Composition containing plant extract and rice fermentation product filtrate and use thereof

Abstract

The present application relates to a composition containing plant extracts and rice fermentation product filtrate and its application. The composition contains Artemisia Princeps leaf extract, Morinda Citrifolia fruit juice and yeast / rice fermentation product filtrate. Compared with the prior art, the present application verifies the moisturizing mechanism and synergistic effect of the combination of plant extracts and yeast / rice fermentation product filtrate at the gene level and cell level, and quickly screens out the combination containing Artemisia Princeps leaf extract, Morinda Citrifolia fruit juice and yeast / rice fermentation product filtrate, which can increase the expression of skin hydration and barrier function related genes FLG, IVL, LOR, KLK5, KLK7 and SPINK5, and is beneficial to improve the moisturizing and barrier function of the skin.

Description

Technical Field

[0001] This invention belongs to the field of daily chemical products technology, and in particular relates to compositions containing plant extracts and rice fermentation product filtrate and their applications. Background Technology

[0002] Currently, commercially available cosmetics generally suffer from poor moisturizing longevity. Although water can be used to replenish moisture in the skin, its efficacy in cosmetics is relatively poor, and the moisturizing effect is not long-lasting enough to achieve the desired effect.

[0003] Chinese patent CN116035959A discloses a composition containing algae extract and yeast / rice fermentation product filtrate, and its application. The composition contains algae extract and yeast / rice fermentation product filtrate, with a mass ratio of 40-70:20-40. However, this method suffers from several drawbacks: firstly, the amount of algae extract added is very high; secondly, it does not verify its moisturizing mechanism and synergistic effect at the gene and cellular levels. Summary of the Invention

[0004] Given the poor moisturizing properties of existing compositions or formulations, this invention provides a composition containing plant extracts and rice fermentation product filtrate, and its application.

[0005] The objective of this invention can be achieved through the following technical solutions:

[0006] The present invention provides a composition containing plant extracts and rice fermentation product filtrate, wherein the composition contains Morinda Citrifolia juice and yeast / rice fermentation product filtrate.

[0007] In one embodiment of the present invention, the mass ratio of sea morinda juice to yeast / rice fermentation product filtrate in the composition is 0.001-1:5-50, preferably 0.005-0.1:5-30, more preferably 0.005-0.05:5-25, and even more preferably 0.005-0.05:20.

[0008] The present invention also provides a composition containing plant extracts and rice fermentation product filtrate, wherein the composition contains Artemisia princeps leaf extract and yeast / rice fermentation product filtrate.

[0009] In one embodiment of the present invention, the mass ratio of Artemisia princeps leaf extract to yeast / rice fermentation product filtrate in the composition is 0.001-1:5-50; preferably 0.1-1:20-40.

[0010] In one embodiment of the invention, the composition contains Artemisia Princeps leaf extract, Morinda Citrifolia juice, and yeast / rice fermentation product filtrate.

[0011] In one embodiment of the present invention, the mass ratio of Artemisia argyi leaf extract, Morinda officinalis juice, and yeast / rice fermentation product filtrate in the composition is 0.001-1:0.001-1:5-97; preferably 0.1-1:0.1-1:20-92, more preferably 0.1-1:0.1-1:30-70, and even more preferably 0.1-1:0.1-1:30-40.

[0012] The present invention further provides a method for preparing a composition containing plant extracts and yeast / rice fermentation product filtrate, wherein two or three of the following are mixed in a certain proportion: Artemisia princeps leaf extract, Morinda citrifolia juice, and yeast / rice fermentation product filtrate.

[0013] This invention further provides the application of a composition containing plant extracts and rice fermentation product filtrate, wherein the composition is used to prepare skincare products. In the preparation of these skincare products, the composition containing plant extracts and yeast / rice fermentation product filtrate replaces part or all of the water in the formulation. The composition of this invention containing plant extracts and yeast / rice fermentation product filtrate is a more highly effective moisturizing ingredient, capable of replacing part or all of the water in skincare product formulations, and achieving better skincare results.

[0014] The present invention further provides a skin care product, the composition of which and the mass fraction of each ingredient are as follows:

[0015] Glycerin 2-10%

[0016] Butanediol 1-5%

[0017] Yeast / rice fermentation product filtrate 5-95%

[0018] Sea ginseng juice 0.001-1%

[0019] Phenoxyethanol 0.1-1%

[0020] Ethylhexylglycerin 0.01-0.1%

[0021] Artemisia leaf extract 0.001-1%

[0022] Add water to 100%.

[0023] The present invention further provides an anhydrous skin care product, the composition of which and the mass fraction of each component are as follows:

[0024] Glycerin 2-10%

[0025] Butanediol 1-5%

[0026] Add yeast / rice fermentation product filtrate to 100%.

[0027] Sea ginseng juice 0.001-1%

[0028] Phenoxyethanol 0.1-1%

[0029] Ethylhexylglycerin 0.01-0.1%

[0030] Artemisia leaf extract: 0.001-1%.

[0031] This invention enhances the basic moisturizing and barrier repair effects of yeast / rice fermentation product filtrate by combining it with plant extracts.

[0032] This invention evaluated the effects of using yeast / rice fermentation product filtrate alone and in combination with plant extracts on the mRNA expression of genes involved in skin hydration and barrier function, as well as epidermal keratinocyte proliferation, through in vitro experiments. The results showed that the combination of yeast / rice fermentation product filtrate and Artemisia princeps leaf extract increased the mRNA expression of all examined genes, exceeding the levels achieved by either gene alone. Furthermore, the combination of yeast / rice fermentation product filtrate and Morinda citrifolia juice significantly increased epidermal keratinocyte proliferation, exceeding the levels achieved by either gene alone. This demonstrates that the combination of plant extracts with yeast / rice fermentation product filtrate alone has a synergistic effect on skin moisturizing and barrier repair. Moreover, the combination of yeast / rice fermentation product filtrate with Morinda citrifolia juice and Artemisia princeps leaf extract exhibits even better technical effects.

[0033] Compared with existing technologies, this invention verifies the moisturizing mechanism and synergistic effect of the combination of plant extracts and yeast / rice fermentation product filtrate at the gene and cellular levels. It rapidly screens out combinations containing Artemisia Princeps leaf extract, Morinda Citrifolia juice and yeast / rice fermentation product filtrate that can increase the expression of genes related to skin hydration and barrier function, such as FLG, IVL, LOR, KLK5, KLK7 and SPINK5, which is beneficial to improving skin moisturization and barrier function. Attached Figure Description

[0034] Figure 1 The effects of different combinations of RFL solid contents and APLE in Examples 1-4 on the mRNA expression of genes related to skin hydration and barrier function in NHEKs.

[0035] Figure 2 Combined effects of different RFL and MCFJ combinations on NHEKs cell proliferation in Examples 5-9 and 11 (MTT assay).

[0036] Figure 3 Synergistic effects of different RFL and MCFJ combinations on NHEKs cell proliferation in Examples 5-7 and 9-11 (cell counting method).

[0037] Figure 4 Synergistic effect of different RFL and MCFJ combinations on NHEKs cell proliferation in Examples 6, 7, 10, and 11 (nuclear staining). Detailed Implementation

[0038] The meanings of key terms or technical abbreviations used in this invention are as follows:

[0039] RFL: Rice Fermented Liquid, Filtrate from Yeast / Rice Fermentation Products

[0040] RFL solid contents: are freeze-dried solids obtained by freeze-drying yeast / rice fermentation product filtrate (freeze-drying conditions: temperature reduced to -20℃~-50℃ in a freeze dryer, vacuum degree controlled at 5~20Pa);

[0041] APLE: Artemisia Princeps Leaf Extract

[0042] MCFJ: Morinda Citrifolia Fruit Juice

[0043] NHEKs: Normal human epidermal keratinocytes.

[0044] KBM: keratinocyte basal medium.

[0045] KGM: keratinocyte growth medium.

[0046] FLG: filaggrin

[0047] IVL: involucrin, inner lining protein

[0048] LOR: loricrin, loricin

[0049] KLK5: kallikrein 5, kallikrein 5

[0050] KLK7: kallikrein 7, kallikrein 7

[0051] SPINK5: Serine peptidase inhibitor, Kazal type 5. The functions or effects of each active ingredient in this invention are described below:

[0052] Rice fermentation product filtrate, also known as yeast / rice fermentation product filtrate, is generally prepared through the following steps: preparing rice flour, adding water, heat sterilization (to sterilize and disinfect the material and avoid microbial contamination), inoculation, fermentation, heat sterilization (to kill yeast and ensure the product meets microbial standards), filtration, and concentration to obtain the yeast / rice fermentation product filtrate.

[0053] Its ingredients include fermentation metabolites obtained by yeast fermenting rice, such as polysaccharides, proteins, and organic acids. These ingredients are beneficial to the skin, which can maintain skin moisture, eliminate free radicals, renew skin cells, fight skin inflammation, reduce cell damage, provide nutrients, and maintain healthy skin.

[0054] Optionally, the method for preparing the yeast / rice fermentation product filtrate includes the following steps:

[0055] Rice powder is mixed with water, heated, and stirred to form rice slurry. β-glucanase is added to the rice slurry for enzymatic hydrolysis to obtain rice slurry hydrolysate.

[0056] Yeast strains were inoculated into rice milk hydrolysate, and fermentation was carried out to obtain fermentation broth;

[0057] The fermentation broth is concentrated using reverse osmosis technology to efficiently enrich active substances, then heated for sterilization and cooled for filtration.

[0058] In this embodiment, the yeast strain in the yeast solution is Saccharomyces veronae.

[0059] Optionally, the rice slurry has a mass concentration of 40-60%. For example, a mass concentration of 50%.

[0060] Optionally, the amount of enzyme added is 0.001-1% of the mass of the rice milk. For example, the amount added is 0.5%.

[0061] Optionally, the enzyme activity is 100-200 U / g. For example, the enzyme activity is 150 U / g.

[0062] Optionally, the fermentation temperature is between 20°C and 40°C. For example, the fermentation temperature is 37°C.

[0063] Optionally, the fermentation time is 24 hours to 72 hours. For example, the fermentation time is 48 hours.

[0064] Artemisia princeps leaf extract is an active ingredient extracted from the leaves of Artemisia princeps. Artemisia princeps is an herbaceous plant, and its leaf extract is currently a chemical raw material used in skincare products. This extract can enhance skin cell metabolism, providing skin revitalization and anti-aging effects; it can also be used as an antioxidant, skin whitening agent, and anti-inflammatory agent. Artemisia princeps leaf extract can enhance the expression of FLG and LOR through the aryl hydrocarbon receptor (AHR) and its downstream transcription factor OVO-like1 (OVOL1) pathway.

[0065] Morinda citrifolia, commonly known as noni fruit, has been used in Polynesian traditional medicine for over 2000 years as a folk remedy to treat various ailments and diseases, including fever, pain, infection, high blood pressure, fatigue, diabetes, cancer, and inflammation. Currently, Morinda citrifolia extract is being used as an active ingredient in skincare products.

[0066] Genes related to skin hydration and barrier function include FLG, IVL, LOR, KLK5, KLK7, and SPINK5.

[0067] FLG is degraded into free amino acids, which help generate natural moisturizing factor (NMF) to maintain epidermal hydration. Low FLG biosynthesis can induce the pathogenesis of xerosis. IVL and LOR are components of the epidermal keratinocyte capsule (CE), which is an important part of the epidermal barrier. Kallikrein-associated peptidase (KLK) is a serine protease expressed from the granular layer of the epidermis to the stratum corneum. KLK5 and KLK7 play an important role in physiological peeling by degrading keratinocytes that adhere to each other. In addition, serine protease inhibitors, such as Kazal-associated lymphoid epithelium 5 (LEKTI; encoded by the SPINK5 gene), regulate KLK activity to achieve balanced desquamation. Therefore, increasing the expression of these genes is beneficial to improving skin hydration and barrier function.

[0068] The sources of the raw materials used in the following embodiments of the present invention are explained as follows:

[0069] Artemisia Princeps Leaf Extract: Hyundai Bioland (Korea), trade name: NEWAPLE

[0070] Morinda Citrifolia Juice: Provital, Spain, trade name: NONI EXTRAC™.S.

[0071] Yeast / rice fermentation product filtrate, homemade, method as follows:

[0072] (1) Rice powder is heated and stirred with water to make rice slurry, and a rice slurry with a mass concentration of 60% is prepared. β-glucanase is added to the rice slurry at a mass of 0.5% of the rice slurry mass and an enzyme activity of 150 U / g. Enzymatic hydrolysis is performed to obtain rice slurry hydrolysate.

[0073] (2) Inoculate rice slurry hydrolysate with yeast Saccharomyces veronae. The fermentation conditions are: fermentation temperature 40℃, fermentation time 48 hours, and pH controlled between 5.5 and 6.5 during the fermentation process. Rice fermentation liquid is obtained after fermentation.

[0074] (3) The fermented rice broth is concentrated 10 times by reverse osmosis technology to efficiently enrich active substances. After heating and sterilization, it is cooled and filtered to obtain concentrated yeast / rice fermentation product filtrate.

[0075] In Examples 1-12 and Application Examples 1-3 of this invention, RFL, APLE, and MCFJ, if the dosage unit is ug / ml, represent that the above raw materials are further prepared into solid contents by the following methods and added in the form of solid contents; if the dosage unit is %, it represents that the above raw materials are added directly.

[0076] Yeast / rice fermentation product filtrate solid contents, abbreviated as RFL solid contents, are obtained by freeze-drying yeast / rice fermentation product filtrate. The freeze-drying conditions are: cooling to -30°C in a freeze dryer, controlling the vacuum degree to 20a, and the weight of RFL solid contents is 1% of the weight of the original yeast / rice fermentation product filtrate.

[0077] The solid contents of Artemisia argyi leaf extract were obtained by freeze-drying the Artemisia argyi leaf extract solution under the following conditions: the temperature was lowered to -30°C in a freeze dryer and the vacuum degree was controlled at 20°C. The weight of the solid contents of Artemisia argyi leaf extract was 5% of the weight of the original Artemisia argyi leaf extract solution.

[0078] The solid contents of Morinda officinalis juice are obtained by freeze-drying Morinda officinalis juice solution. The freeze-drying conditions are: cooling to -30°C in a freeze dryer, controlling the vacuum degree to 20°C, and the weight of the solid contents of Morinda officinalis juice is 5% of the original weight of Morinda officinalis juice.

[0079] The experimental methods used in the following examples are described below:

[0080] Cell culture

[0081] Normal human epidermal keratinocytes (NHEKs) were cultured at 37°C in basal keratinocyte culture medium without growth additives (CC-3101, KBM, Clonetics, Inc.) or in keratinocyte growth medium containing growth additives such as insulin, hydrocortisone, gentamicin / amphotericidal B, bovine pituitary extract, and human epidermal growth factor (CC-3001, KGM; Clonetics, Inc.) with an air containing 5% CO2.

[0082] Evaluation of the promoting effect of skin hydration and barrier function-related gene mRNA expression.

[0083] NHEKs were seeded into 6-well plates and cultured overnight in KGM (37°C, 5% CO2 atmosphere). The medium was then changed to KBM, and the cells were cultured for 24 hours (37°C, 5% CO2 atmosphere). The cells were then treated with RFL solid contents alone (0-400 μg / ml) and APLE (0-50 μg / ml) or a combination of both for 24 hours (see Table 1).

[0084] The mRNA expression of filaggrin (FLG), inner laminarin (IVL), lobe laminarin (LOR), kallikrein 5 (KLK5), kallikrein 7 (KLK7), and serine peptidase inhibitor kazal type 5 (SPINK5) in normal human epidermal keratinocytes (NHEKs) was detected by RT-qPCR, as shown below.

[0085] RNA extraction

[0086] Remove the culture medium from each cell culture plate, wash twice with 1 mL of PBS, and then remove the PBS; 10 cells per well. 8 ~10 9 Add 1 mL of TRIZOL (total RNA extraction reagent) to the cells and carefully scrape them from the culture plate using a pipette tip; mix well by pipetting and transfer to a clean 1.5 mL EP tube, incubate on ice for 5 min; add 200 μL of chloroform, vortex vigorously for 15 s until the system becomes a pink emulsion, incubate on ice for 5 min, and centrifuge at 12000 rpm for 15 min at 4 °C; at this point, the cells will separate into three layers, with RNA in the upper aqueous layer. Carefully transfer the top aqueous layer to a new 1.5 mL tube and calculate the volume; add an equal volume of isopropanol, mix thoroughly, incubate at -20 °C for 45 min to aid precipitation, then centrifuge at 12000 rpm for 15 min at 4 °C; discard the supernatant completely, add 1 mL of ice-cold water... Use 75% RNA-specific ethanol (prepared with DEPC water) to gently tap the bottom of the tube to suspend the precipitate and wash away salt ions from the RNA precipitate. Centrifuge at 12,000 rpm for 10 min at 4°C. Let stand at room temperature for 10-15 min to allow the RNA to dry moderately. Dissolve the RNA in 20 μL of DEPC water and take 2 μL to determine the concentration. Finally, take 1 μL of sample RNA, 1 μL of 10× loading buffer, and 8 μL of DEPC ddH2O to make up to 10 μL for loading. Electrophoresis is performed at 120 V for 18 min.

[0087] Reverse transcription

[0088] The extracted RNA was used to synthesize the first strand of cDNA using a reverse transcription reagent. The reaction conditions and reagent composition are shown below.

[0089] The degenomic system is shown in Table 1, the reverse transcription reaction system is shown in Table 2, and the reverse transcription reaction conditions are shown in Table 3.

[0090] Table 1 Genome-free system

[0091]

[0092] Table 2 Reverse Transcription Reaction System

[0093]

[0094] Table 3 Reverse transcription reaction conditions

[0095] temperature time 42℃ 2min 37℃ 30min 85℃ 5 seconds

[0096] cDNA from each group of cells was used as a template, and GAPDH was used as an internal control. The expression of skin barrier structure-related genes (FLG, IVL, LOR, KLK5, KLK7, SPINK5) was detected by RT-qPCR. The RT-qPCR reaction conditions were: 95℃, 30s; 95℃, 10s; 60℃, 30s; 44 cycles. The primer sequences used are as follows:

[0097] GAPDH Forward Primer: 5'-CCTCTGACTTCAACAGCGAC-3'

[0098] GAPDH Reverse Primer: 5'-TCCTCTTGTGCTCTTGCTGG-3'

[0099] FLG Forward Primer: 5'-TGACAGTCAGGGACACTCAGA-3'

[0100] FLG Reverse Primer: 5'-GGTGTCTGGAGCCATCTCTT-3'

[0101] IVL Forward Primer: 5'-GCAGGTAGGACAGCCAAAG-3'

[0102] IVL Reverse Primer: 5'-AGCGGACCCGAAATAAGT-3'

[0103] LOR Forward Primer: 5'-CAGGTCACTCAGACCTCGT-3'

[0104] LOR Reverse Primer: 5'-CCTCCAGAGGAACCACCT-3'

[0105] KLK5 Forward Primer: 5'-CGTCCCACTAAAGATGTCAGACC-3'

[0106] KLK5 Reverse Primer: 5'-TCAAGCACTGGAGGACCTTAGG-3'

[0107] KLK7 Forward Primer: 5'-CAGGCTGTCATCCATGGTGAAG-3'

[0108] KLK7Reverse Primer: 5'-ATCCACGCACATGAGGTCAGAG-3'

[0109] SPINK5Forward Primer: 5'-ATAGCCACAGTGTCAGTGCTT-3'

[0110] SPINK5Reverse Primer: 5'-TGTTGCGTAAGGCTTGTGTTC-3'

[0111] Evaluation of its promoting effect on epidermal keratinocyte proliferation

[0112] (1) MTT method

[0113] NHEKs were seeded in 96-well plates and cultured overnight in KGM. Cells were then treated for 3 days with KGM, KGM containing 20% ​​water, KGM containing 20% ​​RFL, or a combination of 20% RFL and MCFJ (3.125–50 μg / ml) (Table 5). The cell proliferation-promoting effect was evaluated using the MTT assay [3-(4,5-dimethylthiazolyl-2-yl)-2,5-diphenyltetrazolium bromide].

[0114] The operation method of MTT [3-(4,5-dimethylthiazolyl-2-yl)-2,5-diphenyltetrazolium bromide] can be found in the following reference: Peng L, Wang B, Ren P. Reduction of MTT by flavonoids in the absence of cells[J]. Colloids and Surfaces B: Biointerfaces, 2005, 45(2):108-111.

[0115] (2) Cell counting

[0116] NHEKs were seeded in 96-well plates and cultured overnight in KGM. Cells were then treated for 3 days with a combination of 20% water, 20% RFL, and 20% RFL with MCFJ (3.125–50 μg / ml) (Table 5). Cells were fixed in 10% neutral buffered formalin and washed with phosphate-buffered saline. Nuclei were then stained with Hoechst 33342 (Dojindo) solvent and photographed using a fluorescence microscope (BZ-X810; Keyence). Cell counts were calculated based on the number of stained nuclei using image analysis software (BZ-X800 analyzer; Keyence). The degree of increase in cell count was used to evaluate the promoting effect on cell proliferation.

[0117] Statistical analysis

[0118] Using the bell curve in Excel, the data were statistically analyzed using the Dunnett test. A p-value less than 5% was considered statistically significant.

[0119] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments.

[0120] Examples 1-4

[0121] First, normal human epidermal keratinocytes (NHEKs) were cultured. Then, the cells were treated with different combinations of RFL solid contents and APLE to examine the promoting effect of different combinations on epidermal keratinocyte proliferation. The specific methods are described in the above section on the evaluation of the promoting effect on epidermal keratinocyte proliferation.

[0122] The different combinations of RFL solid contents and APLE in Examples 1-4 are shown in Table 4. In Table 4, RFL (μg / ml) refers to the content of RFL solid contents, and APLE (μg / ml) refers to the content of Artemisia Princeps leaf extract solid contents.

[0123] Table 4 shows the different combinations of RFL solid contents and APLE solid contents in Examples 1-4.

[0124] RFL (μg / ml) APLE (μg / ml) Example 1 0 0 Example 2 400 0 Example 3 0 50 Example 4 350 50

[0125] NHEKs were treated for 24 hours with different combinations of RFL solid contents and APLE solid contents as described in Examples 1-4. The mRNA expression of FLG (A), IVL (B), LOR (C), KLK5 (D), KLK7 (E), and SPINK5 (F) was analyzed using RT-qPCR. The RT-qPCR analysis method was performed according to Hatano Y, Adachi Y, Elias PM, et al. The Th2cytokine, interleukin-4, abrogueates the cohesion of normal stratum corneumin mice: implications for pathogenesis of atopic dermatitis[J]. Experimental dermatology, 2013, 22(1):30-35. Figure 1 The effects of different combinations of RFL solid contents and APLE solid contents in Examples 1-4 on the promoting effect on the mRNA expression of genes related to skin hydration and barrier function are presented. Each bar represents the mean ± SEM, n=3, ***p<0.001, compared with the untreated group (Example 1) using the Dunnett test.

[0126] from Figure 1 It can be seen that RFL tends to increase the expression of FLG, LOR, KLK7, and SPINK5 mRNA, but has no activity on the expression of IVL and KLK5 mRNA. The combination of RFL and APLE increased the mRNA expression of all the examined genes, higher than the levels achieved by using them individually.

[0127] Examples 5-11

[0128] First, normal human epidermal keratinocytes (NHEKs) were cultured. Then, the cells were treated with different combinations of RFL and MCFJ from Examples 5-11 to investigate the promoting effect of different combinations on epidermal keratinocyte proliferation. Specific methods were described above using the MTT assay and cell counting method. The different combinations of RFL and MCFJ in Examples 5-11 are shown in Table 5. In Table 5, 20% water refers to adding 20% ​​water (by weight of KGM) to the KGM culture medium; 20% RFL refers to adding 20% ​​rice fermentation filtrate (by weight of KGM) to the KGM culture medium; and MCFJ (μg / ml) refers to the amount of Morinda officinalis juice solids added to the KGM culture medium.

[0129] Table 5 shows different combinations of RFL and MCFJ in Examples 5-11.

[0130]

[0131]

[0132] The concentration of MCFJ, converted to mass percentage, is: 1 μg / ml = 0.0001%.

[0133] The effects of using yeast / rice ferment broth (RFL) alone or in combination with Morinda officinalis juice (MCFJ) on promoting the proliferation of epidermal keratinocytes are as follows: Figures 2-4 As shown.

[0134] NHEKs were treated with test samples (different compositions from Examples 5-9, 11) for 3 days. Cell proliferation was analyzed using an MTT assay. Figure 2 The combined effects of RFL and MCFJ on NHEK cell proliferation are presented (MTT assay). Each bar represents mean ± SEM, n = 6, ***p < 0.001 compared with 20% water, #p < 0.001 compared with 20% RFL, using Dunnett's test. Figure 2 It can be seen that 20% RFL + 3.125 μg / ml MCFJ or 20% RFL + 12.5 μg / ml MCFJ both promoted cell proliferation more than 20% RFL + 50 μg / ml MCFJ. The reason is that, from the perspective of promoting cell proliferation, the added concentrations of MCFJ at 3.125 μg / ml and 12.5 μg / ml are more suitable. Adding MCFJ to 50 μg / ml may inhibit cell proliferation. Furthermore, from... Figure 2 It can be seen that there is no significant difference between 20% RFL + 50 ug / ml MCFJ and 20% RFL alone.

[0135] NHEKs were treated with test samples (different compositions from Examples 5-7, 9-11) for 3 days. The number of cells per well was analyzed using image analysis software on images taken under a fluorescence microscope. Figure 3 The synergistic effects of different RFL and MCFJ combinations on NHEKs cell proliferation in Examples 5-7 and 9-11 are presented (cell counting method). Each bar represents mean ± SEM, n = 6, ***p < 0.001, #p < 0.05 compared to 20% water, and Dunnett's test was used compared to 20% RFL. Figure 3 It can be seen that 20% RFL + 25 μg / ml MCFJ or 20% RFL + 50 μg / ml MCFJ both promoted cell proliferation more than 20% RFL + 12.5 μg / ml MCFJ, but there was no significant difference between 20% RFL + 25 μg / ml MCFJ and 20% RFL + 50 μg / ml MCFJ.

[0136] The effects of different combinations of RFL and MCFJ on NHEKs cell proliferation were verified using the MTT assay and cell counting assay, respectively. The results are as follows: Figure 2 , Figure 3 As shown, due to the different experimental principles of the two testing methods, the optimal concentration of MCFJ was found to vary during the experiment. The experimental results showed that adding a certain amount of plant extract MCFJ promoted cell proliferation and counting. However, more MCFJ is not necessarily better; excessive MCFJ addition may inhibit cell proliferation.

[0137] Figure 4 The synergistic effects of different RFL and MCFJ combinations on NHEKs cell proliferation in Examples 6, 7, 10, and 11 are presented (nuclear staining). From Figure 4 It can be seen that 20% RFL + 25 μg / ml MCFJ has a better synergistic effect on NHEKs cell proliferation.

[0138] Compared to NHEKs treated with 20% water, treatment with 20% RFL had a higher effect on cell proliferation. The effect of 20% RFL was significantly enhanced when used in combination with MCFJ. Figure 2 , 3 ).and Figure 3 The results were parallel, and nuclear staining observations also showed that the number of cells treated with 20% RFL increased compared to those treated with 20% water, and the effect was further enhanced when 20% RFL was combined with MCFJ. Figure 4 ).

[0139] Example 12

[0140] Based on Examples 1-4, RFL was combined with APLE and MCFJ, and RT-qPCR was used to analyze the mRNA expression of FLG, IVL, LOR, KLK5, KLK7 and SPINK5.

[0141] When 300 μg / ml of RFL solids, 50 μg / ml of APLE, and 50 μg / ml of Morinda officinalis juice were added to KGM culture medium, the FLG / GAPDH ratio was 517.23 ± 13.54. However, when 350 μg / ml of RFL solids and 50 μg / ml of APLE were added to KGM culture medium, the FLG / GAPDH ratio was 363.12 ± 23.25%. # P<0.001;

[0142] When 300 μg / ml of RFL solids, 50 μg / ml of APLE, and 50 μg / ml of Morinda officinalis juice were added to KGM culture medium, the IVL / GAPDH ratio was 372 ± 16.14. However, when 350 μg / ml of RFL solids and 50 μg / ml of APLE were added to KGM culture medium, the IVL / GAPDH ratio was 223.13 ± 6.17%. # P<0.001;

[0143] Test results showed that the combination of the three resulted in significantly higher mRNA expression of FLG and IVL compared to the combination of RFL and APLE. The mRNA expression of the remaining genes was comparable to that of the combination of RFL and APLE, with no significant difference, but was significantly better than using 400ug / ml RFL alone. *** P<0.001).

[0144] Based on Examples 5-11, RFL was combined with APLE and MCFJ at concentrations of 20%, 25 μg / ml, and 25 μg / ml, respectively. The promoting effect of the combination on epidermal keratinocyte proliferation was then tested using the MTT assay and cell counting method. The results showed that the combination of the three substances significantly promoted keratinocyte proliferation more than 20% RFL alone or 50 μg / ml of MCFJ. # P<0.001), and using 20.005% RFL alone ( *** P<0.001).

[0145] Application Examples of Compositions Containing Plant Extracts and Rice Fermentation Product Filtrate for the Preparation of Skin Care Water Application Example 1

[0146] The formula for the skin care lotion is shown in Table 3.

[0147] Table 3 shows the formula for the skin care water in Application Example 1.

[0148]

[0149]

[0150] The preparation process is as follows:

[0151] a. Place raw material 1 in a container equipped with a stirring device.

[0152] b. Mix and moisten ingredients 2, 3, 4, and 5.

[0153] c. While keeping the water from step a stirred, add the moistened and mixed ingredients 2, 3, 4, and 5, and stir thoroughly until completely dissolved.

[0154] d. Add ingredients 6 and 7 to the above mixture and stir thoroughly until completely dissolved.

[0155] Application Example 2

[0156] The formula for the skin care lotion is shown in Table 4.

[0157] Table 4 shows the formula for the skin care water in Application Example 2.

[0158]

[0159] The preparation process is as follows:

[0160] a. Place raw material 1 in a container equipped with a stirring device.

[0161] b. Mix and moisten ingredients 2, 3, 4, and 5.

[0162] c. While keeping the water from step a stirred, add the moistened and mixed ingredients 2, 3, 4, and 5, and stir thoroughly until completely dissolved.

[0163] d. Add ingredients 6 and 7 to the above mixture and stir thoroughly until completely dissolved.

[0164] Application Example 3: Anhydrous Formulation

[0165] The formula for the fruit juice skin care water is shown in Table 5.

[0166] Table 5 shows the formula for the skincare water in Application Example 3.

[0167]

[0168] The preparation process is as follows:

[0169] a. Place raw material 1 in a container equipped with a stirring device.

[0170] b. Mix and moisten ingredients 2, 3, 4, and 5.

[0171] c. While keeping the water from step a stirred, add the moistened and mixed ingredients 2, 3, 4, and 5, and stir thoroughly until completely dissolved.

[0172] d. Add ingredients 6 and 7 to the above mixture and stir thoroughly until completely dissolved.

[0173] The skin care water obtained from the above application examples 1, 2, and 3 has excellent skin barrier repair and moisturizing effects.

[0174] The above description of the embodiments is provided to enable those skilled in the art to understand and use the invention. It will be apparent to those skilled in the art that various modifications can be made to these embodiments, and the general principles described herein can be applied to other embodiments without inventive effort. Therefore, the present invention is not limited to the above embodiments, and any improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the invention should be within the protection scope of the present invention.

Claims

1. A composition containing plant extracts and rice fermentation product filtrate, characterized in that, The composition comprises Artemisia argyi leaf extract, Morinda officinalis juice, and yeast / rice fermentation product filtrate; the mass ratio of Artemisia argyi leaf extract, Morinda officinalis juice, and yeast / rice fermentation product filtrate in the composition is 0.001-1: 0.001-1：5-97； The method for preparing the yeast / rice fermentation product filtrate is as follows: Rice powder is mixed with water, heated, and stirred to form a rice slurry with a mass concentration of 40-60%. β-glucanase is added to the rice slurry for enzymatic hydrolysis. The amount of β-glucanase added is 0.001-1% of the mass of the rice slurry, and the enzyme activity is 100-200 U / g, yielding a rice slurry hydrolysate. Saccharomyces veronae yeast is inoculated into the rice slurry hydrolysate for fermentation at a temperature of 20℃~40℃ for 24h~72h, yielding a fermentation broth. The fermentation broth is concentrated using reverse osmosis technology to efficiently enrich active substances, then heated for sterilization, cooled and filtered to obtain the yeast / rice fermentation product filtrate.

2. The composition containing plant extracts and rice fermentation product filtrate according to claim 1, characterized in that, In the composition, the mass ratio of Artemisia argyi leaf extract, Morinda officinalis juice, and yeast / rice fermentation product filtrate is 0.1-1:0.1-1:20-92.

3. The composition containing plant extracts and rice fermentation product filtrate according to claim 2, characterized in that, In the composition, the mass ratio of Artemisia argyi leaf extract, Morinda officinalis juice, and yeast / rice fermentation product filtrate is 0.1-1:0.1-1:30-70.

4. The composition containing plant extracts and rice fermentation product filtrate according to claim 3, characterized in that, In the composition, the mass ratio of Artemisia argyi leaf extract, Morinda officinalis juice, and yeast / rice fermentation product filtrate is 0.1-1:0.1-1:30-40.

5. A method for preparing the composition containing plant extracts and rice fermentation product filtrate according to any one of claims 1-4, characterized in that, Artemisia argyi leaf extract, Morinda officinalis juice, and yeast / rice fermentation product filtrate were mixed in a specific ratio.

6. The use of the composition containing plant extracts and rice fermentation product filtrate according to any one of claims 1-4, characterized in that, The composition containing plant extracts and rice fermentation product filtrate is used to prepare skin care products. In the process of preparing skin care products, the composition containing plant extracts and rice fermentation product filtrate replaces part or all of the water in the formula.

7. A skincare product, characterized in that, The composition and the mass fraction of each component are as follows: Glycerin 2-10% Butylene glycol 1-5% Yeast / rice fermentation product filtrate 5-95% Sea peony juice 0.001-1% Phenoxyethanol 0.1-1% Ethylhexylglycerin 0.01-0.1% Artemisia leaf extract 0.001-1% Add water to 100%; The method for preparing the yeast / rice fermentation product filtrate is as follows: Rice powder is mixed with water, heated, and stirred to form a rice slurry with a mass concentration of 40-60%. β-glucanase is added to the rice slurry for enzymatic hydrolysis. The amount of β-glucanase added is 0.001-1% of the mass of the rice slurry, and the enzyme activity is 100-200 U / g, yielding a rice slurry hydrolysate. Saccharomyces veronae yeast is inoculated into the rice slurry hydrolysate for fermentation at a temperature of 20℃~40℃ for 24h~72h, yielding a fermentation broth. The fermentation broth is concentrated using reverse osmosis technology to efficiently enrich active substances, then heated for sterilization, cooled and filtered to obtain the yeast / rice fermentation product filtrate.

8. A skincare product, characterized in that, The composition and the mass fraction of each component are as follows: Glycerin 2-10% Butylene glycol 1-5% Add yeast / rice fermentation product filtrate to 100%. Sea peony juice 0.001-1% Phenoxyethanol 0.1-1% Ethylhexylglycerin 0.01-0.1% Artemisia argyi leaf extract 0.001-1%; The method for preparing the yeast / rice fermentation product filtrate is as follows: Rice powder is mixed with water, heated, and stirred to form a rice slurry with a mass concentration of 40-60%. β-glucanase is added to the rice slurry for enzymatic hydrolysis. The amount of β-glucanase added is 0.001-1% of the mass of the rice slurry, and the enzyme activity is 100-200 U / g, yielding a rice slurry hydrolysate. Saccharomyces veronae yeast is inoculated into the rice slurry hydrolysate for fermentation at a temperature of 20℃~40℃ for 24h~72h, yielding a fermentation broth. The fermentation broth is concentrated using reverse osmosis technology to efficiently enrich active substances, then heated for sterilization, cooled and filtered to obtain the yeast / rice fermentation product filtrate.

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