CN111166773A - Application of extract of Cordyceps sinensis in improving gene expression and resisting aging - Google Patents
Application of extract of Cordyceps sinensis in improving gene expression and resisting aging Download PDFInfo
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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- A61K2236/30—Extraction of the material
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- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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Abstract
The invention relates to the field of plant extracts, in particular to application of an extract of cordyceps sinensis in improving gene expression and resisting aging. The extract of the cordyceps sinensis can improve the activity of telomerase by improving the gene expression quantity of telomerase reverse transcriptase genes and telomerase RNA components, thereby achieving the anti-aging effect.
Description
Technical Field
The invention relates to the field of plant extracts, in particular to application of an extract of cordyceps sinensis (Hirsutella sinensis) in improving gene expression quantity and resisting aging.
Background
Since ancient mankind pursues the ever-standing and long-lived way of youth, with the development and progress of medical science and biotechnology in recent years, not only can the disease be combated by the medical technology at the front end, but also many products requiring anti-aging are continuously researched and developed, in recent years, the anti-aging tide slowly spreads to the world, for example, japan, people up to seven or more have anti-aging consciousness, and the concern of taiwan people on anti-aging is continuously increased, the anti-aging trend drives the market sale of anti-aging related products, and the global market is expected to be continuously expanded.
Telomerase (telomerase) is a ribonucleoprotein complex composed of RNA and protein, and is closely related to the regulation mechanism of telomeres. Telomerase can prolong telomere repair ability, reduce loss of telomeres due to cell division, and increase cell division and replication frequency. Telomerase activity is also associated with cell proliferation and survival, cells continue to lose telomerase activity, and during DNA repair, telomeric DNA is also lost, thus telomerase activity is associated with cell aging.
The most common means currently used to delay the aging problem is by means of cosmetics, care products applied to the skin surface, or health foods declared to have anti-aging effect orally. However, conventional cosmetics, health products and health foods are mostly made of chemical components, and are not harmful to human health even after long-term use, and these products are often expensive and not affordable for general users.
In order to solve the above problems, those skilled in the art need to develop a novel pharmaceutical composition or food product with the effects of promoting telomerase activity and anti-aging to benefit the broad population in need thereof.
Disclosure of Invention
In view of the above, the present invention provides a use of an extract of cordyceps sinensis (Hirsutella sinensis) for preparing a composition for increasing the expression level of telomerase reverse transcriptase (TERT) gene and telomerase RNA component (TERC) gene.
Another object of the present invention is to provide a use of an extract of Cordyceps sinensis for preparing an anti-aging composition.
In one embodiment of the present invention, the extract of Cordyceps sinensis is obtained by extracting Cordyceps sinensis with water, alcohols, aqueous alcohols or a combination thereof as an extraction solvent.
In an embodiment of the present invention, the volume ratio of the cordyceps sinensis to the extraction solvent is 1-5: 5-20.
In one embodiment of the present invention, the temperature of the extraction is between 60 ℃ and 90 ℃.
In one embodiment of the present invention, the composition is a pharmaceutical composition or a food product.
In conclusion, the extract of cordyceps sinensis has the following effects: the telomerase activity can be improved by improving the expression of telomerase reverse transcriptase gene and telomerase RNA component gene, thereby achieving the anti-aging effect.
The following examples are presented to illustrate the present invention and are not to be construed as limiting the scope of the invention, which is intended to be limited only by the appended claims.
Drawings
FIG. 1 is a graph showing the efficacy of the extract of Cordyceps sinensis berk of the present invention in regulating the expression of telomerase reverse transcriptase (TERT) gene, wherein ". x" indicates that p is < 0.05 compared to a control group;
FIG. 2 is a data diagram showing the efficacy of the extract of Cordyceps sinensis of the present invention in promoting the expression of the RNA component (TERC) gene of telomerase.
Detailed Description
Definition of
As used herein, the numerical values are approximations and all numerical data are reported to be within the 20 percent range, preferably within the 10 percent range, and most preferably within the 5 percent range.
According to the invention, the cordyceps sinensis (Hirsutella sinensis) is also called cordyceps sinensis, cordyceps sinensis or Hirsutella sinensis, is a fungal organism of Hirsutella (Hirsutella) in Clavicipitaceae, and is also a traditional rare Chinese medicinal material in Chinese history. Cordyceps sinensis is mainly produced in alpine zone and snowy mountain grassland in Qinghai, Tibet, Sichuan, Yunnan, Gansu, Guizhou province and autonomous region in China. Cordyceps sinensis is known to be used for anti-tumor, blood sugar lowering, blood fat lowering, improvement of chronic kidney disease and liver protection.
As used herein, the term "anti-aging" means preventing, slowing down the appearance of aging in human skin, such as: the generation of wrinkles and loss of elasticity. The degree of evaluation to achieve this will be determined by a number of factors known to those skilled in the art, such as the general state of the consumer, age, sex, and the like.
In accordance with the present invention, the pharmaceutical composition may be formulated into a dosage form suitable for parenteral (parenteral) or oral (oral) administration using techniques well known to those skilled in the art, including, but not limited to, injections (e.g., sterile aqueous solutions (solutions) or dispersions (dispersions)), sterile powders (tablets), troches (troche), pills (pill), capsules (capsules), and the like.
According to the present invention, the pharmaceutical composition may be administered parenterally in a route selected from the group consisting of: intraperitoneal injection (intraperitoneal injection), subcutaneous injection (subcutaneous injection), intramuscular injection (intramuscular injection), intravenous injection (intravenous injection), sublingual administration (sublingual administration), and transdermal administration (transdermal administration).
According to the present invention, the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier (pharmaceutically acceptable carrier) which is widely used in pharmaceutical manufacturing technology. For example, the pharmaceutically acceptable carrier may comprise one or more agents selected from the group consisting of: solvents (solvent), buffers (buffer), emulsifiers (emulsifying), suspending agents (suspending agent), disintegrating agents (disintegrant), disintegrating agents (disintegrating agent), dispersing agents (dispersing agent), binding agents (binding agent), excipients (excipient), stabilizers (stabilizing agent), chelating agents (chelating agent), diluents (diluent), gelling agents (gelling agent), preservatives (preserving), wetting agents (wetting agent), lubricants (lubricating), absorption delaying agents (absorption delaying agent), liposomes (liposome) and the like. The selection and amounts of such agents are within the skill and routine skill of those skilled in the art.
According to the present invention, the pharmaceutically acceptable carrier comprises a solvent selected from the group consisting of: water, normal saline (normal saline), Phosphate Buffered Saline (PBS), aqueous alcohol-containing solutions (aqueous solution linking alcohol), and combinations thereof.
According to the invention, the composition can be used as a food additive (food additive) to be added during the preparation of the raw material or during the preparation of food by conventional methods, and formulated with any edible material into a food product for human and non-human animal consumption.
According to the present invention, the types of food products include, but are not limited to: beverages (leafages), fermented foods (fermented foods), bakery products (bakery products), health foods (health foods) and dietary supplements (dietary supplements).
Example 1 preparation of extract of Cordyceps sinensis
Firstly, homogenizing cordyceps sinensis, and then extracting the homogenized cordyceps sinensis for 1-3 hours by using water, alcohols, aqueous alcohols or a combination of the water, the alcohols and the aqueous alcohols as an extraction solvent, wherein the extraction temperature is 60-90 ℃, and the volume ratio of the extraction solvent to the cordyceps sinensis is 5-20: 1-5. The resulting product was then centrifuged, and the centrifuged supernatant was collected and filtered to obtain a filtrate. Then, the filtrate is concentrated under reduced pressure at 45 to 70 ℃ to obtain the extract of cordyceps sinensis of the present invention.
Example 2 evaluation of Effect of extract of Cordyceps sinensis on increasing expression level of telomerase reverse transcriptase (TERT) gene and telomerase RNA component (TERC) gene and anti-aging
First, human fibroblast cells CCD966SK were cultured in 6-well plates in MEM medium supplemented with 10% FBS, 0.1mM non-essential amino acids, 1.5g/L sodium bicarbonate, 1mM sodium pyruvate at a cell concentration of 1.5X 10 in 2mL of medium5Cells/well, followed by culture at 37 ℃.
Thereafter, the cultured cells were divided into 3 groups including 1 control group and 2 experimental groups (i.e., experimental group 1 and experimental group 2). The aqueous extract of Cordyceps sinensis at a concentration of 0.0625mg/mL was added to the cells of experimental group 1, and the ethanol extract of Cordyceps sinensis at a concentration of 0.0625mg/mL was added to the cells of experimental group 2, and the cells of the control group were not treated. After 24 hours of culture, each cell culture was harvested and used for gene expression analysis.
In this example, the genes analyzed for telomerase activity include telomerase reverse transcriptase (TERT) gene and telomerase RNA component (TERC) gene.
The respective cell cultures obtained above were subjected to RNA extraction using RNA extraction kit (Geneaid). Then, in order
III reverse transcriptase (II)
III ReverseTranscriptase) (Invitrogen) reverse transcribes the extracted RNA to cDNA. Subsequently, using cDNA as a template (template) and TERT, TERC and GAPDH (as internal control groups) as primer pairs for amplifying a target gene, the nucleotide sequences of which are shown in Table 1 below, quantitative real-time polymerase chain reaction (hereinafter referred to as quantitative real-time PCR) was performed using KAPA CYBR FAST qPCR kit (2x) (KAPAbiosystems) to amplify and quantify the target gene. The melting curve (melting curve) of the PCR product was confirmed during the quantitative real-time PCR reaction.
TABLE 1
The relative expression of the target gene is derived from equation 2-ΔΔCtAnd the cycle threshold [ cycle threshold (C) of the GAPDH gene (as an internal control group) and the reference gene (reference gene) was usedt)value]And by standard deviation (standard deviation)on, STDEV) to calculate the relative multiple change, where Δ Ct ═ CtTarget/reference genes-CtGAPDH，ΔΔCt＝ΔCtTarget gene-ΔCtReference geneMultiple change is 2-ΔΔCt Mean value of. The expression level of the target gene in the control group was used as a comparative reference for 1. Statistically significant differences between groups were determined by the single tailed Stirling t-assay (single tailed student's t-test). The results of this example are shown in FIGS. 1 and 2.
FIG. 1 is a data diagram showing the efficacy of the extract of Cordyceps sinensis berk of the present invention in regulating the expression of telomerase reverse transcriptase (TERT) gene. FIG. 2 is a data diagram showing the efficacy of the extract of Cordyceps sinensis of the present invention in promoting the expression of the RNA component (TERC) gene of telomerase. As can be seen from FIG. 1, the relative expression level of TERT gene in the test group 1 was significantly increased compared to the control group. As can be seen from FIG. 2, the relative expression level of TERC gene was significantly increased in both experiment 1 and experiment 2 compared to the control. The results of this example show that the extract of Cordyceps sinensis of the present invention can achieve anti-aging effect by controlling the expression of genes related to telomerase activity.
In conclusion, the extract of Cordyceps sinensis of the present invention can increase telomerase activity by increasing the expression level of telomerase reverse transcriptase gene and telomerase RNA component gene, thereby achieving the anti-aging effect.
The foregoing is by way of example only, and not limiting. It is intended that all equivalent modifications or variations without departing from the spirit and scope of the present invention shall be included in the appended claims.
Claims (10)
1. Use of Cordyceps (Hirsutella sinensis) extract in preparing a composition for increasing expression level of telomerase reverse transcriptase (TERT) gene and telomerase RNA component (TERC) gene is provided.
2. The use of claim 1, wherein the extract of Cordyceps sinensis is obtained by extracting Cordyceps sinensis with water, alcohols, aqueous alcohols or a combination thereof as an extraction solvent.
3. The use of claim 2, wherein the volume ratio of the cordyceps sinensis to the extraction solvent is 1-5: 5-20.
4. Use according to claim 2, characterized in that the temperature of the extraction is between 60 ℃ and 90 ℃.
5. The use according to claim 1, wherein the composition is a pharmaceutical composition or a food product.
6. Use of extract of Cordyceps (Hirsutella sinensis) for preparing an anti-aging composition is provided.
7. The use of claim 6, wherein the extract of Cordyceps sinensis is obtained by extracting Cordyceps sinensis with water, alcohols, aqueous alcohols or a combination thereof as an extraction solvent.
8. The use of claim 7, wherein the volume ratio of the Cordyceps sinensis to the extraction solvent is 1-5: 5-20.
9. Use according to claim 7, characterized in that the temperature of the extraction is between 60 ℃ and 90 ℃.
10. The use according to claim 6, wherein the composition is a pharmaceutical composition or a food product.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| TW107140280A TW202017577A (en) | 2018-11-13 | 2018-11-13 | Use of extract of hirsutella sinensis for inducing expression of telomerase reverse transcriptase gene and telomerase rna component gene, and anti-aging |
| TW107140280 | 2018-11-13 |
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| CN111166773A true CN111166773A (en) | 2020-05-19 |
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| CN106511415A (en) * | 2010-05-24 | 2017-03-22 | Nse产品公司 | Oral formulations for counteracting effects of aging |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106511415A (en) * | 2010-05-24 | 2017-03-22 | Nse产品公司 | Oral formulations for counteracting effects of aging |
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