TW202106330A - Use of phalaenopsis plant extract for anti-glycation and improving skin appearance - Google Patents

Abstract

The present invention provides a use of Phalaenopsis plant extract for anti-glycation and improving skin appearance; wherein, the Phalaenopsis plant extract is a Phaaenopsis Sogo Yukidian ukid extract. The Phaaenopsis Sogo Yukidian ukid extract can effectively improve the ability to eliminate glycation end products, increase the gene expression level of KRT14 and SMAD1, increase the amount of hyaluronic acid secreted by keratinocytes, and/or improve the gloss of individual skin. The Phaaenopsis Sogo Yukidian ukid s extract is prepared by extracting Phaaenopsis Sogo Yukidian ukid using water, alcohols, or mixtures of water and alcohols as solvents.

Description

Phalaenopsis plant extract for anti-glycation and improve skin appearance

The present invention relates to the use of a Phalaenopsis plant to resist glycation and improve the appearance of the skin, in particular, a large white Phalaenopsis extract is used to improve the ability to remove the end product of glycation, increase the expression level of the KRT14 gene and the SMAD1 gene, and improve Keratinocytes secrete the amount of hyaluronic acid and/or increase the gloss of the individual's skin.

The epidermal layer is the outermost layer of the skin. From the outside to the inside, it is the stratum corneum, the granular layer, the spinous layer and the basal layer. The epidermal layer is mainly formed by the undifferentiated cylindrical keratinocytes in the basal layer. This process It is called keratinization. The water content in keratinocytes is high. As the cells are metabolized and differentiated, the shape of keratinocytes will gradually become flat, and the nucleus and organelles will begin to degenerate and shrink, and dead cells without nuclei and organelles will be formed in the stratum corneum. The main function of the epidermis is to keep the skin water and form a skin barrier to resist various external damages. The outermost layer of the epidermis is composed of a weakly acidic sebum membrane and a stratum corneum like a brick wall. This barrier can lock the skin Moisture and grease, resistance to the invasion of bacteria on the skin surface, and resistance to damage from external foreign bodies and ultraviolet light, have a very important protective effect on the human body.

In the stratum corneum in the epidermis, although the keratin cells are dead cells, its main component is keratin. Keratin can absorb water to keep the skin moist, and keratinocytes also secrete substances such as hyaluronic acid as intercellular substance. To maintain the structural integrity of the skin barrier of the epidermal layer to prevent the skin from losing water and form a complete protection. When the skin is exposed to a cold or hot environment and UV light is exposed Stimulation will cause the keratinocytes to be unable to maintain the normal metabolic cycle, and the skin's water retention capacity will also decrease, and cause damage to the skin epidermal barrier, making the skin rough, dry and desquamated, fragile, irritated, and sensitive to redness. The health and water retention capacity of the stratum corneum is very important for resisting external damage.

However, as the age gets older, the skin will gradually age (Ageing). The causes and processes of skin aging are very complicated and involve countless physiological phenomena, including ultraviolet radiation damage, free radical damage, reduction of collagen, slower cell renewal, The appearance of abnormal cells, loss of skin fat, lack of intercellular substance, cessation of cell growth, and decline in hormones are more common factors. But up to now, most of the skin anti-aging products on the market can only start to increase the antioxidant activity, and cannot directly and effectively improve or delay the occurrence of skin aging.

Advanced Glycation End Products (AGEs) are a group of highly oxidized compounds, so they are considered to be a kind of glycotoxin (Glycotoxin). Studies have shown that the glycation end products will bind to receptors on the cell surface to change its structure and function. It also promotes the increase of oxidative stress and inflammation of cells, and is inseparable from the formation of diabetes, arteriosclerosis and chronic kidney diseases. In addition to the glycation end products produced during the normal metabolism of the human body, many processed foods also contain glycation end products. Studies have also pointed out that avoiding the intake of glycation end products from food can help delay the progression of chronic diseases and aging.

Based on the above, in order to improve the skin's fragility and susceptibility caused by damaged keratinocytes, decreased water retention capacity, and skin aging, we have developed a method that can directly and effectively improve or delay the occurrence of skin aging, while being effective It is indeed necessary to maintain the normal physiological metabolism of keratinocytes, maintain the structure of the stratum corneum intact, and increase the composition of active ingredients that increase the anti-glycation activity.

For this reason, one purpose of the present invention is to provide a Phalaenopsis ( Phalaenopsis ) plant extract for preparing Keratin 14, KRT14 gene and mothers against decapentaplegic homolog (Mothers against decapentaplegic homolog, SMAD ) The use of the gene expression composition, wherein the Phalaenopsis plant extract is Phaaenopsis Sogo Yukidian ukid (Phaaenopsis Sogo Yukidian ukid) extract, and the Phaaenopsis Sogo Yukidian ukid extract is obtained by extracting a large Phaaenopsis Sogo Yukidian ukid with a solvent, And the solvent is water, alcohol, or a mixture of alcohol and water.

Another object of the present invention is to provide a Phalaenopsis ( Phalaenopsis ) plant extract for the preparation of anti-glycation and improve skin appearance composition, wherein the Phalaenopsis plant extract is a large white phalaenopsis (Phaaenopsis Sogo Yukidian ukid or Phaaenopsis Sogo Yukidian V3) extract, the great white phalaenopsis extract is obtained by extracting a large white phalaenopsis with a solvent, and the solvent is water, alcohol, or a mixture of alcohol and water.

In an embodiment of the present invention, the weight ratio of the large white phalaenopsis to the solvent is 1-5:5-20; and the extraction temperature is 75-95°C.

In another embodiment of the present invention, the large white phalaenopsis extract maintains the arrangement of skin keratinocytes and completes the structure of the skin stratum corneum.

In another embodiment of the present invention, the large white phalaenopsis extract increases the amount of hyaluronic acid secreted by keratinocytes.

In another embodiment of the present invention, the large white phalaenopsis extract enhances the water retention capacity of the skin.

In another embodiment of the present invention, the large white Phalaenopsis extract improves the gloss of the individual's skin.

In another embodiment of the present invention, the concentration of the great white phalaenopsis extract is at least 0.0625 mg/mL.

The large white phalaenopsis extract of the present invention has a strong activity of inhibiting saccharification reaction, which can effectively improve the ability to remove glycation end products. Anti-glycation can inhibit non-enzymatic browning to avoid the denaturation of functional proteins in the body and help delay The progression of chronic diseases and aging; it can also effectively increase the amount of hyaluronic acid secreted by keratinocytes, effectively increase the expression of KRT14 gene and SMAD1 gene in skin cells, and can effectively maintain the arrangement of skin keratinocytes to make the skin stratum corneum structure intact, and Enhance skin water retention capacity, and enhance skin barrier and defense functions to prevent skin water loss, keep skin in a water-retaining state, and maintain skin youthful, smooth and elastic; in addition, it can also effectively enhance the gloss of individual skin and improve skin The dull and yellow condition makes the skin translucent and shiny. Therefore, the large white phalaenopsis extract of the present invention can be used to prepare a composition for anti-glycation and improving skin appearance, wherein the composition is a medicine, a food or a skin care product, which can be taken orally, applied to the skin, etc. Give a body.

The following will further illustrate the implementation of the present invention in conjunction with the drawings. The following examples are used to illustrate the present invention and are not intended to limit the scope of the present invention. Anyone who is familiar with the art will not depart from the spirit and spirit of the present invention. Within the scope, some changes and modifications can be made. Therefore, the scope of protection of the present invention shall be subject to the scope of the attached patent application.

Fig. 1 is a bar graph showing the anti-glycation activity of the Phalaenopsis phalaenopsis extract in an embodiment of the present invention.

Fig. 2 is a bar graph showing the effect of a large white phalaenopsis extract in promoting the secretion of hyaluronic acid from cells according to an embodiment of the present invention.

Fig. 3 is a bar graph showing the effect of an extract of Great White Phalaenopsis on enhancing the expression of KRT14 gene and SMAD1 gene according to an embodiment of the present invention. * p<0.05; ** p<0.01.

Fig. 4 is a bar graph showing the effect of a large white phalaenopsis extract of an embodiment of the present invention on improving the gloss of an individual’s skin.

The numerical values used herein are approximate values, and all experimental data are expressed in the range of 20%, preferably in the range of 10%, and most preferably in the range of 5%.

Use Excel software for statistical analysis. Data mean ± standard deviation (SD), said difference between the groups in a Student's t-test (student's t -test) analysis.

Phaaenopsis Sogo Yukidian ukid (Phaaenopsis Sogo Yukidian ukid or Phaaenopsis Sogo Yukidian V3) is a uniaxial perennial herb of the genus Phalaenopsis in the orchid family (Orchidaceae ). It is an orchid strain derived from Phal. Yukimai and Phal. Taisuco Kochdian.

As used in this article, the term "Extract of Great White Phalaenopsis" means that the flower and solvent of Great White Phalaenopsis are extracted at a ratio of 1-5:5-20 (w/w) for a specific time and temperature.

The "effective concentration" or "effective amount" mentioned in this article means that it can effectively improve the ability to clear the end products of glycation, effectively increase the amount of hyaluronic acid secreted by keratinocytes, effectively increase the expression of KRT14 gene and SMAD1 gene in skin cells, and / Or the concentration of the great white phalaenopsis extract of the present invention required to effectively improve the gloss of the individual's skin. The effective concentration may vary depending on the target, but the effective concentration can be determined experimentally by, for example, a dose escalation test.

According to the present invention, the medicine can be manufactured into a dosage form suitable for parenterally or topically by using techniques well known to those skilled in the art. This includes, but is not limited to: injections (injection) [e.g., sterile aqueous solution or dispersion], sterile powder, external preparation, and the like.

According to the present invention, the medicine may further include a pharmaceutically acceptable carrier which is widely used in medicine manufacturing technology. For example, the pharmaceutically acceptable carrier may include one or more reagents selected from the group consisting of solvent, buffer, emulsifier, suspending agent, decomposer ), disintegrating agent, dispersing agent, binding agent, excipient, stabilizing agent, chelating agent, diluent , Gelling agent, preservative, wetting agent, lubricant, absorption delaying agent, liposome And the like. The selection and quantity of these reagents fall within the scope of professionalism and routine techniques of those who are familiar with this technology.

According to the present invention, the pharmaceutically acceptable carrier includes a solvent selected from the group consisting of water, normal saline (normal saline), phosphate buffered saline (PBS), Aqueous solution containing alcohol and combinations thereof.

According to the present invention, the drug can be administered by a parenteral route selected from the group consisting of: subcutaneous injection, intraepidermal injection, intradermal injection Injection (intradermal injection) and intralesional injection (intralesional injection).

According to the present invention, the medicine can be manufactured into an external preparation suitable for topical application to the skin using techniques well-known to those skilled in the art. This includes, but is not limited to: emulsion, coagulation Gel, ointment, cream, patch, liniment, powder, aerosol, spray, lotion, milk Serum, paste, foam, drop, suspension, salve and bandage.

According to the present invention, the external preparation is prepared by mixing the medicine of the present invention with a base well known to those skilled in the art.

According to the present invention, the substrate may contain one or more additives selected from the following: water, alcohols, glycols, hydrocarbons (such as petroleum jelly) and White petrolatum (white petrolatum,)], wax (such as paraffin and yellow wax), preserving agents, antioxidants, surfactants, absorption enhancers (absorption enhancers), stabilizers (stabilizing agents), gelling agent (gelling agents) [such as Carbopol ^® 974P (carbopol ^® 974P), microcrystalline cellulose (microcrystalline cellulose) and carboxymethyl cellulose (carboxymethylcellulose)] , Active agents, humectants, odor absorbers, fragrances, pH adjusting agents, chelating agents, emulsifiers, occlusion Occlusive agents, emollients, thickeners, solubilizing agents, penetration enhancers, anti-irritants, colorants, and propellants (Propellants) and so on. The selection and quantity of these additives fall within the scope of professionalism and routine technology of those who are familiar with this technology.

According to the present invention, the skin care product can further include an acceptable adjuvant that is widely used in skin care product manufacturing technology. For example, the acceptable adjuvant may contain one or more agents selected from the group consisting of solvents, gelling agents, active agents, preservatives, antioxidants, screening agents, chelating agents, surfactants, dyes Coloring agent, thickening agent, filler, fragrance and odor absorber. The selection and quantity of these reagents fall within the scope of professionalism and routine techniques of those who are familiar with this technology.

According to the present invention, the skin care product can be manufactured into a form suitable for skincare or makeup by using techniques well known to those skilled in the art. This includes, but is not limited to: aqueous solution, water -Alcohol solution (aqueous-alcohol solution) or oily solution (oily solution), oil-in-water type (oil-in-water type), water-in-oil type (water-in-oil type) or complex emulsion, gel Glue, ointment, cream, mask, patch, pack, liniment, powder, aerosol, spray, lotion, emulsion, paste, foam, dispersion, drops, mousse ( mousse, sunblock, tonic water, foundation, makeup remover products, soap, and other body cleansing products.

According to the present invention, skin care products can also be used in combination with one or more external use agents with known activities selected from the following: whitening agents (such as tretinoin, catechins) (catechin), kojic acid, arbutin and vitamin C), moisturizers, anti-inflammatory agents, bactericides, ultraviolet absorbers absorbers), plant extracts (such as aloe extract), skin nutrients, anesthetics, anti-acne agents, antipruritics , Analgesics, antidermatitis agents, antihyperkeratolytic agents, anti-dry skin agents, antipsoriatic agents, antiaging agents ( antiaging agents, antiwrinkle agents, antiseborrheic agents, wound-healing agents, corticosteroids and hormones. The selection and quantity of these topical agents fall within the scope of professionalism and routine techniques of those who are familiar with this technology.

According to the present invention, a food product can be used as a food additive, which can be added during the preparation of raw materials by a conventional method, or added during the production process of the food, and formulated with any edible material for supply Food products consumed by humans and non-human animals.

According to the present invention, the types of food products include, but are not limited to: beverages, fermented foods, bakery products, health foods, and dietary supplements.

The present invention provides a use of a large white phalaenopsis extract for anti-glycation and improvement of skin appearance. The large white phalaenopsis extract of the present invention is obtained by extracting a large white phalaenopsis with a solvent, the solvent being water, alcohol, or alcohol water The mixture, the large white phalaenopsis extract of the present invention has strong anti-glycation activity, can also effectively increase the amount of hyaluronic acid secreted by keratinocytes, effectively increase the expression of KRT14 gene and SMAD1 gene in skin cells, and/or can effectively increase The gloss of the individual's skin.

At the same time, the large white phalaenopsis extract of the present invention can be used to prepare a composition for anti-glycation and skin appearance improvement, which may contain an effective amount of large white phalaenopsis extract and a pharmaceutically acceptable carrier. The composition is a medicine. Product, a food or a skin care product.

In the following, the detailed extraction method of the great white phalaenopsis extract of the present invention, the test of the anti-glycation activity of the great white phalaenopsis extract, the test of the effect of the great white phalaenopsis extract in promoting the secretion of hyaluronic acid by keratinocytes, the great white phalaenopsis The test of the effect of the extract in enhancing the expression of KRT14 gene and SMAD1 gene, and the test of the effect of the large white phalaenopsis extract in enhancing the skin gloss of individuals, to confirm that the large white phalaenopsis extract of the present invention can effectively improve the elimination of glycation end The ability of the product to effectively maintain the arrangement of skin keratinocytes, and/or effectively enhance the gloss of individual skin, can help delay the progression of chronic diseases and aging, and can complete the skin stratum corneum structure, enhance skin moisture retention, and enhance The skin barrier and defense function can also improve the dullness and yellowing of the skin and make the skin bright and shiny.

Example 1 Preparation method of the great white phalaenopsis extract of the present invention

In an embodiment of the present invention, after the flowers of the large white phalaenopsis are washed and cut into pieces of 12 mm in size, the washed large white phalaenopsis is extracted with water, alcohol, or a mixture of alcohol and water. The extraction solvent Preferably it is water, mixed in a volume ratio of 1-5:5-20, homogenized and extracted in a solvent at 75-95°C for 0.5-3 hours, then the crude extraction is cooled to room temperature and then centrifuged. Centrifugation conditions To obtain the supernatant after centrifugation at 300 g for 10 minutes; filter the supernatant with a 400 mesh filter to obtain the filtrate. Finally, after the filtrate is concentrated under reduced pressure at 50-60°C, the large white phalaenopsis extract of the present invention is obtained.

Example 2 The anti-glycation (AGEs) effect of the Great White Phalaenopsis extract of the present invention

This example is to test the anti-glycation activity of the Great White Phalaenopsis extract of the present invention to inhibit the efficiency of D-fructose to produce collagen (Collagen) glycation, and to quantify the glycation activity; anti-glycation It can inhibit non-enzyme browning to avoid the denaturation of functional proteins in the body. First, take 0.2 mL of the 2.5% concentration of the large white phalaenopsis extract solution of the present invention and 0.2 mL of water as the control solution, and add 0.2 mL of 60 mg/mL collagen solution (with 200 mM phosphate buffer Solution configuration, pH 7.4, which contains 0.06% sodium azide (Sodium azide, NaN ₃ )) and 0.2 mL of 1.5 M D-fructose solution (configured with 200 mM phosphate buffer, pH 7.4) After homogenization, take 0.1mL of the mixed solution and measure the fluorescence value at 360nm of excitation light and 460nm of emission light. This is the zero point before the reaction, and then take 0.1mL of the mixed solution and incubate at 50℃ for 24 hours, then take out 0.1 Measure the fluorescence value in mL, which is the end of the reaction; the extract of the great white phalaenopsis is extracted with water as the solvent. And with the same amount of 3mM aminoguanidine (Aminoguanidine, AG) (with 200mM phosphate buffer, pH7.4) re-dissolved the solvent to the same volume as a positive control group, in which Aminoguanidine (AG) is known to inhibit glycation The effect of the effect. Finally, use the following formula to calculate the efficiency of the ability to remove the final glycation product, which represents the anti-glycation activity of each group of samples.

The results of the efficacy test of the anti-glycation activity of the Great White Phalaenopsis extract of the present invention are shown in FIG. 1. After the action of the large white phalaenopsis extract of the present invention, it can reduce about 20% of the amount of saccharification final product. This result shows that the large white phalaenopsis extract of the present invention has a strong activity to inhibit the saccharification reaction and can effectively improve the elimination of saccharification The ability of the final product helps delay the progression of chronic diseases and aging.

Example 3 The effect of the large white phalaenopsis extract of the present invention in promoting the secretion of hyaluronic acid by keratinocytes

Since it is known that keratinocytes secrete hyaluronic acid and other substances as intercellular substance to maintain the integrity of the epidermal barrier, prevent skin water loss and form complete protection, one embodiment of the present invention uses human primary epidermal keratinocytes. keratinocytes, HPEK) to test the effect of the present invention's large white phalaenopsis extract in promoting the secretion of hyaluronic acid from keratinocytes. This human primary skin keratinocyte cell line was purchased from CELLnTEC (Switzerland) with the number HPEK-50, and the cell line was cultured in serum-free keratinocyte-SFM (purchased from Thermo Company, USA, number #17005042), And cultured in a 37℃ cell incubator _{containing 5% CO 2.}

First, in a 96-well culture plate, add 200μL of the above culture medium to each well and implant 1x10 ⁴ human primary skin keratinocytes, and incubate at 37°C for 16-18 hours, then divide the cells into three groups: (1) Only The control group added with cell culture solution, (2) the experimental group added 0.125mg/mL of the great white phalaenopsis extract of the present invention, and (3) the experimental group added 0.0625mg/mL of the great white phalaenopsis extract of the present invention, and After each group was cultured at 37°C for 24 hours, 100 μL of culture solution was collected from each well without disturbing the attached cells; the large white Phalaenopsis extract was extracted with water as a solvent.

Then, the analysis was performed using the ELISA analysis and detection reagent kit of hyaluronic acid (Human Hyaluronic Acid, HA also known as human hyaluronic acid) (purchased from Cusabio Biotech, China, No. CSB-E04805h). First, in a 96-well culture dish covered with a human hyaluronic acid capture antibody at the bottom, add 100 μL of the culture medium collected in each well, or a standard product dissolved in a phosphate buffer solution containing 1% bovine serum albumin. Binding with the capture antibody at 37°C for 2 hours, remove the liquid when the time is up, and directly add 100μL of Biotin detection antibody to each well. After detecting the capture antibody at 37°C for 1 hour , Remove the solution in each well, and wash each 96-well culture plate with 200μL of washing solution (Washing solution containing 0.05% polyoxyethylene anhydrous sorbitol monolaurate (tween 20) phosphate buffer solution) A total of 3 times for each hole; each time the cleaning solution is added, it must be allowed to stand for 2 minutes, and all the cleaning solution must be completely removed, especially after the last cleaning, aspirating or decanting must be used to remove any remaining The cleaning solution of the 96-well culture plate is turned upside down and the remaining cleaning solution is wiped dry with a clean paper towel to make the subsequent measurement results more accurate. Then, add 100μL of 1x horseradish peroxidase-labeled avidin (1x HRP-avidin) to each well, and react at 37°C for 1 hour to bind to the detection antibody, and then Then add 200μL of washing solution in the above method, rinse each hole in the 96-well culture plate 5 times, and then add 90μL of TMB (3,3',5,5'-Tetramethylbenzidine) coloring solution to each well. Incubate at 37°C for 15-30 minutes and protect from light. Then add 50μL of stop solution to each well to stop the reaction. Tap gently to make the solution evenly mixed. Finally, use an enzyme immunoassay analyzer (ELISA reader, BioTek). ) Measure the absorbance at 450nm within 5 minutes. Then use Excel software to perform student t-test to determine whether the coefficient of variation of each group is statistically significant (* p value <0.05; ** p value <0.01; *** p value <0.001).

The test result of the effect of the large white phalaenopsis extract of the present invention in promoting the secretion of hyaluronic acid by keratinocytes is shown in FIG. 2. After the first generation of human skin keratinocytes were treated with 0.125mg/mL of the large white phalaenopsis extract of the present invention, compared with the control group, it could increase the secretion of hyaluronic acid by 11.83%; and after 0.0625 After mg/mL of the present invention's large white phalaenopsis extract, it can increase the secretion of hyaluronic acid by 24.75% compared to the control group. This result shows that the large white phalaenopsis extract of the present invention can effectively increase the amount of hyaluronic acid secreted by keratinocytes, can effectively increase the water retention capacity of the skin, so as to complete the skin stratum corneum structure and enhance the skin barrier function.

Example 4 The effect of the large white phalaenopsis extract of the present invention in enhancing the expression of moisturizing genes in keratinocytes

In one embodiment of the present invention, human primary skin keratinocytes HPEK-50 were used to test the efficacy of the present invention's large white Phalaenopsis extract to enhance the expression of KRT14 gene and SMAD1 gene. First, ^{culture the 1.5x10 5} human primary skin keratinocytes in a 6-well culture dish containing 2 mL of the above culture medium per well, and incubate at 37°C for 16-18 hours, and then divide the cells into the following two groups: (1) Add only The control group of cell culture solution and (2) the experimental group added 0.25mg/mL of the great white phalaenopsis extract of the present invention, and after the two groups were treated at 37°C for 24 hours, the keratin of the primary human skin of each group was tested The expression level of the target gene in the cell; wherein the large white phalaenopsis extract is extracted with water as a solvent. First, the two groups of cells were recovered with cell lysate (RB buffer, purchased from Geanaid Company, Taiwan, Cat No.RBD300), and then RNA extraction reagent kit (purchased from Geneaid Company, Taiwan, Cat No.RBD300) was used to recover the cells. ) Collect the total RNA in the two groups of cells, and then use SuperScript ^® III reverse transcriptase (purchased from Invitrogene, USA, No. 18080-051) with 2000ng of extracted RNA as a template, and use the combination primers and reversers in Table 1 The transcriptase performs reverse transcription to produce the cDNA products corresponding to the mRNAs of these genes, and then uses the ABI StepOnePlus ^TM Real-Time PCR system (Thermo Fisher Scientific Company, USA) and KAPA SYBR FAST (purchased from Sigma Company, USA) , No. 38220000000) The two sets of reverse transcription products were respectively subjected to the quantitative real-time polymerase chain reaction (qPCR) test with the combination primers in Table 1. The conditions were 95℃ for 1 second and 60℃ for reaction. 20 seconds, a total of 40 cycles. For the quantitative expression levels KRT14 gene and mRNA in the two groups of human First generation of skin keratinocytes SMAD1 gene, wherein the quantitative values based taken by the threshold cycle number (Ct), and mRNA of the target gene relative amounts lines derived from equations 2 ^{- △△Ct} , where △C _T =C _{T comparison group or experimental group target gene/control group target gene-} C _{T TBP} (TATA-binding protein); △△C _T =C _{T comparison group or experimental group Target gene-} C _{T controls the target gene} of the group; the fold change of each gene in each group is 2 ^-△△Ct . Then, use Excel software to determine whether the coefficient of variation is statistically significant (* p value <0.05; ** p value <0.01; *** p value <0.001).

The test results of the efficacy of the large white phalaenopsis extract of the present invention in enhancing the expression of KRT14 gene and SMAD1 gene are shown in FIG. 3. Among them, KRT forms keratin filaments, which are related to maintaining the structure of keratinocytes; while SMAD is a signal transmission factor and is related to regulating the expression of collagen. Compared with the control group, human primary skin keratinocytes treated with 0.25 mg/mL of the present invention's large white phalaenopsis extract can significantly increase the expression levels of KRT14 gene and SMAD1 gene by 1.24 times and 1.28 times that of the control group. . This result shows that the large white phalaenopsis extract of the present invention can effectively enhance the expression of KRT14 gene and SMAD1 gene in skin cells, can effectively maintain the arrangement of skin keratinocytes, make the skin stratum corneum structure complete, and strengthen the skin barrier and defense functions, Prevent skin water loss, keep the skin in a water-retaining state, and keep the skin young, smooth and elastic.

Example 5 The effect of the present invention's large white phalaenopsis extract to enhance the gloss of individual skin

An embodiment of the present invention is to test the effect of the present invention's large white phalaenopsis extract in improving the gloss of the individual's skin. First, prepare a large white phalaenopsis extract mask with the essence of 1% (w/w) of the invention’s large white phalaenopsis extract; and a mask that does not contain the large white phalaenopsis extract of the present invention The facial mask of the essence is used as the control group; among them, the essence of the essence is water, saccharin, hexylene glycol, 1,3-butanediol, trixian gum, thickener and triethanolamine; among them, the great white phalaenopsis The extract is extracted with water as a solvent. Then, a transient effect test was conducted and 8 subjects were recruited. After each subject cleans his face every morning and evening, he applied the mask of the control group and the mask containing the large white phalaenopsis extract of the present invention. Take off the skin of the left and right sides of the face after 15 minutes, massage it with your fingertips to promote absorption, and apply the C+K Cutometer® MPA 580 multi-probe skin analyzer (C+K electronic) before and after applying the mask. , Germany) Glossymeter_GL200 gloss probe for the detection of the skin gloss of the whole face. The reflected light and scattered light hit the skin surface by the instrument can accurately and conveniently test the gloss of the skin. The parallel white light is generated by the LED on the top of the probe; among them, the skin gloss before applying the mask is 100% , Relative to the percentage obtained before applying the mask, the relative skin gloss improvement percentage after applying the mask can be obtained.

The results of the present invention of the great white phalaenopsis extract in improving the gloss of the individual's skin are shown in Figure 4. After using the mask containing the great white phalaenopsis extract of the present invention, the skin gloss increased by 10.4% compared to before use, and the control group without the great white phalaenopsis extract of the present invention only increased by 3.6%. This result shows that the large white phalaenopsis extract of the present invention can effectively improve the gloss of the individual's skin, and has the effect of improving the dull and yellow skin and making the skin bright and shiny.

In summary, the great white phalaenopsis extract of the present invention has potent activity in inhibiting glycation reaction, can effectively improve the ability to clear the end products of glycation, and help delay the progression of chronic diseases and aging; it can also effectively improve keratinocytes Secreting the amount of hyaluronic acid, effectively enhancing the expression of KRT14 gene and SMAD1 gene in skin cells, can effectively maintain the arrangement of skin keratinocytes, so that the skin stratum corneum structure is intact, and skin water retention capacity is improved, and skin barrier and defense functions are improved. To prevent skin water loss, keep the skin in a water-retaining state, and keep the skin young, smooth and elastic; in addition, it can also effectively improve the gloss of the individual's skin, improve the dull and yellow skin, and make the skin bright and shiny. . Therefore, the large white phalaenopsis extract of the present invention can be used to prepare a composition for anti-glycation and improving skin appearance, wherein the composition is a medicine, a food or a skin care product, which can be taken orally, applied to the skin, etc. Give a body.

<110> Dajiang Biomedical Co., Ltd.

<120> The use of large white Phalaenopsis extract for anti-glycation and improving skin appearance

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<170> PatentIn version 3.5

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Claims (11)

A Phalaenopsis ( Phalaenopsis ) plant extract is used to prepare a composition for enhancing the expression level of keratin 14 (Keratin 14, KRT14 ) gene and mothers against decapentaplegic homolog ( SMAD ) gene expression, wherein The Phalaenopsis plant extract is a Phaaenopsis Sogo Yukidian ukid (Phaaenopsis Sogo Yukidian ukid) extract. The Phaaenopsis Sogo Yukidian ukid extract is obtained by extracting a Phaaenopsis Sogo Yukidian ukid with a solvent, and the solvent is water, alcohol, or alcohol. Water mixture. The use as described in item 1 of the scope of patent application, wherein the large white Phalaenopsis extract maintains the arrangement of skin keratinocytes and completes the structure of the skin stratum corneum. A Phalaenopsis ( Phalaenopsis ) plant extract is used to prepare a composition for anti-glycation and skin appearance improvement, wherein the Phalaenopsis plant extract is a large white Phalaenopsis ( Phaaenopsis Sogo Yukidian ukid or Phaaenopsis Sogo Yukidian V3) The extract, the large white phalaenopsis extract is obtained by extracting a large white phalaenopsis with a solvent, and the solvent is water, alcohol, or a mixture of alcohol and water. The use described in item 3 of the scope of patent application, wherein the large white phalaenopsis extract increases the amount of hyaluronic acid secreted by keratinocytes. The use as described in item 3 of the scope of patent application, wherein the extract of the great white phalaenopsis is to enhance the water retention capacity of the skin. The use as described in item 3 of the scope of patent application, wherein the extract of the great white phalaenopsis is to enhance the gloss of the individual's skin. For the purposes described in item 1 or item 3 of the scope of patent application, the weight ratio of the large white phalaenopsis to the solvent is 1-5:5-20. For the purposes described in item 1 or item 3 of the scope of patent application, the extraction temperature is 75-95°C. Such as the use described in item 1 or item 3 of the scope of the patent application, wherein the concentration of the large white phalaenopsis extract is at least 0.0625 mg/mL. The use described in item 1 or item 3 of the scope of patent application, wherein the composition further comprises a pharmaceutically acceptable carrier. Such as the use described in item 1 or item 3 of the scope of patent application, wherein the composition is a medicine, a food or a skin care product.

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