Application of traditional Chinese medicinal material extract in preparation of medicine or non-medicine composition for improving mitochondrial activity

Abstract

The invention relates to application of a traditional Chinese medicinal material extract in preparation of a medicine or a non-medicine composition for improving mitochondrial activity, and belongs to the technical field of medicine. The invention provides application of a traditional Chinese medicinal material extract in preparation of a medicine or a non-medicine composition for improving mitochondrial activity. By reducing the hydrogen ion leakage of the mitochondria, improving the triphosadenine synthesis capability of the mitochondria, improving the pre-storage oxygen consumption capability of the mitochondria, improving the maximum oxygen consumption capability of the mitochondria, improving the maximum oxygen consumption proportion of the mitochondria, improving the triphosadenine coupling efficiency of the mitochondria and improving the biological energy health index of the mitochondria, the mitochondria can be improved. Further, the capability of the mitochondria facing the pressure is improved, so that the function and activity of the mitochondria facing the pressure are maintained, and the normal operation of the cells is ensured and is not affected by the adverse effect of the external or internal pressure.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technologies, and particularly to the use of Chinese herbal medicine extracts for preparing pharmaceutical or non-pharmaceutical compositions for enhancing mitochondrial activity. Background Art

[0002] The mitochondrion is the main site for cells to perform oxidative phosphorylation and synthesize adenosine triphosphate (ATP). Since ATP is the energy source for cell activities, the mitochondrion is also known as the "cell energy factory". In addition to providing energy for cells, mitochondria also participate in processes such as cell differentiation, cell signal transduction, and apoptosis, and have the ability to regulate the cell growth cycle.

[0003] However, some by-products generated during the oxidative phosphorylation reaction of mitochondria are harmful to mitochondria. For example, reactive oxygen species (ROS), including superoxide anion (·O 2 - ), perhydroxyl radical (HOO·), hydrogen peroxide (H 2 O 2 ), etc. ROS has strong biochemical reactivity and is prone to cause oxidative damage to cells or mitochondria. Damaged mitochondria will have adverse effects on cell energy supply, cell growth, etc. Over time, severely damaged mitochondria will release cytochrome c (Cyt c), caspases, proteolytic enzymes, and procaspases (procaspase-2, procaspase-3, procaspase-8, procaspase-9), etc., triggering mitochondrial disintegration, and will also release signal transduction factors related to apoptosis, thereby triggering apoptosis. Therefore, how to enhance mitochondrial activity, improve the ability of mitochondria to face stress, protect and repair mitochondria to maintain their functions and slow down mitochondrial disintegration has become an important issue. Summary of the Invention

[0004] The purpose of the present invention is to provide the use of Chinese herbal medicine extracts for preparing pharmaceutical or non-pharmaceutical compositions for enhancing mitochondrial activity, so as to improve the ability of mitochondria to face stress, and then maintain the functions and activities of mitochondria when facing stress, ensuring the normal operation of cells without being adversely affected by external or internal stress.

[0005] To achieve the above-mentioned invention purpose, the present invention provides the following technical solutions:

[0006] The present invention provides the use of a traditional Chinese medicine extract for preparing a pharmaceutical or non-pharmaceutical composition for enhancing mitochondrial activity.

[0007] Preferably, the traditional Chinese medicine extract is composed of Paeonia lactiflora extract, Acanthopanax senticosus extract, and Atractylodes lancea extract.

[0008] Preferably, the enhancement of mitochondrial activity includes one or more of reducing hydrogen ion leakage of mitochondria, enhancing the adenosine triphosphate synthesis ability of mitochondria, enhancing the pre-existing oxygen consumption ability of mitochondria, enhancing the maximum oxygen consumption ability of mitochondria, enhancing the maximum oxygen consumption ratio of mitochondria, enhancing the adenosine triphosphate coupling efficiency of mitochondria, and enhancing the bioenergy health index of mitochondria.

[0009] Preferably, the components of the Paeonia lactiflora extract include paeoniflorin, oxypaeoniflorin, albiflorin, benzoylpaeoniflorin, paeonol, and gallic acid.

[0010] Preferably, the components of the Acanthopanax senticosus extract include triterpenoid saponins, acanthopanaxoside A, acanthopanaxoside B, acanthopanaxoside C, acanthopanaxoside D, acanthopanaxoside E, acanthopanaxoside F, acanthopanaxoside G, polysaccharides, and syringin.

[0011] Preferably, the components of the Atractylodes lancea extract include atractylodin, atractylone, β-eudesmol, and atractylodiene.

[0012] Preferably, the Paeonia lactiflora extract is obtained by concentrating 3.31 parts by weight of raw Paeonia lactiflora medicine with water to 1 part by weight.

[0013] Preferably, the Acanthopanax senticosus extract is obtained by extracting Acanthopanax senticosus with water;

[0014] The volume ratio of the Acanthopanax senticosus to water is 1:10.

[0015] Preferably, the Atractylodes lancea extract is obtained by concentrating 4.76 parts by weight of raw Atractylodes lancea medicine with water to 1 part by weight.

[0016] Preferably, the concentration of the Paeonia lactiflora extract in the pharmaceutical or non-pharmaceutical composition is 250 - 1000 μg / mL.

[0017] Preferably, the concentration of the Acanthopanax senticosus extract in the pharmaceutical or non-pharmaceutical composition is 250 - 1000 μg / mL.

[0018] Preferably, the concentration of the Atractylodes lancea extract in the pharmaceutical or non-pharmaceutical composition is 250 - 1000 μg / mL.

[0019] Preferably, the enhancement of mitochondrial activity is to enhance the mitochondrial activity of skeletal muscle cells.

[0020] The present invention has the following technical effects and advantages:

[0021] Use of the Chinese medicinal material extract of the present invention for preparing a medicament or non-medicament composition for enhancing bioenergy, wherein the Chinese medicinal material extract is composed of Paeonia lactiflora Pall. extract, Acanthopanax senticosus extract, and Atractylodes lancea extract, and the Chinese medicinal material extract can enhance mitochondrial activity, specifically including reducing mitochondrial H + leakage, enhancing the pre-existing oxygen consumption capacity of mitochondria, enhancing the maximum oxygen consumption capacity of mitochondria, enhancing the ATP synthesis capacity of mitochondria, enhancing the ATP coupling efficiency of mitochondria, enhancing the maximum oxygen consumption ratio of mitochondria, and enhancing one or more of the bioenergy health indices. After the mitochondrial activity is enhanced by the Chinese medicinal material extract, it can maintain the function and activity of mitochondria in the face of stress to ensure the normal operation of cells without being adversely affected by external or internal stress. Description of the Drawings

[0022] Figure 1 It is the determination result of the cytotoxicity of the Chinese medicinal material extract on skeletal muscle cells. Among them, A is the determination result of the cytotoxicity of Paeonia lactiflora Pall. extract on skeletal muscle cells, B is the determination result of the cytotoxicity of Acanthopanax senticosus extract on skeletal muscle cells, and C is the determination result of the cytotoxicity of Atractylodes lancea extract on skeletal muscle cells;

[0023] Figure 2 In, A is the determination result of the basal oxygen consumption of mitochondria in skeletal muscle cells treated with Paeonia lactiflora Pall. extract, B is the determination result of the oxygen consumption for overcoming H + leakage of mitochondria in skeletal muscle cells treated with Paeonia lactiflora Pall. extract, C is the determination result of the maximum oxygen consumption of mitochondria in skeletal muscle cells treated with Paeonia lactiflora Pall. extract, and D is the determination result of the pre-existing oxygen consumption of mitochondria in skeletal muscle cells treated with Paeonia lactiflora Pall. extract; in the figure, * indicates a significant difference compared with the control group (*P < 0.05), ** indicates a significant difference compared with the control group (**P < 0.01), *** indicates a highly significant difference compared with the control group (***P < 0.001), # indicating a significant difference compared with the control group ( # P < 0.05), ## indicates a significant difference compared with the control group ( ## P < 0.01), ### indicating a highly significant difference compared with the control group ( ### P < 0.001), the same below;

[0024] Figure 3 In, A is the determination result of the non-mitochondrial oxygen consumption of mitochondria in skeletal muscle cells treated with Paeonia lactiflora Pall. extract, B is the determination result of the oxygen consumption for synthesizing ATP of mitochondria in skeletal muscle cells treated with Paeonia lactiflora Pall. extract, C is the determination result of the ATP coupling efficiency of mitochondria in skeletal muscle cells treated with Paeonia lactiflora Pall. extract, and D is the determination result of the maximum oxygen consumption ratio of mitochondria in skeletal muscle cells treated with Paeonia lactiflora Pall. extract;

[0025] Figure 4 Among them, A is the measurement result of the basal oxygen consumption of skeletal muscle cell mitochondria after being treated with acanthopanax senticosus extract, B is the measurement result of the oxygen consumption for overcoming H + leakage of skeletal muscle cell mitochondria after being treated with acanthopanax senticosus extract, C is the measurement result of the maximum oxygen consumption of skeletal muscle cell mitochondria after being treated with acanthopanax senticosus extract, and D is the measurement result of the pre-existing oxygen consumption of skeletal muscle cell mitochondria after being treated with acanthopanax senticosus extract;

[0026] Figure 5 Among them, A is the measurement result of the non-mitochondrial oxygen consumption of skeletal muscle cell mitochondria after being treated with acanthopanax senticosus extract, B is the measurement result of the oxygen consumption for synthesizing ATP of skeletal muscle cell mitochondria after being treated with acanthopanax senticosus extract, C is the measurement result of the ATP coupling efficiency of skeletal muscle cell mitochondria after being treated with acanthopanax senticosus extract, and D is the measurement result of the maximum oxygen consumption ratio of skeletal muscle cell mitochondria after being treated with acanthopanax senticosus extract;

[0027] Figure 6 Among them, A is the measurement result of the basal oxygen consumption of skeletal muscle cell mitochondria after being treated with atractylodes lancea extract, B is the measurement result of the oxygen consumption for overcoming H + leakage of skeletal muscle cell mitochondria after being treated with atractylodes lancea extract, C is the measurement result of the maximum oxygen consumption of skeletal muscle cell mitochondria after being treated with atractylodes lancea extract, and D is the measurement result of the pre-existing oxygen consumption of skeletal muscle cell mitochondria after being treated with atractylodes lancea extract;

[0028] Figure 7 Among them, A is the measurement result of the non-mitochondrial oxygen consumption of skeletal muscle cell mitochondria after being treated with atractylodes lancea extract, B is the measurement result of the oxygen consumption for synthesizing ATP of skeletal muscle cell mitochondria after being treated with atractylodes lancea extract, C is the measurement result of the ATP coupling efficiency of skeletal muscle cell mitochondria after being treated with atractylodes lancea extract, and D is the measurement result of the maximum oxygen consumption ratio of skeletal muscle cell mitochondria after being treated with atractylodes lancea extract. Detailed implementation mode

[0029] The present invention provides a use of a traditional Chinese medicine extract for preparing a pharmaceutical or non-pharmaceutical composition for improving mitochondrial activity.

[0030] In the present invention, the traditional Chinese medicine extract is composed of paeonia lactiflora extract, acanthopanax senticosus extract, and atractylodes lancea extract. In other embodiments, the traditional Chinese medicine extract may include paeonia lactiflora extract, acanthopanax senticosus extract, or atractylodes lancea extract.

[0031] In the present invention, the improvement of mitochondrial activity includes reducing the hydrogen ions (H +)One or more of leakage, improvement of mitochondrial adenosine triphosphate (ATP) synthesis ability, improvement of mitochondrial pre-existing oxygen consumption ability, improvement of mitochondrial maximum oxygen consumption ability, improvement of mitochondrial maximum oxygen consumption ratio, improvement of mitochondrial ATP coupling efficiency, and improvement of mitochondrial bioenergy health index. After the mitochondrial activity is enhanced by the Chinese herbal medicine extract, it can maintain the function and activity of mitochondria in the face of stress to ensure the normal operation of cells without being adversely affected by external or internal stress.

[0032] In the present invention, the components of the red peony root extract include paeoniflorin, oxypaeoniflorin, albiflorin, benzoylpaeoniflorin, paeonol, and gallic acid.

[0033] In the present invention, the red peony root extract is obtained by concentrating 3.31 parts by weight of raw red peony root to 1 part by weight with water. Specifically, the raw red peony root is soaked in water for 30 min and then heated to 100 °C for concentration until 3.31 parts by weight of raw red peony root is concentrated to 1 part by weight of red peony root extract. In other embodiments, the red peony root extract can be obtained by concentrating 2.979 - 3.641 parts by weight of raw red peony root to 1 part by weight with water.

[0034] In the present invention, red peony root (Radix Paeoniae Rubra) is a plant of the genus Paeonia in the family Paeoniaceae, and its medicinal material is usually the dried root with skin.

[0035] In the present invention, the components of the acanthopanax extract include triterpenoid saponins, eleutheroside A, eleutheroside B, eleutheroside C, eleutheroside D, eleutheroside E, eleutheroside F, eleutheroside G, polysaccharides, and syringin;

[0036] The content ratio of eleutheroside A, eleutheroside B, eleutheroside C, eleutheroside D, eleutheroside E, eleutheroside F, and eleutheroside G is 8:30:10:12:4:2:1.

[0037] In the present invention, the acanthopanax extract is obtained by extracting acanthopanax with water. Specifically, acanthopanax is extracted with water at 50-100°C for 0.5-2 h. In other embodiments, the acanthopanax extract can be obtained by extracting with a solvent that is non-toxic to organisms, and the solvent that is non-toxic to organisms is preferably alcohol or natural plant glycerol;

[0038] The volume ratio of the acanthopanax to the water is 1:10;

[0039] The temperature of the water is preferably 60°C;

[0040] The extraction time is preferably 0.5-1 h.

[0041] In the present invention, acanthopanax (Eleutherococcus senticosus) is a deciduous shrub of the genus Eleutherococcus in the Araliaceae family, and its roots and rhizomes can be used as medicine.

[0042] In the present invention, the components of the atractylodes extract include atractylon, atractylone, β-eudesmol, and atractylodin.

[0043] In the present invention, the atractylodes extract is obtained by adding water to 4.76 parts by weight of crude atractylodes and concentrating it to 1 part by weight. Specifically, the crude atractylodes is soaked in water for 30 min, and then heated to 100°C for concentration until 4.76 parts by weight of the crude atractylodes is concentrated to 1 part by weight of the atractylodes extract. In other embodiments, the atractylodes extract can be obtained by adding water to 4.284-5.236 parts by weight of the crude atractylodes and concentrating it to 1 part by weight.

[0044] In the present invention, atractylodes (Atractylodes lancea) is a perennial herb of the Compositae family, and its rhizome can be used as medicine.

[0045] In the present invention, the concentration of the paeonia extract in the pharmaceutical or non-pharmaceutical composition is 250-1000 μg / mL. In other embodiments, the concentration of the paeonia extract can be 250-500 μg / mL or 500-1000 μg / mL.

[0046] In the present invention, the concentration of the acanthopanax extract in the pharmaceutical or non-pharmaceutical composition is 250-1000 μg / mL. In other embodiments, the concentration of the acanthopanax extract can be 250-500 μg / mL or 500-1000 μg / mL.

[0047] In the present invention, the concentration of the atractylodes extract in the pharmaceutical or non-pharmaceutical composition is 250-1000 μg / mL. In other embodiments, the concentration of the atractylodes extract can be 250-500 μg / mL or 500-1000 μg / mL.

[0048] In the present invention, the method of providing the Chinese medicinal material extract to cells is oral ingestion in an edible manner. Among them, the effective dose of the Paeonia lactiflora extract is 2.703 - 10.812 g. In other embodiments, the effective dose of the Paeonia lactiflora extract can be 2.703 - 5.406 g or 5.406 - 10.812 g; the effective dose of the Eleutherococcus senticosus extract is 2.703 - 10.812 g. In other embodiments, the effective dose of the Eleutherococcus senticosus extract can be 2.703 - 5.406 g or 5.406 - 10.812 g; the effective dose of the Atractylodes lancea extract is 2.703 - 10.812 g. In other embodiments, the effective dose of the Atractylodes lancea extract can be 2.703 - 5.406 g or 5.406 - 10.812 g;

[0049] The effective dose is obtained by conversion according to the conversion formula of the effective dose in cell experiments and human body weight. The conversion formula is: human effective dose = effective dose in cell experiments × mouse body weight × conversion coefficient × human body weight; where the conversion coefficient is obtained by looking up the table of dose conversion coefficients per kilogram of body weight for animals and humans. For example, when the mouse body weight is 20 g and the human body weight is 60 kg, the conversion coefficient is 9.01.

[0050] To facilitate oral ingestion of the Chinese medicinal material extract in an edible manner, the Chinese medicinal material extract can be made into processed products of the Chinese medicinal material extract in liquid, solid, granular, powder, paste or gel form. In some embodiments, without affecting the efficacy and the purpose that can be achieved by the present invention, the processed products of the Chinese medicinal material extract may also contain other components or additives, such as carriers, diluents, adjuvants, excipients or flavoring agents; excipients can make the preparation convenient to use, and flavoring agents can enhance the flavor of the preparation.

[0051] In the present invention, the excipients are starches such as wheat starch, rice starch, corn starch, potato starch, dextrin, cyclodextrin, etc., crystalline cellulose, sugars such as lactose, glucose, granulated sugar, reduced maltose, maltose, fructose, emulsified oligosaccharides, etc., sugar alcohols such as sorbitol, erythritol, xylitol, lactitol, mannitol, etc.

[0052] In the present invention, the flavoring agents are various fruit juice extracts such as longan extract, litchi extract, pomelo extract, etc., various fruit juices such as apple juice, orange juice, lemon juice, etc., various flavors such as peach flavor, plum flavor, yogurt flavor, etc., various sweeteners such as acesulfame potassium, sucralose, erythritol, oligosaccharides, mannose, xylitol, isomerized sugars, etc., various souring agents such as citric acid, malic acid, tartaric acid, gluconic acid, etc., various tea components such as green tea, oolong tea, Banaba tea, Eucommia ulmoides tea, Tieguanyin tea, coix seed tea, gynostemma pentaphyllum tea, water bamboo shoot tea, kelp tea, etc.

[0053] In the present invention, the composition of the Chinese herbal medicine extract may be a pharmaceutical or non-pharmaceutical composition, such as a health food. The Chinese herbal medicine extract or the composition containing the Chinese herbal medicine extract may also be encapsulated in a capsule to facilitate oral intake of the Chinese herbal medicine extract. The Chinese herbal medicine extract or the composition containing the Chinese herbal medicine extract may be encapsulated in a hard capsule in the form of a dry powder, or may be encapsulated in a soft capsule in the form of a solution, suspension, paste, powder or granule.

[0054] In the present invention, the oils and fats used to dissolve or disperse the Chinese herbal medicine extract in the soft capsule are avocado 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, beef fatty acid, macadamia oil, bilberry oil, brown rice germ oil, rice oil, wheat germ oil, safflower oil, shea butter, illipe 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 oil, meadowfoam oil, cottonseed oil, peanut oil, ghost 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, beef tallow, lard, squalene, squalane, pristane, and hydrides of the above oils and fats.

[0055] In addition, additives that comply with the regulations of relevant authorities, such as colorants, preservatives, thickeners, binders, disintegrants, dispersants, stabilizers, gelling agents, antioxidants, surfactants, pH adjusters, etc., may also be added to the processed products of the composition of the Chinese herbal medicine extract of the present invention according to the dosage standards and processing and production requirements stipulated by relevant authorities.

[0056] In the present invention, the improvement of mitochondrial activity is to improve the mitochondrial activity of skeletal muscle cells.

[0057] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0058] In an embodiment of the present invention, the test cells are skeletal muscle cells (C2C12), and DEME culture medium containing 10% fetal bovine serum (FBS) is used as the skeletal muscle cell culture medium (hereinafter simply referred to as the culture medium). The cell replacement method is as follows: After culturing the skeletal muscle cells to a certain amount, the culture medium is removed, and the skeletal muscle cells are rinsed twice with phosphate buffer solution (PBS). Trypsin is added and allowed to act on the skeletal muscle cells at 37 °C for 5 min. Then, the culture medium is added to terminate the action of trypsin to obtain a mixed solution. The mixed solution containing skeletal muscle cells is centrifuged at 300×g for 5 min. After removing the supernatant, the culture medium is added to redissolve the precipitate. Finally, the skeletal muscle cells are transferred to a cell culture flask (175T flask) for storage for subsequent experiments, where the cell count is 1×10 6 cells.

[0059] Experimental Example 1: Cytotoxicity of Chinese herbal medicine extracts

[0060] Alamar blue is a reagent for detecting cell viability. Resazurin in it is a non-toxic, cell membrane-permeable redox indicator in the form of a dark blue dye with low fluorescence. When resazurin enters healthy cells, it will be reduced to pink and highly fluorescent resorufin due to the reducing environment in living cells. Thus, the cell viability is evaluated by measuring the light absorption value or fluorescence value of resorufin produced in the cells. The higher the light absorption value or fluorescence value of resorufin, the higher the cell viability, the healthier the cells, the stronger the proliferation ability, and the more cells that can proliferate. Therefore, Alamar blue can be used as an indicator of cytotoxicity to measure cell viability and cell proliferation rate. Specifically:

[0061] On the first day, 10,000 skeletal muscle cells are seeded into each culture well of a 96-well plate, and the culture medium is added to a total volume of 200 μL, and the skeletal muscle cells are cultured for one day. On the second day, extracts of Paeonia lactiflora, Eleutherococcus senticosus, and Atractylodes lancea are added to each culture well, so that the concentrations of the Chinese herbal medicine extracts in each culture well are 50 μg / mL, 100 μg / mL, 200 μg / mL, 250 μg / mL, 500 μg / mL, and 1000 μg / mL respectively. The culture well without adding any Chinese herbal medicine extract (i.e., the concentration is 0 μg / mL) is used as a control. Then, each 96-well plate is cultured at 37 °C for one day. On the third day, after removing the culture medium, Alamar blue is added for cytotoxicity determination. Specifically: Alamar blue is configured into a 10% Alamar blue solution in a light-proof environment and added to each culture well, 100 μL is added to each culture well. After culturing at 37 °C for 3 h, a spectrophotometer (ELISAreader) is used to measure the light absorption value (OD 530nm) and fluorescence value (OD 595nm ), and obtain the cell viability of skeletal muscle cells in each culture well, which is the cytotoxicity of the Chinese herbal medicine extract to skeletal muscle cells. The results are as Figure 1 shown.

[0062] The results show that the Paeonia lactiflora extract, Eleutherococcus senticosus extract, and Atractylodes lancea extract with concentrations below 1000 μg / mL have no significant effect on the cell viability of skeletal muscle cells, indicating that when the concentration of the Chinese herbal medicine extract is less than 1000 μg / mL, no cytotoxicity is exhibited. Therefore, Chinese herbal medicine extracts with concentrations of 250 μg / mL, 500 μg / mL, and 1000 μg / mL are selected for subsequent experiments.

[0063] Experimental Example 2: Effect of Chinese herbal medicine extract on improving mitochondrial activity

[0064] tert-Butyl hydroperoxide (tBHP) is used as a substance to induce cell oxidative capacity damage, aging, and inhibit mitochondrial activity. Specifically:

[0065] On the first day, 25,000 skeletal muscle cells are seeded in each culture well of a 24-well plate, and the culture medium is added to a total volume of 100 μL. After culturing the skeletal muscle cells for 4 h, 150 mL of culture medium is added, and the cells are cultured for another 1 day. On the second day, the culture wells are divided into Example 1-1 group, Example 1-2 group, Example 1-3 group, Example 2-1 group, Example 2-2 group, Example 2-3 group, Example 3-1 group, Example 3-2 group, and Example 3-3 group. Chinese herbal medicine extract is added to each culture well to a total volume of 250 μL, so that the final concentration of the Paeonia lactiflora extract in the Example 1-1 group is 250 μg / mL, the final concentration of the Paeonia lactiflora extract in the Example 1-2 group is 500 μg / mL, the final concentration of the Paeonia lactiflora extract in the Example 1-3 group is 1000 μg / mL, the final concentration of the Eleutherococcus senticosus extract in the Example 2-1 group is 250 μg / mL, the final concentration of the Eleutherococcus senticosus extract in the Example 2-2 group is 500 μg / mL, the final concentration of the Eleutherococcus senticosus extract in the Example 2-3 group is 1000 μg / mL, the final concentration of the Atractylodes lancea extract in the Example 3-1 group is 250 μg / mL, the final concentration of the Atractylodes lancea extract in the Example 3-2 group is 500 μg / mL, and the final concentration of the Atractylodes lancea extract in the Example 3-3 group is 1000 μg / mL. Each 24-well plate is cultured for one day. On the third day, the fresh culture medium is replaced, and 1000 μmol tBHP is added to each culture well and allowed to act for 1 h. Then, the culture medium in each 24-well plate is replaced with 675 μL of upper machine culture medium (i.e., culture medium without fetal bovine serum with pH = 7.4), and in the absence of CO 2After culturing for 1 h in an incubator, the oxygen consumption of skeletal muscle cells in each culture well was measured with a Seahorse bioenergetics analyzer. The culture wells without the Chinese herbal medicine extract were used as control group 1, control group 2, and control group 3, and the culture wells without the Chinese herbal medicine extract and not treated with tBHP were used as control group 1, control group 2, and control group 3;

[0066] The measurement principle and process of the Seahorse bioenergetics analyzer are as follows: First, the basal oxygen consumption of skeletal muscle cells in each culture well was measured. Then, an ATP synthase inhibitor was added to inhibit the production of ATP by mitochondria. At this time, the decreased oxygen consumption is the oxygen consumption for ATP synthesis (ATP production). Then, an uncoupler was added to make the mitochondria run at maximum capacity without damaging the electron transport chain in the inner mitochondrial membrane to measure the maximum oxygen consumption (maximal respiration) of the mitochondria. Finally, an electron transport chain inhibitor was added to completely shut down the oxygen consumption of the mitochondria, so as to measure the background value and use it as the non-mitochondrial oxygen consumption (non mitochondrial respiration);

[0067] Basal respiration = Basal oxygen consumption - Non-mitochondrial oxygen consumption;

[0068] Overcoming H + Proton leakage = Basal respiration - Oxygen consumption for ATP synthesis;

[0069] Spare respiratory capacity = Maximal respiration - Basal respiration;

[0070] ATP coupling efficiency = Oxygen consumption for ATP synthesis ÷ Basal respiration;

[0071] Relative value of spare respiratory capacity = Maximal respiration ÷ Basal respiration;

[0072]

[0073] The results are shown in Tables 1 - 3 and Figures 2 to 7 as follows.

[0074] Table 1 The effect of Paeonia lactiflora extract on improving mitochondrial activity

[0075]

[0076] Table 2 The effect of Eleutherococcus senticosus extract on improving mitochondrial activity

[0077]

[0078]

[0079] Table 3 Effect of Atractylodes Extract on Improving Mitochondrial Activity

[0080]

[0081] Overcoming H + The measurement results of the oxygen consumption of leakage showed that the mitochondrial overcoming H of the skeletal muscle cells in Control Group 1 to Control Group 3 + The oxygen consumption of leakage was correspondingly higher than that of Control Group 1 to Control Group 3, indicating that the inner mitochondrial membrane of Control Group 1 to Control Group 3 was damaged and more oxygen was needed to overcome H + Leakage. In contrast, after treatment with the Paeonia lactiflora extracts of Example 1-1 to Example 1-3, the Acanthopanax senticosus extracts of Example 2-1 to Example 2-3, or the Atractylodes extracts of Example 3-1 to Example 3-3, the mitochondrial overcoming H + The oxygen consumption of leakage was respectively lower than that of Control Group 1 to Control Group 3, indicating that the activity of mitochondria was enhanced by the Paeonia lactiflora extracts, Acanthopanax senticosus extracts, or Atractylodes extracts. The Chinese herbal medicine extracts could play a role in protecting and repairing mitochondria under oxidative stress. Therefore, the degree of damage to the inner mitochondrial membrane was smaller;

[0082] The measurement results of the maximum oxygen consumption showed that the maximum oxygen consumption of the mitochondria of the skeletal muscle cells in Control Group 1 and Control Group 2 was correspondingly lower than that of Control Group 1 and Control Group 2, indicating that the activity of mitochondria was lower under oxidative stress and the maximum oxygen consumption capacity decreased. In contrast, after treatment with the Paeonia lactiflora extracts of Example 1-1 to Example 1-3 or the Acanthopanax senticosus extracts of Example 2-1 to Example 2-3, the maximum oxygen consumption of the mitochondria was respectively higher than that of Control Group 1 and Control Group 2, indicating that the activity of mitochondria was enhanced by the Paeonia lactiflora extracts or Acanthopanax senticosus extracts, and the maximum oxygen consumption capacity of mitochondria was also enhanced under oxidative stress;

[0083] The measurement results of the pre-stored oxygen consumption showed that the pre-stored oxygen consumption of the mitochondria of the skeletal muscle cells in Control Group 1 and Control Group 2 was correspondingly lower than that of Control Group 1 and Control Group 2, indicating that the activity of mitochondria decreased and the pre-stored oxygen consumption capacity decreased under oxidative stress. In contrast, after treatment with the Paeonia lactiflora extracts of Example 1-1 to Example 1-3 or the Acanthopanax senticosus extracts of Example 2-1 to Example 2-3, the pre-stored oxygen consumption of the mitochondria was respectively higher than that of Control Group 1 and Control Group 2, indicating that the activity of mitochondria was enhanced by the Paeonia lactiflora extracts or Acanthopanax senticosus extracts, and the pre-stored oxygen consumption capacity of mitochondria was also enhanced under oxidative stress, and the stress response ability of mitochondria was better;

[0084] The measurement results of the oxygen consumption for ATP synthesis showed that the oxygen consumption for the leakage of ATP synthesis by mitochondria in skeletal muscle cells of Control Group 1 to Control Group 3 was correspondingly lower than that of Control Group 1 to Control Group 3, indicating that the mitochondrial activity decreased, the ability to synthesize ATP under oxidative stress decreased, and the energy produced decreased. In contrast, after treatment with the Paeonia lactiflora extracts of Example 1-1 to Example 1-3, the Eleutherococcus senticosus extracts of Example 2-1 to Example 2-3, or the Atractylodes lancea extracts of Example 3-1 to Example 3-3, the oxygen consumption for ATP synthesis by mitochondria was correspondingly higher than that of Control Group 1 to Control Group 3, indicating that the mitochondrial activity was enhanced by the Paeonia lactiflora extracts, the Eleutherococcus senticosus extracts, or the Atractylodes lancea extracts, and the ability to synthesize ATP under oxidative stress was also enhanced. Therefore, sufficient energy can be produced for skeletal muscle cells to use;

[0085] The measurement results of the ATP coupling efficiency showed that the ATP coupling efficiency of mitochondria in skeletal muscle cells of Control Group 1 to Control Group 3 was correspondingly lower than that of Control Group 1 to Control Group 3. After treatment with the Paeonia lactiflora extracts of Example 1-1 to Example 1-3, the Eleutherococcus senticosus extracts of Example 2-1 to Example 2-3, or the Atractylodes lancea extracts of Example 3-1 to Example 3-3, the ATP coupling efficiency of mitochondria was correspondingly higher than that of Control Group 1 to Control Group 3, indicating that the mitochondrial activity was enhanced by the Paeonia lactiflora extracts, the Eleutherococcus senticosus extracts, or the Atractylodes lancea extracts, and the ATP coupling efficiency of mitochondria under oxidative stress was also enhanced;

[0086] The measurement results of the maximum oxygen consumption ratio showed that the maximum oxygen consumption ratio of mitochondria in skeletal muscle cells of Control Group 1 and Control Group 2 was correspondingly lower than that of Control Group 1 and Control Group 2. After treatment with the Paeonia lactiflora extracts of Example 1-1 to Example 1-3 or the Eleutherococcus senticosus extracts of Example 2-1 to Example 2-3, the maximum oxygen consumption ratio of mitochondria was correspondingly higher than that of Control Group 1 and Control Group 2, indicating that the mitochondrial activity was enhanced by the Paeonia lactiflora extracts or the Eleutherococcus senticosus extracts, and the maximum oxygen consumption ratio of mitochondria under oxidative stress was also enhanced. Further considering the basal oxygen consumption of mitochondria to reduce the difference in mitochondrial oxygen consumption among different skeletal muscle cells, it can better represent the ability of cells to cope with stress;

[0087] The measurement results of BHI showed that compared with Control Group 1 to Control Group 3, after treatment with the Paeonia lactiflora extracts of Example 1-1 to Example 1-3, the Eleutherococcus senticosus extracts of Example 2-1 to Example 2-3, or the Atractylodes lancea extracts of Example 3-1 to Example 3-3, BHI increased, indicating that the health of mitochondria and skeletal muscle cells was improved.

[0088] As can be seen from the above embodiments, the present invention provides the use of Chinese herbal medicine extracts for preparing pharmaceutical or non-pharmaceutical compositions for enhancing mitochondrial activity. The present invention points out that the extracts of Paeonia lactiflora, Eleutherococcus senticosus, and Atractylodes lancea with a concentration below 1000 μg / mL are not toxic to skeletal muscle cells, and the extracts of Paeonia lactiflora, Eleutherococcus senticosus, or Atractylodes lancea can all enhance mitochondrial activity, specifically including enhancing the pre-existing oxygen consumption capacity, enhancing the maximum oxygen consumption capacity, enhancing the ATP synthesis capacity, reducing H + leakage, enhancing the ATP coupling efficiency, enhancing the maximum oxygen consumption ratio, and enhancing one or more of the bioenergy health index;

[0089] The use of the Chinese herbal medicine extracts of the present invention for preparing pharmaceutical or non-pharmaceutical compositions for enhancing bioenergy, wherein the Chinese herbal medicine extracts are composed of the extracts of Paeonia lactiflora, Eleutherococcus senticosus, and Atractylodes lancea, and the Chinese herbal medicine extracts can enhance mitochondrial activity, specifically including reducing the H + leakage of mitochondria, enhancing the pre-existing oxygen consumption capacity of mitochondria, enhancing the maximum oxygen consumption capacity of mitochondria, enhancing the ATP synthesis capacity of mitochondria, enhancing the ATP coupling efficiency of mitochondria, enhancing the maximum oxygen consumption ratio of mitochondria, and enhancing one or more of the bioenergy health index. Mitochondria whose activity is enhanced by the Chinese herbal medicine extracts can maintain their functions and activities in the face of stress to ensure the normal operation of cells without being adversely affected by external or internal stress.

[0090] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. Use of a Chinese herbal medicine extract for preparing a medical or non-medical composition for improving mitochondrial activity.

2. The use according to claim 1, characterized in that The Chinese medicinal material extract consists of red peony root extract, acanthopanax senticosus extract and atractylodes lancea extract.

3. The use according to claim 1, characterized in that The improvement of mitochondrial activity includes one or more of reducing mitochondrial hydrogen ion leakage, improving mitochondrial adenosine triphosphate synthesis capacity, improving mitochondrial pre-stored oxygen consumption capacity, improving mitochondrial maximum oxygen consumption capacity, improving mitochondrial maximum oxygen consumption ratio, improving mitochondrial adenosine triphosphate coupling efficiency and improving mitochondrial bioenergetic health index.

4. The use according to claim 2, characterized in that: The components of the red peony root extract include paeoniflorin, oxidized paeoniflorin, paeoniflorin, benzoylpeoniflorin, paeonol and gallic acid.

5. The use according to claim 2, characterized in that: The components of the Acanthopanax senticosus extract include triterpenoid saponins, eleutheroside A, eleutheroside B, eleutheroside C, eleutheroside D, eleutheroside E, eleutheroside F, eleutheroside G, polysaccharides and syringin.

6. The use according to claim 2, characterized in that: The components of the atractylodes extract include atractylodesin, atractylodesone, beta-eudesmol and atractylodes furan.

7. The use according to claim 2, characterized in that: The red peony root extract is obtained by adding water to 3.31 parts by weight of the red peony root herb and concentrating it to 1 part by weight.

8. The use according to claim 2, characterized in that: The Acanthopanax senticosus extract is obtained by extracting Acanthopanax senticosus with water; The volume ratio of the Acanthopanax senticosus to water is 1:

10.

9. The use according to claim 2, characterized in that: The Atractylodes lancea extract is obtained by adding water to 4.76 parts by weight of Atractylodes lancea crude drug and concentrating the amount to 1 part by weight.

10. The use according to claim 2, characterized in that: The concentration of the red peony root extract in the medical or non-medical composition is 250-1000 μg / mL.

11. The use according to claim 2, characterized in that: The concentration of the Acanthopanax senticosus extract in the medical or non-medical composition is 250-1000 μg / mL.

12. The use according to claim 2, characterized in that: The concentration of the Atractylodes lancea extract in the medical or non-medical composition is 250 to 1000 μg / mL.

13. The use according to claim 1, characterized in that The improving mitochondrial activity is improving the mitochondrial activity of skeletal muscle cells.

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