阅读说明：本技术 红龙果胚珠萃取物用于制备推迟皮肤老化的组合物的用途 (Use of extract of ovule of mangosteen for preparing composition for delaying skin aging ) 是由 林咏翔 李姿仪 于 2019-01-04 设计创作，主要内容包括：本发明是关于一种植物萃取物的用途,特别是一种红龙果胚珠萃取物用于制备推迟皮肤老化的组合物的用途。该红龙果胚珠萃取物较佳为以水为溶剂对红龙果胚珠进行萃取而制得,其能透过促进皮肤纤维母细胞的增生而提升胶原蛋白分泌,亦能减少胶原蛋白醣化终产物生成,因此有助于维持皮肤的年轻状态。(The invention relates to application of a plant extract, in particular to application of a red dragon fruit ovule extract in preparing a composition for delaying skin aging. The extract of the ovule of the pitaya is preferably prepared by extracting the ovule of the pitaya with water as a solvent, can promote the secretion of collagen by promoting the proliferation of fibroblasts of the skin, and can also reduce the generation of a collagen glycosylation end product, thereby being helpful for maintaining the young state of the skin.)

1. Use of a red dragon fruit ovule extract for the preparation of a composition for delaying skin aging, wherein the red dragon fruit ovule extract is obtained by extracting a red dragon fruit ovule with a solvent.

2. Use according to claim 1, wherein the weight ratio of said solvent to said red dragon fruit ovule is in the range of 10: 1 to 3: 5.

3. use according to claim 1, characterized in that the extraction is carried out at 50 to 100 ℃.

4. Use according to claim 1, characterized in that the solvent is water.

5. The use according to claim 2, wherein the concentration of the extract of the ovule of the red dragon fruit is 0.5mg/mL to 8 mg/mL.

6. The use according to claim 1, wherein the red dragon fruit ovule extract promotes collagen production.

7. The use of claim 1, wherein said red dragon fruit ovule extract promotes proliferation of a dermal fibroblast.

8. The use according to claim 1, wherein the extract of the ovule of the pitaya inhibits collagen glycosylation.

9. The use according to claim 1, wherein the composition is a food product, a beverage product, a nutritional supplement, or a pharmaceutical product.

10. Use according to claim 1, characterized in that said composition has the form of a powder, granules, solution, colloid, or paste.

Technical Field

The invention relates to a use of a plant extract, in particular to a use of an extract of an ovule of pitaya for preparing a composition for delaying skin aging.

Background

Collagen is the most abundant protein in mammals, accounts for about one third of the total protein in human bodies, and is widely present in human skin, cartilage, cornea, blood vessel wall, internal organs, and the like. It is mainly present in the extracellular matrix of connective tissue and provides the mechanical support and strength required by connective tissue. Collagen in the human body can be classified into different types due to the difference in the composition of polypeptides, among which the types of collagen most relevant to skin texture are type i, type iii, and type iv. The dermis contains fibrillar type I and type III collagen, and the basement membrane below the epithelial tissue contains non-fibrillar type IV collagen. It has been shown that aging and environmental adverse factors (e.g., UV light) in an individual can reduce the ability of dermal fibroblasts to produce collagen. In addition, glycosylation (glycation) is also one of the major causes of collagen destruction in the skin. When the collagen is lost at a rate greater than the rate at which it is produced, the skin becomes wrinkled, flaccid, etc.

Glycosylation refers to the slow chemical reaction of sugar molecules attached to proteins or lipids without enzymes, which ultimately results in the formation of glycosylated end products (AGEs). Glycosylation affects collagen aggregation to form fibers, and glycosylated collagen fibers become stiff and brittle. In addition, accumulation of the glycosylated end products in the extracellular matrix of the skin interferes with proliferation, differentiation, migration, or gene expression of peripheral cells, and even promotes the production of matrix metalloproteinases (matrix metalloproteases) that break down collagen and elastin (elastin), and the secretion of cytokines that can cause inflammatory reactions. These factors ultimately contribute to skin aging.

Therefore, one of the methods for preventing or alleviating skin aging is to supplement collagen or to increase the collagen production ability of cells. Meanwhile, inhibition of collagen glycosylation can ensure elasticity of collagen fibers, not only keep skin tight, but also prevent diseases derived from accumulation of glycosylated proteins, such as vascular sclerosis. The collagen products sold in the market at present mainly require direct collagen supplement, however, the molecular weight of the collagen is large, and the collagen can hardly penetrate through the stratum corneum of the skin to reach the affected dermis by the application mode; if the collagen is directly supplemented by injection, although the fine wrinkles of the skin can be immediately reduced, the collagen injected into the body is easily decomposed by enzymes in the body, the collagen needs to be regularly injected, the process is complicated and the cost is high, and the immune rejection possibly generated by the injection of the collagen is a great risk. If the food rich in collagen is used for supplementing collagen, the human body can naturally synthesize protein by using the amino acids, but not limited to collagen, because the protein is digested into small molecular polypeptides or amino acids after entering the gastrointestinal tract, and therefore, the effect of supplementing collagen by using an edible method is limited.

In view of the above, there is a need to develop a novel composition that not only enhances the collagen production ability of skin fibroblasts, but also inhibits collagen glycosylation.

Disclosure of Invention

Accordingly, an object of the present invention is to provide a use of an extract of ovule of pitaya (Hylocereus undatus) obtained by extracting an ovule of pitaya with a solvent for preparing a composition for delaying skin aging.

In an embodiment of the present invention, the weight ratio of the solvent to the ovule of the red dragon fruit is in a range of 10: 1 to 3: 5, and the extraction is carried out at 50 ℃ to 100 ℃.

In one embodiment of the present invention, the solvent is water, and the concentration of the extract of the red dragon fruit ovule is 0.5mg/mL to 8 mg/mL.

In one embodiment of the present invention, the red dragon fruit ovule extract promotes collagen production. In another embodiment, the red dragon fruit ovule extract promotes skin fibroblast proliferation.

In one embodiment of the present invention, the extract of the ovule of the mangosteen inhibits collagen glycosylation.

The extract of the ovule of the pitaya not only can promote the collagen secretion by promoting the proliferation of skin fibroblasts, but also can reduce the generation of a collagen glycosylation final product. Since collagen is one of the major proteins supporting the skin structure, its stable content can maintain the youthful state of the skin, and the accumulation of glycosylated collagen is closely related to skin aging, the aforementioned extract of the red dragon fruit ovule has the effects of promoting collagen proliferation and inhibiting collagen glycosylation, so that it has the effect of delaying skin aging. Accordingly, the present invention provides the use of an extract of the ovule of the pitaya for the preparation of a composition for delaying skin aging. The composition can be in the form of a powder, granule, solution, gel, or paste, and can be made into food, beverage, pharmaceutical, or nutritional supplement for oral administration, skin application, etc. to an individual.

The following embodiments are provided to illustrate the features and applications of the present invention, rather than to limit the scope of the invention, and it will be apparent to those skilled in the art that various changes and modifications can be made therein without departing from the spirit and scope of the invention.

Drawings

FIG. 1 shows the relative yields of collagen from human skin fibroblasts following treatment with and without an extract of the ovule of the pitaya of an embodiment of the present invention;

FIG. 2 shows the proliferation of human dermal fibroblasts following treatment with or without an embodiment of the invention of an ovule extract of a red dragon fruit; the proliferation of dermal fibroblasts after treatment with 20% fetal calf serum is also shown as a positive control group;

FIG. 3 shows the effect of the extract of the ovule of Hylocereus undulatus on the production of the collagen glycosylation final product according to an embodiment of the present invention.

Detailed Description

The invention provides application of an extract of ovule of pitaya in preparing a composition for delaying skin aging. The extract of the ovule of the pitaya is obtained by extracting an ovule of the pitaya with a solvent, wherein the solvent is preferably water, and the weight ratio of the solvent to the ovule of the pitaya is 10: 1 to 3: 5, and the extraction is carried out at 50 ℃ to 100 ℃. The application of the extract of the ovule of the pitaya was demonstrated to promote the proliferation of fibroblasts and the secretion of collagen in human skin by the following examples. In addition, the extract of the red dragon fruit embryo can inhibit collagen glycosylation.

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.

The term "skin aging" as used herein includes changes in the appearance of the skin, such as wrinkles, fine lines, sagging, depressions, etc., and changes in the skin, such as slow growth of fibroblasts, decreased collagen content, and increased glycated collagen. The foregoing variations can be evaluated by conventional techniques.

Materials and methods

Material

Eagle's Minimal Essential Medium (MEM) containing Earle's balanced salt solution (HBSS), Fetal Bovine Serum (FBS), sodium pyruvate, penicillin/streptomycin, and Phosphate Buffered Saline (PBS) were purchased from Thermo Fischer Scientific.

Cell culture

The following examples were conducted using human skin fibroblast (CCD-966 SK (ATCCCRL-1881) purchased from American Type Culture Collection (ATCC). The cells were cultured in MEM medium supplemented with 10% FBS, 1mM sodium pyruvate, and 1% penicillin/streptomycin at 37 ℃ under 5% carbon dioxide, and hereinafter referred to as cell culture medium.

Collagen secretion test

The Collagen secretion amount of the dermal fibroblasts was measured by using a soluble Collagen Assay Kit (Silcosuble Collagen Assay Kit; Bicolor) according to the manufacturer's instructions, and the procedure thereof is briefly described below. From the cell culture subjected to the designated treatment, 1000. mu.L of the cell culture medium was collected and mixed with 200. mu.L of the collagen separating/concentrating agent, and after standing overnight at 4 ℃, the mixture was centrifuged at 12000rpm for 10 minutes, and about 1000. mu.L of the supernatant was removed. Next, the remaining precipitate was mixed with 1000. mu.L of Sircol stain for 30 minutes, and the stain mixture was centrifuged at 12000rpm for 10 minutes to form a collagen precipitate. After removing the supernatant, the collagen precipitate was washed with 750. mu.L of an acidic salt solution and the excess liquid was removed, 250. mu.L of an alkaline reagent was added to dissolve the collagen precipitate, and then the absorbance at 555nm of the collagen solution was measured using an ELISA plate reader (BioTek). And (5) calculating the collagen content in the cell culture medium to be detected by contrasting the standard curve of the water-soluble collagen standard substance. Statistical analysis was determined using student's t-test with Excel software.

Cell proliferation assay

The proliferation of dermal fibroblasts was determined using the cell proliferation enzyme linked immunosorbent assay kit (Cellproliferation ELISA, BrdU; Sigma-Aldrich) according to the manufacturer's instructions, the procedure of which is briefly described below. Bromodeoxyuridine (BrdU) staining solution (100. mu.M) was added at 10. mu.L/well to the cells subjected to the designated treatment in the 96-well plate, and after incubation at 37 ℃ for 24 hours, the supernatant was removed and 200. mu.L/well of the fixing solution was added, and reacted at room temperature for 30 minutes. Thereafter, the cells were washed with a PBS solution, treated with 100 μ L/well peroxidase (peroxidase) in combination with an anti-BrdU antibody solution at room temperature for 90 minutes, removed again from the antibody solution, and rinsed three times with a washing solution. Finally, 100. mu.L/well of the substrate solution was added to the 96-well plate and reacted at room temperature for 5 to 30 minutes, and then 25. mu.L/well of 1M sulfuric acid was added and reacted with shaking (300rpm) for about 1 minute, whereby the absorbance at 450nm of each well was measured. Statistical analysis was determined using student's t-test with Excel software.