Use of extracts for the preparation of non-medical compositions for increasing bioenergy

Abstract

The present application provides a use of an extract for preparing a pharmaceutical or non-pharmaceutical composition for improving bioenergy. The mitochondria with improved activity can maintain its function and activity when facing stress, to ensure normal cell operation without adverse effects from external or internal stress.

Description

Technical Field

[0001] This invention relates to the use of extracts in the preparation of pharmaceutical or non-pharmaceutical compositions that enhance bioenergy, and more particularly to the use of Ashitaba extracts in the preparation of pharmaceutical or non-pharmaceutical compositions that enhance bioenergy. Background Technology

[0002] Mitochondria are the primary sites of oxidative phosphorylation and ATP synthesis within cells. Because ATP is the energy source for cellular activities, mitochondria are often referred to as the "cell's energy factory." In addition to providing energy, mitochondria participate in processes such as cell differentiation, cell signaling, and apoptosis, and possess the ability to regulate the cell cycle.

[0003] However, some byproducts produced during oxidative phosphorylation in mitochondria are harmful to mitochondria. These include reactive oxygen species (ROS), such as superoxide anion (O2•-), perhydroxyl radical (HO2•), and hydrogen peroxide (H2O2). ROS are highly biochemically reactive and easily cause oxidative damage to cells or mitochondria. Damaged mitochondria have adverse effects on cellular energy supply and cell growth. Over time, severely damaged mitochondria release cytochrome c (Cyt c), apoptotic proteases, proteolytic enzymes, and procaspase-2, 3, 8, and 9, triggering mitochondrial disintegration and releasing apoptosis-related signaling factors, thus triggering apoptosis. Therefore, improving the stress resistance of mitochondria, protecting and repairing mitochondria to maintain their function, and slowing down mitochondrial disintegration has become an important research topic. Summary of the Invention

[0004] This invention provides a method to enhance mitochondrial activity by using extracts, thereby improving the mitochondrial ability to cope with stress and maintaining the function and activity of mitochondria when facing stress.

[0005] This invention provides the use of an extract for preparing pharmaceutical or non-pharmaceutical compositions that enhance bioenergy, wherein the extract is an Ashitaba extract, which is derived from Angelica dahurica (Ashitsuba). Angelica keiskei ).

[0006] The extract provided in this invention is used to prepare pharmaceutical or non-pharmaceutical compositions that enhance bioenergy. The extract is *Ashitaba* extract, which can reduce mitochondrial hydrogen ion leakage, increase mitochondrial pre-existing oxygen consumption capacity, maximum oxygen consumption capacity, and adenosine triphosphate (ATP) synthesis capacity, improve mitochondrial ATP coupling efficiency, and enhance bioenergy health index. Mitochondria with enhanced activity from *Ashitaba* extract can maintain their function and activity under stress, ensuring normal cell function without adverse effects from external or internal stress. Attached Figure Description

[0007] Figure 1 The cytotoxicity of different concentrations of Ashitaba extract was shown.

[0008] Figure 2 This shows the amount of oxygen consumed by mitochondria to overcome hydrogen ion leakage.

[0009] Figure 3 This indicates the oxygen consumption required for mitochondrial synthesis of adenosine triphosphate (ATP).

[0010] Figure 4 This indicates the pre-existing oxygen consumption of mitochondria.

[0011] Figure 5 This shows the maximum oxygen consumption of mitochondria.

[0012] Figure 6 This shows the efficiency of mitochondrial adenosine triphosphate coupling. Detailed Implementation

[0013] The following embodiments describe in detail the features and advantages of the present invention, the content of which is sufficient to enable any person skilled in the art to understand the technical content of the present invention and to implement it accordingly. Furthermore, based on the disclosure of this specification, the claims, and the accompanying drawings, any person skilled in the art can easily understand the related objects and advantages of the present invention. The following embodiments further illustrate the viewpoints of the present invention in detail, but are not intended to limit the scope of the present invention in any way.

[0014] The extract of Ashitaba in this embodiment of the invention is taken from Angelica dahurica (Ashita no Basilica). Angelica keiskei Ashitaba (Tomorrow's Leaf) is native to Hachijojima Island, Japan. It can grow on plains in temperate regions and at altitudes of approximately 500–2000 meters in subtropical regions. Ashitaba is extremely resilient; it gets its name from the fact that "a leaf picked today will sprout a new bud tomorrow." Ashitaba provides abundant chlorophyll, vitamins, dietary fiber, protein, amino acids, and various minerals essential for the human body. Reports indicate that Ashitaba is beneficial to human health; its stems and leaves can be used as health foods, and its roots can be used as a medicinal herb or food additive.

[0015] The main bioactive components of Ashitaba include coumarins and chalcones. Coumarin compounds have antioxidant, anticancer, antidepressant, and acetylcholinesterase inhibitory effects, while chalcone compounds have antioxidant, anticancer, and glucosidase inhibitory effects. The coumarins in Ashitaba leaves mainly include xanthotoxin and laserpitin, while the chalcone compounds mainly include xanthoangelol and 4-hydroxyderricin. The chalcone compounds in Ashitaba leaves exhibit xanthine oxidase inhibitory effects.

[0016] The extract of one embodiment of the present invention is an Ashitaba extract, which can be obtained by pulverizing Ashitaba leaves into powder, soaking the powder in water at a ratio of 1 gram of powder to 25 milliliters of water at room temperature for 1 day, centrifuging the solution, and freeze-drying the resulting supernatant. Specifically, the Ashitaba leaves are cleaned and dried, then pulverized into powder using pneumatic cracking (60 mesh) and graded. The Ashitaba powder is inspected using a metal detector to avoid the introduction of metal impurities during the process. The inspected Ashitaba powder is soaked in water at a ratio of 1 gram of powder to 25 milliliters of water at room temperature for 1 day, i.e., water extraction at room temperature for 1 day. The solution is centrifuged at 3000 g for 30 minutes, and the resulting supernatant is freeze-dried to obtain the Ashitaba extract powder used in this embodiment of the invention. The obtained powder is then formulated into an aqueous solution, which is the Ashitaba extract used in this embodiment of the invention.

[0017] The Ashitaba extract obtained by the above method contains flavotoxin, angelicol, hydrodryline, and laserpitin. The content of flavotoxin can be 0.95 to 1.05 mg / g, the content of angelicol can be 1.05 to 1.15 mg / g, the content of hydrodryline can be 0.55 to 0.65 mg / g, and the content of laserpitin can be 0.75 to 0.85 mg / g.

[0018] In some embodiments of the present invention, providing cells with Ashitaba extract at concentrations ranging from 250 μg / mL to 1000 μg / mL can enhance bioenergy. Bioenergy can be expressed as mitochondrial activity, specifically manifested in improved mitochondrial pre-existing oxygen consumption capacity, maximum oxygen consumption capacity, and adenosine triphosphate (ATP) synthesis capacity; reduced mitochondrial hydrogen ion leakage; improved mitochondrial ATP coupling efficiency; and improved bioenergy health index. In another embodiment, the concentration of Ashitaba extract can be from 250 μg / mL to 500 μg / mL. In other embodiments, the concentration of Ashitaba extract can be from 500 μg / mL to 1000 μg / mL.

[0019] One method of providing Ashitaba extract to cells is, for example, oral ingestion of Ashitaba extract via food. When Ashitaba extract is provided to cells via food, the effective dose of Ashitaba extract is 2.703 g to 10.812 g. The effective dose here is calculated using a conversion formula between the effective dose in cell experiments and human weight (kg). The conversion formula is as follows: Effective human dose = Effective dose in cell experiments × Mouse weight × Conversion factor × Human weight (kg). The conversion factor is obtained from a table of conversion factors per kilogram of body weight for animals and humans. When the mouse weight is 20 g and the human weight is 60 kg, the conversion factor is 9.01. In another embodiment, the effective dose of Ashitaba extract is 2.703 g to 5.406 g. In other embodiments, the effective dose of Ashitaba extract is 5.406 g to 10.812 g.

[0020] To facilitate oral ingestion of Ashitaba extract, Ashitaba extract can be prepared into processed Ashitaba extract products in various forms, such as liquid, solid, granular, powder, paste, or gel. In some embodiments of the present invention, without affecting the efficacy and objectives achieved by the present invention, the processed Ashitaba extract products may also contain other ingredients or additives, such as carriers, diluents, excipients, or flavoring agents. Excipients make the formulation convenient to use, while flavoring agents enhance the flavor of the formulation.

[0021] Excipients include starches such as wheat starch, rice starch, corn starch, potato starch, dextrin, and cyclodextrin; crystalline cellulose; sugars such as lactose, glucose, sucrose, reduced maltose, maltose, fructooligosaccharides, and emulsified oligosaccharides; and sugar alcohols such as sorbitol, erythritol, xylitol, lactitol, and mannitol.

[0022] Flavoring agents include various fruit juice extracts such as longan extract, lychee extract, and grapefruit extract; various fruit juices such as apple juice, orange juice, and lemon juice; various flavorings such as peach flavoring, plum flavoring, and yogurt flavoring; various sweeteners such as acesulfame potassium, sucralose, erythritol, oligosaccharides, mannose, xylitol, and isomerized sugars; various acidulants such as citric acid, malic acid, tartaric acid, and gluconic acid; and various tea components such as green tea, oolong tea, Banaba tea, Eucommia tea, Tieguanyin tea, Coix seed tea, Gynostemma pentaphyllum tea, water chestnut tea, and kelp tea.

[0023] Furthermore, the Ashitaba extract composition of the embodiments of the present invention can be a pharmaceutical composition or a non-pharmaceutical composition, such as a health food. Ashitaba extract or a composition containing Ashitaba extract can also be encapsulated for convenient oral intake. Ashitaba extract or a composition containing Ashitaba extract can be encapsulated in hard capsules in the form of a dry powder, or in soft capsules in the form of a solution, suspension, paste, powder, or granules.

[0024] Oils used in soft capsules to dissolve or disperse Ashitaba extract include, for example, pear oil, almond oil, linseed oil, fennel oil, perilla oil, olive oil, olive squalene, sweet orange oil, orange roughy oil, sesame oil, garlic oil, cocoa butter, pumpkin seed oil, chamomile oil, carrot oil, cucumber oil, tallow fatty acids, macadamia nut oil, blueberry oil, brown rice germ oil, rice oil, wheat germ oil, safflower oil, shea butter, liquid shea butter, perilla oil, soybean oil, evening primrose oil, camellia oil, corn oil, rapeseed oil, saw palmetto extract oil, coix seed oil, peach kernel oil, celery seed oil, castor oil, sunflower seed oil, grape seed oil, borage oil, macadamia nut oil, and meadowsweet oil. Oils, cottonseed oil, peanut oil, turtle oil, mink oil, egg yolk oil, fish oil, palm oil, palm kernel oil, wood wax, coconut oil, long-chain / medium-chain / short-chain fatty acid triglycerides, diglycerides, tallow, lard, squalene, squalane, pterostilbene, and hydrogenated forms of these oils.

[0025] In addition, additives that meet the relevant regulations, such as colorants, preservatives, thickeners, binders, disintegrants, dispersants, stabilizers, gelling agents, antioxidants, surfactants, preservatives, and pH adjusters, may also be added to the processed products of Ashitaba extract compositions in accordance with the dosage standards and processing requirements specified by the relevant authorities.

[0026] The following describes the experiment using Ashitaba extract to enhance mitochondrial activity according to an embodiment of the present invention. The cells used in the experiment were skeletal muscle cells (C2C12). DMEM culture medium containing 10% fetal bovine serum (FBS) was used as the skeletal muscle cell culture medium (hereinafter referred to as the culture medium). The cell subculture method was as follows: First, the skeletal muscle cells were cultured to a certain volume, then the culture medium was removed, and the skeletal muscle cells were rinsed twice with phosphate-buffered saline (PBS). Next, trypsin was added, and the trypsin was reacted with the skeletal muscle cells at 37°C for 5 minutes, followed by the addition of culture medium to stop the trypsin reaction. Next, the solution containing the skeletal muscle cells was centrifuged at 300 g (relative centrifugal force, RCF) for 5 minutes, the supernatant was removed, and the precipitate was reconstituted with culture medium. Finally, the skeletal muscle cells were transferred to a cell culture flask (175T flask) for subsequent experiments, with a cell count of 1×10⁶ cells / year. 6 Each cell.

[0027] [Experiment 1] Cytotoxicity of Ashitaba Extract

[0028] First, the cytotoxicity of Ashitaba extract was tested. Alamar blue was used as a assay reagent to detect cell viability. The test kit included resazurin, a redox indicator, which is a non-toxic, cell-membrane-penetrating, low-fluorescence deep blue dye. When resazurin enters healthy cells, it is reduced to pink, highly fluorescent resorufin due to the reducing environment within the living cells. Cell viability can be assessed by measuring the absorbance or fluorescence value of the resorufin produced in the cells. Higher absorbance or fluorescence values ​​of the resorufin indicate higher cell viability. Higher cell viability indicates healthier cells and stronger proliferative capacity. Stronger cell proliferative capacity indicates a greater cell mass. Therefore, alamar blue can serve as an indicator of cytotoxicity to determine cell viability and proliferation rate.

[0029] The following details the cytotoxicity assay procedure for Ashitaba extract. On day one, skeletal muscle cells were cultured for one day in 96-well plates with a total volume of 200 μL of skeletal muscle cells and culture medium, at a density of 10,000 cells per well. On day two, Ashitaba extract was added to each well at concentrations of 50, 100, 200, 250, 500, and 1000 μg / mL. The Ashitaba extract and skeletal muscle cells were co-cultured at 37°C for one day. On day three, cytotoxicity was assessed using alamar blue. Specifically, a 10% (w / w) alamar blue solution was prepared under light-protected conditions and added to each well at a volume of 100 μL. The mixture was then co-cultured with the skeletal muscle cells at 37°C for 3 to 4 hours. Finally, the absorbance and fluorescence values ​​(OD530 / 590) were measured using an ELISA reader. This was used to obtain the survival rate of skeletal muscle cells treated with Ashitaba extract, which represents the toxicity of Ashitaba extract to cells.

[0030] The experimental results revealed Figure 1 , Figure 1 The cytotoxicity of different concentrations of Ashitaba extract is shown. The control group consisted of skeletal muscle cells untreated with Ashitaba extract (concentration of 0 μg / mL), and the vertical axis represents the fold increase in cell viability relative to the control group.

[0031] like Figure 1 As shown, concentrations of Ashitaba extract below 1000 μg / mL had no effect on cell viability, indicating that Ashitaba extract at concentrations below 1000 μg / mL is non-cytotoxic. Accordingly, Ashitaba extracts at concentrations of 250, 500, and 1000 μg / mL were selected as Examples 1 to 3 of this invention for subsequent experiments.

[0032] [Experiment 2] Ashitaba extract enhances mitochondrial activity

[0033] Next, an experiment was conducted to enhance mitochondrial activity using Ashitaba extract. This experiment used tert-butyl hydroperoxide (tBHP) as a substance that induces cellular oxidative stress damage, aging, and inhibits mitochondrial activity.

[0034] The experimental procedure for enhancing mitochondrial activity with Ashitaba extract is detailed below. On day one, skeletal muscle cells were cultured for 4 hours in 24-well hippocampal discs at a total volume of 100 μL of culture medium and 25,000 cells per well. Then, 150 μL of culture medium was added, and the cells were cultured for another day. On day two, Ashitaba extract was added to each well at concentrations of 250, 500, and 1000 μg / mL, with a total solution volume of 250 μL. Skeletal muscle cells were cultured with Ashitaba extract under these conditions for another day. On day three, the culture medium was replaced with fresh medium, and tBHP was added to each well. Skeletal muscle cells were reacted with 100 μM tBHP for 1 hour. Then, the culture medium in the discs was replaced with 675 μL of DMEM culture medium (without fetal bovine serum, pH 7.4). After culturing for 1 hour in a carbon dioxide-free incubator, the oxygen consumption of the skeletal muscle cells in each well was measured using a hippocampal bioenergy meter.

[0035] The measurement principle and procedure of the hippocampal bioenergy grading instrument are as follows: First, the basal oxygen consumption of the cells in the well is detected. Next, an adenosine triphosphate synthase inhibitor is added to inhibit mitochondrial production of adenosine triphosphate; the reduced oxygen consumption at this point is the oxygen consumption for adenosine triphosphate synthesis (ATP production). Then, an appropriate concentration of anticoupling agent is added, allowing the mitochondria to idle under extreme conditions without disrupting the electron transport chain of the mitochondrial inner membrane to assess the mitochondrial maximum oxygen consumption (Maximal Respiration). Finally, an electron transport chain inhibitor is added to completely shut down mitochondrial oxygen consumption, thereby confirming the background value of the measurement, i.e., non-mitochondrial oxygen consumption. The basal oxygen consumption of mitochondria equals the basal oxygen consumption of the cell minus the non-mitochondrial oxygen consumption. The basal oxygen consumption of mitochondria minus the oxygen consumption for adenosine triphosphate synthesis equals the oxygen consumption to overcome hydrogen ion leakage (Proton Leakage). The maximum oxygen consumption of mitochondria minus the basal oxygen consumption of mitochondria equals the mitochondrial spare respiratory capacity. The coupling efficiency of mitochondria with adenosine triphosphate (ATP) is equal to the oxygen consumption for ATP synthesis divided by the basal oxygen consumption of mitochondria.

[0036] The experimental results are shown in Table 1 and Figures 2 to 6 , Figure 2 This shows the amount of oxygen consumed by mitochondria to overcome hydrogen ion leakage. Figure 3 This indicates the oxygen consumption required for mitochondrial synthesis of adenosine triphosphate (ATP). Figure 4 This indicates the pre-existing oxygen consumption of mitochondria. Figure 5 This shows the maximum oxygen consumption of mitochondria. Figure 6This demonstrates the efficiency of mitochondrial adenosine triphosphate (ATP) coupling. The control group consisted of skeletal muscle cells untreated with tBHP and untreated with Ashitaba extract; the control group consisted of skeletal muscle cells treated with tBHP but not with Ashitaba extract; and the experimental group consisted of skeletal muscle cells treated with Ashitaba extract as described in the examples and then treated with tBHP. Figures 2 to 5 In the diagram, the vertical axis represents oxygen consumption, expressed in picomoles per minute (pmole / min). Figure 6 In the figure, the vertical axis represents the coupling efficiency of adenosine triphosphate (ATP) as a percentage (%). "*" and "**" indicate significant differences relative to the control group (*P<0.05, **P<0.01), respectively, while "#", "##", and "###" indicate significant differences relative to the control group (#P<0.05, ##P<0.01, ###P<0.001).

[0037] Table 1

[0038]

[0039] like Figure 2 As shown, in the measurement results of mitochondria overcoming hydrogen ion leakage, the oxygen consumption of the comparison group was higher than that of the control group, indicating that the inner membrane of the mitochondria in the comparison group was damaged and required more oxygen to overcome hydrogen ion leakage. In contrast, the oxygen consumption of the experimental groups treated with Ashitaba extract in Examples 1 to 3 was lower than that of the comparison group, indicating that the activity of mitochondria was enhanced by Ashitaba extract. This is equivalent to Ashitaba extract having the function of protecting and repairing mitochondria under oxidative stress, thus the damage to the mitochondrial inner membrane was less.

[0040] like Figure 3 As shown, in the measurement results of mitochondrial synthesis of adenosine triphosphate (ATP), the oxygen consumption of the comparison group was lower than that of the control group, indicating that the ability of mitochondria to synthesize ATP under oxidative stress is reduced, and the energy produced by mitochondria is less. In contrast, the oxygen consumption of the experimental groups treated with Ashitaba extract in Examples 1 to 3 was higher than that of the comparison group, indicating that the activity of mitochondria was enhanced by Ashitaba extract, and the ability to synthesize ATP under oxidative stress was also enhanced, thus enabling the generation of sufficient energy for cellular use.

[0041] like Figure 4 As shown, in the measurement results of pre-existing oxygen consumption, the oxygen consumption of the comparison group was lower than that of the control group, indicating that the pre-existing oxygen consumption capacity of mitochondria decreased under oxidative stress. In contrast, the oxygen consumption of the experimental groups treated with Ashitaba extract in Examples 1 to 3 was higher than that of the comparison group, indicating that the activity of mitochondria was enhanced by Ashitaba extract, and the pre-existing oxygen consumption capacity of mitochondria was also enhanced under oxidative stress. The enhanced pre-existing oxygen consumption capacity indicates that mitochondria have better stress resistance.

[0042] like Figure 5 As shown, in the measurement results of maximum oxygen consumption, the oxygen consumption of the comparison group was lower than that of the control group, indicating that the maximum oxygen consumption capacity of mitochondria was reduced under oxidative stress. In contrast, the oxygen consumption of the experimental groups treated with Ashitaba extract in Examples 1 to 3 was higher than that of the comparison group, indicating that the activity of mitochondria was enhanced by Ashitaba extract, and the maximum oxygen consumption capacity of mitochondria was also enhanced under oxidative stress.

[0043] like Figure 6 As shown, in the results of adenosine triphosphate coupling efficiency, the values ​​of the comparison group were lower than those of the control group, while the values ​​of the experimental group treated with Ashitaba extracts from Examples 1 to 3 were higher than those of the comparison group. This indicates that the activity of mitochondria was enhanced by Ashitaba extracts, and the adenosine triphosphate coupling efficiency of mitochondria was also enhanced under oxidative stress.

[0044] Based on measurements from a hippocampal bioenergy analyzer, the Bioenergetic Healthy Index (BHI) can be calculated from mitochondrial oxygen consumption. The BHI is an energy metabolism assessment index calculated using mitochondrial energy metabolism data as parameters. BHI = log[Oxygen consumption for adenosine triphosphate synthesis × Pre-stored oxygen consumption] / [Oxygen consumption to overcome hydrogen ion leakage × Non-mitochondrial oxygen consumption]. A high BHI indicates high mitochondrial activity and a strong ability of the cell to cope with stress. Therefore, the BHI can serve as an indicator for assessing the health of mitochondria and cells.

[0045] The bio-health energy index calculated from the above experimental results is shown in Table 2. As shown in Table 2, compared with the control group, the treatment with Ashitaba extract in the examples improved the bio-health energy index, indicating that the health of mitochondria and skeletal muscle cells was enhanced.

[0046] Table 2

[0047]

[0048] The experimental results above show that Ashitaba extract at concentrations below 1000 μg / mL is not toxic to cells. Furthermore, Ashitaba extract can enhance mitochondrial activity, specifically by increasing mitochondrial pre-oxygen consumption capacity, maximum oxygen consumption capacity, and adenosine triphosphate (ATP) synthesis capacity; reducing mitochondrial hydrogen ion leakage; improving mitochondrial ATP coupling efficiency; and enhancing the bioenergetic health index.

[0049] The extract provided in this invention is used to prepare pharmaceutical or non-pharmaceutical compositions that enhance mitochondrial activity. The extract is *Ashitaba* extract, which can reduce mitochondrial hydrogen ion leakage, increase mitochondrial pre-existing oxygen consumption capacity, maximum oxygen consumption capacity, and adenosine triphosphate (ATP) synthesis capacity, improve mitochondrial ATP coupling efficiency, and enhance bioenergetic health index. Mitochondria with enhanced activity from *Ashitaba* extract can maintain their function and activity under stress, ensuring normal cell function without adverse effects from external or internal stress. In addition, *Ashitaba* extract also has effects such as inhibiting tumor growth, improving inflammation, obesity, diabetes and hypertension, anti-ulcer, anti-aging, and lowering blood pressure, blood lipids, blood sugar, and cholesterol.

Claims

1. The use of an extract in the preparation of a non-pharmaceutical composition for enhancing mitochondrial activity in skeletal muscle cells, characterized in that, The extract is an Ashitaba extract, which is derived from Angelica dahurica. The Ashitaba extract was obtained by crushing Ashitaba leaves into powder, soaking them in water at a ratio of 1 gram of powder to 25 milliliters of water for 1 day at room temperature, centrifuging the solution, and freeze-drying the resulting supernatant.

2. The use according to claim 1, wherein enhancing mitochondrial activity in skeletal muscle cells comprises reducing hydrogen ion leakage from mitochondria.

3. The use according to claim 1, wherein enhancing mitochondrial activity in skeletal muscle cells comprises enhancing the mitochondrial capacity for adenosine triphosphate (ATP) production.

4. The use according to claim 1, wherein enhancing mitochondrial activity in skeletal muscle cells comprises enhancing the pre-existing oxygen consumption capacity of mitochondria.

5. The use according to claim 1, wherein enhancing mitochondrial activity in skeletal muscle cells comprises enhancing the mitochondrial maximum oxygen consumption capacity.

6. The use according to claim 1, wherein improving mitochondrial activity in skeletal muscle cells comprises improving the mitochondrial adenosine triphosphate coupling efficiency.

7. The use according to claim 1, wherein enhancing mitochondrial activity in skeletal muscle cells comprises enhancing the bioenergy health index of mitochondria.

8. The use according to claim 1, wherein the Ashitaba extract comprises xanthotoxin, angelicol, hydrodryline, and laserpitin.

9. The use according to claim 1, wherein the concentration of the Ashitaba extract is from 250 μg / mL to 1000 μg / mL.

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