Nymphaea tetragona and rhodomyrtus tomentosa composite beautifying gel for repairing UVB damage and preparation method thereof

Abstract

The invention discloses water lily and rhodomyrtus tomentosa composite beautifying gel for repairing UVB damage and a preparation method of the water lily and rhodomyrtus tomentosa composite beautifying gel. The invention provides an application of a rhodomyrtus tomentosa extract or a rhodomyrtus tomentosa extract and a water lily polysaccharide extract in preparation of cosmetics or medicines for whitening, sunscreen and skin repair. The beauty makeup gel prepared by the invention can significantly improve the survival rate of HaCaT cells damaged by UVB, effectively inhibit cell apoptosis, has a repairing effect on medium-wave ultraviolet (UVB) induced HaCaT cell damage, and also has excellent moisturizing and skin moistening effects. The nymphaea tetragona polysaccharide and the rhodomyrtus tomentosa fruit extract used in the nymphaea tetragona and rhodomyrtus tomentosa composite beautifying gel for repairing UVB damage are derived from natural plant resources with homology of medicine and food, and the nymphaea tetragona and rhodomyrtus tomentosa composite beautifying gel has small irritation to skin lesion parts, low toxicity and few side effects, and has very good development and application prospects in preparation of beautifying products.

Description

Technical Field

[0001] The present invention belongs to the field of cosmetics or biopharmaceuticals, and specifically relates to an extraction or preparation method of water lily polysaccharides, myrtle fruit extracts, and a composite beauty gel prepared by synergistically preparing the two. The beauty gel prepared by the method has a repairing effect on HaCaT cell damage induced by medium-wave ultraviolet (UVB) and has excellent moisturizing and emollient effects. Background Art

[0002] As the largest biological organ in the human body, the skin mainly undertakes functions such as protection, regulation, buffering and resistance. As an organ in direct contact with the outside world, the skin will be affected by external factors and changes in its own body, leading to aging, which is divided into natural aging and light damage. Ultraviolet radiation belongs to electromagnetic radiation. According to the wavelength, it is divided into UVA, UVB and UVC. Studies have shown that at the same dose, the biological effect ratio of UVA to UVB is 1:1000. The main effect of ultraviolet rays on the skin is oxidative damage. Ultraviolet radiation stimulates the body to produce oxidative stress, induces a large amount of ROS, and causes cell or tissue damage. UVB can act on cells by inducing the production of ROS or causing DNA damage, promoting cell apoptosis, accelerating cell proliferation, and inducing skin cancer. In addition, exposure of the skin to ultraviolet rays can cause excessive secretion of MMP in the body, resulting in the decomposition of collagen, causing pigment accumulation and skin leatherization and aging. Therefore, how to effectively protect and repair skin damage caused by UVB is a crucial research direction in the field of skin care and medical beauty health.

[0003] Finding and developing functional substances that protect and repair skin damage caused by UVB has always been a research hotspot for scientists in the fields of medicine and daily chemicals. At present, inorganic UVB protection ingredients such as TiO2, ZnO, PbO, CaCO3, and talcum powder have been found. They shield UVB by reflecting and scattering ultraviolet rays, thereby achieving the effect of protecting against ultraviolet damage. However, such inorganic ingredients are usually only used as ultraviolet shielding agents in the field of cosmetics and do not have the function of repairing ultraviolet damage. At the same time, researchers have also found that some organic compounds with conjugated double bonds can also play the role of ultraviolet shielding and repairing ultraviolet damage, such as benzophenones, benzotriazoles and other compounds, but most of these organic chemical ingredients have strong toxic side effects on the human body, easily causing skin allergies or chapped skin, and such ingredients are often expensive. At present, most of the functional ingredients commonly used in the market for repairing ultraviolet damage are artificial synthetic products, and as functional raw materials for repairing ultraviolet damage, they all have the aforementioned natural disadvantages; therefore, the development of natural, green, and efficient functional raw materials from plant sources for repairing ultraviolet damage has become an inevitable trend in the future daily chemical and pharmaceutical industries.

[0004] Rhodomyrtus tomentosa, also known as Gangshi and Shanshizi, mainly grows in tropical and subtropical areas in southeastern and southwestern my country. Rhodomyrtus tomentosa is a species endemic to my country and is also a very good wild strategic plant resource. The utilization value of Rhodomyrtus tomentosa is very high. Its roots, leaves and fruits can be used as medicine, and have significant pharmacological effects of astringency and antidiarrhea, blood stasis and antibacterial, as well as regulating qi and stagnating the fetus. In addition, Rhodomyrtus tomentosa fruit is often used as a tonic wild fruit with both medicinal and edible properties to enhance immunity, and has extremely strong antioxidant activity. Studies have shown that the main chemical components of Rhodomyrtus tomentosa include natural active ingredients such as phloroglucinol, triterpenes, flavonoids, tannic acid, polysaccharides, etc., which have good anti-tumor, anti-inflammatory, antibacterial, antioxidant and liver protection. However, there have been no research reports on the repair of UV damage by Rhodomyrtus tomentosa at home and abroad so far, so it has very important application potential in the development of cosmetic raw materials for repairing UV damage.

[0005] Water lily is a perennial aquatic herbaceous flower of the genus Nymphaea in the family Nymphaeaceae, widely distributed in temperate, subtropical and tropical regions. This genus of plants not only has rich ornamental and medicinal value, but some water lilies are also food that many people like to eat. They can be eaten directly, processed into different delicacies or used as scented tea. Water lily is an important medicinal plant used in traditional medicine, but there is little exploration of its functional activity. We previously found that water lily (perfume lotus) contains a large amount of polysaccharides with moisturizing, firming, antibacterial and whitening functions. Recent studies have found that as the concentration of this type of polysaccharide molecules increases, the viscosity will gradually increase, and a gel with a uniform appearance and smooth texture can be formed. The natural gel has good air permeability to achieve moisturizing and water locking, allowing the skin to maintain long-lasting skin-friendliness. Therefore, water lily polysaccharides also have huge potential economic value and broad market space in the preparation of medical beauty gel products. Summary of the invention

[0006] The purpose of the present invention is to provide a water lily and myrtle composite cosmetic gel for repairing UVB damage and a preparation method thereof, so as to solve key industry problems in the cosmetics market, such as the shortage of natural functional cosmetics for whitening, sun protection and skin repair, the easy allergic reactions of common UVB products, the dehydration and cracking of the skin caused by long-term use, and the high cost of raw materials, and at the same time meet the urgent demand of the modern medical beauty industry for natural, green and safe products with the function of repairing UVB damage.

[0007] The invention provides application of myrtle extract, or myrtle extract and water lily polysaccharide extract in preparing cosmetics or medicines for resisting ultraviolet damage and repairing skin.

[0008] Preferably, the skin repair is repair after UV damage.

[0009] Preferably, the preparation method of the myrtle extract is: crush fresh myrtle fruit, extract with anhydrous ethanol, concentrate the extract with a rotary evaporator to obtain a crude extract; disperse the crude extract in ultrapure water, extract with petroleum ether, ethyl acetate and n-butanol in sequence, concentrate the extract with reduced pressure to obtain extracts from four parts, and freeze-dry to obtain crude extracts from four parts, namely, petroleum ether extract, ethyl acetate extract, n-butanol extract and water extract, which are all myrtle extracts.

[0010] More preferably, the myrtle extract is an ethyl acetate extract.

[0011] Preferably, the water lily polysaccharide is prepared by the following method: weigh fresh water lily flowers, crush them, first add ethanol to them for extraction, add sterile distilled water to the treated water lily flowers, extract at room temperature, then filter using a 500-mesh filter membrane, concentrate the filtered extract by reduced pressure, then add ethanol to the water lily flower concentrate, stir evenly and wait for it to precipitate, filter the precipitate through gauze, and vacuum dry to obtain a polysaccharide crude extract, dissolve the above polysaccharide crude extract in water, dialyze using a dialysis membrane with a cutoff molecular weight of 20KD, add ethanol to the dialyzed macromolecular water lily polysaccharide aqueous solution, stir until the precipitate is completely precipitated and then filter, and the filtered macromolecular polysaccharide is further freeze-dried to obtain water lily polysaccharide.

[0012] Preferably, the myrtle extract and water lily polysaccharide extract are made into water lily and myrtle composite beauty gel.

[0013] Preferably, the specific preparation method is to dissolve the myrtle extract with ethanol, then add it to the water lily polysaccharide solution, and stir evenly to obtain the water lily and myrtle composite beauty gel.

[0014] Preferably, the content of myrtle extract in the water lily and myrtle composite cosmetic gel is 0.5%-4%, and the content of water lily polysaccharide is 5%.

[0015] The invention also provides a water lily and myrtle composite beauty gel.

[0016] The present invention patent discovered for the first time that myrtle extract has significant pharmacological effects in repairing UV damage, among which the UV damage repair ability of PE extract, EA extract and nBU extract of myrtle fruit is much higher than that of the positive drug ATRA; in particular, myrtle fruit EA extract can effectively repair UVB-damaged human epidermal HaCaT cells at 3.125μg / mL, and significantly improve the survival rate of human epidermal HaCaT cells by inhibiting cell apoptosis, and the effect is significantly better than the positive drug ATRA. At the same time, this project reported for the first time a method for preparing moisturizing and firming medical beauty gel by using water lily polysaccharide. In addition, based on the synergistic effect of water lily polysaccharide and myrtle fruit extract, this patent innovatively developed a method for preparing a water lily and myrtle composite beauty gel with the effect of repairing UVB damage. The water lily and myrtle composite beauty gel prepared by the present invention can be widely used in natural anti-ultraviolet damage, moisturizing and firming cosmetics and medical beauty products, and has important economic potential and application value.

[0017] Technological innovation

[0018] This patent discovered for the first time that myrtle fruit extract has a significant effect on repairing UVB-damaged cells, and reported for the first time a method for preparing a moisturizing beauty gel using water lily polysaccharides; at the same time, based on the above findings, this patent prepared for the first time a water lily and myrtle composite beauty gel, which has a repairing effect on HaCaT cell damage induced by medium-wave ultraviolet (UVB) and has excellent moisturizing and emollient effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 It is the repairing effect of myrtle extract on UVB damage;

[0020] Figure 2 It is an evaluation of the activity of myrtle extract in reducing UVB damage;

[0021] Figure 3 It is the repair activity of water lily and myrtle composite beauty gel against UVB cell damage;

[0022] Figure 4 This is an evaluation of the activity of water lily and myrtle composite beauty gel in reducing UVB damage. DETAILED DESCRIPTION

[0023] The following examples are provided to further illustrate the present invention, rather than to limit the present invention.

[0024] Embodiment 1:

[0025] 1. Preparation of Myrtle Extract

[0026] The myrtle fruit of the present invention was picked from the South China Botanical Garden in July 2023. The specific crude extract preparation method is as follows: 700g of fresh fruit is crushed, and three times the volume of anhydrous ethanol is extracted for 24h, repeated 3 times, and the extract is concentrated by rotary evaporator to obtain a crude extract; the crude extract is dispersed in a small amount of ultrapure water, extracted with petroleum ether, ethyl acetate, and n-butanol in sequence, and each extraction is repeated 3 times; after the extract is concentrated under reduced pressure, four parts of the extract extract are obtained, and after freeze-drying, four parts of the crude extract are obtained: petroleum ether (PE) extract (2.0g), ethyl acetate (EA) extract 2.9g, n-butanol (nBU) extract 3.2g and water extract 2.3g; the four parts of the myrtle fruit extract are prepared into a 20mg / mL stock solution with cell-grade dimethyl sulfoxide (DMSO) and frozen in a -20℃ refrigerator for experimental use.

[0027] 2. Extraction of Water Lily Polysaccharides

[0028] Weigh 1.0 kg of fresh water lily and crush it into 100 mesh powder; first add 10 L of ethanol to extract 3 times, each time for 24 hours, so as to remove the pigment and small molecular phenolic acid compounds in the water lily. Then, add 5 L of sterile distilled water to the treated water lily, and extract at room temperature for 24 hours, and then filter it with a 500 mesh filter membrane. The filtered extract is concentrated by 500 mL under reduced pressure. Then, 800 mL of 100% ethanol is added to the water lily concentrate at one time, and it is stirred evenly and then precipitated. After the precipitate is filtered through gauze, it is vacuum dried to obtain a polysaccharide crude extract (127 g). The above polysaccharide crude extract is dissolved in 1.0 L of sterile distilled water and dialyzed for 2 days using a dialysis membrane with a molecular weight cutoff of 20 KD. The dialyzed macromolecular water lily polysaccharide aqueous solution is placed in a 3.0 L beaker, 1.5 L of 100% ethanol is added, and it is stirred until the precipitate is completely precipitated and then filtered. The filtered macromolecular polysaccharide was further freeze-dried and ground at low temperature to a size of less than 300 meshes, thereby obtaining the target macromolecular water lily polysaccharide (77 g).

[0029] 3. Repair effect of myrtle extract on UVB-induced HaCaT cell damage

[0030] 0.8 × 10 4HaCaT cell suspension with a density of cells / well, 90 μL of cell suspension was added to each well; after culturing for 24 hours, the lid of the culture plate was removed for UVB ultraviolet irradiation treatment, and the irradiation time was 30 minutes; after irradiation, 10 μL of drugs of different concentrations were added to the 96-well plate (the positive control was all-trans retinoic acid ATRA, the negative control was PBS, and the drug-treated group was different concentrations of myrtle fruit extract), and continued to be cultured in a 37°C, 5% CO2 incubator for 24 hours; after culturing, 10 μL of MTT solution (5 mg / mL) was added and continued to incubate for 4 hours; after incubation, the supernatant was carefully aspirated, 100 μL of DMSO was added to dissolve the crystals, and then the OD value of each well at 490 nm was detected by a microplate reader. Cell survival rate calculation formula: Cell survival rate = OD 490样品组 / OD 490CK组 ×100%

[0031] like Figure 1 As shown in the figure, compared with the positive drug ATRA (a retinoic acid receptor agonist commonly used in clinical treatment of UV-induced skin problems), the PE extract, EA extract and nBU extract of myrtle fruit have better UV damage repair ability, and the cell survival rate is much higher than that of the positive drug. The crude extract of EA can effectively repair cells after UVB damage at 3.125 μg / mL, which increases the cell survival rate. Therefore, the myrtle EA extract has a significant effect on repairing UVB damage.

[0032] IV. Evaluation of the activity of myrtle extract in reducing apoptosis of UVB-damaged HaCaT cells

[0033] HaCaT cells in the logarithmic growth phase were trypsinized and adjusted to 1×10 6 cells / mL in a 6-well plate, and 2 mL of culture medium was added to each well; when the cell confluence reached 80%, the lid of the culture plate was removed for UVB damage modeling, and the ultraviolet light irradiation time was 30 min. The blank control group was wrapped with tin foil; after irradiation, different concentrations of crude extracts to be tested were added to each well, and the cells were placed in an incubator and continued to be cultured for 24 h; after culturing, the culture medium in each well was transferred to a centrifuge tube, washed twice with PBS, and 200 μL of trypsin was added to each well to digest the adherent cells; after digestion, the cells were mixed with the culture medium to terminate digestion, and then the culture medium was removed by centrifugation and washed 3 times with PBS; after washing, the cells were resuspended with PBS and the concentration was adjusted to 1×10 6 cells / mL; FITC and PI were added to the cell suspension collected in the above steps, stained for 20 minutes in the dark, and then tested on the computer, and the obtained data was processed by Flowjo.

[0034] Flow cytometry was used to detect the apoptosis of HaCaT cells after UVB irradiation. Figure 2 As shown. After UVB irradiation, the survival rate of HaCaT cells was only 51.8%, but the addition of myrtle EA extract can repair the cell damage caused by UVB irradiation. Compared with the positive drug ATRA, the EA extract of myrtle fruit can effectively reduce the apoptosis rate of HaCaT cells. When the action concentration is 12.50μg / mL, the apoptosis rate of cells is 29.06%, which is much lower than the UVB damage model (apoptosis rate is 48.28%); and with the increase of EA extract concentration, the apoptosis rate of HaCaT cells gradually decreased. When the action concentration is 25μg / mL, the apoptosis rate of HaCaT cells is 23.62%, which is significantly better than the positive drug.

[0035] 5. Preparation of Water Lily Beauty Gel

[0036] Example 1: Weigh 0.05 g of water lily polysaccharide and put it into a 100 mL mortar, add 20 mL of sterile deionized water, and grind evenly to obtain water lily beauty gel 1.

[0037] Example 2: Weigh 0.1 g of water lily polysaccharide and put it into a 100 mL mortar, add 20 mL of sterile deionized water, and grind evenly to obtain water lily beauty gel 2.

[0038] Example 3: Weigh 0.2 g of water lily polysaccharide and put it into a 100 mL mortar, add 20 mL of sterile deionized water, and grind evenly to obtain water lily beauty gel 3.

[0039] Example 4: Weigh 0.4 g of water lily polysaccharide and put it into a 100 mL mortar, add 20 mL of sterile deionized water, and grind evenly to obtain water lily beauty gel 4.

[0040] Example 5: Weigh 0.6 g of water lily polysaccharide and put it into a 100 mL mortar, add 20 mL of sterile deionized water, and grind evenly to obtain water lily beauty gel 5.

[0041] Example 6: Weigh 0.8 g of water lily polysaccharide and put it into a 100 mL mortar, add 20 mL of sterile deionized water, and grind evenly to obtain water lily beauty gel 6.

[0042] Example 7: Weigh 1.0 g of water lily polysaccharide and put it into a 100 mL mortar, add 20 mL of sterile deionized water, and grind evenly to obtain water lily beauty gel 7.

[0043] Example 8: Weigh 1.2 g of water lily polysaccharide and put it into a 100 mL mortar, add 20 mL of sterile deionized water, and grind evenly to obtain water lily beauty gel 8.

[0044] Example 9: Weigh 1.4 g of water lily polysaccharide and put it into a 100 mL mortar, add 20 mL of sterile deionized water, and grind evenly to obtain water lily beauty gel 9.

[0045] Example 10: Weigh 1.6 g of water lily polysaccharide and put it into a 100 mL mortar, add 20 mL of sterile deionized water, and grind evenly to obtain water lily beauty gel 10.

[0046]

[0047] Table 1 Morphology of water lily polysaccharide gel

[0048] By configuring different concentrations of water lily polysaccharides, it was found that when the amount of water lily polysaccharides added was less than 2%, a fluid hydrogel could be successfully prepared; when the concentration of water lily polysaccharides added was between 3% and 4%, a non-fluid gel was obtained, and the gel could present different forms to a certain extent; when the concentration of water lily polysaccharides added was 5% or more, the water lily gel obtained was semi-solid and had high-strength viscosity. The low-concentration hydrogel prepared by this patent can be used in essences and lotions of beauty products, medium-concentration water lily polysaccharide gel is suitable for beauty products such as creams, and high-concentration water lily polysaccharide gel can be used in paste-like beauty products.

[0049] VI. Preparation of Water Lily and Myrtle Composite Cosmetic Gel

[0050] Example 1: Weigh 0.05 g of Myrtle EA extract and place it in a 50 mL beaker, then add 0.45 mL of anhydrous ethanol and stir to fully dissolve it. At the same time, weigh 1.0 g of water lily polysaccharide and place it in a 100 mL mortar, add 18.5 mL of sterile deionized water, and grind it evenly; then add the aforementioned Myrtle EA extract ethanol solution and stir evenly to obtain a thick water lily and myrtle composite beauty gel 1 (Poly-EA-1).

[0051] Example 2: Weigh 0.10 g of Myrtle EA extract and place it in a 50 mL beaker, then add 0.40 mL of anhydrous ethanol and stir to fully dissolve it. At the same time, weigh 1.0 g of water lily polysaccharide and place it in a 100 mL mortar, add 18.5 mL of sterile deionized water, and grind it evenly; then add the aforementioned Myrtle EA extract ethanol solution and stir evenly to obtain a thick water lily and myrtle composite beauty gel 2 (Poly-EA-2).

[0052] Example 3: Weigh 0.20 g of Myrtle EA extract and place it in a 50 mL beaker, then add 0.80 mL of anhydrous ethanol and stir to fully dissolve it. At the same time, weigh 1.0 g of water lily polysaccharide and place it in a 100 mL mortar, add 18.0 mL of sterile deionized water, and grind it evenly; then add the aforementioned Myrtle EA extract ethanol solution and stir evenly to obtain a thick water lily and myrtle composite beauty gel 3 (Poly-EA-3).

[0053] Example 4: Weigh 0.40 g of Myrtle EA extract and place it in a 50 mL beaker, then add 1.60 mL of anhydrous ethanol and stir to fully dissolve it. At the same time, weigh 1.0 g of water lily polysaccharide and place it in a 100 mL mortar, add 17.0 mL of sterile deionized water, and grind it evenly; then add the aforementioned Myrtle EA extract ethanol solution and stir evenly to obtain a thick water lily and myrtle composite beauty gel 4 (Poly-EA-4).

[0054] Example 5: Weigh 0.80 g of Myrtle EA extract and place it in a 50 mL beaker, then add 1.20 mL of anhydrous ethanol and stir to fully dissolve it. At the same time, weigh 1.0 g of water lily polysaccharide and place it in a 100 mL mortar, add 17.0 mL of sterile deionized water, and grind it evenly; then add the aforementioned Myrtle EA extract ethanol solution and stir evenly to obtain a thick water lily and myrtle composite beauty gel 5 (Poly-EA-5).

[0055] VII. Moisturizing and emollient activity of water lily and myrtle composite cosmetic gel

[0056] Testing method for moisturizing and emollient activity: The degree of skin dryness or lubricity is directly related to the moisture content of the skin. The loss of moisture in the skin will directly lead to skin sagging, wrinkling, dryness and even cracking. Therefore, by measuring the inhibition rate of water lily polysaccharide on elastase, the purpose of testing the firming and anti-wrinkle effects can be achieved.

[0057] The water lily and myrtle composite beauty gel (sample 1-5) prepared above was freeze-dried and ground to a fine powder below 200 mesh at low temperature to obtain the water lily and myrtle composite beauty gel freeze-dried powder (sample 1-5). A dry and clean 25mm caliber 10mL flat weighing bottle was selected, and 500mg of the water lily and myrtle composite beauty gel freeze-dried powder prepared above was weighed and placed therein, followed by slow addition of 200mg of sterile deionized water, and the weighing bottle was gently shaken to allow the water lily and myrtle composite beauty gel freeze-dried powder to fully absorb the sterile deionized water. Then, the water lily and myrtle composite beauty gel freeze-dried powder after water absorption was placed in a glass dryer with 200g of anhydrous color-changing silica gel at the bottom, and placed in a 25°C biochemical incubator, and then the moisture retention rate of the water lily and myrtle composite beauty gel freeze-dried powder was tested. During the experiment, hyaluronic acid was used as a positive control, and 700 mg of deionized sterile water was used as a blank control. The flat weighing bottle was taken out and weighed at 2, 4, 6, 8, 10, 12 and 24 hours, and the moisturizing rate was calculated according to the following formula: Moisturizing rate (%) = (mo–m) / mo×100%; where mo is the initial water content of the sample, and m is the water content of the sample after dehydration.

[0058] The results show that the water lily and myrtle composite beauty gel freeze-dried powder has very superior moisturizing activity. Compared with the blank control of sterile deionized water, the water lily and myrtle composite beauty gel freeze-dried powder can show a very significant moisturizing effect for more than 4 hours. At the same time, the water lily and myrtle composite beauty gel freeze-dried powder shows better moisturizing activity when the proportion of water lily polysaccharide content is higher; especially the composite gel freeze-dried powder 1, which has a moisturizing effect similar to hyaluronic acid. The above research results show that the water lily and myrtle composite beauty gel has a very outstanding effect in moisturizing and emollient.

[0059] Table 2 Moisturizing effect of water lily and myrtle composite beauty gel freeze-dried powder

[0060]

[0061] 8. Repair ability of water lily and myrtle composite beauty gel on UVB damage model

[0062] 0.8 × 10 4HaCaT cell suspension with a density of cells / well, add 80μL of cell suspension to each well; after culturing for 24 hours, remove the lid of the culture plate and irradiate with UVB ultraviolet light for 30 minutes; after irradiation, add 20μL of different concentrations of water lily and myrtle composite beauty gel to the 96-well plate (positive control is all-trans retinoic acid ATRA, negative control is PBS), and continue to culture in a 37°C, 5% CO2 incubator for 24 hours; after culturing, add 10μL MTT solution (5mg / mL) and continue incubation for 4 hours; after incubation, carefully aspirate the supernatant, add 100μL DMSO to dissolve the crystals, and then use a microplate reader to detect the OD value of each well at 490nm. Cell survival rate calculation formula: Cell survival rate = OD 490 Sample group / OD 490 CK group × 100%.

[0063] As above Figure 3 As shown, after 20 minutes of UVB irradiation, the cell survival rate of the PBS-treated group was significantly reduced to 28.74%, indicating that UVB caused serious damage to HaCaT cells. Under the treatment of different concentrations of Poly-EA gel, the cell survival rate showed a trend of gradual improvement with increasing concentration. Among them, the 20mg / mL (Water Lily and Myrtle Composite Beauty Gel 4, Poly-EA-4) and 40mg / mL (Water Lily and Myrtle Composite Beauty Gel 5, Poly-EA-5) groups showed significant repair effects, and the survival rates recovered to 98.03% and 218.89%, respectively, which were significantly higher than other low-concentration groups. The results of the study show that high concentrations of Poly-EA have a very prominent protective and repair effect on UVB-damaged cells.

[0064] IX. Evaluation of the activity of myrtle extract in reducing apoptosis of HaCaT cells damaged by UVB

[0065] HaCaT cells in the logarithmic growth phase were trypsinized and adjusted to 1×10 6 cells / mL were inoculated in a 6-well plate, and 2 mL of culture medium was added to each well; when the cell confluence reached 80%, the lid of the culture plate was removed for UVB damage modeling, and the ultraviolet light irradiation time was 30 min. The blank control group was wrapped with tin foil; after irradiation, the water lily and myrtle composite beauty gel prepared above was diluted 100 times, and then 20 μL was added to each well, and the cells were placed in an incubator and continued to be cultured for 24 h; after culturing, the culture medium in each well was transferred to a centrifuge tube, washed twice with PBS, and 200 μL of trypsin was added to each well to digest the adherent cells; after digestion, the cells were mixed with the culture medium to terminate digestion, and then the culture medium was removed by centrifugation and washed 3 times with PBS; after washing, the cells were resuspended with PBS and the concentration was adjusted to 1×10 6cells / mL; FITC and PI were added to the cell suspension collected in the above steps, stained for 20 minutes in the dark, and then tested on the computer, and the obtained data was processed by Flowjo.

[0066] Flow cytometry was used to detect the effect of Poly-EA gel on the apoptosis rate of HaCaT cells damaged by UVB. Figure 4 As shown. After UVB irradiation, the apoptosis rate of HaCaT cells was 64.95%, but the addition of Poly-EA gel can repair the cell damage caused by UVB irradiation and reduce the apoptosis rate. When the action concentration was 10 mg / mL (Water Lily and Myrtle Composite Beauty Gel 3, Poly-EA-3), the apoptosis rate of cells was 62.64%. Compared with the UVB model group, Poly-EA gel showed a weak repair ability; when the action concentration was 20 mg / mL (Water Lily and Myrtle Composite Beauty Gel 4, Poly-EA-4), the damaged HaCaT cells were repaired, and the apoptosis rate dropped to 38.94%. As the action concentration increased, the apoptosis rate of HaCaT cells gradually decreased. When the action concentration was 40 mg / mL (Water Lily and Myrtle Composite Beauty Gel 5, Poly-EA-5), the apoptosis rate of HaCaT cells was 23.62%, which significantly reduced the cell apoptosis caused by UVB damage and could effectively restore cell growth.

[0067] In summary, this patent discovered for the first time that myrtle fruit extract has a significant effect on repairing UVB-damaged cells, and reported for the first time a method for preparing a moisturizing beauty gel using water lily polysaccharides; at the same time, based on the above findings, this patent prepared for the first time a water lily and myrtle composite beauty gel, which has a repairing effect on HaCaT cell damage induced by medium-wave ultraviolet (UVB) and has excellent moisturizing and emollient effects.

Claims

1. Use of myrtle extract, or myrtle extract and water lily polysaccharide extract in the preparation of cosmetics or medicines for resisting ultraviolet damage and repairing skin.

2. The use according to claim 1, characterized in that: The skin repair described is the repair after ultraviolet damage.

3. The use according to claim 1, characterized in that: The preparation method of the myrtle extract is as follows: fresh myrtle fruit is crushed, extracted with anhydrous ethanol, and the extract is concentrated by a rotary evaporator to obtain a crude extract; the crude extract is dispersed in ultrapure water, extracted with petroleum ether, ethyl acetate and n-butanol in sequence, and the extract is concentrated under reduced pressure to obtain extracts of four parts, and after freeze-drying, four parts crude extracts are obtained, namely, petroleum ether extract, ethyl acetate extract, n-butanol extract and water extract, which are all myrtle extracts.

4. The use according to claim 2, characterized in that: The myrtle extract is an ethyl acetate extract.

5. The use according to claim 1, characterized in that: The water lily polysaccharide is prepared by the following method: weighing fresh water lily flowers, crushing them, first adding ethanol to the water lily flowers for extraction, adding sterile distilled water to the treated water lily flowers, extracting at room temperature, then filtering with a 500-mesh filter membrane, concentrating the filtered extract by reduced pressure, then adding ethanol to the water lily flower concentrate, stirring evenly and waiting for precipitation, filtering the precipitation with gauze, vacuum drying to obtain a polysaccharide crude extract, dissolving the polysaccharide crude extract in water, dialyzing with a dialysis membrane with a cut-off molecular weight of 20KD, adding ethanol to the dialyzed macromolecular water lily polysaccharide aqueous solution, stirring until the precipitation is completely precipitated and then filtering, and further freeze-drying the filtered macromolecular polysaccharide to obtain the water lily polysaccharide.

6. The use according to claim 1, characterized in that: The myrtle extract and water lily polysaccharide extract are made into water lily and myrtle composite beauty gel.

7. The use according to claim 6, characterized in that: The specific preparation method is to dissolve the myrtle extract in ethanol, then add it to the water lily polysaccharide solution, and stir evenly to obtain the water lily and myrtle composite beauty gel.

8. The use according to claim 1, characterized in that: The content of myrtle extract in the water lily and myrtle composite beauty gel is 0.5%-4%, and the content of water lily polysaccharide is 5%.

9. A water lily and myrtle composite beauty gel, characterized in that: The preparation method is as follows: dissolving the myrtle extract in ethanol, then adding the solution into a water lily polysaccharide solution, and stirring evenly to obtain a water lily and myrtle composite beauty gel.

10. The water lily and myrtle composite cosmetic gel according to claim 9, characterized in that: The content of myrtle extract in the water lily and myrtle composite beauty gel is 0.5%-4%, and the content of water lily polysaccharide is 5%.

Images