A plant light protection composition, use, and cosmetic

Abstract

The present invention belongs to the field of new material technologies, and discloses a plant light protection composition. By weight, it comprises the following components: 0.01 - 0.5 parts of prickly ash fruit extract; 0.04 - 1.2 parts of epigallocatechin gallate glucoside; 0.1 - 2 parts of cherry blossom extract; 20 - 30 parts of cottonseed extract; 20 - 30 parts of pomegranate seed extract; 0.1 - 1 part of fructooligosaccharide. The above-mentioned active ingredients cooperate with each other and have excellent synergistic effects in aspects of light protection such as anti-UV light damage, antioxidant, anti-inflammatory and repair. At the same time, the present invention also discloses the application of this composition and cosmetics.

Description

Technical Field

[0001] The present invention belongs to the technical field of new materials, and particularly relates to a plant light protection composition, uses, and cosmetics. Background Art

[0002] Light protection refers to protecting the human body or objects from the harm of ultraviolet rays and other harmful rays through various measures. People can achieve light protection for parts such as hair, eyes, and skin by wearing protective caps or glasses, wearing protective clothing, and applying skin care products with light protection effects. Among them, skin care products with light protection effects are favored by more and more consumers, and developing excellent light protection products has great application prospects and economic benefits.

[0003] The wavelength range of the ultraviolet spectrum in sunlight is 200 - 400 nm, including medium-wave ultraviolet (UVB) with a wavelength of 290 - 320 nm and long-wave ultraviolet (UVA) with a wavelength of 320 - 400 nm. Ultraviolet rays are the main light sources causing skin damage such as sunburn, tanning, sunspots, and photoaging. In addition, high-energy visible light (HEV) with a wavelength of 400 - 500 nm also exists in sunlight, electronic products, artificial light sources, etc. There have been many studies showing that HEV can stimulate the generation of reactive oxygen free radicals, causing a series of reactions such as cell damage, thereby causing light damage to the skin.

[0004] For cosmetic products to achieve light protection effects, on the one hand, sunscreen agent components that can reflect or absorb ultraviolet rays (mainly UVB and UVA) can be added to reduce the damage of ultraviolet rays to the skin, etc.; on the other hand, functional raw material components with the effects of resisting UV stimulation, scavenging oxygen free radicals caused by ultraviolet rays and HEV and other harmful rays, promoting the healing ability of damaged cells, and inhibiting the activity of inflammatory reactions can be added, so as to improve the skin's own resistance and repair ability to enhance the resistance and protection ability against external harmful rays.

[0005] CN106726975A discloses a blue light protection composition composed of lutein, cocoa seed extract, black rose extract, rosehip oil, and xylitol, which has the effect of absorbing blue light, but has a weak repair ability for light-damaged skin.

[0006] Therefore, in order to more comprehensively protect the skin against the damage of harmful rays, it is necessary to study light protection products that can improve the skin's own resistance and repair ability in multiple aspects to enhance the resistance and protection ability against external harmful rays. Summary of the Invention

[0007] Aiming at the deficiencies of the prior art, the purpose of the present invention is to provide a plant light protection composition, in which the active ingredients cooperate with each other and have excellent synergistic effects in aspects of light protection such as anti-UV light damage, antioxidant, anti-inflammatory and repair.

[0008] Meanwhile, the present invention also discloses the application of the composition and cosmetics.

[0009] To achieve the purpose of this invention, the following technical solutions are adopted: A plant light protection composition, calculated by weight, includes the following components:

[0010]

[0011] Preferably, calculated by weight, it includes the following components:

[0012]

[0013] In the present invention, the main active ingredient of the zanthoxylum fruit extract is sanshool, and preferably the zanthoxylum fruit extract with a sanshool content of ≥ 15% by weight;

[0014] The cherry blossom extract is preferably a flower extract of Cerasus sp. with a total flavonoid content of ≥ 25% by weight, a polyphenol content of ≥ 3% by weight, and a polysaccharide content of ≥ 10% by weight;

[0015] The cottonseed extract is preferably a raw material with a polyphenol content of ≥ 0.7% by weight;

[0016] The pomegranate seed extract is preferably a raw material with a polyphenol content of ≥ 0.7% by weight.

[0017] Fructooligosaccharide is also known as oligofructose or sucrose triose family oligosaccharides.

[0018] The zanthoxylum fruit extract is a raw material with both medicinal and edible uses. Research shows that its main active ingredient is sanshool, which has effects such as anesthesia and analgesia, intestinal protection and blood sugar lowering in pharmacology. In the application of skin care products, research shows that sanshool can inhibit the production of reactive oxygen species (ROS), with antioxidant effects; it can induce autophagy and enhance the self-repair ability of cells; it can reduce the skin inflammatory response caused by UVB irradiation and has good anti-light damage effects. In the plant light protection composition of the present invention, the zanthoxylum fruit extract with a sanshool content of ≥ 15% by weight is preferred.

[0019] Epigallocatechin Gallatyl Glucoside (EGCG) is the main component of tea polyphenols and is a well-known component with strong antioxidant activity. Some studies have also shown that EGCG can prevent the inflammatory response of the Nuclear Factor kappa-B (NF-κB) pathway, prevent vasodilation, and can also prevent the expression of matrix metalloproteinase MMP-1 in the skin under light irradiation, thereby protecting the skin barrier.

[0020] The main components of cherry blossom extract include polysaccharides, quercetin, polyphenols, and prunasin, and also contain substances such as a small amount of vitamins and arbutin, etc., which have the ability to scavenge ROS and have antioxidant effects; it can also inhibit the inflammatory response of the skin and has anti-inflammatory effects. CN112089660B discloses that cherry blossom extract also has the effects of maintaining the balance of skin flora and anti-blue light. In the plant light protection composition described in the present invention, the cherry blossom extract is preferably a Cerasus sp. flower extract with a total flavonoid content ≥ 25% by weight, a polyphenol content ≥ 3% by weight, and a polysaccharide content ≥ 10% by weight;

[0021] The main components of cottonseed extract include oil components such as linoleic acid, vitamin E, and caryophyllene, and also contain components soluble in alcohols or water such as polyphenols, saponins, alkaloids, and flavonoid compounds, and have good emollient and antioxidant effects. In the plant light protection composition described in the present invention, the cottonseed extract is preferably a raw material with a polyphenol content ≥ 0.7% by weight.

[0022] The main components of pomegranate seed extract include oil components such as fatty acids, vitamin E, and sterol components, and also contain components soluble in alcohols or water such as polyphenols, polysaccharides, and proteins. Among them, pomegranate seed polyphenols mainly include flavonoids, tannins, ellagic acid, and anthocyanins, etc., and have good antioxidant effects. In the plant light protection composition described in the present invention, the pomegranate seed extract is preferably a raw material with a polyphenol content ≥ 0.7% by weight.

[0023] Fructooligosaccharides are widely present in many edible fruits and vegetables and are recognized by international nutritionists as "water-soluble dietary fibers with excellent indigestibility". They can bidirectionally regulate the balance of the human microecosystem and are one of the prebiotics with the most research and the widest application range. Fructooligosaccharides are a type of carbohydrate with a relatively low molecular weight and have good water solubility, showing excellent moisturizing and repair effects when used in the skin. The structures of natural fructooligosaccharides and those obtained by the microbial enzymatic method are almost linear, with the molecular formula G-F-Fn (n = 1, 2, 3, G is glucose, and F is fructose). Currently, there are mainly two major production processes for fructooligosaccharides. One is using sucrose as the raw material and generating it through microbial fermentation; the other is using inulin as the raw material and generating it through enzymatic hydrolysis. Both of the above raw materials can be applied to the present invention and achieve satisfactory results.

[0024] Through the mutual matching in a reasonable ratio, the above composition can achieve the purpose of synergistic enhancement of anti-UV light damage; from the results of several implementation cases and comparative cases of the present invention, when one or more components are missing, the survival performance of HaCaT cells is significantly reduced.

[0025] At the same time, through the test results of DPPH free radical scavenging rate, hyaluronidase inhibition rate, and cell scratch healing rate, it can be seen that the above 6 components also showed excellent synergistic performance in the above three tests.

[0026] Preferably, the present invention further includes a carrier; the zanthoxylum fruit extract, epigallocatechin gallate glucoside, cherry blossom extract, cottonseed extract, pomegranate seed extract, and fructooligosaccharides are active ingredients; the active ingredients are loaded on the carrier or loaded in the carrier.

[0027] In the above plant photoprotective composition, the carrier is one of liposomes, solubilization systems, and hydrogel networks.

[0028] In the field of cosmetics, liposomes are usually composed of phospholipids as the skeleton membrane material and additives. The phospholipids used for preparing liposomes include two categories: natural phospholipids and synthetic phospholipids. Natural phospholipids are mainly lecithin, including lecithin, soybean lecithin, egg yolk lecithin, phosphatidylcholine, etc.; synthetic phospholipids mainly include dilauroyl phosphatidylcholine, dimyristoyl phosphatidylcholine, dipalmitoyl phosphatidylcholine, distearoyl phosphatidylcholine, dioleoyl phosphatidylcholine, etc. The additives mainly include cholesterol, emulsifiers (such as Tweens, polyglycerol esters, etc.), alkanols (such as butanediol, propylene glycol, dipropylene glycol, etc.), and salts (such as sodium chloride, etc.) or buffer salts as solvents or stabilizers.

[0029] In the field of cosmetics, a solubilization system refers to a system formed by dissolving lipophilic raw materials in water through solubilizers. Commonly used solubilizers include polysorbates (such as Tween 20, Tween 80, etc.), polyoxyethylene ethers (such as PPG-26-butanol polyether-26, etc.), polyoxyethylene castor oils (such as PEG-40 hydrogenated castor oil, PEG-60 hydrogenated castor oil), etc.

[0030] In the field of cosmetics, a hydrogel network can be formed by a thickener to form a hydrogel network, such as acrylic thickeners like carbomer; it can also be formed by a multiple network structure composed of boric acid molecules. For details, see the invention patent CN115044150A applied by Sichuan University, a three-component supramolecular hydrogel with shape self-adaptability, its preparation method and application.

[0031] In the above-mentioned plant light protection composition, by weight, the content of the carrier in the composition is 0.05-12 wt%, and the composition also contains water, and the weight of the water added to the composition is 100 parts by weight.

[0032] Meanwhile, the present invention also discloses the use of the plant light protection composition as described above in the preparation of cosmetics.

[0033] The dosage form of the cosmetics of the present invention can be selected from one of cream, milk, gel, powder, block, mud, wax-based, spray, aerosol, patch, film, substrate-containing, freeze-dried and other dosage form categories; preferably one or more of cream, lotion, essence, gel, powder, wax-based dosage form, spray, aerosol, patch, film.

[0034] Preferably, the cosmetics are used for anti-UV light damage.

[0035] Preferably, the cosmetics are used for scavenging DPPH free radicals.

[0036] Preferably, the cosmetics are used for inhibiting hyaluronidase.

[0037] Preferably, the cosmetics are used for promoting cell wound healing.

[0038] Furthermore, the plant light protection composition of the present invention can not only be used in cosmetics, but also has certain potential as an active ingredient of a drug. When the plant light protection composition of this application is a pharmaceutical composition, in addition to the above components, the plant light protection composition of this application can also optionally contain common components of pharmaceutical compositions, including any components known in the art such as drug active ingredients, and the type and dosage can be selected according to specific needs. The drug active ingredient is a component known in the art for makeup and skin care. For example, topical medications for the skin, and these medications can be selected as ointments, dressings, etc.;

[0039] Finally, the present invention also discloses a cosmetic containing the above-described plant light protection composition.

[0040] In the above cosmetic, the plant light protection composition is contained in an amount of 0.01 to 10 wt%.

[0041] In some embodiments of the present invention, the dosage of the plant light protection composition can be 0.01 wt%, 0.05 wt%, 0.1 wt%, 0.5 wt%, 1 wt%, 3 wt%, 5 wt% or 10 wt%.

[0042] Compared with the prior art, the present invention has the following beneficial effects:

[0043] After the above active ingredients with good skin care effects are combined with each other to form a composition, an excellent synergistic effect is achieved in terms of light protection such as anti-UV light damage, antioxidant, anti-inflammatory and repair. Tests verify that the composition can protect cells against UV light-induced damage, effectively scavenge reactive oxygen species induced by blue light, promote cell healing after scratching, and inhibit the activity of hyaluronidase, thereby improving the skin's own resistance and repair ability to enhance the resistance and protection ability against harmful external light. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1 is a photograph of the blank group at 0 h in the cell scratch healing experiment;

[0045] Figure 2 is a photograph of the blank group at 15 h in the cell scratch healing experiment;

[0046] Figure 3 is a photograph of Example 1 at 0 h in the cell scratch healing experiment;

[0047] Figure 4 is a photograph of Example 1 at 15 h in the cell scratch healing experiment;

[0048] Figure 5 is a photograph of Example 2 at 0 h in the cell scratch healing experiment;

[0049] Figure 6 is a photograph of Example 2 at 15 h in the cell scratch healing experiment;

[0050] Figure 7 is a photograph of Example 3 at 0 h in the cell scratch healing experiment;

[0051] Figure 8 is a photograph of Example 3 at 15 h in the cell scratch healing experiment;

[0052] Figure 9It is a photo taken at 0 h in Example 4 during the cell scratch healing experiment;

[0053] Figure 10 It is a photo taken at 15 h in Example 4 during the cell scratch healing experiment;

[0054] Figure 11 It is a photo taken at 0 h in Example 5 during the cell scratch healing experiment;

[0055] Figure 12 It is a photo taken at 15 h in Example 5 during the cell scratch healing experiment;

[0056] Figure 13 It is a photo taken at 0 h in Example 6 during the cell scratch healing experiment;

[0057] Figure 14 It is a photo taken at 15 h in Example 6 during the cell scratch healing experiment;

[0058] Figure 15 It is a photo taken at 0 h in Example 7 during the cell scratch healing experiment;

[0059] Figure 16 It is a photo taken at 15 h in Example 7 during the cell scratch healing experiment;

[0060] Figure 17 It is a photo taken at 0 h in Example 8 during the cell scratch healing experiment;

[0061] Figure 18 It is a photo taken at 15 h in Example 8 during the cell scratch healing experiment;

[0062] Figure 19 It is a photo taken at 0 h in Example 9 during the cell scratch healing experiment;

[0063] Figure 20 It is a photo taken at 15 h in Example 9 during the cell scratch healing experiment;

[0064] Figure 21 It is a photo taken at 0 h in Example 10 during the cell scratch healing experiment;

[0065] Figure 22 It is a photo taken at 15 h in Example 10 during the cell scratch healing experiment;

[0066] Figure 23 It is a photo taken at 0 h in Comparative Example 1 during the cell scratch healing experiment;

[0067] Figure 24 It is a photo taken at 15 h in Comparative Example 1 during the cell scratch healing experiment;

[0068] Figure 25It is a photo of Comparative Example 2 at 0 h in the cell scratch healing experiment;

[0069] Figure 26 It is a photo of Comparative Example 2 at 15 h in the cell scratch healing experiment;

[0070] Figure 27 It is a photo of Comparative Example 3 at 0 h in the cell scratch healing experiment;

[0071] Figure 28 It is a photo of Comparative Example 3 at 15 h in the cell scratch healing experiment;

[0072] Figure 29 It is a photo of Comparative Example 4 at 0 h in the cell scratch healing experiment;

[0073] Figure 30 It is a photo of Comparative Example 4 at 15 h in the cell scratch healing experiment;

[0074] Figure 31 It is a photo of Comparative Example 5 at 0 h in the cell scratch healing experiment;

[0075] Figure 32 It is a photo of Comparative Example 5 at 15 h in the cell scratch healing experiment;

[0076] Figure 33 It is a photo of Comparative Example 6 at 0 h in the cell scratch healing experiment;

[0077] Figure 34 It is a photo of Comparative Example 6 at 15 h in the cell scratch healing experiment;

[0078] Figure 35 It is a photo of Comparative Example 7 at 0 h in the cell scratch healing experiment;

[0079] Figure 36 It is a photo of Comparative Example 7 at 15 h in the cell scratch healing experiment;

[0080] Figure 37 It is a photo of Comparative Example 8 at 0 h in the cell scratch healing experiment;

[0081] Figure 38 It is a photo of Comparative Example 8 at 15 h in the cell scratch healing experiment;

[0082] Figure 39 It is a photo of Comparative Example 9 at 0 h in the cell scratch healing experiment;

[0083] Figure 40 It is a photo of Comparative Example 9 at 15 h in the cell scratch healing experiment;

[0084] Figure 41It is a photo of Comparative Example 10 at 0 h in the cell scratch healing experiment;

[0085] Figure 42 It is a photo of Comparative Example 10 at 15 h in the cell scratch healing experiment;

[0086] Figure 43 It is a photo of Comparative Example 11 at 0 h in the cell scratch healing experiment;

[0087] Figure 44 It is a photo of Comparative Example 11 at 15 h in the cell scratch healing experiment. Detailed implementation manners

[0088] The technical solutions of the present invention will be further described below through specific implementation manners. Those skilled in the art should understand that the embodiments are only for helping to understand the present invention and should not be regarded as specific limitations on the present invention.

[0089] Before elaborating on the method of the present invention, the sources of the reagents used in the present invention will be described first;

[0090] The sanshools content of the prickly ash fruit extract is 16 - 18%; epigallocatechin gallate glucoside is a sample with product number P0119321 from Beijing Lingbao Technology Co., Ltd.; the cherry extract is obtained according to the relevant preparation method of CN112089660B, with the total flavonoids content of 30 - 35%, the polyphenol content of 4 - 5%, and the polysaccharide content of 15 - 20%;

[0091] The polyphenol content of the cottonseed extract is 0.7 - 0.8%;

[0092] The polyphenol content of the pomegranate seed extract is 0.7 - 0.8%;

[0093] Fructooligosaccharide is a sample with product number F861451 from Xidian Experiment;

[0094] Vitamin E is Trolox water-soluble vitamin E with product number A486164-1kit from Shanghai Aladdin Biochemical Technology Co., Ltd.;

[0095] Glycerol is a sample with the trade name Glycerine 99.5% USP from Edenor Oleochemicals (M) Sdn. Bhd.;

[0096] Butylene glycol is a sample with the trade name 1,3Butylene glycol from Daicel Corporation;

[0097] Boric acid is a sample with the product number 10004817 from Sinopharm Reagent;

[0098] Lecithin is a sample with a purity greater than 95% from Shanghai Aiweituo Medical Technology Co., Ltd.

[0099] Cholesterol was an analytically pure sample from Shanghai Macklin Biochemical Co., Ltd.

[0100] Polyglycerol-10 stearate was a sample from Nikko Chemicals Co., Ltd.

[0101] Part 1: Examples and Comparative Examples

[0102] Preparation methods for Examples 1-7 and Comparative Examples 1-11: First, weigh the prickly ash fruit extract and mix it with the components other than water in the other solvent components, heat to 50-60 °C, mix evenly, then cool to room temperature, sequentially add the other components in the examples and comparative examples, and finally add water to 100 wt% and mix evenly to obtain the sample of the composition to be tested.

[0103] Preparation method for Example 8: At room temperature, first weigh the prickly ash fruit extract and mix it evenly with the other solvent components, then sequentially add the other components of the composition components, and stir for 12 hours to obtain the sample of the composition to be tested.

[0104] Preparation methods for Examples 9 and 10: First, weigh the prickly ash fruit extract and mix it with lecithin, cholesterol, and polyglycerol-10 stearate, heat to 80-90 °C, mix evenly, then sequentially add the other components, and obtain the sample of the composition to be tested after high-pressure homogenization.

[0105] The component ratios of Examples 1-10 and Comparative Examples 1-11 are shown in Table 1.

[0106] Table 1 Component ratios of Examples 1-10 and Comparative Examples 1-11

[0107]

[0108]

[0109] Part 2: Performance Tests

[0110] Before the following tests on each sample, dilution is required. Specifically, for the samples of the composition components in Examples 1-10 and Comparative Examples 1-11 plus the other solvent components, they are further prepared into a 1% solution sample with pure water (prepared and used immediately), that is, take 1 g of the solution prepared in Part 1 and dilute it with 99 g of water to become the solution sample.

[0111] 2.1 Safety Test

[0112] Patch test method: Select a suitable patch tester. Using the closed patch test method, apply approximately 0.03 ml of the test substance to the patch tester, and then apply a special tape externally to the back of the subject. After 24 hours, remove the test substance. Observe the skin reaction at 0.5, 24, and 48 hours after the patch test respectively, and record the results according to the skin reaction grading standard in the "Technical Specifications for Cosmetics Safety" (2015 Edition).

[0113] 2.2 Anti-UV light damage effect experiment and results

[0114] In the blank group of this experiment, HaCaT cells were not pretreated with any sample. After pretreating HaCaT cells with 1 μL of the above solution sample for 24 h, then irradiating the blank group and each solution sample with UVB at a dose of 5 mJ / cm 2 The survival rate of HaCaT cells in each test group can be obtained, and the specific test results are shown in Table 2.

[0115] Long-term or excessive exposure to UV in sunlight can cause serious damage to skin cells. Research shows that human keratinocytes (KC) are important target cells for UVB acting on human skin. HaCaT cells are human immortalized keratinocytes, which are derived from keratinocytes and retain the differentiation ability of keratinocytes. They are commonly used to study skin physiological and pathological processes, including wound healing, inflammatory response, and cell proliferation, etc.

[0116] 2.3 DPPH free radical scavenging experiment and results

[0117] Take 2 mL of the above solution sample and mix it thoroughly with 2 mL of DPPH solution prepared with absolute ethanol at a concentration of 2×10 -4 mol / L. After reacting in the dark at room temperature for 30 min, measure the absorbance at 517 nm to obtain A1; and measure the absorbance A2 of the mixture of 2 mL of the above solution sample and 2 mL of absolute ethanol and the absorbance A0 of the mixture of 2 mL of absolute ethanol according to the same operation. Calculate the DPPH free radical scavenging rate according to the following formula: DPPH free radical scavenging rate = [1 - (A1 - A2) / A0] × 100%,

[0118] 2.4 Hyaluronidase inhibition experiment and results

[0119] Reagent preparation: Measure 1155 μL of glacial acetic acid and dilute it to 100 mL to obtain acetic acid solution. Take 4.8 mL of acetic acid solution as solution A. Weigh 2.72 g of sodium acetate crystal, dissolve it in water and make up the volume to 100 mL to obtain sodium acetate solution. Take 45.2 mL of sodium acetate solution as solution B. Mix solution A and solution B and make up the volume to 100 mL with water to prepare an acetic acid buffer solution with pH = 5.6. Use the prepared acetic acid buffer solution as a solvent to prepare hyaluronidase solution and sodium hyaluronate solution (the final working concentrations of the two solutions are 1250 u / mL and 0.5 mg / mL); Prepare the calcium chloride solution with a concentration of 0.25 mmol / L; Prepare the sodium hydroxide solution with a concentration of 0.4 mol / L; Prepare the sodium carbonate solution with a concentration of 1.0 mol / L. Add 3.5 mL of acetylacetone to 50 mL of sodium carbonate solution to prepare acetylacetone solution; Finally, weigh 0.8 g of p-dimethylaminobenzaldehyde and dissolve it in 15 mL of concentrated hydrochloric acid and 15 mL of absolute ethanol to prepare Ehrlich reagent.

[0120] Test procedure: Adopt the in vitro inhibition test method of hyaluronidase. Take 0.1 mL of calcium chloride solution and hyaluronidase solution and incubate them at a constant temperature for 20 min; Add 0.5 mL of sample solution and continue to incubate at 37 °C for 20 min; Add 0.5 mL of sodium hyaluronate solution and incubate for 30 min, then leave it at room temperature for 5 min; Add 0.1 mL of 0.4 mol / L NaOH solution and 0.5 mL of acetylacetone solution, heat it in a boiling water bath for 15 min and then immediately cool it with ice water for 5 min; Add 1.0 mL of Ehrlich reagent and dilute it with 3.0 mL of absolute ethanol, let it stand for 20 min to develop color, and measure its absorbance value with a spectrophotometer.

[0121] The inhibition rate of hyaluronidase is calculated according to the following formula:

[0122] Inhibition rate of hyaluronidase = {(A - B) - (C - D)} / (C - D) × 100%

[0123] In the formula: A is the absorbance of the control solution (using acetic acid buffer solution instead of the sample solution), B is the absorbance of the control blank solution (using acetic acid buffer solution instead of the sample solution and enzyme solution), C is the absorbance value of the test solution, and D is the absorbance of the test blank solution (using acetic acid buffer solution instead of the enzyme solution). During the test, first perform wavelength scanning on the control group samples to determine the maximum absorption wavelength, and then use deionized water as the reference and measure the absorbance at this maximum absorption wavelength respectively.

[0124] 2.5 Cell scratch healing experiment and results

[0125] In the blank group of this experiment, after pre-treating HaCaT cells with 1 μL of pure water for 24 h, 1 μL of the above solution sample was added to pre-treat HaCaT cells for 24 h. Then, a cell scratch was made on the blank group and each sample group with a 10 μL pipette tip, washed three times with phosphate-buffered saline (PBS), and then added to a serum-free medium. Immediately after adding the sample, it was placed under a microscope for photography to record the image at 0 h of the scratch. Then, it was placed in an incubator and cultured for another 15 h, and then taken out and placed under a microscope for photography to record the image of the scratch at 15 h. The ImageJ software was used to process the images obtained by microscope photography to obtain the scratch area data of each image.

[0126] The cell scratch healing rate was calculated according to the following formula:

[0127] Cell scratch healing rate (%) = [(S 0h - S 15h ) / S 0h × 100%

[0128] In the formula: S 0h is the cell scratch area at 0 h, and S 15h is the cell scratch area at 15 h.

[0129] Table 2 Photoprotection test results of Examples 1-10 and Comparative Examples 1-11

[0130]

[0131]

[0132] Result analysis:

[0133] 1. In terms of skin irritation: The test results of the compositions of Examples 1-10 and Comparative Examples 1-11 showed no adverse reactions.

[0134] 2. In terms of anti-UV light damage: The higher the survival rate of HaCaT cells, the better the anti-UV light damage effect of the test group. From the HaCaT cell survival rate test results in Table 2, it can be seen that the photoprotective plant compositions prepared in Examples 1-10 had a significant protective effect on HaCaT cells, and the cell survival rate after UV light irradiation was above 90.72%, which could effectively counteract the damage of UV light to the skin and achieve the effect of anti-light damage. However, the compositions prepared in Comparative Examples 1-11 had a lower cell survival rate after UV light irradiation than the photoprotective plant compositions described in this application. This shows that the components of the photoprotective plant composition described in the present invention cooperate with each other, have a synergistic effect, can effectively inhibit the cell damage caused by UV light, and thus inhibit the skin damage caused by UV light to achieve the photoprotection effect.

[0135] Specifically, through the comparison between Example 3 and Comparative Example 1, it can be seen that even a trace amount of Chinese prickly ash fruit extract and epigallocatechin gallate glucoside can exhibit the effect of resisting UV light damage;

[0136] Through the comparison between Example 3 and Comparative Examples 2 - 4 and Comparative Example 1, it can be seen that when any one of the Chinese prickly ash fruit extract, epigallocatechin gallate glucoside, and cherry blossom extract is missing, it can exhibit a certain effect of resisting UV light damage, but the effect is not as good as that of Example 3; at the same time, through the comparison between Comparative Example 4 and Comparative Example 1, it can be seen that cottonseed extract, pomegranate seed extract, and fructooligosaccharide have a certain synergistic effect on the active ingredients for resisting UV light damage, but the effect is limited; at the same time, the results of Comparative Examples 2 - 4 can further prove that among the Chinese prickly ash fruit extract, epigallocatechin gallate glucoside, and cherry blossom extract, the first two are more important in the process of resisting UV light damage.

[0137] Horizontal comparison with Example 6 shows that even when the dosage of the active ingredients for resisting UV light damage is very small, cottonseed extract, pomegranate seed extract, and fructooligosaccharide can exhibit a very good synergistic effect; this shows that in actual formula applications, if the requirement for resisting UV light damage is not particularly high, when cottonseed extract, pomegranate seed extract, and fructooligosaccharide are used, the total amount of Chinese prickly ash fruit extract, epigallocatechin gallate glucoside, and cherry blossom extract can be as low as 0.2%.

[0138] Through Comparative Examples 5 - 7, it can be seen that when any one of the cottonseed extract, pomegranate seed extract, and fructooligosaccharide is missing, the anti - UV effect of the Chinese prickly ash fruit extract, epigallocatechin gallate glucoside, and cherry blossom extract cannot be fully enhanced, which indicates that the anti - UV damage of cells is not solely determined by aspects such as UV shielding and inhibition of cell pathways, and appropriate antioxidant effects equal to cell - activity - related factors will affect the efficacy of the active ingredients for resisting UV light damage.

[0139] Through Comparative Examples 8 - 10, it can be seen that when a part of the components is replaced with VE with strong antioxidant properties, there is no obvious improvement in the anti - UV light damage, although there is a slight improvement compared with Comparative Examples 4 - 6, but it is far less than that of Example 3. The possible reason is that the factors affecting the improvement of anti - UV active ingredients are not only related to antioxidant functions, but also related to components such as flavonoids and alkaloids in cherry blossom extract, cottonseed extract, and pomegranate seed extract in terms of factors such as inflammatory pathways and the expression of matrix metalloproteinase MMP - 1;

[0140] Through Comparative Example 11, it can be seen that when glycerol with better moisturizing effect is used to replace fructooligosaccharide, there is no obvious improvement in the anti - UV light damage function.

[0141] 3. In terms of the DPPH free radical scavenging rate: Research shows that cells can produce a large amount of reactive oxygen free radicals under blue light induction, causing cell damage. The specific test results are shown in Table 3.

[0142] From the DPPH free radical scavenging rate test results in Table 2, it can be seen that the photo-protective plant compositions prepared in Examples 1-10 have a significant scavenging effect on DPPH free radicals, with the DPPH free radical scavenging rate above 91.13%. They can effectively inhibit the skin damage caused by blue light, achieving the effects of antioxidant and anti-aging. However, the DPPH free radical scavenging rates of the compositions prepared in Comparative Examples 1-11 are far lower than those of the photo-protective plant compositions described in this application. This shows that the components of the photo-protective plant composition described in the present invention cooperate with each other, having a synergistic effect, and can effectively inhibit the generation of free radicals caused by blue light, thereby inhibiting the skin damage caused by blue light and achieving the effects of antioxidant and photo-protection.

[0143] Specifically:

[0144] By comparing Examples 3, Comparative Example 1, and Comparative Examples 2-3, it can be seen that Comparative Example 1 can prove that the pericarpium zanthoxyli extract and epigallocatechin gallate glucoside have good DPPH free radical scavenging effects. Their relatively low DPPH free radical scavenging rates are due to the relatively small amount used. By comparing Comparative Examples 2-3 with Comparative Example 1, it can be proved again that the DPPH free radical scavenging effects of the pericarpium zanthoxyli extract and epigallocatechin gallate glucoside are excellent.

[0145] By comparing Examples 3, Comparative Example 1, and Comparative Examples 4-6, it can be seen that the importance of the Gossypium herbaceum seed extract and Punica granatum seed extract in scavenging DPPH free radicals is greater than that of the Prunus serrulata extract. Of course, it may also be due to the relatively small amount of the Prunus serrulata extract used. However, based on the conventional understanding of the Gossypium herbaceum seed extract and Punica granatum seed extract, even if the amount of the Prunus serrulata extract is significantly increased, the improvement in DPPH free radical scavenging may be limited.

[0146] By comparing Comparative Example 7 with Comparative Example 4, it can be seen that in terms of the DPPH free radical scavenging rate, the Prunus serrulata extract and fructooligosaccharide have similar effects.

[0147] From the results of Comparative Examples 8-10, it can be seen that the Prunus serrulata extract contributes less to the DPPH free radical scavenging rate.

[0148] By comparing Comparative Example 11 with Comparative Example 7, it can be seen that fructooligosaccharide contributes less to the DPPH free radical scavenging rate.

[0149] 4. In terms of hyaluronidase inhibition: The hyaluronidase inhibition rate test results of each test group are shown in Table 4.

[0150] Hyaluronidase is a component in the tissue matrix that restricts the diffusion of water and other extracellular substances. After being hydrolyzed by hyaluronidase, the intercellular space becomes non-viscous, causing cell degranulation and the exudation of newly synthesized mediators, resulting in the occurrence of inflammation or allergic reactions.

[0151] The higher the hyaluronidase inhibition rate, the stronger the anti-inflammatory effect of the test group. From the hyaluronidase inhibition rate test results in Table 2, it can be seen that the photo-protective plant compositions prepared in Examples 1-10 have a significant inhibitory effect on hyaluronidase, with a hyaluronidase inhibition rate of more than 90.82%. They can effectively inhibit the damage to the skin caused by the inflammatory reaction induced by histamine release, so as to achieve the anti-inflammatory effect. However, the inhibitory effect of the compositions prepared in Comparative Examples 1-11 on the activity of hyaluronidase is much lower. This shows that the components of the photo-protective plant composition of the present invention cooperate with each other, have a synergistic effect, can effectively inhibit the skin inflammatory reaction caused by the hydrolysis of hyaluronidase, improve the skin's own anti-inflammatory ability, and thus inhibit the skin damage caused by sunlight or other harmful light rays, so as to achieve the photo-protective effect of anti-inflammatory.

[0152] In terms of hyaluronidase, the trends shown by each comparative example are similar to those of the DPPH free radical scavenging rate.

[0153] 5. In terms of cell scratch healing performance: The specific test results of the cell scratch healing rates of each test group are shown in Table 2 and Figures 1 - 44 the cell scratch pictures.

[0154] Skin wound healing includes the proliferation and migration of skin cells. At present, many studies have shown that the migration ability of cells can be tested through the HaCaT cell scratch experiment, so as to verify the repair ability of the skin barrier. From the cell scratch healing rate test results in Table 2 and Figures 1 - 44 the cell scratch pictures, it can be seen that the photo-protective plant compositions prepared in Examples 1-10 can significantly promote the cell healing speed after scratching, with a scratch cell healing rate of more than 76.29%. They can effectively improve the skin's own repair ability, so as to achieve the effect of anti-photo-damage. However, the healing effect of the compositions prepared in Comparative Examples 1-11 on the scratched cells is relatively weak. This shows that the components of the photo-protective plant composition of the present invention cooperate with each other, have a synergistic effect, can effectively promote the cell healing speed after scratching, improve the skin's own repair ability, and thus inhibit the skin damage caused by sunlight or other harmful light rays, so as to achieve the photo-protective effect of repairing the skin.

[0155] In summary, the formula of the present invention has excellent synergistic effects in photo-protection aspects such as anti-UV light damage, antioxidant, anti-inflammatory and repair.

Claims

1. A plant light protection composition, characterized in that, By weight, it comprises the following components: Zanthoxylum fruit extract 0.1 - 0.4 parts; Epigallocatechin gallate glucoside 0.2 - 0.8 parts; Cherry blossom extract 0.2 - 1.6 parts; Gossypium herbaceum seed extract 22 - 28 parts; Pomegranate seed extract 22 - 28 parts; Fructooligosaccharide 0.2 - 0.8 parts; The content of sanshools in the Zanthoxylum fruit extract is 16 - 18% by weight; The total flavonoid content of the cherry blossom extract is 30 - 35% by weight, the polyphenol content is 4 - 5% by weight, and the polysaccharide content is 15 - 20% by weight; The polyphenol content of the Gossypium herbaceum seed extract is 0.7 - 0.8% by weight; The polyphenol content of the pomegranate seed extract is 0.7 - 0.8% by weight.

2. The plant light protection composition according to claim 1, characterized in that, It further comprises a carrier; the Zanthoxylum fruit extract, epigallocatechin gallate glucoside, cherry blossom extract, Gossypium herbaceum seed extract, pomegranate seed extract, and fructooligosaccharide are active ingredients; the active ingredients are loaded on the carrier or loaded in the carrier.

3. The plant light protection composition according to claim 2, characterized in that, The carrier is one of liposomes, microcapsules, and hydrogel networks.

4. The plant light protection composition according to claim 2, characterized in that, By weight, the weight of the carrier is 0.05 - 5 parts.

5. Use of the plant light protection composition as claimed in claim 1 for preparing cosmetics.

6. The use according to claim 5, characterized in that, The cosmetics are sunscreen products.

7. The use according to claim 6, characterized in that, The cosmetics are used for anti-UV light damage; and / or, the cosmetics are used for scavenging DPPH free radicals; and / or, the cosmetics are used for inhibiting hyaluronidase; and / or, the cosmetics are used for promoting cell wound healing.

8. A cosmetic, characterized in that, Containing the plant light protection composition as claimed in claim 1.

9. The cosmetic according to claim 8, characterized in that, Containing 0.01 - 10 wt% of the plant light protection composition.

10. The cosmetic according to claim 8, characterized in that, The cosmetics are sunscreen products, and the cosmetics further contain chemical sunscreen agents.

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