CN112825990A

CN112825990A - Use of red grape fermented juice for preparing composition for improving skin condition

Info

Publication number: CN112825990A
Application number: CN202011200111.9A
Authority: CN
Inventors: 林咏翔; 庄伟秀
Original assignee: TCI Co Ltd
Current assignee: TCI Co Ltd
Priority date: 2019-11-01
Filing date: 2020-10-30
Publication date: 2021-05-25
Also published as: CN112773825A; TWI767387B; TW202118480A; TW202118503A

Abstract

Use of red grape fermented juice for preparing a composition for improving skin condition is provided. The red grape fermented juice is prepared by fermenting red grape leaching liquor with yeast and lactobacillus. Thus, the red grape fermented juice can improve the oxidation resistance in the human body, improve the resistance of cells to light damage and shrink skin pores.

Description

Use of red grape fermented juice for preparing composition for improving skin condition

Technical Field

The invention relates to a fermentation liquid drink, in particular to an application of red grape fermentation juice in preparing a composition for improving skin conditions.

Background

The food fermented by microorganism has the advantages of easier digestion, increased flavor, improved storage time, etc. For example: kimchi, natto, fruit vinegar, and the like are popular fermented foods.

In the beverage world, fermented beverages are becoming more and more popular with consumers. The fermented beverage is prepared by using milk, vegetable, fruit, bean, etc. as material base and through microbial action.

Disclosure of Invention

In view of this, the present invention provides an application of red grape fermented juice, which can further improve the function of red grapes. Here, red grapes are used as a substrate, highly potent strains are selected, and a red grape fermented juice is prepared through a fermentation process.

In one embodiment, the use of red grape fermented juice for preparing a composition for improving skin condition, wherein the red grape fermented juice is prepared from grape leaching liquor of red grapes (vitas vinifera) by fermentation with yeast and lactobacillus.

In one embodiment, the red grape fermentation broth is used to promote the resistance of skin fibroblasts to oxidation caused by light damage.

In one embodiment, red grape fermentation juice has the ability to increase the expression of a gene associated with DNA repair.

In one embodiment, the red grape fermented juice has the ability to increase the expression level of the MPG gene and the XPA gene.

In one embodiment, red grape fermentation juice has the ability to increase the expression of free radical-related genes.

In one embodiment, the red grape fermented juice has the ability to increase the expression level of the SOD2 gene.

In one embodiment, the fermented red grape juice is used to increase the levels of glutathione S-transferase and superoxide dismutase in humans.

In one embodiment, the red grape fermented juice is used to shrink skin pores.

In one embodiment, the red grape fermented juice has a total polyphenol content of 350 ppm.

In one embodiment, the superoxide dismutase activity of the red grape fermented juice is increased by a factor of 3.2 relative to the grape extract.

In summary, a red grape fermented juice according to any of the embodiments, has at least one of the following capabilities: improving the oxidation resistance in human body, improving the resistance of cells to light injury, shrinking pores on human skin and further improving the skin state.

Drawings

Figure 1 is a data plot of total polyphenol content test results.

FIG. 2 is a graph showing relative expression rate results of superoxide dismutase.

FIG. 3 is a graph of relative ROS content data.

FIG. 4 is a graph of relative expression rate data for genes.

Fig. 5 is a diagram of the results of the pore ratio in human experiments.

FIG. 6 is a diagram showing the relative ratio of glutathione S-transferase content in human experiments.

FIG. 7 is a graph showing the relative ratio of superoxide dismutase content in human body experiment.

Detailed Description

In one embodiment, the red grape fermented juice is prepared from grape leaching liquor of red grapes of Sangioves species (Sangioves) by Yeast (Yeast) and Lactobacillus (Lactobacillus) fermentation.

In one embodiment, the grape extract may be a red grape juice obtained by squeezing red grape fruit, a concentrated grape juice obtained by concentrating red grape fruit, a grape juice dilution obtained by diluting red grape fruit juice or concentrated grape fruit juice, or a grape juice obtained by extracting red grape fruit with a solvent. For example, red grape fruit is directly crushed and filtered to form grape leaching solution. In one embodiment, the red grape fruit comprises a peel, a pulp, and a seed. In one embodiment, the grape extract can be prepared by blending commercially available concentrated grape juice with water. In one embodiment, the grape extract is prepared by blending concentrated grape juice, water and glucose. In one embodiment, the grape extract is prepared by heating concentrated grape juice, water and glucose to above 95 deg.C for 30 min. Wherein the volume ratio of the concentrated grape juice to the water can be 1: 8. Also, the concentration of glucose may be 3% (W/V), or an amount of glucose added such that the Brix (Brix) of the glucose leach solution is greater than or equal to 8. That is, the Brix of the solution is measured simultaneously during the glucose addition process, and the glucose addition is stopped when the Brix of the glucose leaching solution reaches 8 or exceeds 8. In one embodiment, the red grape is red grape of the species sanguioerse (sangioves) or the species lubrusque (Lambrusco) or a mixture thereof.

In some embodiments, the red grape fermented juice can be obtained by fermenting grape leaching solution with 0.1% (W/V) yeast and 0.05% (W/V) Lactobacillus (Lactobacillus). In one embodiment, the fermented red grape juice is prepared by inoculating 0.1% (W/V) yeast and 0.05% (W/V) lactobacillus into the grape leaching solution, and standing for 72 hr. In one embodiment, the fermented red grape juice is prepared by culturing grape leaching solution, yeast and lactobacillus for 72 hr, and concentrating under reduced pressure. In one embodiment, the fermented red grape juice is prepared by culturing grape leaching solution, yeast and lactobacillus for 72 hr, concentrating under reduced pressure, and filtering. Wherein the temperature during the reduced pressure concentration is 55-65 ℃.

In one embodiment, the yeast may be Saccharomyces cerevisiae. For example, the lager brewing yeast may be the lager brewing yeast deposited in the center for biological resources conservation and research (BCRC) of the food industry development and research institute, deposited under BCRC20271 strain, which is also deposited under ATCC and International deposit number ATCC26602, or other commercially available lager brewing yeast. In one embodiment, the Lactobacillus may be Streptococcus thermophilus, Lactobacillus helveticus (Lactobacillus helveticus), or Lactobacillus plantarum (Lactobacillus plantarum). For example, the lactobacillus can be streptococcus thermophilus TCI633 (also deposited at DSMZ and international deposit No. DSM28121) deposited at BCRC and deposit No. BCRC910636, streptococcus thermophilus TCI357 (also deposited at DSMZ and international deposit No. DSM33107) deposited at BCRC and deposit No. BCRC910846, streptococcus thermophilus TCI028 (also deposited at DSMZ and international deposit No. DSM33108) deposited at BCRC910805, streptococcus thermophilus TCI378 (also deposited at DSMZ and international deposit No. DSM 331451) deposited at BCRC910760, other commercially available streptococcus thermophilus, other commercially available lactobacillus helveticus, or other commercially available lactobacillus plantarum.

In one embodiment, the red grape fermentation broth is used to promote the resistance of skin fibroblasts to oxidation caused by light damage. In one embodiment, photodamage is the oxidative state of the cell caused by Ultraviolet (Ultraviolet). In one embodiment, the ultraviolet light is light having a wavelength of 320-400 nanometers (nm).

In one embodiment, red grape fermentation juice has the ability to increase the expression of a gene associated with DNA repair. In one embodiment, the fermented red grape juice has the ability to increase the expression level of MPG (N-methyl purine DNA glycosylase) gene (gene ID: 4350) and XPA (DNA damagerecognition and repair factor) gene (gene ID: 7507). Herein, the MPG gene is a repair ability with respect to DNA single strand breaks, and can maintain DNA stability. The XPA gene is about the repair ability of DNA structural abnormality, and the coded XPA protein can be combined with the damaged part of DNA, and stabilize the Nucleotide excision repair (Nucleotide excision repair) process of the DNA.

In one embodiment, red grape fermentation juice has the ability to increase the expression of free radical-related genes. In one embodiment, the red grape fermented juice has the ability to increase the expression level of SOD2(superoxide dismutase 2) gene (ID: 6648). Therefore, the increase of the expression amount of the SOD2 gene can promote the content of manganese superoxide dismutase in the gland, and further improve the removal of free radicals in the gland. The manganese superoxide dismutase can catalyze the disproportionation reaction of superoxide radicals to remove superoxide ions generated in mitochondria so as to reduce the peroxidation damage of mitochondrial DNA.

In one embodiment, the fermented red grape juice is used to increase the content of glutathione S-transferase (GST) and superoxide dismutase (SOD) in human body.

In one embodiment, the red grape fermented juice is effective in shrinking skin pores in human experiments.

In one embodiment, the red grape fermented juice has a total polyphenol content of greater than 200 μ g/mL. In one example, the total polyphenol content of red grape fermented juice may be 350 ppm.

In some embodiments, the aforementioned composition can comprise a food product comprising a specified amount of red grape fermented juice.

In some embodiments, the food product may be a general food, a health food, or a dietary supplement. In other words, the general food, health foods (health foods) or dietary supplements contain an effective amount of red grape fermented juice.

In some embodiments, the food product described above can be manufactured into a dosage form suitable for oral administration using techniques well known to those skilled in the art. In some embodiments, the aforementioned general food product may be the food product itself. In some embodiments, the generic food product may be, but is not limited to: beverages (leafages), fermented foods (fermented foods), bakery products (bakery products) or sauces.

In some embodiments, the resulting red grape fermented juice may be further used as a food additive (food additive) to produce a food composition containing red grape fermented juice. Here, the red grape fermented juice according to any of the examples can be added to a raw material during preparation of a food product (i.e., a food composition) by a method of the related art, or can be added to a red grape fermented juice according to any of the examples during preparation of a food product, and can be formulated with any edible material into a food product for human and non-human animal ingestion (i.e., a food composition).

In some embodiments, the composition may include a cosmetic product comprising a specific amount of red grape fermentation broth.

In some embodiments, the cosmetic product can be made suitable for external use by techniques well known to those skilled in the art. In some embodiments, the cosmetic product may be the product itself or an additive to another product. In some embodiments, the cosmetic product may be, but is not limited to: lotions, gels, jellies, mud masks, lotions, creams, lipsticks, foundations, pressed powders, honey powders, make-up removers, facial cleansers, shower gels, shampoos, hair tonics, sun blocks, hand creams, nail polishes, perfumes, essences, and facial masks. In some embodiments, the cosmetic or care product may optionally further comprise an external acceptable ingredient. In some embodiments, the topical acceptable ingredient can be, for example, an emulsifier, a penetration enhancer, a softener, a solvent, an excipient, an antioxidant, or a combination thereof.

The first embodiment is as follows: preparation method of red grape fermented juice

First, concentrated juice of red grape fruit produced from Cedrus mullerata, Italy, is added with water to prepare a grape juice solution. Wherein, the ratio of the concentrated fruit juice to the water is 1: 8. here, the concentrated juice was purchased from a supplier diana food, product number CC 01460001.

Next, 3% (W/V) glucose was added to the grape juice solution to obtain a grape-based solution.

Heating the grape base solution to about 95 deg.C (tank temperature) for 30 min, and cooling to below 40 deg.C to obtain grape leaching solution.

Adding 0.1% (W/V) yeast and 0.05% (W/V) lactobacillus into grape leaching solution, and standing and culturing at 30 deg.C for 72 hr to obtain grape fermentation primary solution. Here, as the yeast, Saccharomyces cerevisiae deposited under the accession number BCRC20271 was used, and as the lactic acid bacterium, Streptococcus thermophilus TCI633 deposited under the accession number BCRC910636 was used. Here, the initial liquid of grape fermentation had a Brix of 4.0. + -. 0.5(20 ℃ C.) and a pH of 3.0. + -. 0.5.

Concentrating under reduced pressure at 60 deg.C, and filtering with 400mesh pore size to obtain red grape fermented juice.

Example two: preparation method of red grape fermented juice

Concentrating under reduced pressure at 60 deg.C, and filtering with 400mesh pore size to obtain fermented filtrate.

In addition, the red grape fermented filtrate obtained above was added with red grape flavored vinegar (available from VARVELLO GIOVANNI & c.l' ACETO real s.r.l., product name ACETO Balsamico di model na i.g.p) and isomalto-oligosaccharide, mixed, and sterilized at 100 ℃ for 70 minutes to prepare red grape fermented juice B having brix of 40 ± 2 and pH of 3.3 ± 0.5. Herein, the volume ratio of the red grape fermented filtered juice, the red grape flavored vinegar and the isomalto-oligosaccharide is 100:35: 80. Wherein the red grape flavored vinegar is used for adjusting flavor.

Example three: total polyphenol content test

The grape leaching liquid and the red grape fermented juice obtained in the second example are taken as samples respectively. Each sample was diluted with water and 100. mu.L of the diluted sample was placed in a centrifuge tube. Then, 500. mu.L of Folin-Ciocalteu phenol reagent (Folin-Ciocalteu's phenol reagent, Merck) was added to the centrifuge tube, mixed with the diluted sample and left stand for 3 minutes, and then 400. mu.L of 7.5% (W/V) sodium carbonate (Sigma 31432) was added thereto, mixed and left stand for 30 minutes to obtain a reaction solution to be measured. After confirming that no bubble exists in the reaction solution to be measured, 200. mu.L of the reaction solution to be measured is taken into a 96-well plate, and the absorbance at 750nm of the reaction solution to be measured is measured. Each of the above samples was subjected to three replicates and their rounded average values were taken.

Then, a calibration curve was prepared using Gallic acid (Gallic acid) as a standard. Here, gallic acid (powder, Sigma G7384) and pure water were used to prepare 0. mu.L/mL, 20. mu.L/mL, 40. mu.L/mL, 60. mu.L/mL, 80. mu.L/mL, and 100. mu.L/mL of gallic acid standard solutions, and 100. mu.L of each concentration of the standard solution was taken out into a 10mL centrifuge tube. The 500. mu.L of LFolin-Ciocalteu phenol reagent was added to the centrifuge tube to mix with the standard solution and allowed to stand for 3 minutes, and then 400. mu.L of 7.5% sodium carbonate was added to mix and allowed to stand for 30 minutes to obtain a standard reaction solution. A200. mu.L standard reaction solution was taken into a 96-well plate, and its absorbance at 750nm was measured. Then, a standard curve was drawn by the concentration of gallic acid and its corresponding absorbance value.

And then, converting the light absorption value of the reaction solution to be detected into concentration by an interpolation method by using a standard curve, and multiplying the concentration by the dilution ratio to obtain the total polyphenol content of each sample.

In this way, a total polyphenol content of 120ppm in the grape leach solution and 350ppm in the red grape fermented juice B were obtained, as shown in fig. 1.

According to the experimental result, the total polyphenol content of the red grape fermented juice is improved by 2.9 times compared with the total polyphenol content of the grape leaching liquor. Accordingly, it is known that the fermentation with yeast and lactic acid bacteria is more advantageous for the extraction of the effective components.

Example four: cell antioxidation experiment

Here, human dermal fibroblast cells CCD-966sk (accession number BCRC 60153), hereinafter referred to as CCD-966sk cells, were used. The medium used was MEM (minimum essential medium) medium containing 0.1M of nonessential amino acids (Gibco; Cat.11140050), 1.5G/L of sodium bicarbonate (Sigma; Cat.S5761-500G), 1mM of sodium pyruvate (Gibco; Cat.11360-070), 10% (V/V) of fetal bovine serum (Gibco; Cat.10437-028) and 1% (V/V) of Earle's balanced salt type of penicillin-streptomycin (Gibco; Cat 1514. 0122) by additional addition.

Fluorescent dye DCFH-DA (purchased from Sigma/SI-D6883-50MG) was formulated with Dimethyl sulfoxide (DMSO) as a solvent to an active oxidizing species dye at a concentration of 5 MG/mL.

First, at 1.5X 10 per hole⁵Cell number of individual cells, CCD-966sk cells were seeded into each well of a 6-well culture plate containing 2mL of a medium and cultured at 37 ℃ for 24 hours.

Here, the experiment was performed in four groups. After 24 hours of culture, each group was changed in medium and then cultured at 37 ℃ for another 1 hour. Of these, the first group was supplemented with medium only, and no samples were applied (i.e., blank). The second group was supplemented with medium only, and no samples were applied (i.e., control). The third group of the added culture medium contained the grape extract solution prepared in the first example (i.e., the first experimental group), and the concentration of the added grape extract solution was 0.03125% (V/V) relative to the volume of the culture medium. The fourth group of the added culture medium contained the red grape fermented juice prepared in example two (i.e., the second experimental group), and the concentration of the red grape fermented juice added was 0.03125% (V/V) relative to the volume of the culture medium.

After 1 hour of incubation, 5. mu.g/mL of an active oxidizing substance dye was added and reacted at 37 ℃ for 15 minutes to perform staining. Next, 10. mu.L of 100mM H was added to each well of each group in addition to the first group₂O₂The aqueous solution was treated at 37 ℃ for 1 hour.

After treatment, the liquid in each well was removed, and the cells were rinsed twice with 1mL of 1X DPBS (Dulbecco's phosphate-buffered saline). After removing the liquid from each well in the dark, 200. mu.L of 1 Xtrypsin was added and allowed to act for 5 minutes. Then, the cells of each well were individually collected as a culture medium into a 1.5mL microcentrifuge tube, and centrifuged at 400Xg for 10 minutes. The supernatant in the tube was removed, and the pellet was then resuspended at 1 XDPBS and centrifuged again at 400Xg for 10 minutes. After removing the supernatant in the tube again, the cells were suspended in 1mL of 1X DPBS to obtain the cell fluid to be tested.

Next, the DCFH-DA fluorescence signals (excitation light: 450-490 nm; scattered light: 510-550nm) of the cell fluids to be detected in each well were detected by a flow cytometer (brand Beckman) to obtain the relative Reactive Oxygen Species (ROS) expression of each group, and the relative reactive oxygen species expression of each group was converted with the relative reactive oxygen species expression of the blank group as 1, as shown in FIG. 2.

In FIG. 3, the two groups were analyzed for statistically significant differences, i.e., P-values, by a single Student t-test (Student t-test). # # # represents that the P value of the control group is less than 0.001 relative to the blank group, represents that the P value of the first experimental group is less than 0.01 relative to the control group, and represents that the P value of the second experimental group is less than 0.001 relative to the control group.

As can be seen from fig. 3, when the relative active oxygen expression amount of the blank group was 1, the relative active oxygen expression amount of the control group was 5.21, the relative active oxygen expression amount of the first experimental group was 4.37, and the relative active oxygen expression amount of the second experimental group was 2.69.

If the oxidative damage pressure is higher, the active oxygen expression amount is higher, whereas if H can be properly suppressed₂O₂The lower the relative active oxygen expression is for the damage caused by the oxidative stress. Thus, comparing the blank with the control, at H₂O₂The relative active oxygen expression of the CCD-966sk cells after treatment is obviously increased, namely H₂O₂Can cause oxidative stress damage. As can be seen by comparing the control group with the experimental group, the antioxidant damage capability of the CCD-966sk cells treated by the grape leaching solution and the antioxidant damage capability of the CCD-966sk cells treated by the red grape fermented juice are obviously superior to the antioxidant damage capability of the CCD-966sk cells which are not treated. The antioxidant damage capacity of the CCD-966sk cells treated by the red grape fermented juice is better than that of the CCD-966sk cells treated by the grape leaching solution. Therefore, the fermentation of yeast and lactic acid bacteria is more favorable for improving the anti-oxidative damage capability.

Example five: experiment of related Gene expression

Here, human dermal fibroblast cells CCD-966sk (ATCC, CRL-1881), hereinafter referred to as CCD-966sk cells, were used. The medium used was MEM (minimum essential medium) medium of the type containing 0.1mM of non-essential amino acids, 1.5g/L of sodium bicarbonate, 1mM of sodium pyruvate, 0.1mM of Earle's balanced salt of 10% (V/V) fetal bovine serum (Gibco) by additional addition.

At a rate of 1.5X 10 per hole⁵Cell number of individual cells, CCD-966sk cells were seeded into each well of a 6-well culture plate containing 2mL of a medium and cultured at 37 ℃ for 24 hours.

Here, the experiment was performed in four groups (i.e., blank group, control group, first experimental group, and second experimental group). After 24 hours of culture, each group was changed in medium and then cultured at 37 ℃ for another 1 hour. The blank and control were supplemented with medium only, and no samples were applied. The first experimental group was supplemented with a culture medium containing the grape extract solution prepared in the first example, wherein the concentration of the added grape extract solution was 0.03125% (V/V) relative to the volume of the culture medium. The second experimental group added the culture medium containing the fermented red grape juice prepared in the second example, wherein the concentration of the fermented red grape juice added was 0.03125% (V/V) relative to the volume of the culture medium.

Culturing the above groups at 37 deg.C for 6 hr, and irradiating the control group, the first experimental group and the second experimental group with energy unit of 5J/cm²To perform UVA treatment. The blank was not subjected to UVA treatment.

Next, each group of CCD-966sk cells was subjected to RNA extraction using an RNA extraction kit (RNA extraction kit, available from Geneaid corporation, Taiwan, Lot No. FC24015-G). Then, 2000 nanograms (ng) of the extracted RNA were used as templates in each group

III reverse transcriptase (from Invitrogene, USA, No. 18080-051) was used to perform reverse transcription to generate the corresponding cDNA. Subsequently, the corresponding cDNAs were subjected to quantitative Real-Time reverse transcription polymerase chain reaction (quantitative Real-Time reverse transcription polymerase chain reaction) using ABI StepOnePlusTM Real-Time PCR system (Thermo Fisher Scientific Co., U.S.A.), KAPA SYBR FAST qPCR kit (Sigma Co., U.S.A. No. 38220000000) and a combination Primer (Primer) corresponding to the target Primer (Table I below) to detect each setExpression level of the target gene of CCD-966sk cell (1). Here, the gene expression level was measured by the 2-. DELTA.Ct method.

Then, the detection results of the blank groups are regarded as 1 (i.e., the relative expression rate is 1), and the detection results of each group are converted into the relative expression rate. The expression rates of the MPG gene, SOD2 gene and XPA gene in each group are shown in FIG. 4.

Watch 1

R is REVERSE and F is FORWARD.

In FIG. 4, the two groups were analyzed for statistically significant differences, i.e., P-values, by a single Student t-test (Student t-test). In the figure, "x" represents that the P value is less than 0.05, and "x" represents that the P value is less than 0.01.

As can be seen from fig. 4, in the case where the relative expression rate of the MPG gene of the CCD-966sk cells of the blank group is 1, the relative expression rate of the MPG gene of the control group is 0.96, that is, the expression amount of the MPG gene of the CCD-966sk cells is decreased by the irradiation of UVA. In the case where the relative expression rate of the MPG gene of the CCD-966sk cells of the blank group is 1, the relative expression rate of the MPG gene of the CCD-966sk cells of the first experimental group is 1.11, and the relative expression rate of the MPG gene of the CCD-966sk cells of the second experimental group is 1.29, i.e., the relative expression rate of the MPG gene of the cells after being treated with the grape leaching solution or red grape fermented juice is too high 1 (i.e., higher than that of the blank group) even after being irradiated with the same dose of UVA. That is, the grape leaching solution and the red grape fermentation juice can improve the DNA repair capacity of the CCD-966sk cells. Wherein, the relative expression rate of the MPG gene of the cell treated by the red grape fermented juice is better than that of the MPG gene of the cell treated by the grape leaching liquor. That is, the effect of red grape fermented juice is superior to that of grape leaching solution in enhancing DNA repair ability.

As can be seen from FIG. 4, in the case that the relative expression rate of SOD2 gene in CCD-966sk cell of blank group is 1, the relative expression rate of SOD2 gene in CCD-966sk cell of control group is 0.92, that is, the expression amount of SOD2 gene in CCD-966sk cell is decreased under UVA irradiation. On the other hand, in the case that the relative expression rate of SOD2 gene of CCD-966sk cell of blank group is 1, the relative expression rate of SOD2 gene of CCD-966sk cell of the first experimental group is 1.17, and the relative expression rate of SOD2 gene of CCD-966sk cell of the second experimental group is 1.22, namely, the relative expression rate of SOD2 gene of cell is still over 1 (i.e. higher than blank group) even though the cell is irradiated by same dose of UVA after being treated by grape leaching liquor or red grape fermentation juice. Wherein, no matter the red grape fermentation juice of the grape leaching liquor, the expression quantity of SOD2 gene of CCD-966sk cell can be improved. In particular, the red grape fermented juice has better effect of improving the expression amount of SOD2 gene.

As can also be seen from FIG. 4, in the case that the relative expression rate of the XPA gene of the CCD-966sk cell of the blank group is 1, the relative expression rate of the XPA gene of the CCD-966sk cell of the control group is 0.92, and the relative expression rate of the XPA gene of the cell of the first experimental group is 1.83, which means that the expression level of the XPA gene of the CCD-966sk cell is reduced under the irradiation of UVA, and the expression level of the XPA gene of the CCD-966sk cell is more significantly reduced after the treatment of the grape leaching solution. In contrast, the relative expression rate of the MPG gene of the CCD-966sk cells in the second experimental group was 1.13, which means that the relative expression rate of the XPA gene of the cells after being treated with red grape fermented juice was significantly improved even though the cells were irradiated with the same dose of UVA. That is, red grape fermented juice can improve the ability of CCD-966sk cells to repair DNA structures.

Example six: human body experiment for skin condition change

The experiment was conducted with 8 subjects, who were allowed to take a bottle of 5ml of the fermented red grape juice obtained in example two each day for 4 weeks. Furthermore, blood of each subject was collected before the first administration (i.e., control group) and after 4 weeks of administration (i.e., experimental group) to measure the contents of glutathione S-transferase and superoxide dismutase (measured by clinical test), and the facial skin of the subject was measured using a digital skin test apparatus.

First, a VISIA advanced digital skin texture measuring instrument sold by Canfield, USA, which measures the expression of each pore group on the facial skin of a subject through a high resolution camera lens. The number of pores and their area can be detected by the shadow generated in the facial pore depressions caused by the standard white light illumination, which is darker than the surrounding skin color. And then, software is utilized to analyze according to the number and the area of the pores to obtain numerical pore expression quantity, and the higher the numerical value is, the larger the number and the larger the area of the pores are displayed. After measurement, the relative expression (%) of the pores on the skin of the experimental group was calculated using the expression of the pores of the control group as a reference (i.e., the relative expression of the pores on the skin of the control group was 100%).

As can be seen from fig. 5, in the case where the relative pore expression rate of the control group was 100, the relative pore expression rate of the experimental group was 92.6, i.e., when the subjects continuously drunk red grape fermented juice for four weeks, the pores on the facial skin of the subjects were reduced as compared to before use. That is, the red grape fermented juice can reduce pores on the skin to improve the rough state of the skin, thereby making the skin more delicate and glossy.

In this case, the collected blood was subjected to content detection of glutathione S-transferase and superoxide dismutase in blood by a clinical laboratory. After the detection, the relative content (%) of the experimental group was calculated using the content of the control group as a reference (i.e., the relative content of the control group was 100%).

Referring to fig. 6, in the case where the relative content of glutathione transsulfurase of the control group was 100, the relative content of glutathione transsulfurase of the experimental group was 106.62. It is known that the glutathione S-transferase content in the blood of the subject was significantly increased by 6.62% after the subject continuously drunk the red grape fermented juice for four weeks, compared to before drinking. That is, the red grape fermented juice can increase the content of glutathione S-transferase in human body.

Referring to fig. 7, in the case where the relative content of superoxide dismutase in the control group was 100, the relative content of superoxide dismutase in the experimental group was 104.65. It is known that the superoxide dismutase content in the blood of the subject is significantly increased by 4.65% after the subject continuously drinks the red grape fermented juice for four weeks, compared with the content before drinking. That is, the red grape fermented juice can increase the content of superoxide dismutase in human body.

In FIGS. 6 and 7, the two groups were analyzed for statistically significant differences, i.e., P-values, by a single Student t-test (Student t-test). In the figure, "x" represents that the P value is less than 0.05, and "x" represents that the P value is less than 0.01.

Therefore, the red grape fermented juice can improve the content of glutathione S-transferase and superoxide dismutase in the human body, so that the capability of the human body for removing free radicals is increased, and the human body can resist the damage of oxidative pressure.

The present invention is capable of other embodiments, and various changes and modifications may be made by one skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

SEQUENCE LISTING

<110> Dajiang biomedical corporation Ltd

<120> use of red grape fermented juice for preparing composition for improving skin condition

<130> NA

<150> 62/929375

<151> 2019-11-01

<160> 6

<170> PatentIn version 3.5

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<213> Artificial Sequence

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Claims

1. Use of red grape fermented juice for preparing a composition for improving skin conditions, wherein said red grape fermented juice is prepared by fermenting a grape leaching solution of red grapes with yeast and lactobacillus.

2. The use according to claim 1, wherein the red grape fermented juice is used to promote the resistance of skin fibroblasts to oxidation caused by light damage.

3. The use according to claim 2, wherein the red grape fermented juice has the ability to increase the expression of a gene associated with DNA repair.

4. The use according to claim 3, wherein the red grape fermented juice has an ability to increase the expression of MPG gene and XPA gene.

5. The use according to claim 3, wherein the red grape fermented juice has an ability to increase the expression of free radical scavenging-related genes.

6. The use of claim 5, wherein said red grape fermented juice has the ability to increase the expression level of SOD2 gene.

7. The use of claim 1, wherein said red grape fermented juice is used to increase the levels of glutathione S-transferase and superoxide dismutase in humans.

8. The use according to claim 1, wherein the red grape must is used to shrink skin pores.

9. Use according to any one of claims 1 to 8, wherein the red grape fermented juice has a total polyphenol content of 350 ppm.

10. The use according to any one of claims 1 to 8, wherein the superoxide dismutase activity of the red grape fermented juice is increased by a factor of 3.2 relative to the grape extract.