Use of sakura extract in health promotion of ovary

Abstract

To provide a novel use of Sakura extract for the purpose of promoting health of ovary because it was found through experiments that the extract component has the action of promoting proliferation of human KGN cells, and it can be considered that the component has the action of protecting human ovary and the action of promoting health of the ovary.SOLUTION: Sakura extract is used as an active ingredient in the preparation of a composition that promotes proliferation of human KGN cells.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to the field of plant extracts, and primarily to a novel use of a cherry blossom extract, and more particularly to a novel use of a cherry blossom aqueous extract for promoting human ovarian health. [Background technology]

[0002] Some research has been done on the components of cherry blossoms, particularly their extracts.

[0003] Cherry blossom extract is known to have antioxidant, anti-inflammatory, and skin-whitening properties (see Reference 1). This is primarily due to the presence of two flavonoids, sakuranin and sakuranetin, which have strong antioxidant properties. [ka]

[0004] Currently, cherry blossom components or their extracts are used mainly in the fields of cosmetics, food, and health foods for the purpose of protecting human skin and promoting health.

[0005] For example, cherry blossom extract is used as an active ingredient in skin care products and cosmetics, and can effectively block the invasion of cells by sunlight, external factors, stress, etc., inhibit melanin secretion by melanocytes, and suppress melanin production and deposition. It can also improve dullness of the skin and achieve the ideal skin care effect of fair and pink skin like cherry blossoms.

[0006] Furthermore, cherry blossom components, especially their extracts, can be used as food ingredients if permitted by relevant laws and regulations. For example, the extract extracted from cherry blossom petals that have been harvested to 70% bloom has strong antioxidant and anti-glycation effects, reduces the degree of glycation in fibrocytes, and promotes collagen production, making it suitable for use as a food additive.

[0007] In addition, in another embodiment, for example, cited reference 2 also reports an AGE production inhibitor containing an extract of cherry blossoms (preferably flowers or leaves) and / or a processed product thereof as an active ingredient, which effectively inhibits the production of AGEs, which are advanced glycation end products, and has improved safety for the living body.

[0008] For a similar document, see Reference 3, which discloses a method for producing cherry blossom powder with AGE production inhibitory activity, including low-temperature processes such as washing, countercurrent extraction, filtration, concentration, sterilization, freeze-drying, and sieving.

[0009] Furthermore, Reference 3 discloses the use of naringenin and its pharmaceutically acceptable salts in the manufacture of a medicament for the treatment or prevention of toxoplasmosis. The effective concentration of naringenin is 50 to 120 μg / mL. It also discloses the use of a pharmaceutical composition containing naringenin and its pharmaceutically acceptable salts in the preparation of a medicament for the treatment or prevention of toxoplasmosis. Naringenin is the aglycone of naringin, a flavanone compound, and is naturally derived primarily from the buds of cherry blossoms (Prunus yedoensis Mate), a plant of the Rosaceae family, and the pits of the fruit of Amacardi-umoccidentale L, a plant of the Anacardiaceae family.

[0010] Thus, while some research has been conducted in the industry on the functions of some active ingredients derived from cherry blossoms, there is still room for further research and development on the effects of ingredients such as cherry blossom extracts. [Prior art documents] [Patent documents]

[0011] [Patent Document 1] (Citation 1) "Protective skin aging effects of cherry blossom extract (Prunus Yedoensis) on oxidative stress and apoptosis in UVB-irradiated HaCaT cells", Yaning Wang et al., Cytotechnology (IF 2.2) Pub Date: 2019-03-16, DOI: 10.1007 / s10616-018-0215-7 [Patent Document 2] (Cited document 2) CN102883733B [Patent Document 3] (Cited document 3) CN114794473A [Patent Document 4] (Cited document 4) CN112843046A Summary of the Invention [Problem to be solved by the invention]

[0012] As mentioned above, the industry is increasingly aware of the beneficial effects of cherry blossom compounds, but the development of new uses for them, especially those aimed at human health benefits, is still worthy of further exploration.

[0013] Therefore, the present invention provides a novel use of cherry blossom components, particularly cherry blossom extracts. Furthermore, the components are thought to have the effect of promoting the proliferation of human KGN cells, and to have the effect of protecting the human ovaries and promoting ovarian health.

[0014] A further object of the present invention is to provide a novel use of cherry components, particularly cherry extracts, in the preparation of a (non-therapeutic) composition for promoting the proliferation of human KGN cells.The present invention also provides a novel use of cherry components, particularly cherry extracts, in the preparation of a (non-therapeutic) composition for promoting ovarian health and protecting the ovaries.

[0015] The present invention also provides foods containing cherry blossom extract as an active ingredient and health foods having health benefits.

[0016] As mentioned above, the inhibitory effect of cherry blossom extract on AGEs (advanced glycation end products) has been studied, for example, in the above-mentioned references 2 and 3, and it has been reported that this improves physical health. However, the mechanism of the present invention is different from this, and actually achieves the effect of promoting the health of human ovaries through cell proliferation and improving the health of the skin. [Means for solving the problem]

[0017] The above problems can be solved by implementing the following aspects. The present invention provides the use of a cherry extract as an active ingredient in the preparation of a composition for promoting the proliferation of human KGN cells. The present invention further provides the use of a cherry blossom extract as an active ingredient in the preparation of a composition for promoting human ovarian health. The present invention also provides the use of a cherry blossom extract as an active ingredient in the preparation of a composition for improving human skin conditions by promoting the proliferation of human KGN cells. The present invention also provides the non-therapeutic use of a composition comprising a cherry blossom extract as an active ingredient for promoting human ovarian health. The present invention also provides non-therapeutic uses of a composition containing a cherry blossom extract as an active ingredient, which improves skin conditions by promoting the proliferation of human KGN cells. The above use, wherein the cherry blossom extract is an extract of cherry blossom petals. The above-mentioned use, wherein the cherry blossom extract is a water extract of cherry blossom petals. The above-mentioned use, wherein the water extract is a water extract of cherry blossom petals at a temperature range of 25 to 45°C. The above use, wherein the composition is an edible composition. The use, wherein the composition comprises one or more of a beverage, a powder, a solid beverage, a capsule, a tablet, an oral liquid, a gummy, a jelly, or a gel candy. [Effects of the Invention]

[0018] By implementing the above embodiment, the following effects can be obtained. 1) The cherry blossom extract of the present invention, especially the water extract, is easy to extract and does not require special conditions, so it can meet the needs of large-scale industrial production and maximize the activity of the extract. 2) By providing an edible composition containing cherry blossom extract, particularly a water extract, as an essential active ingredient, the active ingredient can promote KGN cell proliferation, thereby promoting the health of human ovaries and improving the health of human skin. [Brief explanation of the drawings]

[0019] [Figure 1] Measurement results of the administration experiment of two groups (where ** indicates P<0.01, *** indicates P<0.001, the data on the far left is the data of the blank control group, and the data on the far right is the data of the positive control group). [Figure 2] Measurement results of the administration experiment of two groups (where ** indicates P<0.01, *** indicates P<0.001, the data on the far left is the data of the blank control group, and the data on the far right is the data of the positive control group). DETAILED DESCRIPTION OF THE INVENTION

[0020] Various embodiments, configurations, and aspects of the present invention are described in detail below. As used herein, the term "exemplary" means "serving as an example, instance, or illustration." Any example described herein as "exemplary" should not be construed as more suitable or superior than other examples.

[0021] Furthermore, in the following specific embodiments, numerous specific details are provided to better explain the present invention. It should be understood by those skilled in the art that the present invention can be practiced without certain specific details. In other embodiments, methods, means, devices, and steps well known to those skilled in the art are not described in detail in order to emphasize the essence of the present invention.

[0022] Unless otherwise specified, all units used herein are international standard units, and it should be understood that the numerical values ​​and numerical ranges appearing in the present invention include systematic errors that are unavoidable in industrial production.

[0023] In this specification, the term "may" includes both cases where some processing is performed and cases where some processing is not performed.

[0024] In this specification, a numerical range expressed as "numerical value A to numerical value B" means a range including the limit values ​​A and B.

[0025] In the specification, unless otherwise specified, "%" means percent by weight or mass.

[0026] In this specification, unless otherwise specified, the content is expressed as mass % based on the dry weight of the solid content.

[0027] Unless otherwise specified, "room temperature" as used herein generally means a temperature of 23±2°C.

[0028] As used herein, references to "some specific / preferred embodiments," "other specific / preferred embodiments," "embodiments," etc., mean that the particular elements (e.g., features, structures, properties, and / or characteristics) described in connection with an embodiment are included in at least one embodiment described herein and may or may not be present in other embodiments. Furthermore, it is to be understood that such elements may be combined in any suitable manner in the various embodiments.

[0029] The present invention mainly provides a novel non-therapeutic use of a cherry blossom extract, particularly a cherry blossom water extract, which is based on the following findings:

[0030] Comparative experiments have shown that cherry blossom extract, especially the aqueous extract, has the effect of promoting the proliferation of human KGN cells (human ovarian granulosa cells), and furthermore, through this mechanism, cherry blossom extract, especially the aqueous extract, has the effect of promoting the health of human ovaries, promoting the stability of ovarian physiological functions, and protecting the ovaries, and furthermore, through this mechanism, it has been found that cherry blossom extract can also improve the health of human skin.

[0031] Sakura In the present invention, there is no particular limitation in principle on the type or origin of the cherry tree, but for example, cherry trees grown all over the world can be used.

[0032] In some preferred embodiments, the cherry tree may be Kanzan (Cerasus Sato-zakura Group 'Sekiyama' Koidz.).

[0033] The cherry blossom extract of the present invention mainly refers to an extract of cherry blossom petals. Such petals may be freshly picked or may be preserved petals that have been frozen or dried after picking. Preferably, the cherry blossom extract of the present invention is an extract of freshly picked cherry blossom petals.

[0034] In principle, there are no particular restrictions on the time to pick cherry blossom petals, but from the perspective of excellent extraction effects, they can be harvested when the cherry blossoms are 60% to 80% in bloom.

[0035] Cherry Blossom Extract The cherry blossom extract of the present invention is not particularly limited in principle, but may be, for example, a solvent (aqueous solvent) extract or other types of extract such as a supercritical fluid extract. In some specific embodiments of the present invention, from the viewpoint of efficient extraction of components and maintaining activity, the cherry blossom extract of the present invention is preferably a water extract of cherry blossoms (petals).

[0036] Furthermore, in some preferred embodiments, the extraction process for the cherry blossom extract of the present invention comprises: The process may include a water extraction step, a concentration step, and, if necessary, a petal pretreatment step, a water extract filtration step, a concentrate drying step, a sterilization step, etc.

[0037] The petal pretreatment process of the present invention is not particularly limited in principle, but in some specific embodiments, it may include one or more of the steps of sorting, washing, crushing, drying or wetting the petals, etc., and this pretreatment process can result in a pretreated product or a cherry blossom raw material that can be extracted with water.

[0038] In the water extraction step of the present invention, the raw material is brought into contact with water through the above-mentioned pretreatment, thereby extracting the water-soluble components into the aqueous phase. The water used in the water extraction step may usually be deionized water or distilled water.

[0039] In some specific embodiments, the ratio of the amount of water to the pretreated material (cherry raw material) in the water extraction step is not particularly limited in principle. However, from the viewpoint of the effectiveness and efficiency of the water extraction, the mass of water is preferably at least 5 times the mass of the pretreated material, more preferably 6 to 20 times, for example, 8 times, 10 times, or 15 times. Furthermore, from the viewpoint of the effectiveness and efficiency of the water extraction, the temperature during the water extraction may be 25 to 45°C, preferably 28 to 42°C. Furthermore, other auxiliary devices or means used during the water extraction are not particularly limited, and the water extraction can be assisted by means such as stirring or ultrasonic waves. Furthermore, in some specific embodiments, the time for the water extraction can be 0.5 to 5 hours, preferably 0.5 to 2 hours. The water extraction step allows for the production of an extract-containing liquid system.

[0040] In some other specific embodiments, in principle, only water may be used as the extraction solvent in the aqueous extraction step of the present invention, but if necessary, a certain amount of polarity adjuster may be added to the water to assist the extraction step. Such polarity adjuster may typically be an aqueous alcoholic solvent such as ethanol, and the amount of such adjuster used may generally be controlled to 20% by mass or less, preferably 15% by mass or less, more preferably 10% by mass or less, based on the total mass of the extraction solvent.

[0041] Furthermore, the filtration step of the aqueous extract system of the present invention mainly involves solid-liquid separation of the extract system described above. The filtration method is not particularly limited, but in some specific embodiments, treatment can be performed using one or more devices such as a mesh filter or a filtration membrane. In some preferred embodiments, solid particles in the system with particle diameters of 1000 nm or more, 800 nm or more, 600 nm or more, 400 nm or more, or 200 nm or more can be removed by filtration.

[0042] In the concentration step of the present invention, the system after filtration can be concentrated by evaporation, membrane concentration, etc. Preferably, the concentration treatment of the present invention can be carried out using a reverse osmosis membrane.

[0043] The drying step of the present invention is not particularly limited in principle and can be carried out by spray drying, heat drying, freeze drying, etc., and is preferably carried out by freeze drying. In some specific embodiments of the present invention, a solid extract powder is obtained after drying.

[0044] The method for preparing the water extract of the present invention will be described in more detail below. Dried Kansan flowers or fresh dried Kansan flowers are crushed by crushing or grinding, or moistened by adding water, and the cherry blossom flowers are made into a slurry by crushing or grinding, the mass ratio of water to the cherry blossom raw material is 5:1 to 20:1, the extraction temperature is controlled at 25 to 45°C, and an extract is obtained after 0.5 hours.

[0045] The extract is filtered through an 80-150 mesh filter to remove coarse particles, and the resulting crude filtrate is then filtered through a microfiltration membrane such as a 200-800 nm ceramic membrane. The resulting retentate is concentrated through a reverse osmosis (RO) membrane to obtain a solid concentrate. The concentrate is then freeze-dried to obtain a powdered product.

[0046] Active ingredient The active ingredients in the cherry blossom extract of the present invention may generally include water-soluble substances of cherry blossoms that are easily soluble in water, and may also include other ingredients dispersed in water by the above-mentioned extraction method.

[0047] In some specific embodiments, the cherry extract of the present invention comprises one or a mixture of two or more of the following ingredients or derivatives thereof as active ingredients:

[0048] The physiologically active ingredients are 1-O-(E)-Caffeoyl-β-D-glucopyranoside, 1-O-(E)-Coumaroyl-β-D-glucopyranoside, 1-O-(E)-Cinnamoyl-β-D-glucopyranoside, Kaempferol 3-O-β-D-glucopyranoside, and Quercetin 3-O-β-D-glucopyranoside. At least one of the following is used: kaempferol 3-O-(6''-malony)-β-D-glucopyranoside, Kaempferol 3-O-(6''-malony)-β-D-glucopyranoside, and Quercetin 3-O-(6''-malony)-β-D-glucopyranoside.

[0049] In addition to the above-mentioned active ingredients, the extract may also contain other monosaccharides, polysaccharides, water-soluble cellulose components, vitamin components, etc. that are extracted at the same time.

[0050] composition The composition of the present invention uses the cherry blossom extract as an essential active ingredient, and in some specific embodiments, the composition is an edible composition.

[0051] The composition may be in a liquid, solid or semi-solid form, preferably in the form of a liquid or semi-solid form such as various drinks, pastes, etc. Most preferably, the composition of the present invention is in the form of an oral drink.

[0052] Furthermore, in some preferred embodiments of the present invention, the beverage of the present invention may be a homogeneous aqueous solution system or a microemulsion system (such an emulsion system can have good system stability even when left at room temperature).

[0053] In addition to the cherry blossom extract contained as an active ingredient, other nutritional ingredients may be added to the composition of the present invention as needed, as long as they do not affect the effects of the present invention. Such ingredients include proteins, fats, necessary carbohydrates, dietary fiber, trace elements, vitamins, other plants or plant extracts, etc.

[0054] Here, the protein is at least one selected from whey protein powder, soy protein isolate, whole milk powder, whole egg powder, lactoferrin, bovine colostrum, amino acids, and protein peptides, and the amino acids are at least one selected from L-lysine-L-glutamic acid, L-glutamic acid, L-arginine, L-tryptophan, L-glutamine, taurine, L-valine, L-isoleucine, and L-leucine, and the protein peptides are at least one selected from soybean oligopeptides, wheat protein peptides, silkworm pupa protein peptides, marine fish oligopeptide powder, cola peptides, amino peptides, and ovalbumin peptides.

[0055] The fat may include at least one of saturated fatty acids, polyunsaturated fatty acids, monounsaturated fatty acids, DHA, EPA, ARA, and phospholipids.

[0056] The carbohydrates include starch or modified starch.

[0057] The dietary fiber includes one or more of inulin, konjac flour, galactooligosaccharides, fructooligosaccharides, isomaltooligosaccharides, soybean polysaccharides, cyclodextrin, indigestible dextrin, and soybean dietary fiber (soy fiber).

[0058] The trace elements are one or more selected from organic acid-based metal ion salts such as calcium citrate, calcium L-lactate, calcium hydrogen phosphate, potassium gluconate, sodium citrate, ferrous gluconate, potassium iodide, zinc gluconate, sodium selenite, copper gluconate, chromium sulfate, manganese gluconate, and magnesium gluconate.

[0059] The vitamin is one or more selected from vitamin A, β-carotene, vitamin D3, vitamin E, vitamin K1, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, pantothenic acid, folic acid, niacin, choline, inositol, and biotin.

[0060] Furthermore, the other plants or plant extracts mentioned above are not particularly limited in principle, and may be various components or extracts derived from fruits, vegetables, etc.

[0061] In some specific embodiments of the present invention, the fruits are not particularly limited in principle, and examples thereof include apple, wild apple, Japanese maple, aronia berry (Aronia melanocarpa), loquat, medlar, hawthorn, pear (such as Shanli (fragrant pear) and snow pear), quince, rose hip, apricot, cherry, peach (such as water peach, snow pear, and peaches), plum, plum (green plum), European plum, Japanese toad, blackberry, raspberry, chamomile, loganberry, wineberry, strawberry, pineberry, citrus fruits, mandarin (Shatangju), orange, lemon, lime, pomelo, kumquat, grapefruit, citron, bush quince, finger lime, wampi, watermelon, Hami melon, melon, honeydew melon, bitter melon, banana, plantain, red banana, date, grape, blueberry, cranberry, Berries, lingonberries, huckleberries, mangoes, kiwifruit (Actinidia chinensis), gold kiwi, arguta, red kiwi, pineapple, bayberry, persimmon, Japanese bean oak, Japanese black persimmon, papaya, mulberry, fig, jackfruit, paper mulberry, star apple, dragon fruit, yellow pitaya, red dragon fruit, prickly pear, lychee, longan, rambutan, durian, baobab fruit (fruit of the baobab tree), Oxalidaceae, star fruit, bilimbi, pomegranate, coconut, arecanut, date palm, salak, acai berry, mangosteen, myrtle, jaboticaba, guava, feijoa, pitanga, passion fruit Examples include peanuts, avocado, custard apple, gyushinri (beef heart pear), wolfberry, pepino, lantern fruit, cherry tomatoes, sea buckthorn, and autumn jasmine.

[0062] The vegetables are not particularly limited in principle, but include melons, green vegetables, nightshades, Chinese cabbages, tubers, root vegetables, kale, legumes, perennial vegetables, aquatic vegetables, and the like. Further specific examples include radish, turnip, Chinese cabbage (including Chinese cabbage and Chinese cabbage subspecies), kale (including cabbage, kohlrabi, cauliflower, broccoli, etc.), mustard (including Daitosai and mizuna varieties), celery, carrot, tomato, cucumber, zucchini, pumpkin, squash, wax gourd, loofah, bottle gourd, bitter melon, chayote, kidney beans (including dwarf kidney beans and climbing kidney beans), cowpea, pea, broad bean, edamame (i.e., soybean), hyacinth beans, sword beans, peanuts, mung beans, asparagus, lily, spinach, radish, carrot, burdock, jicama, sweet potato, kudzu, stem lettuce, wild rice, stem mustard, kohlrabi, and komatsuna.

[0063] The amount of the cherry blossom extract active ingredient in the composition is determined in principle by the range of content stipulated by the laws and regulations of each country. Preferably, the amount of the cherry blossom extract active ingredient added in the composition is an amount sufficient to provide 10 mg to 500 mg, for example, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 500 mg, etc. of the cherry blossom extract per day during the administration period (using a predetermined method as necessary).

[0064] Effects and uses The composition of the present invention, containing the cherry blossom extract as an essential active ingredient, can promote the proliferation of human KGN cells, and through this mechanism, can achieve the goal of promoting the health of human ovaries, for example, by maintaining good physiological function of the human ovaries and protecting the physiological level of the ovaries. Furthermore, through this mechanism, it can contribute to improving human skin conditions, such as preventing sagging skin, preventing the occurrence of blemishes, preventing skin dullness, and alleviating unpleasant symptoms such as enlarged pores and dryness.

[0065] Based on the above mechanism, the present invention can further have the following applications: Use of the cherry extract of the present invention or a composition comprising the same in the preparation of a composition or product that promotes the proliferation of human KGN cells. 1. Use of the cherry blossom extract of the present invention as an active ingredient in the preparation of a composition that promotes human ovarian health by promoting cell proliferation. Use of the cherry blossom extract of the present invention as an active ingredient in the preparation of a composition for improving human skin conditions by promoting the proliferation of human KGN cells. 1. A non-therapeutic use of the cherry blossom extract of the present invention or a composition comprising the same for promoting human ovarian health, wherein the composition comprises a cherry blossom extract as an active ingredient and is capable of promoting cell proliferation. 2. A non-therapeutic use of the cherry blossom extract of the present invention or a composition containing the same to improve human skin conditions by promoting the proliferation of human KGN cells. Example

[0066] Hereinafter, embodiments of the present invention will be described in detail with reference to examples. However, those skilled in the art will understand that the following examples are merely for the purpose of illustrating the present invention and are not intended to limit the present invention. In the examples, unless specific conditions are specified, the experiments were carried out under standard conditions or conditions recommended by the manufacturer. Reagents and equipment used without a manufacturer's name are all commercially available standard products.

[0067] Experimental conditions: (1) Raw materials: Kanzan flower petal water extract. (2) Preparation of mother liquor: The above cherry blossom water extract was dissolved in water to prepare a 10 mg / mL stock solution. Estradiol powder was weighed out and prepared into a 10 mM stock solution in DMSO (positive control). Phenol red-free medium without drug (blank control group). (3) Screening for ovarian protection groups: (Cell culture conditions) Medium: DMEM / 12 medium containing 10% FBS and 1% PS Culture conditions: 37℃, 5%CO2 (inoculation) KGN cells in the logarithmic growth phase were selected, trypsinized, and then inoculated into 96-well plates at a concentration of 5000 cells / well, 90 μL / well. (Administration) After 24 hours of culture, during which the cells adhered to the wall, three replicate wells were set up for each group. Treatment group: 10 μL of water extract solution (cherry blossom water extract and estradiol positive control) Blank control group (Con): 10 μL phenol red-free medium (Cell viability measurement) After culturing for 24 hours, 10 μL of CCK-8 reagent was added per well, and the plate was cultured for 4 hours, and the absorbance at 450 nm was measured.

[0068] Example 1: The treatment group contained a water extract of kanzan flower petals as the active ingredient, and was compared with a positive control group and a blank control group. Two batches of cells were cultured under the same conditions, and a parallel comparison test was conducted. In other words, two groups of cultured cells were used to conduct two measurements, and the control conditions for the measurements in each group were the same.

[0069] The specific measurement conditions are as follows: 1 material 1.1 Drugs Kanzan flower petal water extract, estradiol valerate 1.2 cells Human ovarian granulosa cell line (KGN cells) purchased from Wuhan Pricella Biotechnology Co., Ltd. 1.3 Reagents Fetal bovine serum, penicillin / streptomycin, DMEM / F12 medium, 0.25% trypsin, CCK-8, DMSO. 2 methods 2.1 Drug Preparation 10 mg of Kanzan petal water extract powder was weighed out and dissolved in 1 mL of autoclaved purified water and filtered through a microporous filter membrane. Estradiol valerate tablets were crushed into powder, and 3.5651 mg of the powder was weighed out and dissolved in 1 mL of DMSO. 2.2 Cell culture KGN cells were cultured in DMEM / F12 medium containing 10% fetal bovine serum and 1% penicillin / streptomycin in a 37°C incubator containing 5% CO2 and saturated humidity. The medium was changed every other day, and cell growth was monitored under an inverted microscope. Once the cells had proliferated and formed a dense monolayer (70-80% coverage), they were routinely digested with 0.25% trypsin and centrifuged at 1200 rpm for 3 minutes. They were passaged once every 2-3 days at a ratio of 1:3 or 1:4, and two groups of cells were prepared under the same conditions.

[0070] 2.3 Measurement of cell viability by CCK-8 In this experiment, KGN cells were treated with different concentrations of Kansan petal water extract (0.1, 1, 10, 100 μg / mL) and 10-6 M estradiol for 24 hours, and the cell proliferation effect was observed. The specific steps are as follows: KGN cells in the logarithmic growth phase were harvested, and a single cell suspension was prepared in phenol red-free DMEM / F12 medium containing 10% fetal bovine serum. 90 μL of the suspension was inoculated into a 96-well plate at a concentration of 5,000 cells / well. The experiment was divided into a blank group, a blank control group, and different drug groups. The blank group was not inoculated with cells, but instead received the same volume of culture medium as the drug group. The blank control group was inoculated with cells and received the same volume of culture medium as the drug group. After the cells attached to the wall, 10 μL of culture medium was added to the blank and blank control groups. Each drug group received 10 μL of 10x concentrated water extract of kansan petals or estradiol. Three replicate wells were used for each group. The final volume of each well was the same, 100 μL. After 24 hours of drug intervention, 10 μL of CCK-8 reagent was added to each well in the dark and incubated in a 5% CO incubator at 37°C for 4 hours. The absorbance (OD value) of each well at a wavelength of 450 nm was measured using a microplate reader, and cell viability was calculated using the formula: (OD value of treated wells - OD value of blank wells) / (OD value of blank control wells - OD value of blank wells). Each experiment was independently repeated three times.

[0071] 2.4 Statistical analysis All data were statistically analyzed using the statistical software GraphPad Prism 9.5.0. Experimental results were expressed as mean ± standard deviation (x ± s) and analyzed by one-way analysis of variance (ANOVA). P < 0.05 indicated statistical significance.

[0072] Please refer to Figures 1 and 2 for the measurement results of the two groups (the rightmost group is the positive control group).

[0073] When Kansan powder was administered at 0.1 to 100 μg / mL for 24 hours, it was found to be able to significantly promote the proliferation of KGN cells, with the greatest promoting effect being observed at 100 μg / mL.

[0074] Although the present invention has been described with reference to specific embodiments, those skilled in the art will understand that the present invention is not limited to these. Although various embodiments of the present invention have been described above, the above descriptions are illustrative and not exhaustive, and are not limited to the disclosed embodiments. It will be apparent to those skilled in the art that various modifications and variations are possible without departing from the scope and spirit of the described embodiments. The terms used in this specification have been selected to best explain the principles, actual use, or technical improvements in the market of each embodiment, or to enable other ordinary skilled in the art to understand each embodiment disclosed herein.

Claims

1. Use of a cherry blossom extract as an active ingredient in the preparation of a composition that promotes the proliferation of human KGN cells.

2. Use of cherry blossom extract as an active ingredient in the preparation of a composition for promoting human ovarian health.

3. Use of cherry blossom extract as an active ingredient in the preparation of a composition for improving human skin conditions by promoting the proliferation of human KGN cells.

4. 1. A non-therapeutic use of a composition containing cherry blossom extract as an active ingredient for promoting human ovarian health.

5. 1. A non-therapeutic use of a composition for improving skin conditions by promoting the proliferation of human KGN cells, said composition comprising cherry blossom extract as an active ingredient.

6. The use according to any one of claims 1 to 5, characterized in that the cherry blossom extract is an extract of cherry blossom petals.

7. The use according to any one of claims 1 to 5, characterized in that the cherry blossom extract is a water extract of cherry blossom petals.

8. The use according to claim 7, characterized in that the water extract is a water extract of cherry blossom petals at a temperature ranging from 25 to 45°C.

9. The use according to any one of claims 1 to 5, characterized in that the composition is an edible composition.

10. The use according to any one of claims 1 to 5, characterized in that the composition comprises one or more of a beverage, a powder, a solid beverage, a capsule, a tablet, an oral liquid, a gummy, a jelly or a gel candy.

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