TWI830057B - Use of ramie extract in preparing pharmaceutical or non-pharmaceutical composition for improving the activity of mitochondria and improving the activity of telomerase - Google Patents

Abstract

The present disclosure provides a use of ramie extract in preparing pharmaceutical or non-pharmaceutical composition for anti-aging. The ramie extract can improve the function and activity of mitochondria and telomerase. The ramie extract can protect mitochondria from oxidative stress so as to maintain the function and activity of mitochondria including a decrease of proton leakage, an increase of ATP production, an increase of spare respiratory capacity, an increase of maximal respiration or an increase of coupling efficiency.

Description

Use of ramie root extract for preparing pharmaceutical or non-pharmaceutical compositions that increase mitochondrial activity and increase telomerase activity

The present invention relates to the use of ramie root extract for preparing anti-aging compositions.

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. Therefore, mitochondria are considered to be closely related to the aging of organisms.

Telomeres are DNA repeating sequences at the ends of eukaryotic chromosomes. Their function is to maintain the integrity of chromosomes and control the cell division cycle. Due to limitations in the DNA replication mechanism, the telomeres at the end of the chromosome cannot be completely replicated during chromosome replication. Once telomeres are exhausted, cells will immediately initiate apoptosis and die.

Telomerase is a complex composed of RNA and protein that can repair and lengthen telomeres so that they are not worn out by cell division. Highly active telomerase is usually only detected in hematopoietic cells, stem cells and germ cells. Therefore, telomerase is also considered to be closely related to the aging of organisms.

Because the status of mitochondria and telomeres is related to the aging of cells and even organisms. Therefore, how to protect and repair mitochondria to maintain mitochondrial function and slow down mitochondrial disintegration, as well as increase telomerase activity to slow down the rate of telomere loss, has become an important anti-aging issue.

Embodiments of the present invention provide the use of ramie root extract to increase mitochondrial activity and the use of ramie root extract to increase telomerase activity to achieve the purpose of anti-aging.

One embodiment of the present invention provides a use of ramie root extract for preparing pharmaceutical or non-pharmaceutical compositions that improve mitochondrial activity.

One embodiment of the present invention provides a use of ramie root extract for preparing pharmaceutical or non-pharmaceutical compositions that increase telomerase activity.

According to the use of ramie root extract provided by embodiments of the present invention for preparing anti-aging pharmaceutical or non-pharmaceutical compositions, it can protect mitochondria from oxidative stress damage to maintain the function and activity of mitochondria, including reducing the Hydrogen ion leakage, increased ability to synthesize adenosine triphosphate, increased pre-stored oxygen consumption capacity, increased maximum oxygen consumption capacity, improved adenosine triphosphate matching efficiency or increased BHI value to achieve anti-aging effects. According to the use of ramie root extract provided in embodiments of the present invention for preparing anti-aging compositions, it can also increase telomerase activity to repair telomeres and slow down the rate of telomere wear, so as to achieve anti-aging effects.

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.

Ramie root is the root of Boehmeria nivea (L.) Gaud, also known as Ramie, a perennial herbaceous plant of the Urticaceae family. In traditional Chinese medicine, it has the effects of stopping bleeding, anti-fetal, and detoxifying. Ramie root mainly contains polyphenols and flavonoids such as p-coumaric acid, caffeic acid, chlorogenic acid, hyperin and rutin.

The following is a brief description of the extraction process of the ramie root extract used in the embodiments of the present invention. First, the ramie root raw material is crushed, soaked in water for 2 hours, and extracted with water for 12 hours at 60°C, three times. Then, it is concentrated 10 times, filtered, spray drying, mixed and powdered, filtered, and encapsulated to complete the ramie root extract. In one embodiment of the present invention, the components of the ramie root extract include p-coumaric acid and caffeic acid, and are stored in a refrigerated and dry environment. When used, the ramie root extract is prepared into a solution with water. In another embodiment of the present invention, the ramie root extract consists essentially of p-coumaric acid and caffeic acid. In other embodiments of the present invention, in addition to p-coumaric acid and caffeic acid, the ramie root extract further contains polyphenols and flavonoid components such as chlorogenic acid, hyperoside, and rutin.

In one embodiment of the present invention, the weight ratio of p-coumaric acid to caffeic acid is 15:1 to 25:1, but is not limited to this. In other embodiments of the present invention, the weight ratio of p-coumaric acid to caffeic acid is 10:1 to 30:1. In still other embodiments of the present invention, the weight ratio of p-coumaric acid to caffeic acid is 19:1 to 21:1.

In one embodiment of the present invention, when ramie root extract is provided to cells at a concentration of 50 μg/ml to 700 μg/ml, the ramie root extract entering the cells can protect and repair the inner membrane of mitochondria to enhance Mitochondrial activity. In this way, the efficiency of the oxidative phosphorylation reaction in the inner mitochondrial membrane to synthesize adenosine triphosphate is improved. In detail, the amount of adenosine triphosphate synthesized by oxidative phosphorylation in mitochondria treated with ramie root extract increased, the basal oxygen consumption of mitochondria increased, and the leakage of hydrogen ions in the mitochondrial inner membrane decreased. The maximum oxygen consumption capacity of the mitochondria is increased, the pre-existing oxygen consumption capacity of the mitochondria is increased, and the adenosine triphosphate matching efficiency of the mitochondria is increased. In a preferred embodiment, the concentration of ramie root extract may be 100 μg/ml to 650 μg/ml. In a more preferred embodiment, the concentration of ramie root extract may be 180 μg/ml to 520 μg/ml. In another more preferred embodiment, the concentration of ramie root extract may be 200 μg/ml to 500 μg/ml.

In another embodiment of the present invention, when ramie root extract is provided to cells at a concentration of 50 μg/ml to 700 μg/ml, the ramie root extract entering the cells can increase the activity of telomerase. In this way, the increased activity of telomerase can repair telomeres, thereby slowing down the rate of telomere wear. In a preferred embodiment, the concentration of ramie root extract may be 100 μg/ml to 650 μg/ml. In a more preferred embodiment, the concentration of ramie root extract may be 180 μg/ml to 520 μg/ml. In another more preferred embodiment, the concentration of ramie root extract may be 250 μg/ml to 500 μg/ml.

Therefore, by providing ramie root extract to cells, mitochondrial activity and telomerase activity can be increased to achieve anti-aging effects.

A method of providing the ramie root extract to the cells is, for example, oral ingestion of the ramie root extract in the form of food. When ramie root extract is provided to cells in the form of food, the effective dose of ramie root 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 order to facilitate oral ingestion of the ramie root extract in an edible form, the ramie root extract can be made into ramie root extract processed products, such as liquid, solid, granular, powder, paste or gel. In some embodiments of the present invention, ramie root extract processed products may also contain other ingredients or additives, such as carriers, diluents, and auxiliaries, without affecting the effects and purposes achieved by 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 ramie root extract composition according to the embodiment of the present invention can be a medical or non-medical composition or a health food. Ramie root extract or a composition containing ramie root extract can also be encapsulated in a capsule to facilitate oral ingestion of the ramie root extract. Ramie root extract or a composition containing ramie root extract can be coated in a hard capsule in the form of dry powder. Ramie root extract or a composition containing ramie root extract can also be coated in a soft capsule in the form of solution, suspension, paste, powder or granule.

Oils and fats used to dissolve or disperse ramie root extract in soft capsules include calyx pear oil, almond oil, linseed oil, cumin oil, perilla oil, olive oil, olive squalene, sweet orange oil, and spinach. 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, bilberry 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, shark fin Alkanes, and hydrogenated compounds of such oils and fats, etc. Among them, borage oil and evening primrose oil contain a large amount of gamma-linolenic acid (GLA). Gamma-linolenic acid is an essential fatty acid for the human body. It can moisturize, promote cell regeneration and increase the activity of brown fat. To promote fat burning function.

In addition, colorants, preservatives, tackifiers, binders, disintegrants, dispersants, stabilizers, gelling agents, antioxidants, surfactants, preservatives, pH adjusters, etc. are added in compliance with government regulations. It can also be added to ramie root extract processed products in accordance with the dosage standards and processing and production requirements stipulated by government agencies.

The following describes the efficacy of pharmaceutical or non-pharmaceutical compositions prepared using the ramie root extract according to embodiments of the present invention for improving mitochondrial activity.

Example 1 is a ramie root extract solution with a concentration of 200 μg/ml, Example 2 is a ramie root extract solution with a concentration of 250 μg/ml, and Example 3 is a ramie root extract solution with a concentration of 500 μg/ml.

The cells used in the following experiments were skeletal muscle cells (C2C12). Use DMEM culture medium containing 10% Fetal Bovine Serum (FBS) for culture. The method of cell subculture is explained below. First, culture the cells to a certain volume, remove the culture medium, and rinse the cells twice with phosphate buffered saline (PBS). Next, add trypsin and react with the cells for 5 minutes at 37°C. Then add culture medium to stop the trypsin action. Then, centrifuge at 300 g (relative centrifugal force, RCF) for 5 minutes, remove the supernatant, and redissolve in culture medium. Finally, the cell count in a cell culture flask (175T flask) was 1 × 10 ⁶ cells.

First, the cytotoxicity of ramie root extract was tested. 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 evaluated by measuring the light absorption value or fluorescence value of rufin. 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 ramie root 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, ramie root extract was added so that the concentration of ramie root extract in the wells was 25, 50, 100, 150, 200, 250, 500, and 1000 μg/ml, and the cells were cultured at 37°C for one day. The ramie root extract used contains p-coumaric acid and caffeic acid, and the weight ratio of p-coumaric acid to caffeic acid is 20.9:1. On the third day, Alamar blue was used to detect cytotoxicity. Specifically, Alma blue was prepared into a 10% (weight percent concentration) solution in a light-proof environment, added to the well plate at a volume of 100 μl per well, and incubated with cells at 37°C. Incubate them together for 3 to 4 hours, and finally measure the absorption value and fluorescence value (OD530/590) with a spectrophotometer (ELISA reader) to obtain the cell survival rate after treatment with ramie root extract to represent the effect of ramie root extract on cells. toxicity.

The following illustrates the experimental results of the cytotoxicity of ramie root extract.

Please refer to Figure 1, which shows the cytotoxicity test results of ramie root extracts at different concentrations. The control group is the cells that have not been treated with ramie root extract. The vertical axis is the cell survival rate relative to the control group. ** (P<0.01) indicates a significant difference relative to the control group.

In the test of cytotoxicity of ramie root extract, as shown in Figure 1, only ramie root extract with a concentration of 1000 μg/ml caused a decrease in cell survival rate, while ramie root extract with a concentration of less than 500 μg/ml had There was no effect on cell viability, indicating that ramie root extract at a concentration below 500 μg/ml is not cytotoxic to cells. Accordingly, 200, 250, and 500 micrograms/ml of ramie root extracts were selected as Examples 1 to 3 of the present invention for subsequent experiments.

Next, experiments using ramie root extract to increase mitochondrial activity were conducted. In the experiment, tert-butyl hydroperoxide (tBHP) was used as a substance that induces cellular oxidative stress damage, aging, and inhibits mitochondrial activity.

The following describes in detail the experimental procedure for using ramie root 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 ramie root extract so that the concentration of ramie root extract in the well is 200, 250, 500 μg/ml and the total volume of the solution in the well is 250 μl. Under these conditions, the cells and ramie root extract Cultivate for a day. On the third day, 100 μM tert-butyl hydroperoxide (tBHP) was added to each well to react with the cells for 1 hour, and then the culture medium in the well plate was replaced with 675 μl of the upper machine culture medium ( DMEM culture medium without fetal bovine serum), after culturing for 1 hour in a carbon dioxide-free incubator, the oxygen consumption of the cells in the wells was measured with a hippocampal bioenergetic 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, a nucleoside triphosphate synthase inhibitor is added to inhibit the mitochondrial production of nucleoside triphosphate. The reduced oxygen consumption at this time is the oxygen consumption for the synthesis of nucleoside triphosphate. Then, an appropriate concentration of anti-coupling agent was added to allow the mitochondria to idling at their limit without damaging the electron transport chain of the mitochondrial inner membrane to evaluate the maximum oxygen consumption capacity of the mitochondria. Finally, an electron transport chain inhibitor is added to completely shut down mitochondrial oxygen consumption, thereby confirming the measured background value, which is non-mitochondrial oxygen consumption. Mitochondrial basal oxygen consumption is equal to the cellular basal oxygen consumption minus non-mitochondrial oxygen consumption. The basal oxygen consumption of mitochondria minus the amount of oxygen consumed for the synthesis of nucleoside triphosphate is equal to the oxygen consumption to overcome the leakage of hydrogen ions. The maximum oxygen consumption of mitochondria minus the basal oxygen consumption of mitochondria is equal to the pre-stored oxygen consumption of mitochondria. Mitochondrial nucleoside triphosphate compatibility efficiency is equal to the oxygen consumption of synthesizing nucleoside triphosphate divided by the basal oxygen consumption of mitochondria.

BHI (Bioenergetic Healthy Index) is an energy metabolism evaluation index calculated by using the Seahorse Bioscience XFe Analyzer to detect the energy metabolism data of mitochondria in cells and using the energy metabolism data of mitochondria as parameters. BHI = [Oxygen consumption for synthesizing nucleoside triphosphate × prestored oxygen consumption]/[Oxygen consumption to overcome hydrogen ion leakage × non-mitochondrial oxygen consumption]. The higher the BHI of the cell, the stronger the function of the mitochondria in the cell, and the more powerful the mitochondria are usually the younger the mitochondria. Therefore, BHI is used as an indicator to evaluate the degree of mitochondrial aging.

The following describes the experimental results of using pharmaceutical or non-pharmaceutical compositions prepared from the ramie root extract of the present invention to improve mitochondrial activity.

Please refer to Figures 2 to 6 and Table 1. Figure 2 shows the experimental results of the oxygen consumption of mitochondria to overcome the leakage of hydrogen ions. Figure 3 shows the experimental results of the oxygen consumption of mitochondria for synthesizing nucleoside triphosphate. Figure 4 is the experimental result of mitochondrial pre-stored oxygen consumption. Figure 5 is the experimental result of mitochondrial maximum oxygen consumption. Figure 6 is the experimental result of mitochondrial adenosine triphosphate compatibility efficiency. Table 1 shows the experimental results of each experiment. data. In Figures 2 to 5, the vertical axis represents the number of picomoles of oxygen consumed per minute. In Figure 6, the vertical axis represents the nexoside triphosphate merging efficiency in percentage (%). The control group is normal cells that have not been treated with tertiary butanol peroxide and ramie root extract. The comparison group is damaged cells treated with 100 μM tertiary butanol peroxide. Experimental groups 1 to 3 are cells treated with 100 μM tertiary butanol peroxide. The damaged cells were treated with 200 μg/ml, 250 μg/ml of Example 2, and 500 μg/ml of Example 3, and then treated with 100 μM of tertiary butanol peroxide. #(P＜0.05), ##(P＜0.01) indicate significant differences compared with the control group, *(P＜0.05), **(P＜0.01), ***(P＜0.001) indicate that the comparison group has significant differences. Significant difference.

Table 1 Ramie root extract (µg/mL) Oxygen consumption to overcome hydrogen ion leakage (picomoles/min) Oxygen consumption for the synthesis of linear triphosphate (pimol/min) Prestored oxygen consumption (pimol/min) Maximum oxygen consumption (pimol/min) Adenosine triphosphate matching efficiency (%) BHI control group - 12.53 73.78 200.25 286.57 85 2.03 comparison group - 30.76 53.13 136.7 220.58 63 1.17 Experimental group one 200 20.07 65.33 178.59 263.99 76 1.6 Experimental group two 250 19.34 67.08 184.05 270.47 77 1.68 Experimental group three 500 15.91 70.31 212.12 298.34 81 1.84

In experiments using ramie root extract to improve mitochondrial activity, mitochondrial function and activity were judged through mitochondrial oxygen consumption. As shown in Figure 2, in terms of the oxygen consumption of mitochondria to overcome hydrogen ion leakage, the comparison group is higher than the control group, while the experimental group treated with the ramie root extract of Examples 1 to 3 is lower than the comparison group and close to the control group. group. As shown in Figure 3, in terms of mitochondrial oxygen consumption for synthesizing nucleoside triphosphate, the comparison group was lower than the control group, while the experimental group treated with the ramie root extract of Examples 1 to 3 was higher than the comparison group and close to the control group. group. As shown in Figure 4, in terms of mitochondrial pre-existing oxygen consumption, the comparison group was lower than the control group, while the experimental group treated with the ramie root extract of Examples 1 to 3 was higher than the comparison group and close to the control group. As shown in Figure 5, in terms of the maximum oxygen consumption of mitochondria, the comparison group was lower than the control group, while the experimental group treated with the ramie root extract of Examples 1 to 3 was higher than the comparison group and close to the control group. As shown in Figure 6, in terms of mitochondrial nucleoside triphosphate compatibility efficiency, the comparison group is lower than the control group, while the experimental group treated with the ramie root extract of Examples 1 to 3 is higher than the comparison group and close to the control group. group. As shown in Table 1, in terms of mitochondrial BHI value, the comparison group was lower than the control group, while the experimental group treated with the ramie root extract of Examples 1 to 3 was higher than the comparison group and close to the control group.

According to the above experimental results, it was shown that the inner membrane of the mitochondria in the comparison group was damaged, requiring more oxygen to be consumed to overcome the leakage of hydrogen ions, and the mitochondria's ability to synthesize adenosine triphosphate, pre-stored oxygen consumption capacity, and maximum oxygen consumption capacity were all reduced. , the matching efficiency of adenosine triphosphate is also reduced. Compared with the comparison group, the activity of mitochondria in the experimental group treated with the ramie root extract of Examples 1 to 3 was increased by the ramie root extract, which is equivalent to the ability of the ramie root extract to protect and repair mitochondria under oxidative stress. Therefore, the inner membrane of mitochondria is less damaged. Smaller damage means that the mitochondria need less oxygen to overcome the leakage of hydrogen ions. Therefore, the proportion of oxygen consumption used to synthesize adenosine triphosphate in the basic oxygen consumption increases, which increases the mitochondrial ability to synthesize adenosine triphosphate. The efficiency is also improved as a result. The maximum oxygen consumption capacity of mitochondria is the maximum output that mitochondria can achieve under oxidative pressure. Compared with the comparison group, the maximum oxygen consumption of mitochondria in the experimental group was treated with the ramie root extract of Examples 1 to 3. The ability was improved and was close to the maximum oxygen consumption capacity of normal cells in the control group. The difference between the maximum oxygen consumption and the basal oxygen consumption is the pre-stored oxygen consumption. Compared with the comparison group, the pre-stored oxygen consumption capacity of mitochondria treated with the ramie root extract of the embodiment in the experimental group has improved and is close to The pre-existing oxygen consumption capacity of normal cells in the control group represents the increased ability of mitochondria to cope with oxidative stress when it is subjected to oxidative stress. An increase in the BHI value represents an increase in the overall anti-aging ability of the mitochondria. In summary, in cells treated with the ramie root extract according to the embodiment of the present invention, the mitochondrial ability to synthesize adenosine triphosphate, pre-existing oxygen consumption capacity, and maximum oxygen consumption capacity are all improved, and the adenosine triphosphate merging efficiency and BHI value are also improved.

The experimental procedure for improving telomerase activity using ramie root extract is described in detail below. On the first day, cells were cultured in 6-well plates at 200,000 cells per well for one day. The next day, ramie root extract was added so that the concentration of ramie root extract in the wells was 250 or 500 μg/ml, and the cells were cultured at 37°C for one day. On the third day, use TeloTAGGG™ Telomerase PCR ELISA kit (Roche) to collect cellular RNA, and perform polymerase chain reaction (PCR) according to the instructions attached to TeloTAGGG™ Telomerase PCR ELISA kit (Roche). The reaction result The cDNA was stored in a 4°C refrigerator. On the fourth day, Telomerase activity was measured using the TeloTAGGG™ Telomerase PCR ELISA kit. Specifically, PCR was first used to amplify the telomeric sequence of TAGGG as a PCR product. Then, use the hybridization probe and antibody attached to the TeloTAGGG™ Telomerase PCR ELISA kit to react with the PCR product and develop color, and measure the absorbance value at 450 nm. The larger the absorbance value, the more telomere sequences protected by telomerase, and the stronger the activity of telomerase.

The following describes the experimental results of using pharmaceutical or non-pharmaceutical compositions prepared from the ramie root extract of the present invention to improve telomerase activity.

Please refer to Figure 7 and Table 2. Figure 7 shows the experimental results of telomerase activity. In Figure 7, the vertical axis is the multiple of the absorbance value compared to the control group to represent the multiple of the telomerase activity compared to the control group. *(P＜0.05), **(P＜0.01) indicate significant differences compared with the control group. The control group is cells that have not been treated with ramie root extract. Experimental groups 4 to 5 are cells that have been treated with ramie root extract of 250 μg/ml in Example 2 and 500 μg/ml in Example 3, respectively.

Table 2 Ramie root extract (µg/mL) Telomerase activity (times) control group - 1 Experimental group four 250 2.461 Experimental group five 500 5.388

According to the above experimental results, compared with the control group, the telomerase activity of the experimental group treated with the ramie root extract of Example 2 and Example 3 was increased, indicating that the ability of telomerase to repair telomeres was improved, which in turn can delay The rate of telomere loss.

According to the use of ramie root extract provided by embodiments of the present invention for preparing anti-aging pharmaceutical or non-pharmaceutical compositions, it can protect mitochondria from oxidative stress damage to maintain the function and activity of mitochondria, including reducing the Hydrogen ion leakage, increased ability to synthesize adenosine triphosphate, increased pre-stored oxygen consumption capacity, increased maximum oxygen consumption capacity, improved adenosine triphosphate matching efficiency or increased BHI value to achieve anti-aging effects. According to the use of ramie root extract provided in embodiments of the present invention for preparing anti-aging compositions, telomerase activity can also be increased to repair telomeres and slow down the rate of telomere wear to achieve anti-aging effects.

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 shows the cytotoxicity test results of ramie root extracts at different concentrations. Figure 2 shows the experimental results of mitochondria overcoming the oxygen consumption of hydrogen ion leakage. Figure 3 shows the experimental results of oxygen consumption for mitochondrial synthesis of nucleoside triphosphate. Figure 4 shows the experimental results of mitochondrial pre-existing oxygen consumption. Figure 5 shows the experimental results of the maximum oxygen consumption of mitochondria. Figure 6 shows the experimental results of mitochondrial adenosine triphosphate compatibility efficiency. Figure 7 shows the experimental results of telomerase activity.

Claims (8)

A use of a ramie root extract for preparing pharmaceutical or non-pharmaceutical compositions that improve mitochondrial activity. The ramie root extract is obtained from the roots of ramie (Boehmeria nivea (L.) Gaud), wherein the ramie root extract is The root of the ramie was crushed, soaked in water, and extracted with water at 60° C. for 12 hours. A use of a ramie root extract for preparing pharmaceutical or non-pharmaceutical compositions that improve telomerase activity. The ramie root extract is obtained from the roots of ramie (Boehmeria nivea (L.) Gaud), wherein the ramie root extract is The root of the ramie was crushed, soaked in water, and extracted with water at 60° C. for 12 hours. The use as described in claim 1, wherein the ramie root extract reduces mitochondrial hydrogen ion leakage, improves mitochondrial ability to synthesize adenosine triphosphate, increases mitochondrial pre-existing oxygen consumption capacity, and increases mitochondrial maximum consumption. Oxygen capacity may improve mitochondrial adenosine triphosphate compatibility efficiency. The use as described in claim 3, wherein the ramie root extract increases the BHI value (Bioenergetic Healthy Index). The use as described in claim 1 or 2, wherein the mitochondria are mitochondria of skeletal muscle cells. The use as described in claim 1 or 2, wherein the ramie root extract contains p-coumaric acid and caffeic acid. The use as described in claim 6, wherein the weight ratio of p-coumaric acid to caffeic acid is 10:1 to 30:1. The use as described in claim 1 or 2, wherein the concentration of the ramie root extract is 50 micrograms/ml to 700 micrograms/ml.

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