on a slide glass to prepare tissue sections, and various evaluations were performed. The tissue sections were stained with hematoxylin and eosin (HE staining), the number of hairs per hair follicle was counted, and the mean ± standard error (SEM) of the measured values for each group was calculated. The results were tested using the Tukey-Kramer method (p<0.05).

Furthermore, in order to verify the effect of cell proliferation on hair follicles, skin tissue sections were immunostained using an anti-Ki67 antibody in order to examine the expression level of Ki67, a cell proliferation marker. In addition, the total cell number and ki67-positive cells in the hair follicle were counted under a microscope, the percentage of positive cells per total cell number was calculated, and the assay was performed in the same manner as above. In addition, the expression of each protein was examined by fluorescent immunostaining using anti-cytokeratin 19 (KRT19) antibody, anti-cytokeratin 15 (KRT15) antibody, and anti-CD34 antibody.

(1-6) Evaluation Results Table 1 shows the upstream factors of genes whose expression changed through BCE, as predicted by analysis using microarray and IPA.

The factors shown in Table 1 are factors that can bring about changes in gene expression in human dermal papilla cells due to BCE. A z score of 2.0 or more or -2.0 or less indicates that there is a significant difference. As shown in Table 1, β-estradiol has the highest z-score, so we decided to focus on the downstream genes of β-estradiol.

Among the downstream genes of β-estradiol, genes whose expression levels increased when the test medium containing BCE was used compared to when the control medium was used are shown in Table 2. As shown in Table 2, the expression levels of 17 types of genes increased by using the test medium containing BCE.

FIG. 2 shows the results of measuring the expression level of KRT19, which was the gene whose expression level increased the most based on the results of the microarray analysis, by real-time PCR. FIG. 2 shows the relative expression level when using the test medium containing BCE, when the expression level of KRT19 when using the control medium is set to 1. As shown in FIG. 2, the expression level of KRT19 increased about 15 times when the test medium containing BCE was used compared to when the control medium was used.

Figure 3 shows the results of H-E staining of hair follicles in rat skin tissues (A to C; scale bar is 20 μm) and the results obtained by counting the number of hairs per hair follicle (D). show. When compared with the OVX control group, a significant difference was observed between the OVX BCE group and the Sham group, with the number of hairs increasing by about 0.5 on average. Furthermore, the number of hairs in the OVX BCE group was comparable to that in the Sham group, and no significant difference was observed between the two.

The results of immunostaining using an anti-Ki67 antibody (A to D; scale bar is 100 μm) and the measurement results of the percentage of Ki67-positive cells per total cell number of hair follicles (E) are shown in FIG. As shown in Figure 4, the intensity of color development increased in the OVX BCE and Sham groups compared to the OVX control group, and the Ki67-positive cell rate also increased significantly, confirming an increase in the expression level of Ki67. Ta.

The results of fluorescent immunostaining using anti-KRT19 antibody (scale bar is 100 μm) are shown in FIG. 5. Furthermore, the results of fluorescent immunostaining using antibodies for KRT15 and CD34, which are stem cell markers similar to KRT19, are shown in FIG. 6 (scale bar is 100 μm). An increase in fluorescence intensity was confirmed not only for KRT19 but also for KRT15 and CD34 in the OVX BCE group and Sham group compared to the OVX control group.

The increased expression levels of KRT19, KRT15, CD34, and Ki67 showed that BCE is involved in the maintenance of dermal papilla stem cells and proliferation of dermal papilla cells, and thus has an effect on improving hair thinning in menopausal models. . Furthermore, like BCE, blueberry fruit powder containing β-estradiol and 17α-estradiol was also found to have an effect on improving hair thinning in menopausal models.

[Example 2 Evaluation of the weight gain suppressing effect and fat gain suppressing effect of blackcurrant extract and blueberry fruit powder on a menopausal model]
(2-1) Evaluation of the suppressive effect on weight gain and the suppressive effect on blood neutral fat rise using ovariectomized rats Same as above (1-5) except that 11-week-old ovariectomized rats and Sham rats were used. The rats were divided into groups and fed with different diets for 3 months. The body weights of rats in each group were measured before and after rearing.

After raising the rats, blood was collected and then dissected, and the weights of intraperitoneal adipose tissue and liver were measured.

Using intraperitoneal adipose tissue, tissue sections with a thickness of 3 μm were prepared in the same manner as in (1-5) above, and various evaluations were performed. The tissue sections were stained with HE, and the diameters of five adipocytes on the images of the HE-stained specimens of intraperitoneal adipose tissue of rats in each group were measured. The average of the measured values for each group was calculated, and the OVX control group was tested using the Tukey-Kramer method (p<0.05).

(2-2) Gene analysis by real-time PCR In order to analyze the expression of hepatitis-related genes, real-time PCR was performed in the same manner as in (1-4) above using mRNA extracted from the liver. TNFα and IL-6 were targeted as hepatitis-related genes.

(2-3) Evaluation results Figure 7 shows the changes in body weight before and after rearing for 3 months. There were no individual differences in body weight before feeding. However, after 3 months of food intake, the body weights of the OVX BCE and Sham groups decreased statistically significantly compared to the OVX control group (p<0.05). Furthermore, the body weight of the OVX BCE group was not significantly different from that of the Sham group.

The results of intraperitoneal fat and liver weight after 3 months of rearing are shown in FIGS. 8 and 9, respectively. The OVX BCE group and the Sham group showed a statistically significant decrease in intra-abdominal fat compared to the OVX control group. However, no significant variation in liver weight was observed between these groups. Furthermore, the weight of intraperitoneal fat in the OVX BCE group was slightly larger than that in the Sham group.

FIG. 10 shows a photograph obtained by HE staining intraperitoneal adipose tissue after 3 months of rearing. Furthermore, the measurement results of the diameter of adipocytes in intraperitoneal adipose tissue are shown in FIG. 11. As these figures show, the OVX BCE group and the Sham group had statistically significant reductions in adipocyte diameter compared to the OVX control group. Furthermore, the diameter of adipocytes in the OVX BCE group was slightly larger than that in the Sham group.

The results of real-time PCR analysis of the expression levels of hepatitis-related genes TNFα and IL-6 in liver cells are shown in FIGS. 12 and 13, respectively. The OVX BCE group and the Sham group statistically significantly suppressed the increase in the expression level of hepatitis-related genes compared to the OVX control group. However, the effect of suppressing the increase in the expression level of hepatitis-related genes in the OVX BCE group was slightly smaller than that in the Sham group.

From the above results, it was found that BCE has a weight gain suppressing effect, a fat increase suppressing effect, a fat cell expansion suppressing effect, and a hepatitis suppressing effect on a menopausal model. Furthermore, like BCE, blueberry fruit powder containing β-estradiol and 17α-estradiol was also found to have these effects on menopausal models.

[Example 3 Evaluation of blood vessel hardening effect of blackcurrant extract and blueberry fruit powder on menopausal model]
(3-1) Gene analysis using microarray and IPA Microarray analysis was performed in the same manner as in (1-3) above, except that HUVEC (Promocell), which is a human vascular endothelial cell, was used as the cell. In addition, analysis was performed using IPA software in the same manner as in (1-3) above, except that genes whose expression increased by 1.5 times or more were used.

(3-2) RT-qPCR
Endothelial nitric oxide synthase (eNOS) mRNA expression was assessed by RT-qPCR analysis. EA. which is a human vascular endothelial cell like HUVEC. hy926 cells (ATCC) were used. HUVEC and EA. hy926 cells were seeded in a 12-well culture plate and cultured until confluent. The medium was changed and cells were then treated with or without anthocyanins (10 μM), BCE (0.5 or 1.0 μg/mL) or 17β-estradiol (E2) (10 nM) for 24 hours. In the fulvestrant experimental system, after 24 hours of incubation in the presence or absence of 100 nM fulvestrant, the treated cells were further incubated with anthocyanin and BCE for 24 hours. After washing twice with PBS, total RNA was extracted using "RNeasy mini kit".

cDNA was reverse transcribed from total RNA (200 ng) using "PrimeScript (registered trademark) RT Master Mix" (Takara). The level of specific mRNA was quantified by qPCR using "SYBR (registered trademark) Premix ExTaq (trademark) II" (Takara Bio Inc.). Transcript levels were normalized using glyceraldehyde 3-phosphate dehydrogenase (GAPDH) levels. The PCR conditions and primers used were those described in the literature of the present inventors (Molecules 2019, 24(7), 1259). PCR specificity was confirmed by melting curve analysis. All samples were analyzed in triplicate and relative gene expression was calculated by the 2 ^-ΔΔCt method.

(3-3) Nitric oxide (NO) measurement EA. hy926 cells were seeded in a 12-well culture plate and cultured until confluent. The medium was changed and cells were then incubated for 24 h in the presence or absence of 100 nM fulvestrant followed by anthocyanins (10 μM), BCE (0.5 or 1.0 μg/mL) or E2 (10 nM). After incubation of the cells for an additional 5 days in the absence or presence of phthalate, the medium was collected and concentrated in a SpeedVac concentrator (“SPD1010”; Thermo Fisher Scientific).

The amount of NO released was quantified using the "QuantiChrom (trademark) nitric oxide assay kit" (Funakoshi). Specifically, the culture medium was deproteinized with ZnSO ₄ and NaOH, followed by addition of working reagents from the kit, and then incubated at 60° C. for 10 minutes. The samples were transferred to a 96-well plate, and the optical density (OD) was then measured using a microplate reader (“Benchmark”; Bio-Rad) at a wavelength of 570 nm. The data were converted into concentrations based on a standard curve prepared using standard samples.

(3-4) Immunohistochemical staining of eNOS protein using ovariectomized rats Each rat was divided into groups in the same manner as in (1-5) above, and raised for 3 months using the respective diet, and then the celiac artery tissue was was excised and tissue sections were prepared.

For tissue sections, endogenous peroxidase in the specimen was reacted for 5 minutes at room temperature using a peroxidase removal solution (Dako Cytomation A/S), followed by a serum-free protein block reagent ("Protein Block Serum-Free"; After incubation using Dako Cytomation A/S), an anti-eNOS antibody (pre-diluted; Abcam) was applied as a primary antibody.

After incubating for 90 minutes at room temperature and then washing, the tissue sections were incubated with EnVision™/HRP rabbit/mouse secondary antibody (Dako Cytomation A/S) for 30 minutes at room temperature and then chromogen-treated. Incubated with 3,3'-diaminobenzidine. Nuclei were counterstained using Myers hematoxylin (Wako Pure Chemical Industries, Ltd.). Tissue sections not treated with primary antibody were used as negative controls. The staining intensity of the tissue samples was measured semi-quantitatively (0: none, 1: weak, 2: moderate, 3: strong).

(3-5) Statistical analysis Measurement results were expressed as the mean±standard error (SEM) of at least three independent experiments. Statistical analysis was performed using "BellCurve Excel version 2.13 software" (Social Information Service, Inc.), steel post hoc test; Kruskal-Wallis analysis with p < 0.05 determined statistical significance. It was assumed that

(3-6) Evaluation results Table 3 shows the upstream factors of genes related to the vascular system whose expression changed throughout BCE as determined by microarray and IPA.

The z-score of ERα was 2.7, indicating that ERα is involved in the phytoestrogenic effect of BCE on HUVEC.

Raloxifene showed a z-score of 2.3, and the downstream gene of raloxifene was upregulated by BCE. Raloxifene is a tissue-selective ER modulator (SERM) that is clinically effective in preventing postmenopausal osteoporosis. Raloxifene is known to behave similarly to estrogen and has been shown to induce NO production through ER-dependent enhancement of eNOS enzyme activity in human endothelial cells.

Since the z-score for NO was 2.1, we separately evaluated the expression changes associated with the gene sets regulated by estradiol and NO, and found that BCE could affect the expression levels of genes involved in pathways related to NO synthesis. significantly changed.

RT-qPCR analysis was performed to evaluate the effect of anthocyanins and BCE on the expression of eNOS. The results are shown in FIG.

As shown in FIGS. 14A and 14B, eNOS mRNA levels were significantly increased in BCE- or anthocyanin-treated HUVEC and EA. A significant increase was observed in hy926 cells. eNOS mRNA expression increased in a dose-independent manner at both 0.5 and 1.0 μg/mL BCE. eNOS mRNA expression levels were not significantly different between concentrations of BCE, further indicating that the optimal concentration of BCE differs between cell tumors. Anthocyanins contained in BCE significantly increased the eNOS mRNA expression level, but there was no significant difference between the four types of anthocyanins.

As Figure 14C shows, eNOS mRNA expression levels were decreased in fulvestrant-treated cells, but were not significantly different compared to control levels. Fulvestrant is a synthetic ER antagonist. Therefore, it is speculated that BCE exerts phytoestrogenic activity mediated through the ER and influences eNOS expression.

Next, BCE or anthocyanin is added to EA. The induction of NO synthesis in hy926 cells was evaluated by measuring NO. The results are shown in FIG. As Figure 15A shows, NO levels were significantly higher in cells treated with BCE or anthocyanins than in control cells. However, there were no significant differences between BCE concentrations and between the four anthocyanins.

As Figure 15B shows, NO synthesis levels were decreased in fulvestrant-treated cells, but were not significantly different from control levels. These results are consistent with eNOS mRNA expression, indicating that BCE and anthocyanins may control NO synthesis via eNOS expression.

As shown in Table 4, the results of total transcription microarray analysis of HUVECs contacted with BCE (2.0 μg/mL) showed that the expression of NO-related genes VEGFA, HSP90AB1, PIK3CA, and AKT1 was enhanced. Expression of the VEGFA gene increased 1.7-fold, while expression of other VEGF genes remained virtually unchanged. However, although VEGFC expression increased 1.2 times, VEGFB expression was not significantly different from the control level.

BCE may regulate VEGF expression and thereby promote NO release through a mechanism involving tyrosine and PI3K kinases. The levels of gene expression of PIK3CA (1.4-fold) and AKT1 (1.1-fold) based on microarray were not significantly increased (see Table 4).

Furthermore, the expression of PI3K and phospho-Akt proteins was examined by immunohistochemical staining of vascular endothelial cells from BCE-treated OVX rats. However, neither protein was clearly detected. These results showed that BCE could induce NO synthesis in endothelial cells not only through phytoestrogenic activity but also through other pathways.

To verify the effect of BCE on eNOS protein expression in vivo, we evaluated whether dietary BCE increases eNOS protein expression by subjecting vascular endothelial cells of menopausal model rats to immunohistochemical staining. did. The results are shown in FIG.

As shown in FIG. 16, eNOS protein expression was statistically significantly higher in the OVX BCE group and the Sham group to which 3% BCE by weight was administered than in the OVX Control group. However, the expression level of the OVX BCE group was lower than that of the Sham group. Therefore, we showed that dietary intake of BCE improves eNOS protein levels in blood vessels of OVX rats.

From the above results, it was found that BCE strongly increases eNOS mRNA expression and NO production in human endothelial cells through phytoestrogen activity. Furthermore, dietary BCE increased eNOS protein expression in a menopausal model. These results indicate that BCE may have beneficial health effects on vascular symptoms, such as vascular stiffness, observed in postmenopausal female individuals. Furthermore, like BCE, blueberry fruit powder containing β-estradiol and 17α-estradiol was also suggested to have these effects on menopausal models.

The composition of one aspect of the present invention can be used for female individuals suffering from menopausal symptoms such as hair thinning, vascular stiffness, weight gain, and fat gain.

Claims (2)

Contains an active ingredient that is a blackcurrant extract, and has the effect of improving menopausal symptoms for postmenopausal females other than skin deterioration due to decreases in collagen, elastin, and hyaluronic acid. An improvement composition comprising:
The composition, wherein the menopausal symptom is hair thinning . 2. The composition of claim 1 , wherein the composition is an oral composition.