Composition for prevention and treatment of aging containing extract of edible insect Cordyceps sinensis

Abstract

The present invention relates to a composition for preventing or treating aging containing an extract of Cordyceps sinensis from edible insects as an active ingredient. Since the composition of the present invention exhibits an effect of preventing or treating aging, it can be effectively used to prevent or improve various diseases and health problems caused by aging. In particular, the composition of the present invention can be usefully used to prevent or treat aging-related diseases such as sarcopenia, cognitive decline, cardiovascular disease, cancer, and diabetes. The extract of Cordyceps sinensis from edible insects according to the present invention can be obtained by extracting Cordyceps sinensis fruiting bodies, obtained by culturing Cordyceps sinensis fungi on edible insects such as crickets, mealworm larvae, white-spotted flower beetle larvae, rhinoceros beetle larvae, and grasshoppers, using solvents such as water, ethanol, and methanol. The composition of the present invention can be prepared in various forms, such as food compositions, health functional food compositions, pharmaceutical compositions, and cosmetic compositions.

Description

Technology Field

[0001] The present invention relates to a composition for preventing or treating aging containing an extract of Cordyceps sinensis from edible insects. More specifically, the invention relates to a composition for preventing or treating aging containing, as an active ingredient, an extract of a Cordyceps sinensis fruiting body obtained by culturing Cordyceps sinensis fungi on edible insects. Background Technology

[0003] As average human life expectancy increases, interest in anti-aging and extending healthy life expectancy is growing. Aging is a natural phenomenon in which biological functions decline over time, leading to various diseases and health problems. Aging-related conditions include sarcopenia, cognitive decline, cardiovascular disease, cancer, and diabetes, which not only reduce an individual's quality of life but also increase the socioeconomic burden.

[0004] Aging is known to be caused by complex factors such as oxidative stress, inflammation, cell damage, and hormonal changes. Therefore, effectively controlling these factors is crucial for preventing or treating aging. Recently, research on anti-aging and treatment utilizing naturally derived ingredients has been actively underway, and among them, Cordyceps is attracting attention as a natural material possessing various physiological activities.

[0005] Cordyceps is a mushroom that has traditionally been used for medicinal purposes and is known to possess various benefits, including boosting immunity, anticancer, anti-inflammatory, and antioxidant effects. In particular, with the recent advancement of cultivation technology using edible insects, edible insect Cordyceps is garnering attention as a new functional ingredient. The problem to be solved

[0007] The present invention aims to provide a composition having the effect of preventing or treating aging. In particular, it aims to provide a composition containing an extract of the edible insect Cordyceps as an active ingredient, thereby providing a safe and effective effect of preventing or treating aging. means of solving the problem

[0009] The present invention provides a composition for preventing or treating aging containing an extract of the edible insect Cordyceps as an active ingredient.

[0011] The edible insect Cordyceps used in the present invention refers to a Cordyceps fruiting body obtained by culturing Cordyceps fungi on edible insects such as crickets, mealworm larvae, white-spotted flower beetle larvae, rhinoceros beetle larvae, and grasshoppers.

[0013] The edible insect Cordyceps extract of the present invention can be obtained by extracting the fruiting body using a solvent such as water, ethanol, or methanol, and the extraction method is not particularly limited. For example, methods such as hot water extraction, cold maceration extraction, ultrasonic extraction, and reflux cooling extraction may be used.

[0015] The composition of the present invention can be manufactured in various forms, such as food compositions, health functional food compositions, pharmaceutical compositions, and cosmetic compositions, having anti-aging or therapeutic effects. Effects of the invention

[0017] The composition containing the edible insect Cordyceps extract of the present invention as an active ingredient exhibits an effect of preventing or treating aging, and thus can be effectively used to prevent or improve various diseases and health problems caused by aging. In particular, the composition of the present invention can be usefully used to prevent or treat aging-related diseases such as sarcopenia, cognitive decline, cardiovascular disease, cancer, and diabetes. Specific details for implementing the invention

[0019] The present invention provides a composition for preventing or treating aging, containing an extract of the edible insect Cordyceps as an active ingredient. The present invention will be described in detail below by way of examples. However, these examples are intended to illustrate the present invention, and the scope of the present invention is not limited to these examples.

[0021] 6.1 Materials and Methods

[0023] 6.1.1 Culture of Edible Insect Cordyceps

[0024] The edible insect Cordyceps used in the present invention was cultured by inoculating a two-spotted cricket with Cordyceps militaris. The two-spotted cricket was obtained from the Rural Development Administration, and the Cordyceps fungus was obtained from the Korea Research Institute of Biotechnology.

[0026] Cordyceps cultivation was performed using the following method.

[0027] Two-spotted crickets were placed in a sterilized container and inoculated with Cordyceps fungus.

[0028] Inoculated two-spotted crickets were cultured for 4 weeks in a culture room at 25℃ and 70% humidity.

[0029] The cultured Cordyceps fruiting bodies were harvested and dried.

[0031] 6.1.2 Preparation of Edible Insect Cordyceps Extract

[0032] Dried Cordyceps fruiting bodies were ground into a powder, placed in 10 times (w / v) 70% ethanol, and extracted by reflux cooling at 80°C for 24 hours. The extract was filtered through filter paper, concentrated under reduced pressure, and freeze-dried.

[0034] 6.1.3 Cell Culture

[0035] Human fibroblasts (HDF), human keratinocytes (HaCaT), mouse muscle cell line (C2C12), and mouse neuronal cell line (HT22) were obtained from the Korean Cell Line Bank and used. HDF cells and C2C12 cells were cultured in DMEM medium containing 10% FBS and 1% penicillin-streptomycin, while HaCaT cells and HT22 cells were cultured in MEM medium containing 10% FBS and 1% penicillin-streptomycin. All cells were cultured at 37°C under 5% CO2 conditions.

[0037] 6.1.4 Induction of Cellular Aging

[0038] Cellular senescence was induced in HDF cells by treating them with 200 μM H2O2 for 2 hours.

[0040] 6.1.5 Measurement of Cell Viability

[0041] Cell viability was measured using the MTT assay. Cells were seeded in a 96-well plate and cultured for 24 hours, after which various concentrations of Cordyceps extract were applied. After 24 hours, MTT solution was added and reacted for 4 hours, after which DMSO was added to dissolve the formazan crystals. Absorbance was measured at 540 nm using a microplate reader.

[0043] 6.1.6 Measuring ROS Generation

[0044] The amount of intracellular reactive oxygen species (ROS) produced was measured using the DCFH-DA assay. Cells were seeded in a 96-well plate and cultured for 24 hours, after which various concentrations of Cordyceps extract were applied. After 24 hours, DCFH-DA was added and reacted for 30 minutes, after which fluorescence intensity was measured at 485 nm excitation and 535 nm emission using a fluorescence microplate reader.

[0046] 6.1.7 Measurement of Antioxidant Enzyme Activity

[0047] Intracellular antioxidant capacity was evaluated by measuring the activities of catalase, superoxide dismutase (SOD), and glutathione peroxidase (GPx).

[0049] 6.1.8 Measurement of Telomerase Activity

[0050] Telomerase activity was measured using a telomerase activity kit (TRAPeze® RT Telomerase Detection Kit, Millipore).

[0052] 6.1.9 β-galactosidase staining

[0053] β-galactosidase activity was measured using the Senescence β-Galactosidase Staining Kit (Cell Signaling Technology).

[0055] 6.1.10 Western blot analysis

[0056] Cells were lysed using lysis buffer, and protein concentration was measured. Proteins were electrophoresed on an SDS-PAGE gel and transferred to a PVDF membrane. After blocking the membrane with blocking buffer, the reaction was carried out using the primary and secondary antibodies. Protein expression was confirmed using ECL solution.

[0058] 6.1.11 Animal testing

[0059] 8-week-old male C57BL / 6 mice were purchased from Orient Bio and used. The mice were housed in an animal housing room maintained at a temperature of 22±2℃, humidity of 55±5%, and a 12-hour light-dark cycle (07:00–19:00).

[0061] 6.1.12 Induction of Skin Aging

[0062] Skin aging was induced by irradiating the back of mice with UVB.

[0064] 6.1.13 Skin Wrinkle Measurement

[0065] Mouse skin replicas were produced, and the depth and number of wrinkles were measured.

[0067] 6.1.14 Measurement of Collagen Content

[0068] Collagen content was measured in mouse skin tissue.

[0070] 6.1.15 Statistical Analysis

[0071] All experimental results were expressed as mean ± standard deviation, and statistical significance was analyzed using Student's t-test or one-way ANOVA. A p-value of less than 0.05 was considered statistically significant.

[0073] 6.2 Experimental Results

[0075] 6.2.1 Effects of Edible Insect Cordyceps Extract on HDF Cell Viability

[0076] The effect of edible insect Cordyceps extract on the viability of HDF cells was confirmed using an MTT assay. As a result, it was found that Cordyceps extract did not affect cell viability up to a concentration of 100 μg / mL.

[0078] 6.2.2 Effects of Edible Insect Cordyceps Extract on HDF Cell Aging

[0079] The effects of Cordyceps extract were confirmed in an H2O2-induced HDF cell aging model. As a result, Cordyceps extract was found to inhibit cell aging in a concentration-dependent manner. Specifically, Cordyceps extract was found to decrease β-galactosidase activity and increase telomerase activity.

[0081] 6.2.3 Effects of Edible Insect Cordyceps Extract on ROS Production and Antioxidant Enzyme Activity in HDF Cells

[0082] The effect of edible insect Cordyceps extract on ROS production in HDF cells was confirmed using the DCFH-DA assay. As a result, it was found that the Cordyceps extract inhibited ROS production in a concentration-dependent manner.

[0083] In addition, the effect of Cordyceps extract on the antioxidant enzyme activity of HDF cells was examined. As a result, Cordyceps extract was found to increase catalase, SOD, and GPx activities.

[0085] 6.2.4 Effects of Edible Insect Cordyceps Extract on Viability, ROS Production, and Antioxidant Enzyme Activity of HaCaT, C2C12, and HT22 Cells

[0086] The effects of edible insect Cordyceps extract on the viability, ROS production, and antioxidant enzyme activity of HaCaT, C2C12, and HT22 cells were confirmed using the same method as above. As a result, it was found that Cordyceps extract inhibited ROS production and increased antioxidant enzyme activity without affecting cell viability.

[0088] 6.2.5 Effects of Edible Insect Cordyceps Extract on Mouse Skin Aging

[0089] The effects of Cordyceps extract were confirmed in a UVB-induced mouse skin aging model. As a result, Cordyceps extract was found to reduce the depth and number of skin wrinkles and increase collagen content.

[0091] 6.3 Conclusion

[0092] Through the experimental results described above, it was confirmed that the edible insect Cordyceps extract exhibited effects of inhibiting cellular aging, suppressing ROS production, and increasing antioxidant enzyme activity in various cells without affecting cell viability. Furthermore, animal experiments confirmed that the edible insect Cordyceps extract inhibits skin aging.

[0093] Therefore, the edible insect Cordyceps extract of the present invention can be usefully used as an active ingredient in a composition for preventing or treating aging.

Claims

Claim 1 A composition for preventing or treating aging containing an extract of edible insect Cordyceps as an active ingredient. The composition is characterized in that the edible insect Cordyceps is a fruiting body obtained by culturing Cordyceps fungi on one or more insects selected from the group consisting of crickets, mealworm larvae, white-spotted flower beetle larvae, rhinoceros beetle larvae, and grasshoppers.

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