TWI820598B - Use of extract in manufacturing a pharmaceutical or non-pharmaceutical composition for enhancing the activity of mitochondria - Google Patents

Abstract

The embodiment according to the present disclosure provides a use of extract in manufacturing a pharmaceutical or non-pharmaceutical composition for enhancing the activity of mitochondria. The activity-enhanced mitochondria can maintain its function and activity when encountering stress to ensure normal work of cells without affecting by external or internal stress.

Description

Use of extracts for the preparation of pharmaceutical or non-pharmaceutical compositions that enhance mitochondrial activity

The present invention relates to the use of extracts for preparing pharmaceutical or non-pharmaceutical compositions that improve mitochondrial activity, and in particular to the use of honeysuckle extracts for preparing pharmaceutical or non-pharmaceutical compositions that improve mitochondrial activity.

Mitochondria are the main site for oxidative phosphorylation and synthesis of adenosine triphosphate (ATP) in cells. Since adenosine triphosphate is the energy source for cellular activities, mitochondria are also known as "cell energy factories." In addition to providing energy to cells, mitochondria are also involved in processes such as cell differentiation, cell messaging, and apoptosis, and have the ability to regulate the cell growth cycle.

However, some by-products produced by mitochondria during oxidative phosphorylation reactions are harmful to mitochondria, such as reactive oxygen species (ROS), including superoxide anions (O ₂ •- ), perhydroxyl radical (perhydroxyl radical, HO ₂ •), hydrogen peroxide (hydrogen peroxide, H ₂ O ₂ ), etc. ROS has strong biochemical reactivity and can easily cause oxidative damage to cells or mitochondria. Damaged mitochondria will have adverse effects on cell energy supply and cell growth. Over a long period of time, severely damaged mitochondria will release cytochrome c (Cyt c), apoptotic protease (caspase), proteolytic enzymes and apoptotic protease progenitors (procaspase-2, 3, 8, 9). etc., will trigger mitochondrial disintegration; it will also release apoptosis-related signaling factors, including proteases of the B cell lymphoma/leukemia-2 (Bcl-2) protease family, Apoptotic protease activating factor-1 (Apaf-1), p53 gene and serine protease Omi/HtrA2 (serine protease, Omi/HtrA2), etc., thereby triggering cell apoptosis. Therefore, how to 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 topic.

Embodiments of the present invention provide the use of extracts to increase the activity of mitochondria to enhance the ability of mitochondria to face stress, thereby maintaining the function and activity of mitochondria when facing stress.

Embodiments of the present invention provide the use of an extract for preparing pharmaceutical or non-pharmaceutical compositions that improve mitochondrial activity, wherein the extract is a Lonicera Japonica extract.

The extract provided according to the embodiment of the present invention is used for preparing pharmaceutical or non-pharmaceutical compositions that improve mitochondrial activity, wherein the extract is honeysuckle extract, and the honeysuckle extract can improve the pre-existing oxygen consumption capacity of mitochondria. , maximum oxygen consumption capacity and adenosine triphosphate synthesis capacity, reduce mitochondrial hydrogen ion leakage, improve mitochondrial adenosine triphosphate compatibility efficiency and improve mitochondrial bioenergy health index, mitochondrial activity increased by honeysuckle extract It maintains its function and activity in the face of stress, ensuring that cells function normally without being adversely affected by stress from the outside or inside.

The detailed features and advantages of the present invention are described in detail in the following embodiments. The content is sufficient to enable anyone skilled in the relevant art to understand the technical content of the present invention and implement it accordingly. Based on the content disclosed in this specification, the patent scope and the drawings, , anyone familiar with the relevant arts can easily understand the relevant objectives and advantages of the present invention. The following examples further illustrate the present invention in detail, but do not limit the scope of the present invention in any way.

Honeysuckle ( Lonicera Japonica ) is a perennial semi-evergreen vine of the genus Lonicera in the Caprifoliaceae family. It is distributed in the temperate and subtropical regions of North America, Europe, Asia and northern Africa. The buds and petals of honeysuckle contain chlorogenic acid, isochlorogenic acid, ginnol, β-sitosterol, stigmasterol, and β-sitosterol. -D-glucoside (β-sitosterol-β-D-glucoside), stigmasterol-D-glucoside (stigmasterol-D-glucoside) and other ingredients, as well as volatile oil ingredients, including linalool (linalool), cis-2, 6,6-trimethyl-2-vinyl-5-hydroxytetrahydrofuran (cis-2,6,6-trimethyl-2-vinyl-5-hydroxytetrahydrofuran), ethylpalmitate, 1,1- Bicyclohexane (1,1-bicyclohexyl), 3-methyl-2-(2-pentenyl)-2-cyclopenten-1-one (3-methyl-2-(2-pentenyl)- 2-cyclopenten-1-one), trans-trans-farnesol, ethyllinolemate, β-cubebene, cis-3-hexane En-1-ol (cis-3-hexen-1-ol), α-terpineol (α-erpineol), benzylbenzoate (benzylbenzoate), 2-methyl-1-butanol (2-methyl- 1-butanol), benzyl alcohol, phenethyl alcohol, cis-linalool oxide, eugenol, carvacrol.

The main active ingredient of honeysuckle is chlorogenic acid. Its biological activities include inhibiting hyaluronidase and glucose-6-phosphatase, scavenging free radicals, resisting lipid peroxidation, anti-mutagenic, hepatoprotective, choleretic, antibacterial, and antiviral. and antispasmodic effects. In traditional Chinese medicine, honeysuckle has the functions of clearing away heat and detoxifying, cooling and dissipating wind and heat, and is used for carbuncles, sore throat, erysipelas, dysentery, wind-heat colds, plague and other symptoms. It has antibacterial, anti-inflammatory, antiviral, Pharmacological effects such as lowering blood lipids.

An extract according to one embodiment of the present invention is a honeysuckle extract, which can be obtained by extracting the petals of honeysuckle with water and drying them. Specifically, first, wash the honeysuckle petals three times with water. Next, extract the petals of honeysuckle three times with water at 60°C to 80°C. The volume ratio of petals to water is about 1:3, and each extraction takes about three hours. The extracted solution was cooled to room temperature. Next, the cooled extraction solution was sterilized and spray-dried at 80°C for 20 hours to obtain a powdered extract. Finally, filter the powdered extract using an 80-mesh screen to obtain honeysuckle extract with a yellow-brown powder appearance. In other embodiments, the honeysuckle extract can also be extracted by using other polar solvents, or the petals can be chopped or ground before extraction.

The honeysuckle extract obtained by water extraction in the above manner contains chlorogenic acid. The concentration of chlorogenic acid may be 20% to 30% by weight. In some embodiments, the weight percentage concentration of chlorogenic acid in the honeysuckle extract may be 25%. In other embodiments, the honeysuckle extract extracted with water may include a weight concentration of about 25% chlorogenic acid, a weight concentration of about 17.7% secologanic acid, and a weight concentration of about 25%. About 0.5% Galuteolin, use HPLC to analyze this honeysuckle extract, you can get Table 1 and Figure 1. Figure 1 is the HPLC chromatogram of the honeysuckle extract, 1 to 3 in Table 1 correspond to Figure 1 The signal peaks are 1~3. Using an infrared spectrometer to analyze the honeysuckle extract and processing it in the Savitsky-Golay smoothing mode, Figure 2 can be obtained. Figure 2 is the near-infrared spectrum of the honeysuckle extract processed in the Savitsky-Golay smoothing mode.

Table 1 No. Element average concentration weight percentage 1 strychnosic acid 0.3795 mg/mL 17.7% 2 Chlorogenic acid 0.5340 mg/mL 25% 3 Luteolin 10.69 µg/mL 0.5%

In some embodiments of the present invention, providing honeysuckle extract with a concentration of 200 μg/ml (μg/mL) to 500 μg/ml to cells can increase mitochondrial activity, specifically by increasing mitochondrial activity. Pre-stored oxygen consumption capacity, maximum oxygen consumption capacity and adenosine triphosphate synthesis ability, reduce mitochondrial hydrogen ion leakage, improve mitochondrial adenosine triphosphate compatibility efficiency and improve mitochondrial bioenergy health index. In another embodiment, the concentration of the honeysuckle extract may be 200 μg/ml to 250 μg/ml. In other embodiments, the concentration of the honeysuckle extract may be from 250 μg/ml to 500 μg/ml.

A method of providing the honeysuckle extract to the cells is, for example, taking the honeysuckle extract orally in the form of food. When honeysuckle extract is provided to cells in the form of food, the effective dose of honeysuckle extract is 2.162 grams to 5.406 grams. The effective dose here is calculated based on the conversion formula between the effective dose in cell experiments and the kilogram of the human body. The conversion formula is as follows: Human effective dose = Effective dose of cell experiment × mouse weight × conversion factor × human body kilograms. The conversion coefficient is obtained by looking up the conversion coefficient table of dose per kilogram of body weight for animals and humans. When the mouse weighs 20 grams and the human body weighs 60 kilograms, the conversion factor is 9.01. In another embodiment, the effective dosage of honeysuckle extract is 2.162 grams to 2.703 grams. In other embodiments, the effective dosage of honeysuckle extract ranges from 2.703 grams to 5.406 grams.

In order to facilitate the oral intake of honeysuckle extract in an edible form, the honeysuckle extract can be made into a processed honeysuckle extract product such as liquid, solid, granular, powder, paste or gel. In some embodiments of the present invention, honeysuckle extract processed products may also contain other ingredients or additives, such as carriers, diluents, and auxiliaries, without affecting the effects and purposes of the present invention. , excipients or flavoring agents. Excipients can make the preparation convenient and practical, while flavoring agents can enhance the flavor of the preparation.

Examples of excipients include starches such as wheat starch, rice starch, corn starch, potato starch, dextrin, and cyclodextrin; crystalline cellulose; lactose, glucose, sugar, reduced maltose, maltose, fructooligosaccharides, and emulsified oligosaccharides. Sugar alcohols such as sorbitol, erythritol, xylitol, lactitol, mannitol and other sugar alcohols.

Examples of 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 spices such as peach flavor, plum flavor, and yogurt flavor; acetyl sulfonamide Potassium acid, sucralose, erythritol, oligosaccharides, mannose, xylitol, isomerized sugars and other sweeteners; citric acid, malic acid, tartaric acid, gluconic acid and other sour agents; green tea, oolong tea, Various tea ingredients such as Banaba tea, Eucommia ulmoides tea, Tieguanyin tea, Job's tears tea, Aescin tea, wild rice tea, kelp tea, etc.

Furthermore, the honeysuckle extract composition according to the embodiment of the present invention can be a pharmaceutical composition or a non-medical composition, or it can also be a health food. Honeysuckle extract or a composition containing honeysuckle extract can also be encapsulated in a capsule to facilitate oral ingestion of the honeysuckle extract. The honeysuckle extract or the composition containing the honeysuckle extract can be coated in the hard capsule in the form of dry powder, or can be coated in the form of solution, suspension, paste, powder or granule. In soft capsules.

Oils and fats used to dissolve or disperse honeysuckle extract in soft capsules, such as calyx oil, almond oil, linseed oil, cumin oil, perilla oil, olive oil, olive squalene, sweet orange oil, chestnut oil, etc. Orange roughy oil, sesame oil, garlic oil, cocoa butter, pumpkin seed oil, chamomile oil, carrot oil, courgette oil, butter fatty acid, macadamia stone fruit oil, lingonberry 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 oil , peach kernel oil, parsley oil, castor oil, sunflower oil, grape seed oil, borage oil, macadamia walnut oil, meadowfoam oil, 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, Pristane, and hydrogenated compounds of these oils and fats, etc.

In addition, colorants, preservatives, tackifiers, binders, disintegrants, dispersants, stabilizers, gelling agents, antioxidants, surfactants, preservatives, pH adjusters and other additives that comply with the regulations of relevant units The substance can also be added to the processed products of the honeysuckle extract composition in accordance with the dosage standards specified by the relevant units and the needs of processing and production.

The following describes the experiment of using the honeysuckle extract according to the embodiment of the present invention to improve mitochondrial activity. The honeysuckle extract used in the experiment was obtained by extracting the petals of honeysuckle with water and drying them as described above, and the honeysuckle extract at least contains chlorogenic acid. The cells used in the experiment were skeletal muscle cells (C2C12). Cell culture was performed using DMEM culture medium containing 10% Fetal Bovine Serum (FBS). The method of cell subculture is as follows: first, culture the cells to a certain amount and then remove the culture medium, and then rinse the cells twice with phosphate buffered saline (PBS). Next, trypsin was added, and trypsin was allowed to interact with the cells at 37°C for 5 minutes, and then culture medium was added to stop the trypsin action. Next, the solution containing cells was centrifuged at 300 g (relative centrifugal force, RCF) for 5 minutes, the supernatant was removed, and the precipitate was redissolved with culture medium. Finally, the cells were moved to a cell culture flask (175T flask) for storage until subsequent experiments, where the cell count was 1 × 10 ⁶ cells.

[Experiment 1] Cytotoxicity of honeysuckle extract

First, the cytotoxicity test of honeysuckle extract was conducted. Alamar blue is a detection reagent used to detect cell viability. Resazurin in the test kit is a redox indicator, which is a non-toxic, cell membrane-penetrating 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. The survival rate of cells can be assessed 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 survival rate. The higher the cell survival rate, the healthier the cells and the stronger their ability to proliferate. The stronger the cell proliferation ability, the greater the number of cells. Therefore, Alma Blue can be used as an indicator of cytotoxicity to determine cell survival rate and cell proliferation rate.

The following details the testing process for the cytotoxicity of honeysuckle extract. On the first day, the cells were cultured for one day in a 96-well plate with a total volume of cells and culture medium of 200 μl and 10,000 cells per well. The next day, honeysuckle extract was added so that the concentration of honeysuckle extract in each well was 50, 100, 200, 250, 500, and 1000 μg/ml, and the cells were cultured at 37°C for one day. On the third day, Alamar blue was used to detect cytotoxicity. Specifically, Alma blue was prepared into a 10% weight concentration solution in a light-protected environment, and added to the well plate at a volume of 100 μl per well, together with the cells at 37°C. Incubate for 3 to 4 hours, and finally measure the absorbance and fluorescence values (OD530/590) with a spectrophotometer (ELISA reader). The cell survival rate after treatment with honeysuckle extract is obtained in this way, which represents the toxicity of honeysuckle extract to cells.

The experimental results are revealed in Figure 3, which shows the results of the cytotoxicity test of honeysuckle extracts at different concentrations. The control group is the cells that have not been treated with honeysuckle extract, and the vertical axis is the cell survival rate relative to the control group. *** (P<0.001) indicates a very significant difference relative to the control group.

As shown in Figure 3, honeysuckle extracts at concentrations below 500 μg/ml have no effect on cell viability, indicating that honeysuckle extracts at concentrations below 500 μg/ml have no cytotoxicity. Accordingly, honeysuckle extracts at 200, 250, and 500 μg/ml were selected as Examples 1 to 3 of the present invention to conduct subsequent experiments.

[Experiment 2] Honeysuckle extract improves mitochondrial activity

Next, experiments using honeysuckle extract to increase mitochondrial activity were conducted. This experiment uses tert-butyl hydroperoxide (tBHP) as a substance that induces cellular oxidative stress damage, aging, and inhibits mitochondrial activity.

The following is a detailed description of the experimental procedure for using honeysuckle extract to improve mitochondrial activity. On the first day, the cells were cultured for 4 hours in a 24-well seahorse plate with a total volume of cells and culture medium of 100 μl and 25,000 cells per well. Then, 150 μl of culture medium was added and cultured for one day. . The next day, add honeysuckle extract so that the concentration of honeysuckle extract in each well is 200, 250, 500 μg/ml and the total volume of the solution in the well is 250 μl. Under these conditions, cells and honeysuckle are extracted. culture for one day. On the third day, the culture medium was replaced and 100 μM tBHP was added to each well to interact with the cells for 1 hour. Then, the culture medium in the well plate was replaced with 675 μl of upper machine culture medium (DMEM culture medium without fetal bovine serum). ), after culturing for 1 hour in a carbon dioxide-free incubator, measure the oxygen consumption of the cells in the wells with a hippocampal bioenergy meter.

The measurement principle and process of the hippocampal bioenergy meter 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 the mitochondrial production of adenosine triphosphate. The reduced oxygen consumption at this time is the oxygen consumption for the synthesis of adenosine triphosphate (ATP production). Then, an appropriate concentration of anti-coupling agent is added, and the mitochondria are allowed to idling at extreme conditions without damaging the electron transport chain of the mitochondrial inner membrane to evaluate the maximum oxygen consumption (Maximal Respiration) of the mitochondria. Finally, an electron transport chain inhibitor is added to completely shut down mitochondrial oxygen consumption, thereby confirming the measured background value, that is, non-mitochondrial oxygen consumption (Non-mitochondrial Respiration). Mitochondrial basal oxygen consumption (Basal Respiration) is equal to the cell's basal oxygen consumption minus non-mitochondrial oxygen consumption. The basal oxygen consumption of mitochondria minus the oxygen consumption for the synthesis of adenosine triphosphate is equal to the oxygen consumption to overcome hydrogen ion leakage (Proton Leakage). The maximum oxygen consumption of mitochondria minus the basal oxygen consumption of mitochondria is equal to the spare oxygen consumption of mitochondria (Spare Respiratory Capacity). Mitochondrial adenosine triphosphate coupling efficiency (Coupling efficiency) is equal to the oxygen consumption for synthesizing adenosine triphosphate divided by the basal oxygen consumption of the mitochondria.

The experimental results are revealed in Table 2 and Figures 4 to 8. Figure 4 shows the measurement results of the oxygen consumption of mitochondria to overcome the leakage of hydrogen ions. Figure 5 shows the measurement results of the oxygen consumption of mitochondria to synthesize adenosine triphosphate. Figure 6 is the measurement result of mitochondrial pre-stored oxygen consumption, Figure 7 is the measurement result of mitochondrial maximum oxygen consumption, and Figure 8 is the mitochondrial adenosine triphosphate compatibility efficiency. The control group is cells that have not been treated with tBHP and have not been treated with honeysuckle extract. The control group is cells that have been treated with tBHP but have not been treated with honeysuckle extract. The experimental group includes cells that have been treated with the honeysuckle extract of the embodiment and have been treated with tBHP. treated cells. In Figures 4 to 7, the vertical axis represents the number of picomoles of oxygen consumed per minute (pmol/min). In Figure 8, the vertical axis represents the adenosine triphosphate compatibility efficiency in percentage (%). *** (P＜0.001) indicates a very significant difference compared to the control group, ### (P＜0.001) indicates a very significant difference compared to the control group.

Table 2 Honeysuckle Extract basal oxygen consumption Overcome hydrogen ion leakage and oxygen consumption Oxygen consumption for synthesis of adenosine triphosphate maximum oxygen consumption Prestored oxygen consumption non-mitochondrial oxygen consumption Adenosine triphosphate compatibility efficiency unit μg/mL pmol/min % control group - 85.5 ±3.04 11.6 ±2.04 73.9 ±1.55 216.6 ±9.42 131.1 ±9.26 14.5 ±3.04 86.5 ±1.98 control group - 86.1 ±1.13 33.5 ±3.35 52.7 ±2.23 157.6 ±10.83 71.5 ±11.60 13.9 ±1.13 61.2 ±3.39 Embodiment 1 200 85.0 ±2.08 14.2 ±1.45 70.8 ±0.69 228.0 ±7.44 143.0 ±9.17 15.0±2.08 83.4 ±1.32 Embodiment 2 250 85.1 ±1.57 14.2 ±1.12 70.9 ±1.07 230.6 ±18.92 145.5 ±19.27 14.9±1.57 83.4 ±1.09 Embodiment 3 500 83.0 ±1.76 13.1 ±1.27 70.0 ±0.82 249.8 ±9.63 166.7 ±10.20 17.0±1.76 84.3 ±1.24

As shown in Figure 4, in the measurement results of mitochondria overcoming hydrogen ion leakage, the comparison group was higher than the control group, indicating that the inner membrane of the mitochondria in the comparison group was damaged and needed to consume more oxygen to overcome hydrogen ion leakage. . In comparison, the experimental group treated with the honeysuckle extract of Examples 1 to 3 is lower than the comparison group and close to the control group, indicating that the mitochondrial activity is increased by the honeysuckle extract, which is equivalent to the oxidative stress of the honeysuckle extract. It has the function of protecting and repairing mitochondria, so the inner membrane of mitochondria is less damaged.

As shown in Figure 5, in the measurement results of mitochondrial synthesis of adenosine triphosphate, the comparison group was lower than the control group, which means that the ability of mitochondria to synthesize adenosine triphosphate under oxidative stress is reduced, and the energy that mitochondria can produce decreases. In comparison, the experimental group treated with the honeysuckle extract of Examples 1 to 3 is higher than the comparison group and close to the control group, indicating that the activity of mitochondria is improved by the honeysuckle extract, and the ability to synthesize adenosine triphosphate under oxidative stress is also It is improved, so it can produce enough energy for the cells to use, and can maintain a level similar to the control group.

As shown in Figure 6, in the measurement results of pre-stored oxygen consumption, the comparison group was lower than the control group, indicating that the pre-stored oxygen consumption capacity of mitochondria was reduced under oxidative stress. In comparison, the experimental group treated with the honeysuckle extract of Examples 1 to 3 is higher than the comparison group and close to the control group, indicating that the activity of mitochondria is enhanced by the honeysuckle extract and the pre-preservation of mitochondria under oxidative stress. Oxygen consumption capacity was also improved. The increase in pre-stored oxygen consumption capacity indicates that the mitochondria have better adaptability to stress and can be maintained at a level similar to that of the control group.

As shown in Figure 7, in the measurement results of maximum oxygen consumption, the comparison group was lower than the control group, indicating that the maximum oxygen consumption capacity of mitochondria was reduced under oxidative stress. In comparison, the experimental group treated with the honeysuckle extract of Examples 1 to 3 is higher than the comparison group and close to the control group, indicating that the activity of mitochondria is increased by the honeysuckle extract, and the maximum mitochondrial activity under oxidative stress Oxygen consumption capacity was also improved and maintained at a level similar to that of the control group.

As shown in Figure 8, in the results of adenosine triphosphate compatibility efficiency, the comparison group was lower than the control group, while the experimental group treated with the honeysuckle extract of Examples 1 to 3 was higher than the comparison group and close to the control group, indicating that the mitochondria The activity is enhanced by honeysuckle extract, and the mitochondrial adenosine triphosphate compatibility efficiency is also improved under oxidative stress, and can be maintained at a level similar to that of the control group.

According to the measurement results of the hippocampal bioenergetic meter, the Bioenergetic Healthy Index (BHI) can be calculated from the mitochondrial oxygen consumption. The bioenergetic health index is an energy metabolism evaluation index calculated using mitochondrial energy metabolism data as parameters. BHI = log [oxygen consumption to synthesize adenosine triphosphate × prestored oxygen consumption]/[oxygen consumption to overcome hydrogen ion leakage × non-mitochondrial oxygen consumption]. A high bioenergetic health index of a cell means high mitochondrial activity in the cell and a strong ability of the cell to cope with stress. Therefore, the Bioenergetic Health Index can be used as an indicator to evaluate the health of mitochondria and cells.

The biological health energy index calculated from the above experimental results is revealed in Table 3. As shown in Table 3, compared with the control group, treatment with the honeysuckle extract of the embodiment can improve the bioenergetic health index, indicating that the health of mitochondria and cells has been improved.

table 3 Honeysuckle extract (µg/mL) BHI control group - 1.77±0.03154 control group - 0.91±0.08980 Embodiment 1 200 1.68±0.02247 Embodiment 2 250 1.69±0.06771 Embodiment 3 500 1.72±0.03790

It can be seen from the above experimental results that honeysuckle extract with a concentration of less than 500 μg/ml is not toxic to cells. In addition, honeysuckle extract can improve mitochondrial activity, specifically by increasing mitochondrial pre-existing oxygen consumption capacity, maximum oxygen consumption capacity and adenosine triphosphate synthesis capacity, reducing mitochondrial hydrogen ion leakage, and improving mitochondrial activity. The efficiency of adenosine triphosphate compatibility and the improvement of mitochondrial bioenergetic health index.

The extract provided according to the embodiment of the present invention is used for preparing pharmaceutical or non-pharmaceutical compositions that improve mitochondrial activity, wherein the extract is honeysuckle extract, and the honeysuckle extract can improve the pre-existing oxygen consumption capacity of mitochondria. , maximum oxygen consumption capacity and adenosine triphosphate synthesis capacity, reduce mitochondrial hydrogen ion leakage, improve mitochondrial adenosine triphosphate compatibility efficiency and improve mitochondrial bioenergy health index, mitochondrial activity increased by honeysuckle extract It maintains its function and activity in the face of stress, ensuring that cells function normally without being adversely affected by stress from the outside or inside.

Although the present invention is disclosed in the foregoing embodiments, they are not intended to limit the present invention. All changes and modifications made without departing from the spirit and scope of the present invention shall fall within the scope of patent protection of the present invention. Regarding the protection scope defined by the present invention, please refer to the attached patent application scope.

without.

Figure 1 is the HPLC chromatogram of honeysuckle extract.

Figure 2 is the near-infrared spectrum of honeysuckle extract processed by Savitsky-Golay smoothing mode.

Figure 3 shows the results of the cytotoxicity test of honeysuckle extracts at different concentrations.

Figure 4 shows the measurement results of the oxygen consumption of mitochondria to overcome hydrogen ion leakage.

Figure 5 shows the measurement results of oxygen consumption for mitochondrial synthesis of adenosine triphosphate.

Figure 6 shows the measurement results of mitochondrial pre-existing oxygen consumption.

Figure 7 shows the measurement results of the maximum oxygen consumption of mitochondria.

Figure 8 shows the ATP binding efficiency of mitochondria.

without.

Claims (10)

An extract is used to prepare a pharmaceutical or non-pharmaceutical composition for improving mitochondrial activity, wherein the extract is a honeysuckle ( Lonicera Japonica ) extract, and the honeysuckle extract is obtained by dividing the petals of honeysuckle at 60 It is obtained by extracting and drying with water at ℃ to 80℃. The volume ratio of honeysuckle petals to water is 1:3. The use as claimed in claim 1, wherein increasing mitochondrial activity includes increasing the pre-existing oxygen consumption capacity of the mitochondria. The use as described in claim 1, wherein increasing mitochondrial activity includes increasing the maximum oxygen consumption capacity of the mitochondria. The use as claimed in claim 1, wherein increasing mitochondrial activity includes increasing the adenosine triphosphate synthesis ability of the mitochondria. The use as claimed in claim 1, wherein increasing mitochondrial activity includes reducing hydrogen ion leakage from the mitochondria. The use as claimed in claim 1, wherein increasing mitochondrial activity includes increasing the adenosine triphosphate merging efficiency of the mitochondria. The use as claimed in claim 1, wherein increasing mitochondrial activity includes increasing the bioenergetic health index of the mitochondria. The use as described in claim 1, wherein the honeysuckle extract contains chlorogenic acid. The use as described in claim 8, wherein the weight percentage concentration of the chlorogenic acid in the honeysuckle extract is 20% to 30%. The use as claimed in claim 1, wherein in the composition, the concentration of the honeysuckle extract is 200 micrograms/ml to 500 micrograms/ml.