Plant composition with synergistic whitening effect and application thereof

Abstract

The invention discloses a plant composition with a synergistic whitening effect and application thereof, and belongs to the technical field of cosmetics. The plant composition comprises a rhizoma bletillae extract and arbutin. The invention further relates to plant whitening gel containing the plant composition. The plant whitening gel is prepared from the plant composition, sorbitol, carbomer, glycerol and pure water. The plant composition disclosed by the invention is mainly prepared from natural components, and the active plant extract is mainly prepared from rhizoma bletillae extract and is matched with arbutin to achieve a synergistic whitening effect. The two components not only synergistically enhance the effect of the composition for inhibiting synthesis and transmission of melanocytes, but also improve the inhibition effect on melanin synthesis related enzymes, thereby achieving a better whitening effect.

Description

Technical Field

[0001] The present invention belongs to the technical field of cosmetics, and in particular relates to a plant composition with synergistic whitening effects and application thereof. Background Art

[0002] Melanocytes (B16F10) in the basal layer of the human epidermis are the site of melanin production. After production, melanin is transferred from the dendritic tips of melanocytes to the stratum corneum and excreted through metabolism by keratinocytes (HaCaT). Melanin production is induced by various internal or external factors, such as inflammation, hormonal changes, ultraviolet (UV) light, and stress-mediated skin irritation. Excessive melanin production can lead to excessive skin pigmentation. The number of melanocytes, the extent of their dendrites, the activity of related enzymes, and the transfer of melanosomes all affect pigmentation. Excessive production or abnormal accumulation of melanin can cause various skin diseases, such as hyperpigmentation, spots, age spots, freckles, chloasma, and melanoma.

[0003] Melanin is divided into brown-black, alkaline-insoluble eumelanin and reddish-yellow, alkaline-soluble pheomelanin. Tyrosine oxidation is the initial step in melanin synthesis, and the main enzymes involved are tyrosinase (TYR) and tyrosinase-related proteins 1 and 2 (TRP1 and TRP2). TYR is the key rate-limiting enzyme in melanin synthesis. Melanin production mainly goes through two stages. First, tyrosine in melanocytes is hydroxylated to 3,4-dihydroxyphenylalanine (DOPA) under the catalysis of TYR, and then forms dopaquinone under the oxidation of TYR. Starting from dopaquinone, eumelanin and pheomelanin are produced through two different pathways. Melanin production is also related to the regulation of signaling pathways in melanocytes. Under ultraviolet irradiation, keratinocytes stimulate the secretion of α-melanocyte-stimulating hormone (α-MSH), which promotes melanin biosynthesis in epidermal melanocytes.

[0004] As society steadily advances, technology advances rapidly, and living standards continue to rise, people's pursuit of natural, green, healthy, and safe products is growing stronger. In this era, cosmetics with green and functional characteristics are increasingly popular among consumers. Plant extracts are rapidly developing in the cosmetics field, with their application scope continuously expanding and the variety of products involved increasing. Compared to traditional cosmetics, cosmetics formulated with plant extracts as active ingredients offer distinct advantages. On the one hand, these cosmetics break away from the excessive reliance on synthetic chemicals, significantly improving product safety. On the other hand, their natural ingredients are more compatible with the physiological characteristics of the skin and are more easily absorbed by the skin, resulting in superior product efficacy. Furthermore, these cosmetics demonstrate superior performance in specific functions such as moisturizing, whitening, and antioxidant properties. Therefore, the use of plant extracts in cosmetics is not only in line with market trends but also an inevitable trend in the future development of the cosmetics industry. Existing research indicates that most whitening ingredients on the market primarily work by inhibiting melanin synthesis, blocking the signaling pathways associated with melanin production, or interfering with melanin transfer, thereby achieving a skin-whitening effect. Various whitening ingredients exhibit significant differences in how they exert their whitening effects. Therefore, rationally combining substances that act on different whitening targets may be an effective strategy for enhancing product whitening effects, potentially providing new insights and directions for the research and development of whitening products. Summary of the Invention

[0005] Because existing whitening products have a single effect and are unable to simultaneously inhibit the secretion, production, and transport of melanin to achieve a whitening effect, or simultaneously inhibit key melanin synthesis proteins such as TYR, TRP1, and TRP2, the present invention provides a plant composition with a synergistic whitening effect. This composition can both enhance the whitening effect of the product and reduce the amount of a single ingredient (Bletilla striata extract or arbutin) added to the cosmetic, thereby achieving the goals of increasing efficacy and reducing costs.

[0006] The plant composition with synergistic whitening effect provided by the present invention consists of a bletilla striata extract and arbutin, wherein the mass ratio of the bletilla striata extract to the arbutin is 1:10 to 1:20.

[0007] Furthermore, in the plant composition having a synergistic whitening effect of the present invention, the mass ratio of the Bletilla striata extract to arbutin is preferably 1:10 to 1:15.

[0008] The above-mentioned Bletilla striata extract is obtained by the following method: drying the Bletilla striata tuber to constant weight, crushing it with an ultra-high-speed grinder, and passing it through a 60-mesh sieve to obtain Bletilla striata powder; mixing the Bletilla striata powder with pure water at a material-liquid ratio of 1g:30mL, ultrasonicating at a power of 150-300W for 30 minutes, centrifuging, taking the supernatant, concentrating, filtering and sterilizing, and obtaining the Bletilla striata extract.

[0009] The plant composition of the present invention has a significant whitening effect and can be used to prepare cosmetics having a whitening effect. Therefore, the present invention also provides a plant-based whitening gel, which is made from the following ingredients in the following weight percentage ratios: 7% to 10% sorbitol, 8% to 12% carbomer, 5% to 10% glycerin, 8% to 15% of the plant composition, and the balance being pure water.

[0010] The preparation method of the plant whitening gel comprises the following steps:

[0011] S1. Evenly mixing the plant composition, sorbitol, glycerin and purified water to obtain an aqueous phase;

[0012] S2. Evenly mix carbomer and water, and wait until the carbomer fully absorbs water and expands to obtain a plant whitening gel.

[0013] The present invention further provides use of the plant whitening gel in preparing cosmetics with whitening effects.

[0014] The beneficial effects of the present invention are as follows:

[0015] Bletilla striata extract is extracted from the tuber of Bletilla striata through ultrasonic extraction. This extraction method is highly controllable and reproducible, with low cost and easy operation. Its main active ingredients are Bletilla striata polysaccharides, Bletilla striata glycosides, and Bletilla striata polyphenols, which have excellent whitening and antioxidant effects. Arbutin is a natural compound that can inhibit melanin synthesis and free radical activity and promote stratum corneum shedding. The present invention prepares a composition with a synergistic whitening effect by compounding Bletilla striata extract and arbutin in a scientific ratio. These two substances cooperate with each other and, through different protein inhibition pathways, synergistically enhance the composition's effect on melanin production in melanocytes and enhance the inhibitory effect on TYR and TRP1 protein expression. This effectively reduces the production and deposition of melanin in the underlying skin, brightens the skin tone, and achieves a better whitening effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is the effect of the blank control group, positive control group and experimental group on the melanin content of B16F10 cells.

[0017] Figure 2 This is the effect of the blank control group, positive control group and experimental group on the expression of TYR and TRP1 proteins in B16F10 cells.

[0018] Figure 3 Effects of microneedle injection of cosmetic sample solutions on UVB-induced pigmentation in guinea pig skin (Masson-Fontana, 40x). (A) Guinea pig skin changes, (B) melanin content in sections from each group, (C) section staining; a represents the blank control group, b represents the 7.5 μg / mL Bletilla striata extract group, c represents the arbutin group, and d represents the botanical combination group.

[0019] Figure 4 Effects of microneedle injection of a cosmetic sample solution combined with application of a botanical whitening gel on UVB-induced pigmentation in guinea pig skin (Masson-Fontana, 40x). (A) Image of guinea pig skin changes, (B) Melanin content in sections from each group, (C) Stained sections; a represents the blank control group, b represents the 7.5 μg / mL Bletilla striata extract group, c represents the arbutin group, and d represents the botanical combination group.

[0020] In the figures, * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001, and **** indicates p < 0.0001 vs. blank control group; # Indicates p<0.05, ## Indicates p<0.01, ### Indicates p<0.001, #### Indicates p<0.0001 vs. arbutin group. DETAILED DESCRIPTION

[0021] To better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments. Those skilled in the art should understand that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0022] The instruments and equipment used in the embodiments of the present invention are all conventional instruments and equipment in the art and can be replaced by instruments and equipment that meet the corresponding standards.

[0023] Unless otherwise specified, the Chinese herbal medicine raw materials used in the examples of the present invention are all commercially available, and the reagents used in the examples of the present invention are all commercially available analytically pure chemical reagents unless otherwise specified.

[0024] Experimental reagents and equipment: Bletilla striata tubers were purchased from Yunnan Jinggu Forestry Co., Ltd., B16F10 cells were purchased from Wuhan Punuosai Life Science Technology Co., Ltd., Anti-Tyrosinase antibody, Anti-TRP1 antibody, and Anti-GAPDH antibody were purchased from Abcam, horseradish enzyme-labeled goat anti-rabbit IgG (H+L) and horseradish enzyme-labeled goat anti-mouse IgG (H+L) were purchased from Zhongshan Jinqiao, guinea pigs were purchased from Shaanxi Junxing Biotechnology Co., Ltd., Masson-Fontana melanin staining kit was purchased from Solebao, and microneedles were purchased from Shenzhen Peini Medical Technology Co., Ltd.

[0025] Example 1

[0026] 1. Preparation of Bletilla striata extract

[0027] Bletilla striata extract: Bletilla striata tubers were dried to constant weight, crushed in an ultra-high-speed grinder, and passed through a 60-mesh sieve to obtain Bletilla striata powder. The Bletilla striata powder was weighed and mixed with pure water at a material-liquid ratio of 1g:30mL. The mixture was ultrasonically cleaned at 200W for 30 minutes, centrifuged, and the supernatant was collected, concentrated, and filtered for sterilization to obtain Bletilla striata extract. The concentration of Bletilla striata polyphenols (BRP-U) in the Bletilla striata extract was determined using the Folin phenol method. The results showed that the total polyphenol concentration of Bletilla striata was 30.35μg / mL.

[0028] 2. Melanin content detection

[0029] Preparation of plant composition: Bletilla striata extract and arbutin were mixed in a mass ratio of 1:20, 3:40, and 1:10 to obtain a plant composition. Using pure water as the solvent, samples were prepared according to the concentrations of Bletilla striata extract and arbutin in Table 1. In the following experiments, the sample without Bletilla striata extract and arbutin was set as the blank control group, the sample with only arbutin added was set as the positive control group, and the sample with Bletilla striata extract and the sample with the plant composition added were set as the experimental group. B16F10 cells were treated with each group of samples for 48 hours, the cells were collected, 200 μL of cell lysis solution (10% DMSO, 1M NaOH) was added, blown evenly, placed in a water bath and heated at 85°C for 30 minutes, vortexed to mix, 100 μL of cell lysis solution was drawn into a 96-well plate, and the absorbance was measured at 405 nm. The calculation formula for the inhibition rate of melanin production by the test sample is as follows:

[0030] Melanin production inhibition rate (%) = [1-(OD experimental group / OD blank control group)] × 100%

[0031] The results of melanin content detection are shown in Table 1 and Figure 1 shown.

[0032] Table 1 Effects of different samples on melanin production in B16F10 cells (X±S, n=3)

[0033]

[0034] Note: ** indicates p < 0.01, *** indicates p < 0.001, **** indicates p < 0.0001 vs. blank control group; ## Indicates p<0.01, #### Indicates p<0.0001 vs. arbutin group.

[0035] The results showed that compared with the blank control group, both single Bletilla striata extract and arbutin significantly inhibited the secretion of melanin, and the inhibitory effect of high-concentration Bletilla striata extract was stronger than arbutin. Compared with the effect of arbutin alone, the low-concentration plant combination (Bletilla striata extract concentration of 5μg / mL) significantly inhibited the production of melanin (P < 0.01), and the medium and high-concentration plant combinations (Bletilla striata extract concentrations of 7.5μg / mL and 10μg / mL) extremely significantly inhibited the production of melanin (P < 0.0001). This shows that Bletilla striata extract and arbutin have a significant synergistic effect in inhibiting melanin synthesis.

[0036] 3. Detection of TYR and TRP1 protein expression in B16F10 cells

[0037] B16F10 cells were treated with the samples from each group in Experiment 2 for 48 h, and the cells were collected for protein sample preparation. The expression of TYR and TRP1 proteins in B16F10 cells was then detected by Western Blot. The membrane was exposed using an automatic chemiluminescence system, and the optical density of the developed bands was analyzed using Image J software. The expression results of TYR and TRP1 proteins in B16F10 cells are shown in Tables 2 and 3. Figure 2 shown.

[0038] Table 2 Effects of different samples on protein expression in B16F10 cells (X±S, n=3)

[0039]

[0040]

[0041] Note: ** indicates p < 0.01, *** indicates p < 0.001 vs. blank control group; ## Indicates p<0.01, #### Indicates p<0.0001 vs. arbutin group.

[0042] The results showed that compared with the blank control group, both single Bletilla striata extract and arbutin inhibited the expression of TYR and TRP1 proteins. Compared with arbutin alone, the medium and high concentrations of the plant combination (Bletilla striata extract at concentrations of 7.5 μg / mL and 10 μg / mL) significantly inhibited the expression of TYR protein (P < 0.01). Compared with the same concentration of Bletilla striata extract group, the plant combination reduced the expression of TRP1 protein (P < 0.01). This indicates that Bletilla striata extract and arbutin have a significant synergistic effect in inhibiting the expression of TYR and TRP1 proteins.

[0043] 4. Study on the whitening efficacy of plant compositions using a guinea pig model induced by ultraviolet irradiation

[0044] (1) Experimental principles and significance of test indicators

[0045] Guinea pig skin structure and physiological functions are similar to those of human skin. It contains melanocytes, which are activated by ultraviolet (UV) radiation and other stimuli, promoting melanin production. As melanin accumulates and deposits, the guinea pig's skin gradually darkens. This makes the results of whitening studies conducted in the guinea pig model more extrapolable and can better predict the effects of whitening products or drugs in humans.

[0046] Melanin granule distribution: By using Masson-Fontana staining on guinea pig skin sections, the distribution of melanin granules in the tissue can be observed, reflecting the effect of the sample on melanin production and transport.

[0047] Melanin content was estimated using ImageJ software.

[0048] (2) Sample preparation

[0049] Four botanical whitening gels were prepared according to the mass percentage ratios shown in Table 3 below: a blank control group (A), a medium-concentration Bletilla striata extract group (B), an arbutin group (C), and a botanical combination group (D). The specific preparation method was as follows: Bletilla striata extract, arbutin, sorbitol, glycerin, and purified water were uniformly mixed to obtain an aqueous phase; carbomer was uniformly mixed with the aqueous phase, and the carbomer was allowed to fully absorb water and swell to obtain the botanical whitening gel.

[0050] Table 3 Recipe components (wt%)

[0051]

[0052]

[0053] Four cosmetic sample solutions were prepared according to the mass percentage ratios in Table 4 below: a blank control group (A), a medium-concentration Bletilla striata extract group (B), an arbutin group (C), and a plant composition group (D).

[0054] Table 4 Recipe Components (wt%)

[0055]

[0056] (3) Experimental methods

[0057] Six guinea pigs (weighing approximately 300g) were selected. The hair on their backs was first shaved short with an electric shaver, and then the remaining hair was removed daily with Veet Hair Removal Cream. Each guinea pig's back was divided into four sections, one blank control group and three experimental groups, with three independent experiments conducted in each group. The guinea pigs' backs were exposed to UVB radiation (311nm, total energy 1000μW / cm 2 ) for 15 minutes. Guinea pigs were divided into two groups: one microneedle group (after UV irradiation, the cosmetic sample solution was injected) and the other microneedle gel group (after UV irradiation, the cosmetic sample solution was injected and the same plant whitening gel was applied). This was repeated for 15 days. The guinea pig skin epidermis was excised, fixed with 4% paraformaldehyde, dehydrated with anhydrous ethanol, transparentized with xylene, embedded in paraffin, and sectioned with paraffin (4 μm thickness). After xylene dewaxing and hydration with graded ethanol, the sections were stained according to the instructions of the Masson-Fontana melanin staining kit. After sealing, the sections were placed under an upright microscope with a 40x objective lens to observe the distribution of melanin particles. The melanin content was estimated using ImageJ software, and statistics were calculated using Graph Prism software.

[0058] The results of the microneedle group were as follows Figure 3 As shown in the results, compared with the blank control group, the medium concentration of Bletilla striata extract and the plant composition significantly inhibited the hyperpigmentation of the skin. Although the arbutin group had a certain inhibitory effect on skin pigmentation, it was not significant. Further observation of the microneedle gel group ( Figure 4 ) found that the plant composition had a stronger inhibitory effect on skin hyperpigmentation caused by ultraviolet radiation than the white ginseng extract and arbutin when used alone. Based on the above results, it can be concluded that whether the cosmetic sample solution is injected only by microneedle, or the cosmetic sample solution is injected by microneedle and applied with the same group of plant whitening gel, the skin pigmentation problem caused by UVB radiation can be effectively alleviated. It is worth noting that when the cosmetic sample solution is injected by microneedle and applied with the same group of plant whitening gel, the plant composition has a synergistic effect in inhibiting skin hyperpigmentation.

Claims

1. A plant composition with synergistic whitening effect, characterized in that: The composition consists of a bletilla striata extract and arbutin, wherein the mass ratio of the bletilla striata extract to the arbutin is 1:10 to 1:

20.

2. The plant composition with synergistic whitening effect according to claim 1, characterized in that: The mass ratio of the Bletilla striata extract to arbutin is 1:10-15.

3. The plant composition with synergistic whitening effect according to claim 1 or 2, characterized in that: The bletilla striata extract is obtained by the following method: drying the bletilla striata tuber to constant weight, crushing it with an ultra-high-speed grinder, and passing it through a 60-mesh sieve to obtain bletilla striata powder; mixing the bletilla striata powder with pure water at a material-liquid ratio of 1g:30mL, ultrasonically treating the mixture at a power of 150-300W for 30 minutes, centrifuging, taking the supernatant, concentrating, filtering and sterilizing, and obtaining the bletilla striata extract.

4. A plant-based whitening gel, characterized in that: The invention is prepared from the following ingredients in the following mass percentages: 7% to 10% of sorbitol, 8% to 12% of carbomer, 5% to 10% of glycerol, 8% to 12% of the plant composition according to claim 1 or 2, and the balance being pure water.

5. A method for preparing the plant whitening gel according to claim 4, characterized in that: The steps include: S1. Evenly mixing the plant composition, sorbitol, glycerin and purified water to obtain an aqueous phase; S2. Evenly mix carbomer and water, and wait until the carbomer fully absorbs water and swells to obtain a plant whitening gel.

6. Use of the plant whitening gel according to claim 4 in the preparation of cosmetics with whitening effect.

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