Senomorphic composition comprising extract of maackia amurensis as active ingredient

Abstract

The present invention relates to a senomorphic composition comprising an extract of Maackia amurensis as an active ingredient. By using the composition, the condition of senescent cells can be improved to a more youthful state.

Description

Cenomorphic composition containing azuki bean extract as an active ingredient

[0001] The present invention relates to a cenomorphic composition comprising an extract of azuki bean blossom tree as an active ingredient.

[0002] Senescent cells accumulate in the tissues and organs of an individual as they age. This accumulation of senescent cells not only induces functional and structural changes in tissues and organs due to aging, but also plays a significant role in the pathogenesis of various age-related diseases, such as cancer, diabetes, obesity, tissue fibrosis, age-related ophthalmopathy, cardiovascular disease, degenerative brain disease, osteoarthritis, skin aging, and chronic skin wounds. Therefore, delaying or overcoming aging has been suggested as the most effective method for preventing and treating age-related diseases such as cancer, diabetes, and cardiovascular disease.

[0003] Rapamycin, SIRT1 activators, calorie restriction mimics, AMPK activators, and telomerase activators are being suggested as potential aging-control drugs. In addition, senotherapeutics, which target senescent cells, have recently been developed, and their efficacy has been reported at the cellular level and in animal models. Senotherapeutics are divided into senolytics, which selectively kill only senescent cells, and senomorphics, which restore the function or morphology of senescent cells to that of young cells. In aged individuals where tissue regeneration through stem cells is not active, the application of drugs that eliminate senescent cells is of concern, so drugs that suppress the phenotype of senescent cells are relatively more urgently needed.

[0004] For example, Korean Patent No. 10-2161041 discloses a reagent composition for cenomorphic drugs containing salinomycin as an active ingredient, characterized in that it restores the function or morphology of senescent cells to normal cells.

[0005] However, no drug has been reported yet to restore the overall characteristics of senescent cells, and the development of drugs for cenomorphic treatment that improve the characteristics of senescent cells to those of young cells is still necessary.

[0006] In the above circumstances, the inventors of the present invention conducted research to develop a drug for senomorphs and confirmed that the flower bud extract of the azuki bean tree reduces the level of senescence markers in cells that have induced senescence.

[0007] Accordingly, the purpose of the present invention is to provide a composition for senomorphs comprising an extract of azuki bean and / or a fraction thereof, and a use thereof for treating diseases related to cellular aging.

[0008] In order to achieve the above purpose, one aspect of the present invention provides a cenomorphic composition comprising an azuki bean extract or a fraction thereof as an active ingredient.

[0009] The aforementioned Daphne Genkwa is a deciduous shrub native to Korea, China, and Japan, and is planted for ornamental purposes. Its flowers bloom from March to May, with 3 to 7 flowers borne in cymes at the tips of the previous year's branches. They are light purple in color.

[0010] The term "extract" used in the present invention refers to the resultant product, such as a liquid component obtained by immersing a material including a target plant in various solvents, then extracting and fractionating it for a certain period of time at room temperature or at a temperature, and a solid component obtained by removing the solvent from the liquid component. In addition, it can be comprehensively interpreted to include all fractions, dilutions, concentrates, adjustments, and purified products of the resultant product in addition to the resultant product.

[0011] In the present invention, the extraction solvent may be selected from the group consisting of water, alcohols having 1 to 4 carbon atoms, and mixed solvents thereof, and preferably may be ethanol or 70% ethanol.

[0012] According to one example of the present invention, the azuki bean extract may be extracted from one or more parts selected from the group consisting of flower buds, leaves, fruit stems, and roots of the azuki bean.

[0013] In one specific example of the present invention, the flower buds of the azuki bean tree were dried and ground into a fine powder, and then 70% methanol was added to the powder to prepare an extract (Example 1).

[0014] The above azuki bean flower bud extract is not particularly limited, as long as it can obtain an extract having a cenomorphic effect, and can be prepared using general extraction, separation, and purification methods known in the art. The extraction method is not limited thereto, but may include, but is not limited to, hot water extraction, hot water extraction, cold immersion extraction, reflux cooling extraction, or ultrasonic extraction.

[0015] The above ultrasonic extraction is a method of extracting useful ingredients from various natural materials at room temperature using physical force rather than chemical force, using a vibrating element with a vibration frequency of 20,000 times per second or more. Unlike solvent extraction, vegetable oil extraction, or steam distillation extraction, it has the advantage of being able to extract high concentrations of effective ingredients contained in natural materials at room temperature without destroying them.

[0016] As another specific example, the hot water extraction is performed by adding purified water to dried azuki bean flower bud powder and reacting at a high temperature, preferably 100 to 150°C, more preferably 100 to 130°C, even more preferably 110 to 130°C, even more preferably 121°C, for 20 minutes to 1 hour, preferably 20 minutes to 50 minutes, more preferably 20 minutes to 40 minutes, even more preferably 30 minutes.

[0017] The obtained azuki bean flower bud extract may be concentrated under reduced pressure using, but is not limited to, a vacuum decompression concentrator or a vacuum rotary evaporator. In addition, after concentration under reduced pressure, it may be dried using, but is not limited to, freeze-drying, decompression drying, vacuum drying, boiling drying, or spray drying.

[0018] The term "senomorphics" used in the present invention is often used to refer to the senescence effect, and refers to the effect of restoring the function or morphology of senescent cells to that of young cells. Senescent cells secrete SASP (senescence-associated secretory phenotype), and the SASP can induce the activation of SASP genes such as cytokines, chemokines, growth factors, and proteases. Therefore, senescent cells can affect other normal cells in the vicinity.

[0019] According to one specific example of the present invention, the senomorphic composition can improve aging of cells selected from the group consisting of microglia, oligoglia, and astroglia, but is not limited thereto.

[0020] According to one specific example of the present invention, the cenomorphic effect may be a decrease in the level or activity of a marker selected from the group consisting of SA-β-galactosidase, γH2AX, and P16.

[0021] The above β-galactosidase (β-gal) is a lysosomal exoglycosidase that removes galactose residues from various substrates such as gangliosides, glycoproteins, sphingolipids, and keratin sulfate. Because β-galactosidase in the human body is abnormally accumulated in aged tissues and cells, senescent cells have high β-galactosidase activity. Therefore, the activity of β-galactosidase has been used as an important cellular senescence marker, β-gal (senescent-associated β-galactosidase, SA-β-galactosidase, SA-β-gal), for detecting senescent tissues and cells.

[0022] The above γH2AX refers to phosphorylated H2AX (H2A histone family member X). Phosphorylation of H2AX (γH2AX) is one of the key reactions that occurs initially when double-stranded breaks (DSBs) of DNA occur in cells due to causes such as ultraviolet rays and aging. This phosphorylation of H2AX (γH2AX) continues to be maintained until the DNA double-strand breaks are repaired. Therefore, a high level of γH2AX in a cell indicates that DNA damage caused by ultraviolet rays or aging has occurred in the cell, and conversely, a decrease in the level indicates recovery of DNA damage, reversal of aging, etc.

[0023] The above P16 is a cell cycle-regulating protein that primarily inhibits CDK4 / 6 activity, thereby blocking the activity of the E2F transcription factor and regulating the cell cycle. Senescent cells have high levels of P16 because their cell cycle is arrested.

[0024]

[0025] Another aspect of the present invention provides a pharmaceutical composition for preventing or treating cell aging or aging-related diseases, comprising an extract of azuki bean or a fraction thereof as an active ingredient.

[0026] In the present invention, “prevention” means any act of inhibiting the onset of cellular aging or aging-related disease or delaying its progression by administering a pharmaceutical composition according to the present invention.

[0027] In the present invention, “treatment” means any act of improving the symptoms of cellular aging or aging-related diseases by administering a pharmaceutical composition according to the present invention.

[0028] The above cellular aging or aging-related diseases may be selected from the group consisting of, but are not limited to, overall body aging due to aging, depression, anxiety disorder, post-stroke sequelae, epilepsy, migraine, schizophrenia, chronic fatigue syndrome, obesity, and metabolic syndrome.

[0029] Aging of glial cells, particularly astrocytes, can impair their ability to regulate neurotransmitters in the brain. This can lead to neurotransmitter imbalances, which can trigger or worsen mood disorders such as depression and anxiety.

[0030] Aging brain cells can increase inflammatory responses in blood vessels and the brain, hindering recovery after stroke. In particular, aged microglia have limited ability to regulate inflammation, potentially exacerbating brain tissue damage after stroke. Therefore, reverting aged microglia to younger cells could be beneficial in treating stroke sequelae.

[0031] Aging glial cells can weaken their ability to suppress excessive neural activity. Specifically, if astrocytes fail to properly uptake glutamate, this can lead to hyperexcitability in the brain, potentially leading to epileptic seizures.

[0032] Aging astrocytes and microglia can contribute to inflammatory responses and altered nerve conduction in the brain, potentially increasing the frequency and severity of migraines. In particular, neuroinflammation is known to be associated with migraine attacks.

[0033] Aging glial cells can weaken synaptic function in the brain and lead to imbalances in neural connectivity, which may be a risk factor for psychiatric disorders such as schizophrenia.

[0034] Aging glial cells can interfere with brain energy metabolism and increase inflammation, leading to systemic fatigue and pain symptoms such as chronic fatigue syndrome.

[0035] As glial cells in the hypothalamus age, appetite control and metabolic function may decline, which may increase the risk of obesity, insulin resistance, and metabolic syndrome.

[0036] Additionally, the above cellular aging or aging-related diseases may also include infections.

[0037] The content of the azuki bean extract included in the above pharmaceutical composition is not particularly limited thereto, but may be included in an amount of 0.00001 to 100 wt%, preferably 0.0001 to 15 wt%, more preferably 0.001 to 15 wt%, and even more preferably 0.001 to 10 wt%, based on the total weight of the composition.

[0038] The term "pharmaceutical composition" of the present invention means a composition manufactured for the purpose of preventing or treating a disease, and each may be formulated and used in various forms according to conventional methods. For example, it may be formulated as an oral dosage form such as a powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol, etc., and it may be formulated and used as a parenteral dosage form using a diluent or excipient such as a lubricant, wetting agent, flavoring agent, emulsifier, suspending agent, preservative, surfactant, etc. In addition, it may be formulated and used in the form of an external preparation, suppository, external skin preparation, and sterile injection solution. As a specific example, the composition of the present invention may be used in an oral dosage form for oral administration.

[0039] In addition, depending on each formulation, it can be manufactured by additionally including a pharmaceutically acceptable carrier, such as a buffer, analgesic, solubilizer, isotonic agent, stabilizer, or carrier known in the art.

[0040] The term "pharmaceutically acceptable carrier" used in the present invention may refer to a carrier or diluent that does not stimulate a living organism and does not inhibit the biological activity and properties of the active ingredient to be injected. The type of the carrier that can be used in the present invention is not particularly limited, and any carrier that is commonly used in the relevant technical field and is pharmaceutically acceptable may be used. Non-limiting examples of the carrier include saline solution, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, etc. These may be used alone or in combination of two or more. The carrier may be a non-naturally occurring carrier.

[0041] The composition of the present invention can be administered in a pharmaceutically effective amount. The pharmaceutically effective amount refers to an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and not causing side effects. The effective dosage level can be determined based on the patient's health condition, type and severity of the disease, activity of the drug, sensitivity to the drug, administration method, administration time, administration route and excretion rate, treatment period, drugs used in combination or simultaneously, and other factors well known in the medical field. Specifically, the dosage is generally 0.01 mg to 5000 mg per 1 kg of body weight of the administered subject per day, and may be administered once or several times a day at regular intervals depending on the judgment of a doctor or pharmacist, but is not limited thereto.

[0042] In addition, the pharmaceutical composition of the present invention can be used alone or in combination with other pharmaceutically active compounds exhibiting a cell aging improvement effect or in an appropriate set.

[0043] The pharmaceutical composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents. It can be administered singly or in multiple doses. Taking all of the above factors into account, it is important to administer the amount that achieves maximum effect with the minimum amount possible without causing side effects, a determination readily made by those skilled in the art.

[0044] The method of administration of the pharmaceutical composition according to the present invention is not particularly limited and may follow any method commonly used in the art. Non-limiting examples of the above-mentioned administration method include oral administration or parenteral administration.

[0045] The pharmaceutical composition according to the present invention can be prepared in various dosage forms depending on the intended administration method.

[0046]

[0047] Another aspect of the present invention provides a food composition for preventing or improving cell aging or aging-related diseases, comprising an extract of azuki bean or a fraction thereof as an active ingredient.

[0048] The above food composition may be a functional food composition.

[0049] The term "functional food" in the present invention means a food manufactured and processed using raw materials or ingredients having functionality useful to the human body according to the Health Functional Food Act, and "functionality" means consuming it for the purpose of obtaining a useful effect for health purposes such as regulating nutrients for the structure and function of the human body or physiological action.

[0050] In addition, the term "health functional food" of the present invention refers to a food manufactured or processed by using a specific ingredient as a raw material or by extracting, concentrating, refining, mixing, etc. a specific ingredient contained in a food raw material for the purpose of health supplementation, and refers to a food designed and processed so that the above-mentioned ingredient can sufficiently exert biological regulation functions such as biological defense, biological rhythm regulation, disease prevention, and recovery on the body, and the above-mentioned health food composition can perform functions related to disease prevention and disease recovery.

[0051] When the health functional food composition of the present invention is used in the form of a beverage, it may contain various sweeteners, flavoring agents, or natural carbohydrates as additional ingredients, just like conventional beverages. In addition to the above, the health functional food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, and the like. In addition, it may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks.

[0052] The content of the azuki bean extract included in the above composition is not particularly limited thereto, but may be included in an amount of 0.00001 to 100 wt%, preferably 0.0001 to 15 wt%, more preferably 0.001 to 15 wt%, and even more preferably 0.001 to 10 wt%, based on the total weight of the composition.

[0053]

[0054] Another aspect of the present invention provides a method for treating cellular aging or aging-related diseases, comprising administering an extract of azuki bean or a fraction thereof to a subject in need of treatment.

[0055] The description of the above azuki bean extract or its fraction, cell aging or aging-related diseases applies to the description already described.

[0056] By using the senomorphic composition of the present invention, the state of aged cells can be improved to a young state, and diseases related to cell aging can be treated.

[0057] Figure 1 schematically illustrates a method for isolating primary glial cells and inducing cell senescence.

[0058] Figure 2 shows the results of staining senescent microglia with B-gal, senegreen, γH2AX, and P16, which are known senescence markers.

[0059] Figure 3 is a graph showing the results of Figure 2 and the results of confirming the mRNA level of P16 in aging-induced microglia:

[0060] A: Quantitative graph for B-gal staining, a marker of aging;

[0061] B: Quantitative graph for senegrin staining, a marker of aging;

[0062] C: Quantitative graph for γH2aX staining, a senescence marker;

[0063] D: Quantitative graph for P16 staining, a marker of aging; and

[0064] E: Quantitative graph of mRNA levels of P16.

[0065] Figure 4 shows the results of confirming the cenomorphic effect after treating senescent glial cells (MGC) with azuki bean flower bud extract (GFE):

[0066] A: Results of staining with Senegrin of glial cells that were not induced to age (Young MGCs), glial cells that were induced to age (old MGCs+Vehicle), glial cells that were cultured to age (old MGCs+GFE) treated with GFE, and glial cells that were cultured to age (old MGCs+Rapa) treated with rapamycin; and

[0067] B: Quantitative graph for Senegrin staining.

[0068] Figure 5 shows the results of confirming the cenomorphic effect after treating aging-induced microglial cells with Genkwa Daphneflower extract (GFE):

[0069] A: Results of staining with P16 of non-senescent microglia (young microglia), senescent microglia (old microglia), and senescent microglia cultured by treatment with GFE;

[0070] B: Quantitative graph for P16 staining;

[0071] C: Results of staining with Senegrin of non-senescent microglia (Young microglia), senescent microglia (old microglia), and senescent microglia cultured by treatment with GFE; and

[0072] D: Quantitative graph for Senegrin staining.

[0073] Figure 6 shows the results of confirming the cenomorphic effect after treating senescent astrocytic cells with azure flower bud extract (GFE):

[0074] A: Results of staining p16 on senescent astrocytes (old astrocytes + vehicle) and senescent astrocytes cultured by treating GFE (old astrocytes + GFE); and

[0075] B: Quantitative graph for p16 staining.

[0076] C: Results of staining with Senegrin of senescent astrocytes (old astrocytes + vehicle) and senescent astrocytes cultured by treating GFE (old astrocytes + GFE); and

[0077] D: Quantitative graph for Senegrin staining.

[0078] In Fig. 7, A is a schematic diagram illustrating a method for producing senescent BV-2 cells (Sene BV-2) by treating BV-2 cells with doxorubicin, and B shows a photograph of BV-2 cells and senescent BV-2 cells (Sene BV-2) that are large and have many granules compared to BV-2 cells.

[0079] Figure 8 shows the results of confirming the cenomorphic effect after treating BV-2 cells and senescence-induced BV-2 (Sene BV-2) with azuki bean flower bud extract (GFE):

[0080] A: Results of staining with Senegrin after GFE treatment of BV-2 cells and senescent BV-2 cells (Sene BV-2);

[0081] B: Quantitative graph for senegreen staining; and

[0082] C: Quantitative graph of the results of staining with Bodipy after GFE treatment of BV-2 cells and senescence-induced BV-2 cells (Sene BV-2).

[0083] Hereinafter, one or more specific examples will be described in more detail through examples. However, these examples are provided for illustrative purposes only and the scope of the present invention is not limited to these examples.

[0084]

[0085] Example 1: Preparation of extract from azuki bean flower buds

[0086] The flower buds of the Daphne flower tree were dried and ground to produce a fine powder. 3.5 liters of 70% methanol (powder:methanol = 1:7 (w / w)) was added to the powder and refluxed twice. The collected liquid was then filtered and concentrated under reduced pressure through a freeze-drying process. DMSO (dimethyl sulfoxide) was added to the concentrate to produce a Daphne flower bud extract (Genkwa Daphneflower extract (GFE); hereinafter referred to as GFE) at a concentration of 100 mg / ml.

[0087]

[0088] Example 2: Induction and confirmation of cell senescence

[0089] 2-1. Induction of cell aging

[0090] To confirm the cenomorphic effect of GFE prepared in Example 1 on senescent cells, cell senescence was induced by the following method (Fig. 1).

[0091]

[0092] 1) Primary MGC: Doxorubicin treatment

[0093] Primary MGCs (mixed glial cells; a mixture of glial cells, microglia, oligoglia, and astrocytes) were cultured in DMEM (Dulbecco's modified eagle medium; Gibco) containing 10% FBS (Gibco, USA) and 100 U / mL penicillin / streptomycin (Gibco) at 37°C and 5% CO2. The cultured MGCs were treated with doxorubicin, a well-known cellular senescence inducer, for 48 h, and the cells were washed and cultured in the culture medium for an additional 5 days to induce cellular senescence.

[0094]

[0095] 2) Primary MGC: Cultured for 2 weeks in iron-containing medium

[0096] MGCs were cultured using the same method as in 1) above. Afterwards, MGCs were cultured in a medium supplemented with iron to induce cell senescence.

[0097]

[0098] Microglia and astrocytes were isolated from MGCs induced to undergo cellular senescence, and these cells were used to examine the cenomorphic effects of GFE. The microglia and astrocytes induced to undergo cellular senescence are described below.

[0099] Doxorubicin-induced senescent microglia / astrocytic cells; and

[0100] Iron-induced senescence of microglia / astrocytes

[0101]

[0102] 3) BV-2 cells: doxorubicin treatment

[0103] BV-2 cells (a mouse microglial cell line) were cultured in a medium containing 5% FBS and 50 μg / ml gentamicin at 37°C and 5% CO2. Cell senescence was induced in the cultured BV-2 cells by treatment with doxorubicin (Fig. 7A). Compared to normal BV-2 cells, the senescent BV-2 cells were larger and contained more granules (Fig. 7B). Hereinafter, these cells will be referred to as doxorubicin-induced senescent BV-2 cells.

[0104]

[0105] 2-2. Confirmation of whether cell aging is induced

[0106] The senescent microglia isolated in Example 2-1 were expressed using β-gal and Senegrin (CellEvent), which are known as senescence markers. TM Cell senescence was confirmed by staining with γH2AX, P16, and Senescence Green.

[0107] The CellEvent Senescence Green kit detects cellular senescence through β-galactosidase activity. This kit contains a fluorescent substance that specifically targets β-galactosidase and an optimized buffer. β-galactosidase is used as a biomarker for aging cells. This enzyme functions in lysosomes, degrading β-galactosides into simple sugars in an acidic environment. The degraded fluorescent substance binds to intracellular proteins, remains inside the cell, and emits a fluorescent signal at absorption / emission wavelengths of 490 / 514 nm.

[0108] β-Galactosidase exists in lysosomes and converts β-galactosides into simple sugars under acidic pH conditions. This enzyme's activity is optimized at pH 4 in lysosomes, but is typically measured at pH 6. Studies have shown that, regardless of the pH used, normalized β-galactosidase activity in senescent cells is twice as high as in presenescent cells.

[0109] Cell staining results showed that the expression of senescence markers was significantly increased in iron-induced senescent microglia (Iron) compared to doxorubicin-induced senescent microglia (Doxo) (Fig. 2). Figures 3A to 3D are graphical quantifications of the results in Fig. 2: A: quantitative graph for B-gal staining; B: quantitative graph for senegreen staining; C: quantitative graph for γH2aX staining; and D: quantitative graph for P16 staining.

[0110] In addition, when we confirmed the mRNA expression of P16 in iron-induced senescent microglia, we found that the expression was significantly increased by senescence induction (Fig. 3E).

[0111]

[0112] Example 3: Confirmation of the cenomorphic effect of azuki bean extract in senescence-induced MGC cells.

[0113] To determine the cenomorphic effect of azuki bean extract, iron-induced senescent MGCs were cultured in 96-well plates. After treatment with GFE at a concentration of 10 μg / mL for 24 h, immunostaining with senegreen was performed.

[0114] Staining results showed that the number of senegin-positive cells significantly increased in iron-induced senescent MGCs (old MGCs) compared to primary MGCs (young MGCs). However, the number of senegin-positive cells significantly decreased in iron-induced senescent MGCs (GFE) treated with GFE (10 μg / mL). GFE reduced the number of senegin-positive cells more effectively than rapamycin, which was used as a positive control (Figures 4A and 4B).

[0115]

[0116] Example 4: Confirmation of the cenomorphic effects of GFE in senescence-induced microglia.

[0117] Iron-induced senescent microglia were cultured in 96-well plates. After treatment with GFE at a concentration of 10 μg / mL for 24 h, immunostaining with P16 and Senegreen was performed.

[0118] Staining results showed that the number of P16-stained cells was significantly increased in iron-induced senescent microglia (Old microglia+vehicle) compared to microglia isolated from primary MGCs (Young microglia). However, the number of P16-stained cells was significantly reduced in iron-induced senescent microglia (Old microglia+GFE) treated with GFE (10 μg / mL) (Figures 5A and 5B).

[0119] The same trend was observed in the results of senegreen staining, with a significant increase in the number of cells stained with senegreen in iron-induced senescent microglia (Old microglia+vehicle), but a significant decrease in the number of stained cells in iron-induced senescent microglia (Old microglia+GFE) treated with GFE (10 μg / mL) (Figures 5C and 5D).

[0120]

[0121] Example 5: Confirmation of the cenomorphic effect of azuki bean extract in senescent astrocytic cells.

[0122] Iron-induced senescent astroglial cells were cultured in 96-well plates. After treatment with GFE at a concentration of 10 μg / mL for 24 h, immunostaining with senegreen and P16 was performed.

[0123] Staining results showed that the number of P16-positive cells was high in iron-induced senescent astrocytes (old astrocytes+vehicle), but when iron-induced senescent astrocytes (old astrocytes+vehicle) were cultured by treating with GFE (10 μg / mL), the number of P16-positive cells was significantly reduced (Figures 6A and 6B).

[0124] The same trend was observed in the results of senegreen staining, with a large number of senegreen-positive cells in iron-induced senescent astrocytes (old astrocytes+vehicle), but when iron-induced senescent astrocytes (old astrocytes+vehicle) were cultured with GFE (10 μg / mL), the number of stained cells was significantly reduced (Figs. C and 6D).

[0125]

[0126] Example 6: Confirmation of the senomorphic effect of azuki bean extract in senescence-induced BV-2 cells.

[0127] Doxorubicin-induced senescent BV-2 cells were cultured in 96-well plates. After treatment with GFE at a concentration of 10 μg / mL for 24 h, staining was performed with Senegrin and Bodipy.

[0128] Staining results showed that the number of seneggrin-positive cells was high in doxorubicin-induced senescent BV-2 cells (Sene BV-2+Vehicle), but the number of seneggrin-positive cells was reduced in doxorubicin-induced senescent BV-2 cells treated with GFE (old BV-2+GFE) (Figs. 7C and 8A).

[0129] Additionally, in doxorubicin-induced senescent BV-2 cells treated with GFE (old BV-2+GFE), the number of intracellular lipid droplets stained with Bodiphyrin was significantly reduced (Fig. 8B).

Claims

1. A cenomorphic composition comprising an extract of azuki bean or a fraction thereof as an active ingredient.

2. A cenomorphic composition according to claim 1, wherein the azuki bean extract is extracted from at least one part selected from the group consisting of flower buds, leaves, fruit stems, and roots of the azuki bean.

3. A cenomorphic composition according to claim 1, wherein the azuki bean extract is extracted using water, alcohol or a mixture thereof as a solvent.

4. A cenomorphic composition in the third paragraph, wherein the azuki bean extract is extracted using a mixture of water and methanol as a solvent.

5. A cenomorphic composition according to claim 1, wherein the composition improves aging of cells selected from the group consisting of microglia, oligoglia, and astroglia.

6. In the fifth paragraph, the improvement of cell aging is a cenomorphic composition in which the expression of a marker selected from the group consisting of β-galactosidase (SA-β-gal), γH2AX, and P16 is reduced.

7. A pharmaceutical composition for preventing or treating cell aging or aging-related diseases, comprising an extract of azuki bean or a fraction thereof as an active ingredient.

8. A pharmaceutical composition for preventing or treating cell aging or aging-related diseases, wherein the azuki bean extract in paragraph 7 is extracted from at least one part selected from the group consisting of flower buds, leaves, fruit stems, and roots of the azuki bean tree.

9. A pharmaceutical composition for preventing or treating cell aging or aging-related diseases, wherein the azuki bean extract in paragraph 7 is extracted using water, alcohol or a mixture thereof as a solvent.

10. A pharmaceutical composition for preventing or treating cellular aging or aging-related diseases, wherein the cellular aging or aging-related diseases in paragraph 6 are selected from the group consisting of depression, anxiety disorder, post-stroke syndrome, epilepsy, migraine, schizophrenia, chronic fatigue syndrome, obesity, metabolic syndrome, infection, and chronic inflammatory diseases.

11. A food composition for preventing or improving cell aging or aging-related diseases, comprising an extract of azuki bean or a fraction thereof as an active ingredient.

12. A method for treating cellular aging or aging-related diseases, comprising administering an extract of azuki bean or a fraction thereof to a subject in need of treatment.

Images